阅读说明：本技术 具有提高的溶解度和生物利用度的营养组合物 (Nutritional compositions with improved solubility and bioavailability ) 是由 P·琼斯 M·文玛 D·奥尔布赖特 S·蒂米歇尔 G·凯慈 T·莱迪 于 2014-08-28 设计创作，主要内容包括：本发明涉及具有提高的溶解度和生物利用度的营养组合物。本文公开了一种营养组合物,其具有至少一种蛋白质、至少一种脂肪和至少一种亲脂性化合物,所述组合物包含至少一种组装物,所述组装物包含至少一种疏水蛋白质、甘油单酯和甘油二酯(“MDG”)以及至少一种亲脂性化合物,其中在20℃下以100,000x g离心1小时后,所述营养组合物中的总MDG的至少1％仍在水相中。(The present invention relates to nutritional compositions having improved solubility and bioavailability. Disclosed herein is a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising at least one assembly comprising at least one hydrophobic protein, mono-and diglycerides ("MDG") and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃.)

1. A nutritional composition, comprising:

a protected premix comprising:

a premix comprising a lipophilic compound, a surfactant, and mono-and di-glyceride ("MDG") components, wherein the premix is free of oils or fats other than the MDG component and the surfactant, and

a fat-coated protein slurry.

2. The nutritional composition of claim 1, wherein the premix consists of the lipophilic compound, the surfactant, and the MDG component.

3. The nutritional composition of claim 1, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, docosahexaenoic acid (DHA), gangliosides, phospholipids, inositol phosphate, glycophospholipids, or any combination thereof.

4. The nutritional composition of claim 1, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, a source of long chain polyunsaturated fatty acids (LCPUFAs), or a combination thereof.

5. The nutritional composition of claim 1, wherein the surfactant is lecithin, polyglycerol ester, or a combination thereof.

6. The nutritional composition of claim 5, wherein the surfactant is lecithin.

7. The nutritional composition of claim 1, wherein the protein of the fat-coated protein slurry is a protein having a total average hydropathic index (GRAVY) value of-0.5 to 0.

8. The nutritional composition of claim 7, wherein the protein is beta-lactoglobulin, bovine lactoferrin, gamma₂-casein, β -casein, α -lactalbumin or a combination thereof.

9. The nutritional composition of claim 1, wherein the protected premix has a size of 60 kD to 1,000 kD.

10. The nutritional composition of claim 1, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 31,000 x g for at least 1 hour at 20 ℃.

11. A method of making a nutritional composition containing at least one protein, at least one fat, at least one carbohydrate, and at least one lipophilic compound, the method comprising:

mixing the lipophilic compound, surfactant, and MDG component to form a premix, wherein the MDG component comprises monoglycerides, diglycerides, and no more than 20wt.% impurities;

Combining the premix thus formed with the fat-in-protein slurry to form a protected premix; and

the protected premix thus formed is combined with the remaining ingredients of the nutritional composition.

12. The method of claim 11, wherein the protected pre-mixture has a size of 60 kD to 1,000 kD.

13. The method of claim 11, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, a source of long chain polyunsaturated fatty acids (LCPUFAs), or a combination thereof.

14. The method of claim 13, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, docosahexaenoic acid (DHA), or any combination thereof.

15. The method of claim 14, wherein the lipophilic compound is lutein.

16. The method of claim 15, wherein the surfactant is lecithin, polyglycerol ester, or a combination thereof.

17. The method of claim 16, wherein the surfactant is lecithin.

18. The method of claim 11, wherein the surfactant is lecithin, polyglycerol ester, or a combination thereof.

19. The method of claim 18, wherein the surfactant is lecithin.

20. The method of claim 11, wherein said protein is a protein having a total average hydrophilic index (GRAVY) value of-0.5 to 0.

21. The method of claim 11, wherein the lipophilic compound, surfactant, and MDG component are mixed under heating.

22. A nutritional composition, comprising:

an activation assembly comprising at least one hydrophobic protein having a total average hydrophilicity index (GRAVY) value of-0.5 to 0, at least one combination of mono-and diglycerides (MDG), and at least one lipophilic compound,

wherein at least 5% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 31,000 x g for at least 1 hour at 20 ℃.

23. The nutritional composition of claim 22, wherein the assembly is (i) water soluble; and (ii) has a size of 60 kD to 1000 kD.

24. The nutritional composition of claim 22, wherein the hydrophobic protein comprised in the assembly is selected from the group consisting of beta-lactoglobulin, bovine lactoferrin, gamma₂-hydrophobic proteins of casein, β -casein, α -lactalbumin or a combination thereof.

25. The nutritional composition of claim 22, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, a source of long chain polyunsaturated fatty acids (LCPUFAs), or a combination thereof.

26. The nutritional composition of claim 22, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, docosahexaenoic acid ("DHA"), or any combination thereof.

27. The nutritional composition of claim 22, wherein the composition further comprises choline at a concentration of 5 mg/kg to 1 g/kg based on the total weight of the composition.

Technical Field

The present disclosure relates to compositions containing lipophilic compounds with enhanced bioavailability. More specifically, the compositions contain an activation premix or an MDG protected premix comprising mono-and diglycerides. These premixes can provide lipophilic compounds in a form that can be more efficiently absorbed into the body upon consumption.

Background

Adult, child and infant processed nutritional and pharmaceutical products may contain various lipophilic compounds such as carotenoids, fat soluble vitamins, lipophilic antioxidants, or combinations thereof. Current nutritional compositions comprising lipophilic compounds typically provide limited bioavailability upon consumption because the lipophilic compounds do not maintain a stable water soluble form. Thus, products are often over-fortified with lipophilic compounds to ensure that the desired nutritional or pharmaceutical benefits from these compounds can be obtained. In some cases, the over-reinforcement may be about 2 times to about 10 times the amount needed to achieve the desired benefit.

These high fortification rates may result in increased production costs and potential complications in formulating the final product without providing additional consumer benefits. In addition, over-fortified formulations may saturate the digestive tract with insoluble lipophilic compounds, resulting in dyspepsia.

Thus, there is a need for nutritional compositions for infants, toddlers, children, and adults that provide desirable lipophilic compounds in a more stable, soluble form.

Summary of The Invention

In one aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000 kD.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of proteins of the assembly by at least 2% compared to a nutritional composition without MDG.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compounds of the assembly by at least 2% compared to a nutritional composition without MDG.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compounds of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compounds of the composition is increased by at least 15% when administered to a subject, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compounds of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compounds of the composition is increased by at least 15% when administered to a subject, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at about 31,000xg for at least 1 hour at 20 ℃.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compounds of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compounds of the composition when administered to a subject is increased by at least 15%, wherein the increase in bioavailability of 15% is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at least 1 hour at about 31,000x g at 20 ℃, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition is increased by at least 15% when administered to a subject, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is an intact protein, a hydrolyzed protein, or a combination thereof.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is an intact protein, a hydrolyzed protein, or a combination thereof, wherein the assembly is stable at 23 ℃ for at least 12 months.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000xg for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein the MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without the MDG, wherein the MDG increases the conjugated lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition is increased by at least 15% when administered to a subject, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to a nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at about 31,000Xg for at least 1 hour at 20 ℃, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at least 12 months at 23 ℃, wherein the assembly is stable for at least 24 months at room temperature.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at 23 ℃ for at least 12 months, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of from about-0.5 to about 0.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000xg for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the increase in bioavailability of 15% is measured by lymphatic absorption of the composition relative to a nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable at 23 ℃ for at least 12 months, wherein the assembly is stable at room temperature Stable for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of from about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma₂-casein, β -casein, α -lactalbumin or a combination thereof.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000xg for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at 23 ℃ for at least 12 months, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma ₂-casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400 kD.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one amphiphileAn activated assembly of fatty compounds wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at 23 ℃ for at least 12 months, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma ₂-casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compounds of the assembly by at least 2% compared to a nutritional composition without MDG, wherein when administered to a subjectAn increase in bioavailability of the lipophilic compound of the composition by at least 15% when the subject is present, wherein the increase in bioavailability by 15% is measured by lymphatic absorption of the composition relative to a nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at about 31,000x g for at least 1 hour at 20 ℃, wherein at least 5% of the assembly remains in the water phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at least 12 months at 23 ℃, wherein the assembly is stable for at least 24 months at room temperature, wherein the assembly comprises a hydrophobic protein having a vgray value of about-0.5 to about 0, wherein the hydrophobic protein is β -lactoglobulin, bovine lactoferrin, γ, wherein the hydrophobic protein is beta-lactoglobulin, and a pharmaceutically acceptable salt thereof ₂-casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition is increased by at least 15% when administered to a subject, wherein the increase in bioavailability of 15% is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at least 1 hour at about 31,000x g at 20 ℃, wherein at least 5% of the assembly remains after centrifugation An aqueous phase, wherein the protein is an intact protein, a hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at least 12 months at 23 ℃, wherein the assembly is stable for at least 24 months at room temperature, wherein the assembly comprises a hydrophobic protein having a GRAVY value of from about-0.5 to about 0, wherein the hydrophobic protein is β -lactoglobulin, bovine lactoferrin, γ -lactoglobulin, or a combination thereof₂-casein, beta-casein, alpha-lactalbumin or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant or a combination thereof, wherein the lipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitamin K or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, marine oil, cottonseed oil, flaxseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compounds of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compounds of the composition is increased by at least 15% when administered to a subject, wherein the increase in bioavailability of 15% is measured by lymphatic absorption of the composition relative to a nutritional composition without MDG, wherein at 20 ℃ the increase is measured by the concentration of the protein of the composition in the composition, wherein the increase in the concentration of the protein is measured by the concentration of the protein of the composition in the composition, and wherein the increase in the concentration of the protein is measured by the concentration of the protein in the compositionAfter centrifugation at about 31,000x g for at least 1 hour, the assembly remains water soluble, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at least 12 months at 23 ℃, wherein the assembly is stable for at least 24 months at room temperature, wherein the assembly comprises a hydrophobin having a GRAVY value of from about-0.5 to about 0, wherein the hydrophobin is beta-lactoglobulin, bovine lactoferrin, gamma ₂Casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea hare oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof, comprising about 145mg/kg to about 890mg/kg of MDG, based on the total weight of the composition.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compounds of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the relative affinity of the composition when administered to a subject An increase in bioavailability of a lipid compound of at least 15%, wherein the increase in bioavailability of 15% is measured by lymphatic absorption of the composition relative to a nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at about 31,000x g for at least 1 hour at 20 ℃, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is an intact protein, a hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at least 12 months at 23 ℃, wherein the assembly is stable for at least 24 months at room temperature, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is β -lactoglobulin, bovine lactoferrin, γ -lactoglobulin₂Casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea hare oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof, comprising about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprising about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a molecular weight of 100kD to 1000kA size of D, wherein MDG increases the concentration of protein of the assembly by at least 2% as compared to a nutritional composition without MDG, wherein MDG increases the concentration of bound lipophilic compounds of the assembly by at least 2% as compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compounds of the composition when administered to a subject is increased by at least 15%, wherein the increase in bioavailability of 15% is measured by lymphatic absorption of the composition as compared to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is an intact protein, a hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at least 12 months at 23 ℃, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of from about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, or gamma ₂Casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea hare oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof, comprising about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprising about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprising about 1.0 μ g/kg to about 7.0g/kg of lipophilic compounds based on the total weight of the composition.

In another aspect, the present disclosure may relate to nutritional compositions having at least one protein, at least one fat, and at least one lipophilic compound The composition comprises: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at 23 ℃ for at least 12 months, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma ₂Casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea hare oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof, comprising about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition,comprising about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprising about 1.0 μ g/kg to about 7.0g/kg of lipophilic compounds based on the total weight of the composition, comprising about 147.4mg/kg to about 589.6mg/kg of MDG based on the total weight of the composition, and about 1.12mg/kg of lipophilic compounds based on the total weight of the composition.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at 23 ℃ for at least 12 months, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma ₂-casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, soy protein, or a combination thereof,Corn oil, olive oil, safflower oil, high oleic coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, canola oil, marine oil, cottonseed oil, linseed oil, hemp oil, peanut oil, borage oil, algae oil, fungal oil, MCT oil (medium chain triglycerides), and combinations thereof, comprises from about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprises from about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprises from about 1.0 μ g/kg to about 7.0g/kg of lipophilic compounds based on the total weight of the composition, comprises from about 147.4mg/kg to about 589.6mg/kg of MDG and about 1.12mg/kg of lipophilic compounds based on the total weight of the composition, wherein the ratio of MDG to lipophilic compound is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1 and 1: 1 (weight/weight).

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compounds of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compounds of the composition is increased by at least 15% when administered to a subject, wherein the bioavailability of 15% is increasedThe improvement is measured by lymphatic absorption of the composition relative to a nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at about 31,000x g for at least 1 hour at 20 ℃, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is an intact protein, a hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at least 12 months at 23 ℃, wherein the assembly is stable for at least 24 months at room temperature, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is β -lactoglobulin, bovine lactoferrin, γ -gamma ₂Casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea hare oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof, comprises about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprises about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprises about 1.0 μ g/kg to about 7.0g/kg of lipophilic compound based on the total weight of the composition, comprises about 147.4mg/kg to about 589.6mg/kg of MDG based on the total weight of the composition and about 1.12mg/kg of lipophilic compound, wherein the ratio of MDG to lipophilic compound is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 100: 1, 2: 1, and the like, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1 1. 2: 1 and 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound comprises a ratio of about 12000: 1 to about 1: 1 (weight/weight).

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at 23 ℃ for at least 12 months, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma ₂-casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, or a combination thereof,High oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, canola oil, marine oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof, comprising from about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, from about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, from about 1.0 μ g/kg to about 7.0g/kg of lipophilic compounds based on the total weight of the composition, from about 147.4mg/kg to about 589.6mg/kg of MDG based on the total weight of the composition, and from about 1.12mg/kg of lipophilic compounds, wherein the ratio of MDG to lipophilic compounds is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, and MCT/kg, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1 and 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound comprises a ratio of about 12000: 1 to about 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound is 4: 1, 3: 1 or 2: 1.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compounds of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compounds of the composition is increased by at least when administered to a subject15%, wherein the increase in bioavailability of 15% is measured by lymphatic absorption of the composition relative to a nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at about 31,000x g for at least 1 hour at 20 ℃, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is an intact protein, a hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at least 12 months at 23 ℃, wherein the assembly is stable for at least 24 months at room temperature, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is β -lactoglobulin, bovine lactoferrin, γ -lactoglobulin ₂Casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea hare oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof, comprises about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprises about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprises about 1.0 μ g/kg to about 7.0g/kg of lipophilic compound based on the total weight of the composition, comprises about 147.4mg/kg to about 589.6mg/kg of MDG based on the total weight of the composition and about 1.12mg/kg of lipophilic compound, wherein the ratio of MDG to lipophilic compound is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 100: 1, 2: 1, and the like, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1, and 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compounds comprises a ratio of about 12000: 1 to about 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compounds is 4: 1, 3: 1, or 2: 1, comprising about 1.0g/L to about 100g/L of hydrophobic protein based on the total weight of the composition.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at 23 ℃ for at least 12 months, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma ₂-casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, whereinThe fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, marine oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algae oil, fungal oil, MCT oil (medium chain triglycerides), and combinations thereof, comprises about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprises about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprises about 1.0 μ g/kg to about 7.0g/kg of lipophilic compounds based on the total weight of the composition, comprises about 147.4mg/kg to about 589.6mg/kg of MDG and about 1.12mg/kg of lipophilic compounds based on the total weight of the composition, wherein the ratio of MDG to lipophilic compound is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1 and 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound comprises a ratio of about 12000: 1 to about 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound is 4: 1, 3: 1, and 1 to 1 (weight/weight), wherein the composition comprises about 0.1 to about 100g of hydrophobic compound, further comprising choline.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% as compared to a nutritional composition without MDG, wherein MDG increases the concentration of bound lipophilic compounds of the assembly by at least 2% as compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compounds of the composition when administered to a subject is increased by at least 15%, wherein the increase in bioavailability of 15% is measured by lymphatic absorption of the composition as compared to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is an intact protein, a hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at least 12 months at 23 ℃, wherein the assembly is stable for at least 24 months at room temperature, wherein the assembly comprises a hydrophobic protein having a GRAVY value of from about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, or gamma ₂Casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea hare oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof, comprising about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprising about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprising about 1.0 μ g/kg to about 7.0g/kg of lipophilic compounds based on the total weight of the composition, comprising about 147.4mg/kg to about 589.6mg/kg of MDG based on the total weight of the composition, and about 1.12mg/kg of lipophilic compounds, wherein the ratio of MDG to lipophilic compounds is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, 9000: 1, a, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1 and 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compounds comprises a ratio of about 12000: 1 to about 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compounds is 4: 1, 3: 1 or 2: 1, comprising from about 1.0g/L to about 100g/L, based on the total weight of the composition, further comprising a hydrophobic choline protein, comprising about 5mg/kg to about 1g/kg choline, based on the total weight of the composition.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at 23 ℃ for at least 12 months, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein Is beta-lactoglobulin, bovine lactoferrin, gamma₂Casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea hare oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof, comprises about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprises about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprises about 1.0 μ g/kg to about 7.0g/kg of lipophilic compound based on the total weight of the composition, comprises about 147.4mg/kg to about 589.6mg/kg of MDG based on the total weight of the composition and about 1.12mg/kg of lipophilic compound, wherein the ratio of MDG to lipophilic compound is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 100: 1, 2: 1, and the like, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1, and 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound comprises a ratio of about 12000: 1 to about 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound is 4: 1, 3: 1, or 2: 1, comprises about 1.0g/L to about 100g/L of hydrophobic protein based on the total weight of the composition, further comprises choline, comprises about 5mg/kg to about 1g/kg choline based on the total weight of the composition, further comprises lecithin.

In another aspect, the present disclosure may relate to an implementA nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at 23 ℃ for at least 12 months, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma ₂Casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea harp oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (r) (r) ("m) (" g) ("m)" palm oil, palm kernel oil, palm olein oil, rapeseed oil, sea harp oil, cottonseed oil, linseed oil, peanut oil, borage oil, algal oil, fungal oil, and mixtures thereofMedium chain triglycerides) and combinations thereof, comprising about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprising about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprising about 1.0 μ g/kg to about 7.0g/kg of lipophilic compounds based on the total weight of the composition, comprising about 147.4mg/kg to about 589.6mg/kg of MDG and about 1.12mg/kg of lipophilic compounds based on the total weight of the composition, wherein the ratio of MDG to lipophilic compounds is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 1: 1, 50: 1, 25: 1, 20: 1, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1 and 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound comprises a ratio of about 12000: 1 to about 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound is 4: 1, 3: 1, or 2: 1, comprises from about 1.0g/L to about 100g/L of hydrophobic protein based on the total weight of the composition, further comprises choline, comprises from about 5mg/kg to about 1g/kg choline based on the total weight of the composition, further comprises lecithin, comprises from about 10mg/kg to about 5g/kg lecithin based on the total weight of the composition.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compounds of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the composition is lipophillized when administered to a subjectThe bioavailability of the composition is increased by at least 15%, wherein the increase in bioavailability of 15% is measured by lymphatic absorption of the composition relative to a nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at about 31,000x g for at least 1 hour at 20 ℃, wherein at least 5% of the assembly remains in the water phase after centrifugation, wherein the protein is an intact protein, a hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at least 12 months at 23 ℃, wherein the assembly is stable for at least 24 months at room temperature, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma-lactoglobulin ₂Casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea hare oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof, comprises about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprises about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprises about 1.0 μ g/kg to about 7.0g/kg of lipophilic compound based on the total weight of the composition, comprises about 147.4mg/kg to about 589.6mg/kg of MDG based on the total weight of the composition and about 1.12mg/kg of lipophilic compound, wherein the ratio of MDG to lipophilic compound is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 100: 1, 2: 1, and the like, 15: 1, 14: 1, 13: 1, 12: 1 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1, and 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound comprises a ratio of about 12000: 1 to about 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound is 4: 1, 3: 1, or 2: 1, comprises from about 1.0g/L to about 100g/L of hydrophobic protein based on the total weight of the composition, further comprises choline, comprises from about 5mg/kg to about 1g/kg choline based on the total weight of the composition, further comprises lecithin, comprises from about 10mg/kg to about 5g/kg lecithin based on the total weight of the composition, further comprises docosahexaenoic acid ("DHA").

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at 23 ℃ for at least 12 months, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma ₂-casein, beta-casein, alpha-lactalbumin or a mixture thereofA combination, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, marine oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides), and combinations thereof, comprising from about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprises about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprises about 1.0 μ g/kg to about 7.0g/kg of lipophilic compound based on the total weight of the composition, comprises about 147.4mg/kg to about 589.6mg/kg of MDG and about 1.12mg/kg of lipophilic compound based on the total weight of the composition, wherein the ratio of MDG to lipophilic compound is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 10: 1, 8: 1, 75: 1, 2, 1, 2, 1, 2, 1, 2, 1, 2, 1, 2, 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1 and 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound comprises a ratio of about 12000: 1 to about 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound is 4: 1, 3: 1, or 2: 1, comprising about 1.0g/L to about 100g/L of hydrophobic protein based on the total weight of the composition, further comprising choline, comprising about 5mg/kg to about 1g/kg choline based on the total weight of the composition, further comprising lecithin, comprising about 10mg/kg to about 5g/kg lecithin based on the total weight of the composition, further comprising docosahexaenoic acid ("DHA"), wherein the nutritional composition comprises about 10mg/k based on the total weight of the composition. g to about 10g/kg DHA.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at 23 ℃ for at least 12 months, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma ₂-casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea oil, or a combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea oil, or a combination thereofCottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides), and combinations thereof, comprising about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprising about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprising about 1.0 μ g/kg to about 7.0g/kg of lipophilic compounds based on the total weight of the composition, comprising about 147.4mg/kg to about 589.6mg/kg of MDG and about 1.12mg/kg of lipophilic compounds based on the total weight of the composition, wherein the ratio of MDG to lipophilic compounds is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1, and 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compounds comprises a ratio of about 12000: 1 to about 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compounds is 4: 1, 3: 1, or 2: 1, comprises from about 1.0g/L to about 100g/L of hydrophobic protein based on the total weight of the composition, further comprises choline, comprises from about 5mg/kg to about 1g/kg of choline based on the total weight of the composition, further comprises lecithin, comprising about 10mg/kg to about 5g/kg lecithin based on the total weight of the composition, further comprising docosahexaenoic acid ("DHA"), wherein the nutritional composition comprises about 10mg/kg to about 10g/kg DHA based on the total weight of the composition, further comprising high palmitic acid vegetable oil.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) hasA size of 100kD to 1000kD, wherein the MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without the MDG, wherein the MDG increases the bound lipophilic compound of the assembly by at least 2% compared to the nutritional composition without the MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the increase in bioavailability of 15% is measured by lymphatic absorption of the composition relative to the nutritional composition without the MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is an intact protein, a hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at least 12 months at 23 ℃, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, or gamma ₂Casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea hare oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof, comprising about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprising about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprising about 1.0 μ g/kg to about 7.0g/kg of lipophilic compounds based on the total weight of the composition, comprising about 147.4mg/kg to about 589.6mg/kg of MDG based on the total weight of the composition, and about 1.12mg/kg of lipophilic compounds, wherein the ratio of MDG to lipophilic compounds is selected from the group consisting of 12000: 1, 11 000: 1, 10000: 1, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1 and 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound comprises a ratio of about 12000: 1 to about 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound is 4: 1, 3: 1 or 2: 1, comprises about 0.0 to about 100g of hydrophobic protein based on the total weight of the composition, further comprising choline, comprising about 5mg/kg to about 1g/kg choline based on the total weight of the composition, further comprising lecithin, comprising about 10mg/kg to about 5g/kg lecithin based on the total weight of the composition, further comprising docosahexaenoic acid ("DHA"), wherein the nutritional composition comprises about 10mg/kg to about 10g/kg DHA based on the total weight of the composition, further comprising a high palmitic acid vegetable oil, wherein the nutritional composition comprises about 0.5g/kg to about 35g/kg high palmitic acid vegetable oil based on the total weight of the composition.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compounds of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compounds of the composition is increased by at least 15% when administered to a subject, wherein the 15% increase in bioavailability is through lymphatic absorption of the composition relative to the nutritional composition without MDGA measurement wherein the assembly remains water soluble after centrifugation at about 31,000x g for at least 1 hour at 20 ℃, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at least 12 months at 23 ℃, wherein the assembly is stable for at least 24 months at room temperature, wherein the assembly comprises a hydrophobic protein having a GRAVY value of from about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma ₂Casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea hare oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof, comprises about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprises about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprises about 1.0 μ g/kg to about 7.0g/kg of lipophilic compound based on the total weight of the composition, comprises about 147.4mg/kg to about 589.6mg/kg of MDG based on the total weight of the composition and about 1.12mg/kg of lipophilic compound, wherein the ratio of MDG to lipophilic compound is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 100: 1, 2: 1, and the like, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1 and 1: 1 (weight/weight), wherein MDG is for lipophilic compounds The ratio includes a ratio of about 12000: 1 to about 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound is 4: 1, 3: 1, or 2: 1, comprises about 1.0g/L to about 100g/L of hydrophobic protein based on the total weight of the composition, further comprises choline, comprises about 5mg/kg to about 1g/kg choline based on the total weight of the composition, further comprises lecithin, comprises about 10mg/kg to about 5g/kg lecithin based on the total weight of the composition, further comprises docosahexaenoic acid ("DHA"), wherein the nutritional composition comprises about 10mg/kg to about 10g/kg DHA based on the total weight of the composition, further comprises a high palmitic acid vegetable oil, wherein the nutritional composition comprises about 0.5g/kg to about 35g/kg high palmitic acid vegetable oil based on the total weight of the composition, wherein the nutritional composition comprises about 0.2g/L to about 20g/L of beta-casein, based on the total weight of the composition.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at 23 ℃ for at least 12 months, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises A hydrophobic protein having a GRAVY value of from about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma₂Casein, β -casein, α -lactalbumin, or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, sea hare oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides) and combinations thereof, comprises about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprises about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprises about 1.0 μ g/kg to about 7.0g/kg of lipophilic compound based on the total weight of the composition, comprises about 147.4mg/kg to about 589.6mg/kg of MDG based on the total weight of the composition and about 1.12mg/kg of lipophilic compound, wherein the ratio of MDG to lipophilic compound is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 100: 1, 2: 1, and the like, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1, and 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound comprises a ratio of about 12000: 1 to about 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound is 4: 1, 3: 1, or 2: 1, comprises about 1.0g/L to about 100g/L of hydrophobic protein based on the total weight of the composition, further comprises choline, comprises about 5mg/kg to about 1g/kg choline based on the total weight of the composition, further comprises lecithin, comprises choline based on the combination The nutritional composition comprises about 10mg/kg to about 5g/kg lecithin, further comprising docosahexaenoic acid ("DHA"), wherein the nutritional composition comprises about 10mg/kg to about 10g/kg DHA, based on the total weight of the composition, further comprising high palmitic acid vegetable oil, wherein the nutritional composition comprises about 0.5g/kg to about 35g/kg high palmitic acid vegetable oil, based on the total weight of the composition, wherein the nutritional composition comprises about 0.2g/L to about 20g/L beta-casein, based on the total weight of the composition, wherein the nutritional composition comprises carbohydrate.

In another aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000x g for 1 hour at 20 ℃, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000kD, wherein MDG increases the concentration of protein of the assembly by at least 2% compared to a nutritional composition without MDG, wherein MDG increases the bound lipophilic compound of the assembly by at least 2% compared to a nutritional composition without MDG, wherein the bioavailability of the lipophilic compound of the composition when administered to a subject is increased by at least 15%, wherein the 15% increase in bioavailability is measured by lymphatic absorption of the composition relative to the nutritional composition without MDG, wherein the assembly remains water soluble after centrifugation at 20 ℃ for at least 1 hour at about 31,000x g, wherein at least 5% of the assembly remains in the aqueous phase after centrifugation, wherein the protein is intact protein, hydrolyzed protein, or a combination thereof, wherein the assembly is stable for at 23 ℃ for at least 12 months, wherein the assembly is stable for at room temperature for at least 24 months, wherein the assembly comprises a hydrophobic protein having a GRAVY value of about-0.5 to about 0, wherein the hydrophobic protein is beta-lactoglobulin, bovine lactoferrin, gamma ₂-casein, beta-casein, alpha-lactalbumin or a combination thereof, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat-soluble vitamin, a lipophilicAn antioxidant or a combination thereof, wherein the lipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the fat is MDG, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, marine oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides), and combinations thereof, comprising about 145mg/kg to about 890mg/kg of MDG based on the total weight of the composition, comprising about 145mg/kg to about 600mg/kg of MDG based on the total weight of the composition, comprising from about 1.0 μ g/kg to about 7.0g/kg of lipophilic compound based on the total weight of the composition, comprising from about 147.4mg/kg to about 589.6mg/kg of MDG and from about 1.12mg/kg of lipophilic compound based on the total weight of the composition, wherein the ratio of MDG to lipophilic compound is selected from the group consisting of 12000: 1, 11000: 1, 10000: 1, 9000: 1, 8000: 1, 7000: 1, 6000: 1, 5000: 1, 4000: 1, 3000: 1, 2000: 1, 1000: 1, 900: 1, 800: 1, 700: 1, 600: 1, 500: 1, 400: 1, 300: 1, 200: 1, 100: 1, 75: 1, 50: 1, 25: 1, 20: 1, 15: 1, 14: 1, 13: 1, 12: 1, 11: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 3: 1, and 2: 1, wherein the ratio of MDG to lipophilic compound comprises a ratio of about 12000: 1 to about 1: 1 (weight/weight), wherein the ratio of MDG to lipophilic compound is 4: 1, 3: 1, or 2: 1, comprises about 1.0g/L to about 100g/L of hydrophobic protein based on the total weight of the composition, further comprises choline, comprises about 5mg/kg to about 1g/kg choline based on the total weight of the composition, further comprises lecithin, comprises about 10mg/kg to about 5g/kg lecithin based on the total weight of the composition, further comprises docosahexaenoic acid ("DHA"), wherein the nutritional composition comprises about 10mg/kg to about 10g/kg DHA based on the total weight of the composition, further comprises high palmitic acid vegetable oil, wherein the nutritional composition comprises from about 0.5g/kg to about 0.5g/kg based on the total weight of the composition. About 35g/kg high palmitic acid vegetable oil, wherein the nutritional composition comprises about 0.2g/L to about 20g/L beta-casein based on the total weight of the composition, wherein the nutritional composition comprises a carbohydrate, wherein the carbohydrate is maltodextrin, hydrolyzed or modified starch, hydrolyzed or modified corn starch, glucose polymer, corn syrup solids, rice-derived carbohydrate, glucose, fructose, lactose, high fructose corn syrup, honey, sugar alcohol, maltitol, erythritol, sorbitol, or a combination thereof.

In one aspect, the present disclosure may relate to a nutritional composition comprising at least one protein and vitamin D having improved vitamin D bioavailability, wherein the nutritional composition comprises: an assembly comprising a combination of monoglycerides and diglycerides ("MDG") and vitamin D, wherein vitamin D has improved bioavailability when administered to a subject.

In another aspect, the present disclosure may relate to a nutritional composition comprising at least one protein and vitamin D having improved bioavailability of vitamin D, wherein the nutritional composition comprises: an assembly comprising a combination of monoglycerides and diglycerides ("MDG") and vitamin D, wherein vitamin D has improved bioavailability when administered to a subject, comprising from about 0.1mg/kg to about 20mg/kg MDG, based on the total weight of the composition.

In another aspect, the present disclosure may relate to a nutritional composition comprising at least one protein and vitamin D having improved bioavailability of vitamin D, wherein the nutritional composition comprises: an assembly comprising a combination of monoglycerides and diglycerides ("MDG") and vitamin D, wherein vitamin D has improved bioavailability when administered to a subject, comprises from about 0.1mg/kg to about 20mg/kg MDG based on the total weight of the composition, comprises from about 0.1mg/kg to about 10mg/kg MDG based on the total weight of the composition.

In another aspect, the present disclosure may relate to a nutritional composition comprising at least one protein and vitamin D having improved bioavailability of vitamin D, wherein the nutritional composition comprises: an assembly comprising a combination of monoglycerides and diglycerides ("MDG") and vitamin D, wherein vitamin D has improved bioavailability when administered to a subject, comprises from about 0.1mg/kg to about 20mg/kg MDG based on the total weight of the composition, comprises from about 0.1mg/kg to about 10mg/kg MDG based on the total weight of the composition, and comprises from about 10 μ g/kg to about 30 μ g/kg vitamin D based on the total weight of the composition.

In another aspect, the present disclosure may relate to a nutritional composition comprising at least one protein and vitamin D having improved bioavailability of vitamin D, wherein the nutritional composition comprises: an assembly comprising a combination of monoglycerides and diglycerides ("MDG") and vitamin D, wherein vitamin D has improved bioavailability when administered to a subject, comprises from about 0.1mg/kg to about 20mg/kg MDG based on the total weight of the composition, comprises from about 0.1mg/kg to about 10mg/kg MDG based on the total weight of the composition, comprises from about 10 μ g/kg to about 30 μ g/kg vitamin D based on the total weight of the composition, comprises from about 0.1mg/kg to about 10mg/kg MDG and about 20 μ g/kg vitamin D based on the total weight of the composition.

In one aspect, the present disclosure may relate to a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising: an activated assembly comprising at least one hydrophobic protein, at least one combination of monoglycerides and diglycerides ("MDG"), and at least one lipophilic compound, wherein at least 15% of the total lipophilic compounds in the nutritional composition remain in the aqueous phase after centrifugation at 1,000x g for 1 hour at 20 ℃.

In one aspect, the present disclosure may relate to a method of preparing a nutritional composition having at least one lipophilic compound with improved bioavailability, the method comprising the steps of: providing a premix comprising an assembly comprising at least one hydrophobic protein, a combination of mono-and diglycerides ("MDG"), and at least one lipophilic compound, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000 kD; adding the premix to an aqueous solution to form an activated premix; and adding the activated premix to a nutritional composition.

In another aspect, the present disclosure may relate to a method of preparing a nutritional composition having at least one lipophilic compound with increased bioavailability, the method comprising the steps of: providing a premix comprising an assembly comprising at least one hydrophobic protein, a combination of mono-and diglycerides ("MDG"), and at least one lipophilic compound, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000 kD; adding the premix to an aqueous solution to form an activated premix; and adding the activated premix to a nutritional composition, wherein the assembly has a size of 60kD to 400 kD.

In another aspect, the present disclosure may relate to a method of preparing a nutritional composition having at least one lipophilic compound with increased bioavailability, the method comprising the steps of: providing a premix comprising an assembly comprising at least one hydrophobic protein, a combination of mono-and diglycerides ("MDG"), and at least one lipophilic compound, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000 kD; adding the premix to an aqueous solution to form an activated premix; and adding the activated premix to a nutritional composition, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant or a combination thereof.

In another aspect, the present disclosure may relate to a method of preparing a nutritional composition having at least one lipophilic compound with increased bioavailability, the method comprising the steps of: providing a premix comprising an assembly comprising at least one hydrophobic protein, a combination of mono-and diglycerides ("MDG"), and at least one lipophilic compound, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000 kD; adding the premix to an aqueous solution to form an activated premix; and adding the activated premix to the nutritional composition, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K or any combination thereof.

In another aspect, the present disclosure may relate to a method of preparing a nutritional composition having at least one lipophilic compound with increased bioavailability, the method comprising the steps of: providing a premix comprising an assembly comprising at least one hydrophobic protein, a combination of mono-and diglycerides ("MDG"), and at least one lipophilic compound, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000 kD; adding the premix to an aqueous solution to form an activated premix; and adding the activated premix to the nutritional composition, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the premix is heated to a temperature of 120 ° F for about 10 minutes.

In one aspect, the present disclosure may relate to a method of preparing a nutritional composition having at least one lipophilic compound with improved bioavailability, the method comprising the steps of: providing a premix comprising an assembly comprising at least one hydrophobic protein, a combination of mono-and diglycerides ("MDG"), and at least one lipophilic compound, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000 kD; adding the premix to a fat-containing solution to form a protected premix; and adding the protected premix to a nutritional composition.

In another aspect, the present disclosure may relate to a method of preparing a nutritional composition having at least one lipophilic compound with increased bioavailability, the method comprising the steps of: providing a premix comprising an assembly comprising at least one hydrophobic protein, a combination of mono-and diglycerides ("MDG"), and at least one lipophilic compound, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000 kD; adding the premix to a fat-containing solution to form a protected premix; and adding the protected premix to a nutritional composition, wherein the assembly has a size of 60kD to 400 kD.

In another aspect, the present disclosure may relate to a method of preparing a nutritional composition having at least one lipophilic compound with increased bioavailability, the method comprising the steps of: providing a premix comprising an assembly comprising at least one hydrophobic protein, a combination of mono-and diglycerides ("MDG"), and at least one lipophilic compound, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000 kD; adding the premix to a fat-containing solution to form a protected premix; and adding the protected premix to a nutritional composition, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant or a combination thereof.

In another aspect, the present disclosure may relate to a method of preparing a nutritional composition having at least one lipophilic compound with improved bioavailability, the method comprising the steps of: providing a premix comprising an assembly comprising at least one hydrophobic protein, a combination of mono-and diglycerides ("MDG"), and at least one lipophilic compound, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000 kD; adding the premix to a fat-containing solution to form a protected premix; and adding the protected premix to a nutritional composition, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof.

In another aspect, the present disclosure may relate to a method of preparing a nutritional composition having at least one lipophilic compound with increased bioavailability, the method comprising the steps of: providing a premix comprising an assembly comprising at least one hydrophobic protein, a combination of mono-and diglycerides ("MDG"), and at least one lipophilic compound, wherein the assembly is (i) water soluble; and (ii) has a size of 100kD to 1000 kD; adding the premix to a fat-containing solution to form a protected premix; and adding the protected premix to the nutritional composition, wherein the assembly has a size of 60kD to 400kD, wherein the lipophilic compound is a carotenoid, a fat soluble vitamin, a lipophilic antioxidant, or a combination thereof, wherein the lipophilic compound is lutein, vitamin a, vitamin D, vitamin E, vitamin K, or any combination thereof, wherein the premix is heated to a temperature of 120 ° F for about 10 minutes.

Brief Description of Drawings

Figure 1 shows lymph flow rates per hour over 6 hours after lutein administration of different nutritional composition batches.

Figure 2 shows the hourly lutein output in bars within 6 hours after lutein application of different nutritional composition batches.

Figure 3 shows the cumulative lymph lutein absorption over 6 hours after lutein application for different nutritional composition batches.

Figure 4 shows the percent change in lymphatic lutein absorption over a 6 hour period with mono-and diglycerides versus control.

Fig. 5 shows lymph flow rates per hour over 8 hours after administration of a variable nutritional composition containing lutein.

Fig. 6 shows the hourly lutein output in lymph over 8 hours after administration of the variable nutritional composition containing lutein.

Figure 7 shows cumulative lymph lutein absorption over 6 and 8 hours after administration of the lutein containing variable nutritional composition.

Figure 8 shows the percent change in lymphatic lutein absorption after administration of the lutein containing variable nutritional composition versus 6 and 8 hours after administration of the control.

Fig. 9 shows the lymphatic triglyceride output per hour over 6 hours after administration of the nutritional composition.

Fig. 10 shows the hourly lymphophospholipid output over 6 hours after administration of the nutritional composition.

Fig. 11 shows the change in ARA lymphatic absorption from fasting over 6 hours after administration of the nutritional composition.

Fig. 12 shows the change in DHA lymphatic absorption from fasting within 6 hours after administration of the nutritional composition.

Figure 13 shows the high molecular weight lutein assemblies present in the aqueous phase after centrifugation of the variable nutrient composition as a function of lutein concentration.

Figures 14-16 show the high molecular weight lutein assembly in the aqueous fraction after centrifugation of variable nutritional compositions by different centrifugal forces (figure 14); a protein greater than 137kD (FIG. 15); and the presence of conjugated lipophilic substances (figure 16).

Figure 17 shows the effect of choline and lecithin addition to the nutritional composition on the formation of high molecular weight lutein assemblies.

Fig. 18 shows lymphatic output of triglycerides over 6 hours after infusion of variable nutritional compositions.

Fig. 19 shows lymphatic absorption of palmitic acid within 6 hours after infusion of the variable nutritional composition.

Fig. 20 shows lymphatic absorption of linoleic acid within 6 hours after infusion of the variable nutritional composition.

Figure 21 shows lymphatic uptake of alpha-linolenic acid within 6 hours after infusion of variable nutritional compositions.

Fig. 22 shows the DHA lymphatic output as a function of time after administration of the variable nutritional composition.

Fig. 23 shows UV spectra of the aqueous portion of the variable nutritional composition. UF: ultrafiltration with a membrane of 10kD or 100 kD; UC: centrifugation method C (100,000x g); SMP: skim milk powder/skim milk powder; WPC: whey protein concentrate 80.

Fig. 24 shows lymphatic triglyceride output within 6 hours after administration of the variable nutritional composition, where the composition was stratified based on the length of time after production.

Detailed description of the invention

The present invention relates to nutritional compositions for delivering nutrients, including lipophilic compounds in a more soluble form, by using mono-and diglycerides (hereinafter "MDG") assembled with lipophilic proteins and hydrophobic proteins. This assembly of nutritional compositions containing MDG, hydrophobic proteins and lipophilic compounds allows for the concomitant bringing of the lipophilic compounds into a more soluble form to promote digestibility. The formation of stable high molecular weight assemblies (MDG plus hydrophobic and lipophilic proteins, about 60kD to about 400kD) allows for the highest nutrient delivery and bioavailability of lipophilic compounds, as these stable soluble forms are increased compared to lipophilic compound combinations that do not contain MDG and hydrophobic proteins. The increased water-soluble form of the lipophilic compound results in increased bioavailability upon consumption.

Finally, this allows the nutritional composition to deliver lipophilic compounds without being over-fortified, thereby reducing manufacturing costs, limiting complications in the composition, and producing a more desirable product. The nutritional compositions may be ingested by infants, toddlers, children, or adults to provide nutrients needed for proper development, growth, and/or as a nutritional supplement. The nutritional compositions may further be consumed to deliver lipophilic compounds for the treatment of a variety of conditions, including bone health/growth, eye health, cardiovascular health, and the like.

2. Definition of

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document, including definitions, will control. Preferred methods and materials are described below, but methods and materials similar or equivalent to those described herein can be used in the practice or testing of the nutritional compositions. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting.

As used herein, the terms "comprising," "including," "having," "can," "containing," and variations thereof are intended to be open-ended transitional phrases, terms, or words that do not exclude the possibility of additional acts or structures. The singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments that "comprise," "consist of," and "consist essentially of" the embodiments or elements mentioned herein, whether or not explicitly stated.

The term "activated" refers to MDG that has been mixed with water or other aqueous liquid. The term "activation" refers to the step of mixing the MDG with the aqueous liquid. For example, "activated MDG component" means an aqueous liquid into which MDG has been mixed. For the purposes of this disclosure, an activated MDG is typically an aqueous liquid that is substantially free of oils or fats other than MDG.

The term "activating assembly" refers to an assembly comprising at least one MDG, at least one hydrophobic protein and at least one lipophilic compound, wherein the assembly may have been contacted with an aqueous liquid.

As used herein, the terms "adult formulation" and "adult nutritional product" are used interchangeably to refer to formulations used to generally maintain or improve the health of an adult, and include those formulations designed for adults suffering from, susceptible to, or at risk of developing age-related macular degeneration.

The term "bioavailable" as used herein, unless otherwise specified, means that lipophilic compounds are capable of being absorbed from the gastrointestinal tract into the lymph which is then to be drawn into the blood stream of an individual so that the substance can be absorbed into the organs and tissues of the body. As the bioavailability of a compound increases, the compound becomes more likely to enter and remain in the bloodstream where it can be absorbed and used by the body. As the bioavailability of the compound decreases, the compound becomes less likely to be absorbed from the gastrointestinal tract into the lymph and will be excreted from the body before entering the bloodstream.

Unless otherwise indicated, the term "casein" as used herein should be understood to refer to those proteins in milk that will precipitate from milk at a solution pH of 4.6(20 ℃); casein typically comprises about 80% of the whole undiluted milk. In general, any type of milk-derived protein source can be used as a source of bovine beta-casein for the nutritional composition, including whole milk, skim milk powder, milk protein concentrate, whole milk protein, milk protein isolate, acid casein, calcium caseinate, sodium caseinate, magnesium caseinate, purified beta-casein, and combinations thereof, all of which will contain beta-casein. Beta-casein constitutes around 30% of the protein contained in milk and may exist in one of several genetic variants, the two most common being genetic variants a1 and a 2. Bovine beta-casein has been further classified as a genetic variant, including a3, B, C, D, E, F, G, H1, H2 and I.

The term "composition" as used herein, unless otherwise specified, refers to a mixture suitable for enteral administration to a subject. The compositions may be in powder, solid, semi-solid, liquid, gel, and semi-liquid forms. The composition may further comprise vitamins, minerals and other ingredients.

As used herein, unless otherwise specified, the term "dry blending" refers to mixing components or ingredients to form a base nutritional powder or to adding dry powdered or granular components or ingredients to a base powder to form a powdered nutritional composition.

As used herein, unless otherwise specified, the terms "fat," "lipid," and "oil" are used interchangeably to refer to a lipid material derived or processed from a plant or animal. These terms also include synthetic lipid materials, so long as such synthetic materials are suitable for administration to a subject as defined herein.

The term "glyceride" as used herein generally refers to a lipophilic compound comprising a glycerol molecule bound to fatty acid groups. Monoglycerides are molecules of glycerol bound to a single fatty acid group; diglycerides are glycerol molecules bound to two fatty acid groups; and a triglyceride is a glycerol molecule bound to three fatty acid groups. Fats and oils comprise glycerides and typical fats and oils from animal, fish, algae, plant or seed sources consist primarily of triglycerides.

The term "high palmitic acid vegetable oil" or "HPAV oil" refers to a vegetable oil comprising greater than about 22% palmitic acid as a percentage of total fatty acids in HPAV oil. Examples of HPAV oils include, but are not limited to, palm oil and palm olein. For purposes of the present disclosure, HPAV oil may be from sources including, but not limited to, plants, transgenic plants, fungi, algae, bacteria, and other unicellular organisms. For the purposes of this disclosure, the term "HPAV oil" does not include oils and fats from multicellular animal sources, such as beef tallow, chicken fat, fish oil, or milk cream.

The term "human milk fortifier" as used herein, unless otherwise indicated, refers to a nutritional product suitable for mixing with human milk or preterm or infant formula for consumption by preterm or term infants.

The term "infant" as used herein, unless otherwise indicated, refers to a human with an age of about 12 months or less. The term "young child" as used herein, unless otherwise indicated, refers to a human being from about 12 months to about 3 years of age. The term "child" as used herein, unless otherwise specified, refers to a human being from about 3 to about 18 years of age. The term "adult" as used herein, unless otherwise specified, refers to a human being about 18 years of age or older.

As used herein, the term "infant formula" or "infant nutritional product" are used interchangeably to refer to a nutritional composition having an appropriate balance of macronutrients, micronutrients and calories to provide sole or supplemental nutrition for the health of an infant, a toddler, or both, and generally to maintain or improve their health. Infant Formula preferably comprises nutrients for the target consumer or user group according to relevant Infant Formula guidelines, an example of which is the Infant Formula Act, 21u.s.c.section 350 (a).

The term "lipophilic compound" as used herein refers to a component that is more soluble in an organic solvent (such as ethanol, methanol, diethyl ether, acetone, chloroform, benzene, or lipids) than in water. Vitamin D is an example of a lipophilic nutrient. For the purposes of this disclosure, the term "lipophilic nutrient" may apply to other lipophilic compounds, including but not limited to pharmaceutical compounds.

The term "monoglyceride and diglyceride oil" or "MDG oil" as used herein refers to a combination of both monoglycerides and diglycerides present in the premix. Premixes according to the present disclosure may comprise monoglycerides and diglycerides in an amount of at least 12%, including from about 12% to about 98%, and also including from about 20% to about 80%, including from about 20% to about 60%, and including from about 25% to about 50%, based on the weight of the premix (excluding the weight of lipophilic compounds). The premix may comprise at least 12%, from about 12% to about 98%, from about 20% to about 80%, from about 20% to about 60%, or from about 25% to about 50% by weight of the mixture of mono-and diglycerides; the amount of lipophilic compound as described above; and impurities in an amount of less than about 20%, less than about 15%, less than about 10%, or less than about 2% by weight. It will be recognized by those skilled in the art based on the disclosure herein that the premix may include a small portion of impurities such that the mixture of mono-and diglycerides is not 100% mono-and diglycerides by weight. In some embodiments, the impurities may be triglycerides and/or free glycerol. These impurities may generally be less than about 20%, less than about 15%, less than about 10%, or less than about 2% by weight. In embodiments where the premix comprises monoglycerides and impurities, the remainder of the premix (excluding lipophilic compounds) is diglycerides.

The term "protected premix" refers to a mixture comprising a surfactant and MDG. The surfactant may be present in, but is not limited to, the premix, the activation premix, and other components of the nutritional composition. Non-limiting examples of suitable surfactants in the protected premix include lecithin, polyglycerol esters, and combinations thereof. The protected premix may be prepared by mixing the surfactant and MDG in the presence of heat or at ambient temperature and in some embodiments with agitation. The protected premix may be added to a fat-containing solution to form an "MDG protected component".

The term "nutritional composition" as used herein, unless otherwise specified, refers to nutritional powders, solids, semisolids, liquids, gels, and semisolids that comprise at least one of protein, carbohydrate, and lipid and are suitable for enteral administration to a subject. The nutritional composition may further comprise vitamins, minerals and other ingredients, and represents the sole, primary or supplemental source of nutrition.

The term "pharmaceutical composition" as used herein may include any form suitable for administration to a subject. Non-limiting examples of modes of administration include enteral and parenteral administration.

As used herein, unless otherwise indicated, the term "powdered nutritional composition" refers to a spray-dried and/or dry-mixed powdered and/or agglomerated nutritional composition comprising a fat-soluble nutrient (such as lutein) and a mixture of mono-and diglycerides, which is reconstitutable with an aqueous liquid and is suitable for oral administration to humans.

The term "shelf life" as used herein, unless otherwise specified, refers to the stability of the assembly as present in, but not limited to, powders, premixes, activation premixes, protected activation premixes, suspensions, liquids, concentrated liquids, mixtures, or combinations thereof.

The term "subject" as used herein refers to a mammal that ingests a composition, including but not limited to a human (e.g., an infant, a toddler, a child, or an adult), a domesticated livestock (e.g., a cow, a horse, or a pig), or a pet (e.g., a dog or a cat).

As used herein, unless otherwise specified, the terms "susceptible to" or "at risk" are used interchangeably to mean having little resistance to a condition or disease, including being genetically predisposed to the condition or disease, having a family history of the condition or disease, and/or symptoms of the condition or disease.

The term "vegetable oil" as used herein refers to fat derived from a plant, seed, fungal or algal source (e.g., not from a multicellular animal). Vegetable oils may be solid or liquid fats at room temperature. Examples of vegetable oils include, but are not limited to, coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, and combinations thereof.

To the extent that the term "includes," "including," or "containing" is used herein, it is intended to be inclusive in a manner similar to the term "comprising" as "comprising" is interpreted when employed as a transitional word in a claim. Further, to the extent that the term "or" (e.g., a or B) is used, it is intended to mean "a or B or both. When applicants intend to indicate "only a or B but not both," the term "only a or B but not both" will be used. Thus, use of the term "or" herein is the inclusive, and not the exclusive use. Additionally, to the extent that the term "in …" or "into …" is used herein, it is intended to additionally mean "on …" or "on …".

All percentages, parts and ratios used herein are by weight of the total composition, unless otherwise specified. Unless otherwise indicated, all such weights as they pertain to listed ingredients are based on the activity level and, therefore, do not include solvents or by-products that may be included in commercially available materials.

All references to singular features or limitations of the present disclosure are to include the corresponding plural features or limitations and vice versa unless otherwise indicated herein or clearly implied to the contrary by the context in which they are referred to.

All combinations of method or process steps used herein can be performed in any order, unless otherwise indicated herein or otherwise clearly implied to the contrary by the context in which the combination is referred to.

Various embodiments of the nutritional compositions of the present disclosure may also be substantially free of any optional or selected essential ingredients or features described herein, provided that the remaining nutritional composition still contains all the essential ingredients or features described herein. In this regard, unless otherwise indicated, the term "substantially free" means that the selected composition contains less than a functional amount of the optional ingredient, typically less than about 0.1% (by weight), and also including 0% (by weight of such optional or selected base ingredient).

The nutritional and pharmaceutical compositions of the present disclosure, and the corresponding methods of manufacture, may comprise, consist of, or consist essentially of the essential elements of the disclosure as described herein, as well as any additional or optional elements described herein or otherwise useful in the nutritional and pharmaceutical composition applications.

To the extent that numerical ranges are recited herein, each numerical value therebetween is specifically contemplated to the same degree of accuracy. For example, for a range of 6-9, values of 7 and 8 are contemplated in addition to 6 and 9, and for a range of 6.0-7.0, values of 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are expressly contemplated.

3. Nutritional composition

Provided herein is a nutritional composition. The nutritional composition comprises at least one protein, at least one fat, and at least one lipophilic compound. As explained below, the nutritional composition includes an assembly of agglomerated compounds that increases the bioavailability of lipophilic compounds upon consumption. The nutritional composition may include an assembly comprising at least one lipophilic compound, at least one protein, and at least one fat. The combination of fat (e.g., MDG) and protein (e.g., hydrophobic protein) interacts with lipophilic compounds (e.g., lutein and vitamin D), which enhances the solubility and stability of the lipophilic compounds compared to assemblies that do not contain MDG and hydrophobic protein. The nutritional composition may comprise a premix, an activation premix, and a protected premix.

The nutritional composition can be used for food. The nutritional composition may be ingested by an infant and thus provide the infant's nutrients needed for proper development and growth. Nutritional compositions may also be ingested by young children or children for proper delivery of nutrients for sustained development and growth. The nutritional composition may also be ingested by adults as a nutritional supplement. The nutritional compositions may be used to deliver lipophilic compounds for the treatment of a variety of conditions, including bone health/growth, eye health, cardiovascular health, and the like.

The composition can be in any useful form. Non-limiting product forms include: solid, semi-solid, ready-to-drink liquid, concentrated liquid, gel, and powder. In some embodiments, the composition may be in the form of a flowable or substantially flowable particulate composition. In some embodiments, the composition may be easily scooped and measured with a spoon or other similar device so that the composition may be easily reconstituted with a suitable aqueous liquid (such as water) by the intended user to form a liquid nutritional or pharmaceutical composition for immediate oral or enteral use. In this regard, "immediate" use generally means within about 48 hours, most typically within about 24 hours, and in some embodiments immediately followed by rehydration.

In some embodiments, the composition may comprise a spray-dried powder, a dry-blended powder, an agglomerated powder, a combination thereof, or a powder prepared by other suitable methods.

In some embodiments, the composition may be contained in a dosage unit suitable for oral consumption. Suitable dosage units include tablets, hard gelatin capsules, starch capsules, cellulose-based capsules, soft capsules, and elixirs.

In some embodiments, the powdered composition may be compressed into a tablet. In some embodiments, the powdered composition may be contained in a capsule. The capsule comprises a shell surrounding and containing the composition. The capsule shell dissolves or disintegrates in the digestive tract after the capsule is ingested, thereby releasing the composition to be absorbed by the body of the individual ingesting the capsule. Capsule shells are typically made from gelatin, starch, cellulose, or other components that readily dissolve or disintegrate upon ingestion, and the composition, manufacture, and use of capsule shells are well known in the art. In some embodiments, the dosage unit of the composition is a soft gel capsule. Soft capsules are particularly suitable for containing liquid-based ingredients such as nutrients dissolved, dispersed or suspended in a carrier oil. The outer shell of the soft capsule is usually made of gelatin plasticized with glycerin and water, but vegetable soft capsules made of starch or carrageenan can also be utilized. Soft capsule shells are typically made and filled with the composition in a continuous process as is known in the art. Soft capsules are manufactured by manufacturers such as Catalent Pharma Solutions, LLC (Somerset, NJ) and Captek Softgel International (Cerritos, Calif.).

Capsules have a wide range of sizes, and the capsule size should be selected to contain the appropriate volume or weight of the composition and hence the appropriate dose of lipophilic compound. The capsule size may be selected to contain at least about 0.1 grams of the composition, including from about 0.1 grams to about 30 grams, from about 0.2 grams to about 20 grams, from about 0.25 grams to about 15 grams, from about 0.25 grams to about 10 grams, from about 0.25 grams to about 5 grams, from about 0.25 grams to about 3 grams, from about 0.25 grams to about 1.5 grams, or from about 0.25 grams to about 1.0 grams of the composition. In some embodiments, for use in humans, the capsules contain from about 0.1 grams to about 1.5 grams or from about 0.2 grams to about 1.0 gram of the composition, as these capsules are the smallest size convenient for most adults and children to swallow. In some embodiments, for use in large animals, such as domesticated livestock, the capsule contains from about 0.1 grams to about 30 grams, or from about 1.0 gram to about 30 grams, of the composition.

The nutritional compositions may be formulated to have a sufficient variety and quantity of nutrients to provide a sole, primary, or supplemental source of nutrition, or to provide a specialized nutritional composition for individuals with a particular disease or condition.

The nutritional composition may have a caloric density tailored to the nutritional needs of the end user, but in most cases, the nutritional composition may comprise from about 65 to about 800kcal/240 mL. The nutritional compositions as discussed herein provide a method of easily and effectively controlling caloric intake of an individual (e.g., an infant, a toddler, a child, or an adult). It is important to be able to tightly control caloric intake because different individuals have different caloric needs. The nutritional composition may comprise a caloric density of about 75 to about 700kcal/240 mL. The nutritional composition may comprise a caloric density of about 100 to about 650kcal/240 mL. For example, the nutritional composition may comprise about 65kcal/240mL, 70kcal/240mL, 75kcal/240mL, 80kcal/240mL, 85kcal/240mL, 90kcal/240mL, 95kcal/240mL, 100kcal/240mL, 105kcal/240mL, 110kcal/240mL, 115kcal/240mL, 120kcal/240mL, 125kcal/240mL, 130kcal/240mL, 135kcal/240mL, 140kcal/240mL, 145kcal/240mL, 150kcal/240mL, 155kcal/240mL, 160kcal/240mL, 165kcal/240mL, 170kcal/240mL, 175kcal/240mL, 180kcal/240mL, 185kcal/240mL, 190kcal/240mL, 195kcal/240mL, 200kcal/240mL, 205kcal/240mL, 210kcal/240mL, 240kcal/240mL, 220kcal/240mL, 225kcal/240mL, 230kcal/240mL, 235kcal/240mL, 240kcal/240mL, 245kcal/240mL, 250kcal/240mL, 255kcal/240mL, 260kcal/240mL, 265kcal/240mL, 270kcal/240mL, 275kcal/240mL, 280kcal/240mL, 285kcal/240mL, 290kcal/240mL, 295kcal/240mL, 300kcal/240mL, 305kcal/240mL, 310kcal/240mL, 315kcal/240mL, 320kcal/240mL, 325kcal/240mL, 330kcal/240mL, 335kcal/240mL, 340kcal/240mL, 345kcal/240mL, 350kcal/240mL, 355kcal/240mL, 360kcal/240mL, 365/240 mL, 240kcal/240mL, 380kcal/240mL, 240kcal/240mL, 240, 385kcal/240mL, 390kcal/240mL, 395kcal/240mL, 400kcal/240mL, 405kcal/240mL, 410kcal/240mL, 415kcal/240mL, 420kcal/240mL, 425kcal/240mL, 430kcal/240mL, 435kcal/240mL, 440kcal/240mL, 445kcal/240mL, 450kcal/240mL, 455kcal/240mL, 460kcal/240mL, 465kcal/240mL, 470kcal/240mL, 475kcal/240mL, 480kcal/240mL, 485kcal/240mL, 490kcal/240mL, 495kcal/240mL, 500kcal/240mL, 550kcal/240mL, 600kcal/240mL, 650kcal/240mL, 700kcal/240mL, 750kcal/240mL, or 800kcal/240 mL.

Further, non-limiting examples of compositions include human milk fortifiers, preterm infant formulas, elemental and semielemental formulas, pediatric formulas, adult formulas, nutritional supplements, capsules, suppositories, sprays, drops, lotions, ointments, microcapsules, and liposomes.

The at least one lipophilic compound in the nutritional composition may be about 1 μ g/kg to about 10g/kg of lipophilic compound. The at least one lipophilic compound in the nutritional composition may be about 1 μ g/kg to about 7g/kg of lipophilic compound. The at least one lipophilic compound in the nutritional composition may be about 500 μ g/kg to about 5g/kg of lipophilic compound. The at least one lipophilic compound in the nutritional composition may be about 1mg/kg to about 1g/kg of lipophilic compound. The at least one lipophilic compound in the nutritional composition may be from about 100mg/kg to about 1g/kg of lipophilic compound. For example, the at least one lipophilic compound in the nutritional composition may be about 1 μ g/kg, 10 μ g/kg, 50 μ g/kg, 100 μ g/kg, 150 μ g/kg, 200 μ g/kg, 250 μ g/kg, 300 μ g/kg, 350 μ g/kg, 400 μ g/kg, 450 μ g/kg, 500 μ g/kg, 550 μ g/kg, 600 μ g/kg, 650 μ g/kg, 700 μ g/kg, 750 μ g/kg, 800 μ g/kg, 850 μ g/kg, 900 μ g/kg, 950 μ g/kg, 1mg/kg, 10mg/kg, 50mg/kg, 100mg/kg, 150mg/kg, 200mg/kg, 250mg/kg, 300mg/kg, 350mg/kg, 400mg/kg, 450mg/kg, 500mg/kg, 550mg/kg, 600mg/kg, 650mg/kg, 700mg/kg, 750mg/kg, 800mg/kg, 850mg/kg, 900mg/kg, 950mg/kg, 1g/kg, 0.5g/kg, 1g/kg, 1.5g/kg, 2g/kg, 2.5g/kg, 3g/kg, 3.5g/kg, 4g/kg, 4.5g/kg, 5g/kg, 5.5g/kg, 6g/kg, 6.5g/kg, 7g/kg, 7.5g/kg, 8g/kg, 8.5g/kg, 9g/kg, 9.5g/kg or 10 g/kg.

The at least one fat in the nutritional composition may be from about 10g/kg to about 100 g/kg. The at least one fat in the nutritional composition may be from about 20g/kg to about 80 g/kg. The at least one fat in the nutritional composition may be from about 30g/kg to about 70 g/kg. The at least one fat in the nutritional composition may be from about 40g/kg to about 60 g/kg. For example, the at least one fat in the nutritional composition may be about 10g/kg, 10.5g/kg, 11g/kg, 11.5g/kg, 12g/kg, 12.5g/kg, 13g/kg, 13.5g/kg, 14g/kg, 14.5g/kg, 15g/kg, 15.5g/kg, 16g/kg, 16.5g/kg, 17g/kg, 17.5g/kg, 18g/kg, 18.5g/kg, 19g/kg, 19.5g/kg, 20g/kg, 20.5g/kg, 21g/kg, 21.5g/kg, 22g/kg, 22.5g/kg, 23g/kg, 23.5g/kg, 24g/kg, 24.5g/kg, 25g/kg, 25.5g/kg, 26g/kg, 26.5g/kg, 27g/kg, 27.5g/kg, 28g/kg, 28.5g/kg, 29g/kg, 29.5g/kg, 30g/kg, 30.5g/kg, 31g/kg, 31.5g/kg, 32g/kg, 32.5g/kg, 33g/kg, 33.5g/kg, 34g/kg, 34.5g/kg, 35g/kg, 35.5g/kg, 36g/kg, 36.5g/kg, 37g/kg, 37.5g/kg, 38g/kg, 38.5g/kg, 39g/kg, 39.5g/kg, 40g/kg, 40.5g/kg, 41g/kg, 41.5g/kg, 42g/kg, 42.5g/kg, 43g/kg, 43.5g/kg, 44g/kg, 44.5g/kg, 45g/kg, 45.5g/kg, 46g/kg, 46.5g/kg, 47g/kg, 47.5g/kg, 48g/kg, 48.5g/kg, 49g/kg, 49.5g/kg, 50g/kg, 50.5g/kg, 51g/kg, 51.5g/kg, 52g/kg, 52.5g/kg, 53g/kg, 53.5g/kg, 54g/kg, 54.5g/kg, 55g/kg, 55.5g/kg, 56g/kg, 56.5g/kg, 57g/kg, 57.5g/kg, 58g/kg, 58.5g/kg, 59g/kg, 59.5g/kg, 60g/kg, 60.5g/kg, 61g/kg, 61.5g/kg, 62g/kg, 62.5g/kg, 63g/kg, 63.5g/kg, 50g/kg, 50.5g/kg, 56g/kg, 51.5g/kg, 52g/kg, 52.5g/kg, 50.5g/kg, 64g/kg, 64.5g/kg, 65g/kg, 65.5g/kg, 66g/kg, 66.5g/kg, 67g/kg, 67.5g/kg, 68g/kg, 68.5g/kg, 69g/kg, 69.5g/kg, 70g/kg, 70.5g/kg, 71g/kg, 71.5g/kg, 72g/kg, 72.5g/kg, 73g/kg, 73.5g/kg, 74g/kg, 74.5g/kg, 75g/kg, 75.5g/kg, 76g/kg, 76.5g/kg, 77g/kg, 77.5g/kg, 78g/kg, 78.5g/kg, 79g/kg, 79.5g/kg, 80g/kg, 80.5g/kg, 81g/kg, 81.5g/kg, 82g/kg, 82.5g/kg, 69.5g/kg, 70g/kg, 70.5g/kg, 74.5g/kg, and the like, 82.5g/kg, 83.5g/kg, 84g/kg, 84.5g/kg, 85g/kg, 85.5g/kg, 86g/kg, 86.5g/kg, 87g/kg, 87.5g/kg, 88g/kg, 88.5g/kg, 89g/kg, 89.5g/kg, 90g/kg, 90.5g/kg, 91g/kg, 91.5g/kg, 92g/kg, 92.5g/kg, 93g/kg, 93.5g/kg, 94g/kg, 94.5g/kg, 95g/kg, 95.5g/kg, 96g/kg, 96.5g/kg, 97g/kg, 97.5g/kg, 98g/kg, 98.5g/kg, 99g/kg, 99.5g/kg, or 100 g/kg.

The at least one protein in the nutritional composition may be from about 10g/kg to about 100 g/kg. The at least one protein in the nutritional composition may be from about 20g/kg to about 80 g/kg. The at least one protein in the nutritional composition may be from about 30g/kg to about 70 g/kg. The at least one protein in the nutritional composition may be from about 40g/kg to about 60 g/kg. For example, at least one protein in the nutritional composition may be about 10g/kg, 10.5g/kg, 11g/kg, 11.5g/kg, 12g/kg, 12.5g/kg, 13g/kg, 13.5g/kg, 14g/kg, 14.5g/kg, 15g/kg, 15.5g/kg, 16g/kg, 16.5g/kg, 17g/kg, 17.5g/kg, 18g/kg, 18.5g/kg, 19g/kg, 19.5g/kg, 20g/kg, 20.5g/kg, 21g/kg, 21.5g/kg, 22g/kg, 22.5g/kg, 23g/kg, 23.5g/kg, 24g/kg, 24.5g/kg, 25g/kg, 25.5g/kg, 26g/kg, 26.5g/kg, 27g/kg, 27.5g/kg, 28g/kg, 28.5g/kg, 29g/kg, 29.5g/kg, 30g/kg, 30.5g/kg, 31g/kg, 31.5g/kg, 32g/kg, 32.5g/kg, 33g/kg, 33.5g/kg, 34g/kg, 34.5g/kg, 35g/kg, 35.5g/kg, 36g/kg, 36.5g/kg, 37g/kg, 37.5g/kg, 38g/kg, 38.5g/kg, 39g/kg, 39.5g/kg, 40g/kg, 40.5g/kg, 41g/kg, 41.5g/kg, 42g/kg, 42.5g/kg, 43g/kg, 43.5g/kg, 44g/kg, 44.5g/kg, 45g/kg, 45.5g/kg, 46g/kg, 46.5g/kg, 47g/kg, 47.5g/kg, 48g/kg, 48.5g/kg, 49g/kg, 49.5g/kg, 50g/kg, 50.5g/kg, 51g/kg, 51.5g/kg, 52g/kg, 52.5g/kg, 53g/kg, 53.5g/kg, 54g/kg, 54.5g/kg, 55g/kg, 55.5g/kg, 56g/kg, 56.5g/kg, 57g/kg, 57.5g/kg, 58g/kg, 58.5g/kg, 59g/kg, 59.5g/kg, 60g/kg, 60.5g/kg, 61g/kg, 61.5g/kg, 62g/kg, 62.5g/kg, 63g/kg, 63.5g/kg, 50g/kg, 50.5g/kg, 56g/kg, 51.5g/kg, 52g/kg, 52.5g/kg, 50.5g/kg, 64g/kg, 64.5g/kg, 65g/kg, 65.5g/kg, 66g/kg, 66.5g/kg, 67g/kg, 67.5g/kg, 68g/kg, 68.5g/kg, 69g/kg, 69.5g/kg, 70g/kg, 70.5g/kg, 71g/kg, 71.5g/kg, 72g/kg, 72.5g/kg, 73g/kg, 73.5g/kg, 74g/kg, 74.5g/kg, 75g/kg, 75.5g/kg, 76g/kg, 76.5g/kg, 77g/kg, 77.5g/kg, 78g/kg, 78.5g/kg, 79g/kg, 79.5g/kg, 80g/kg, 80.5g/kg, 81g/kg, 81.5g/kg, 82g/kg, 82.5g/kg, 69.5g/kg, 70g/kg, 70.5g/kg, 74.5g/kg, and the like, 82.5g/kg, 83.5g/kg, 84g/kg, 84.5g/kg, 85g/kg, 85.5g/kg, 86g/kg, 86.5g/kg, 87g/kg, 87.5g/kg, 88g/kg, 88.5g/kg, 89g/kg, 89.5g/kg, 90g/kg, 90.5g/kg, 91g/kg, 91.5g/kg, 92g/kg, 92.5g/kg, 93g/kg, 93.5g/kg, 94g/kg, 94.5g/kg, 95g/kg, 95.5g/kg, 96g/kg, 96.5g/kg, 97g/kg, 97.5g/kg, 98g/kg, 98.5g/kg, 99g/kg, 99.5g/kg, or 100 g/kg.

The ratio and amount between the different compounds is critical for the resulting assembly formed during the composition. To form a stable high molecular weight assembly, the weight/weight ratio of MDG to lipophilic compound in the nutritional composition may be from about 12,000: 1 to about 1: 1. The weight/weight ratio of MDG to lipophilic compound in the nutritional composition may be from about 10,000: 1 to about 1: 1. The weight/weight ratio of MDG to lipophilic compound in the nutritional composition may be from about 1,000: 1 to about 1: 1. The weight/weight ratio of MDG to lipophilic compound in the nutritional composition may be from about 100: 1 to about 1: 1. For example, the weight/weight ratio of MDG to lipophilic compound in the nutritional composition may be about 12000: 1, 11500: 1, 11000: 1, 10500: 1, 10000: 1, 9500: 1, 9000: 1, 8500: 1, 8000: 1, 7500: 1, 7000: 1, 6500: 1, 6000: 1, 5500: 1, 5000: 1, 4900: 1, 4800: 1, 4700: 1, 4600: 1, 4500: 1, 4400: 1, 4300: 1, 4200: 1, 4100: 1, 4000: 1, 3900: 1, 3800: 1, 3700: 1, 3600: 1, 3500: 1, 3400: 1, 3300: 1, 3200: 1, 3100: 1, 2900: 1, 2800: 1, 2700: 1, 2600: 1, 2400: 1, 1800: 1, 2200: 1, 2100: 1, 1700: 1, 1400: 1, 3000: 1, 1400: 1, 2000: 1, 3000: 1, 2000: 1, 2000: 1, 2: 1, 2: 1, 2: 1, 2: 1, 2: 1, 850: 1, 800: 1, 750: 1, 700: 1, 650: 1, 600: 1, 550: 1, 500: 1, 450: 1, 400: 1, 350: 1, 300: 1, 250: 1, 200: 1, 150: 1, 100: 1, 90: 1, 80: 1, 70: 1, 60: 1, 50: 1, 40: 1, 30: 1, 20: 1, 10: 1, 9: 1, 8: 1, 7: 1, 6: 1, 5: 1, 4: 1, 3: 1, 2: 1, or 1: 1.

One embodiment of the nutritional composition may comprise about 145mg/kg to about 600mg/kg MDG and about 1.12g/kg lipophilic compounds.

In another embodiment, the nutritional composition may comprise about 0.1mg/kg to about 10mg/kg MDG and about 20 μ g/kg vitamin D.

b. Assembly object

The nutritional composition may comprise an assembly. The assembly may comprise at least one fat, at least one protein and at least one lipophilic compound. The assembly may comprise MDG, at least one hydrophobic protein and at least one lipophilic compound. It is well known in the art that lipophilic compounds have limited water solubility, resulting in reduced bioavailability when consumed as part of a nutritional composition. The assembly increases the water solubility of the lipophilic compound. Thus, the assemblies enhance the bioavailability of lipophilic compounds, as well as their ultimate absorption in the Gastrointestinal (GI) tract relative to controls that do not contain MDG and hydrophobic proteins.

The assembly is water soluble and the interaction between the MDG, the at least one hydrophobic protein and the at least one lipophilic compound allows the stability of the assembly to withstand high speed centrifugal forces. For example, the nutritional composition may be centrifuged at: 1, 50, 100, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10000, 10500, 11000, 11500, 12000, 12500, 13000, 13500, 14000, 14500, 15000, 15500, 16000, 16500, 17000, 17500, 18000, 18500, 19000, 19500, 20000, 20500, 21000, 21500, 22000, 22500, 23000, 23500, 24000, 24569, 45000, 2000, 3000, 200, and/or 4, and/or a pharmaceutically acceptable salt thereof, 25000, 25500, 26000, 26500, 27000, 27500, 28000, 28500, 29000, 29500, 30000, 30500, 31000, 31500, 32000, 32500, 33000, 33500, 34000, 34500, 35000, 35500, 36000, 37000, 37500, 38000, 39000, 40000, 40500, 41000, 41500, 42000, 42500, 43000, 43500, 44000, 44500, 45000, 45500, 46000, 46500, 47000, 47500, 48000, 49000, 49500, 50000, 50500, 47000, 47500, 48000, 49000, 49500, and the like, 51000, 51500, 52000, 53000, 53500, 54000, 55000, 55500, 56000, 57000, 57500, 58000, 59000, 60000, 60500, 61000, 61500, 62000, 62500, 63000, 64000, 65000, 65500, 66000, 66500, 67000, 67500, 68000, 68500, 69000, 69500, 70000, 70500, 71000, 71500, 72000, 72500, 73000, 74000, 75000, 76000, 76500, 53500, 54000, 550500, 56000, 560500, 5600, 600000, and 64000, 77000x g, 77500x g, 78000x g, 78500x g, 79000x g, 79500x g, 80000x g, 81000x g, 82000x g, 82500x g, 83000x g, 84000x g, 85000x g, 86000x g, 87000x g, 87500x g, 89000x g, 90000x g, 91000x g, 92000x g, 92500x g, 93000x g, 94000x g, 95000x g, 95500x g, 96000x g, 97000x g, 98000x g, 98500 983672, 99000x g, x g or 100000x g.

In some embodiments, the assembly remains water soluble after centrifugation at about 31,000x g for at least 1 hour at 20 ℃. In some embodiments, at least 5% of the assembly remains in the aqueous phase after centrifugation.

The nutritional composition may be spun down using a centrifuge to assess the amount of fats, proteins, and lipophilic compounds remaining in the aqueous phase. It is hypothesized, but not bound by any particular theory, that the fatty and lipophilic compounds remaining in the aqueous phase after centrifugation may bind to the assembly due to their limited water solubility. It should be noted that different centrifugation speeds and times may be used to centrifuge the nutritional composition for sedimentation. Thus, it will be appreciated by those skilled in the art that different centrifugation conditions may provide different amounts of the assembly and corresponding compound in the aqueous phase, but will still allow quantification of the desired compound.

In some embodiments, the details of centrifugation are as follows: centrifugal speed: 1000x g; centrifuging time: 15 min; centrifugation temperature: 20 ℃; centrifuging the tube: 50ml Cellstar tubes from Greiner bio-one; pipe diameter: 23 mm; amount of product in tube: 45ml of the solution; a centrifuge: eppendorf 5810R type; and the aqueous phase was separated via the cream layer by syringe.

In some embodiments, the details of centrifugation are as follows: centrifugal speed: 4500x g; centrifuging time: 15 min; centrifugation temperature: 20 ℃; centrifuging the tube: 50ml Cellstar tubes from Greiher bio-one; pipe diameter: 23 mm; amount of product in tube: 45ml of the solution; a centrifuge: eppendorf 5810R type; and the aqueous phase was separated via the cream layer by syringe.

In some embodiments, the details of centrifugation are as follows: centrifugal speed: 100,000x g; centrifuging time: 1 h; centrifugation temperature: 20 ℃; centrifuging the tube: ultra Clear centrifuge tubes from Beckmann Coulter; tube size: 14x89 mm; amount of product in tube: 11.3 g; a centrifuge: beckmann Coulter type L-90K; the aqueous phase was separated by freezing the test tube with the contents in liquid nitrogen and cutting the portion of the tube including the wall of the tube with the slurry phase, followed by thawing the contents of this portion of the test tube.

In some embodiments, the centrifugation procedure may be performed as follows: approximately 35g of the nutritional composition was transferred to a 50mL polypropylene centrifuge tube (capable of withstanding 100,000x g; e.g., VWR catalog number 21007-290). The tubes were capped and placed in a JA-20 rotor in a Beckman Coulter model Avanti J-E centrifuge. The temperature was set at 20 ℃ and the tubes were centrifuged at 31,000x g (16,000rpm) for 1 to 4 hours. The tube was removed from the centrifuge and approximately 0.6 to 1.0g of supernatant was transferred to a 10mL volumetric flask that had been tared and the supernatant sample weight was recorded. A sample of the supernatant in a 10mL volumetric flask was diluted to 7.0g with Milli-Q Plus water and stirred for five minutes. The diluted supernatant sample in the 10mL volumetric flask was then diluted to 10mL with acetonitrile containing 0.33% (v/v) trifluoroacetic acid and stirred for an additional five minutes. The flask was stoppered and inverted repeatedly for thorough mixing. From the flask, 1.0 to 1.5mL aliquots were transferred to HPLC autosampler vials. The vial was capped and the aqueous phase was tested for the presence of different variables (e.g., MDG, lutein, hydrophobic proteins, etc.) for aliquots.

In one embodiment, the nutritional composition may be centrifuged at 1,000x g for 1 hour at 20 ℃. In one embodiment, the nutritional composition may be centrifuged at 31,000x g for 1 hour at 20 ℃. In one embodiment, the nutritional composition may be centrifuged at 100,000x g for 1 hour at 20 ℃.

The MDG in the aqueous phase may be about 0.001% to about 50% of the nutritional composition after centrifugation at 1,000x g for 1 hour at 20 ℃. The MDG in the aqueous phase may be about 0.5% to about 40% of the nutritional composition after centrifugation at 1,000x g for 1 hour at 20 ℃. The MDG in the aqueous phase may be about 5% to about 30% of the nutritional composition after centrifugation at 1,000x g for 1 hour at 20 ℃. The MDG in the aqueous phase may be about 15% to about 25% of the nutritional composition after centrifugation at 1,000x g for 1 hour at 20 ℃. For example, the MDG in the aqueous phase may be about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.25%, 1.5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%, 4.25%, 4.5%, 4.75%, 5%, 5.25%, 5.5%, 5.75%, 6%, 6.25%, 6.5%, 6.75%, 7%, 7.25%, 7.5%, 7.75%, 8%, 8.25%, 8.5%, 8.75%, 9.75%, 10.75%, 11.75%, 10.75%, 11.75%, 14.75%, 10.75%, 11.75%, 14.75%, 10.75%, 10%, 10.75%, 10%, 10.75%, 11.75%, 10%, 10.75%, 10%, 10.75%, 10%, 1.75%, 1% of the nutritional composition after centrifugation at 20%, 14.75%, 15%, 15.25%, 15.5%, 15.75%, 16%, 16.25%, 16.5%, 16.75%, 17%, 17.25%, 17.5%, 17.75%, 18%, 18.25%, 18.5%, 18.75%, 19%, 19.25%, 19.5%, 19.75%, 20%, 20.25%, 20.5%, 20.75%, 21%, 21.25%, 21.5%, 21.75%, 22%, 22.25%, 22.5%, 22.75%, 23%, 23.25%, 23.5%, 23.75%, 24%, 24.25%, 24.5%, 24.75%, 25%, 25.25%, 25.5%, 25.75%, 26%, 26.25%, 26.5%, 26.75%, 27%, 27.25%, 27.5%, 27.75%, 28%, 28.25%, 28.75%, 29%, 29.25%, 29.5%, 29.75%, 30%, 30.5%, 30.25%, 31.25%, 31.75%, 33.75%, 33.35%, 31.75%, 33.25%, 31.75%, 31.25%, 31.75%, 33.75%, 33.35%, 33.25%, 31.25%, 31.75%, 33.75%, 33.25%, 31.25%, 31%, 31.75%, 31.25%, 31.75%, 35%, 31.25%, 35%, 31.25%, 35.25%, 35%, 35.25%, 35% or more, 36.25%, 36.5%, 36.75%, 37%, 37.25%, 37.5%, 37.75%, 38%, 38.25%, 38.5%, 38.75%, 39%, 39.25%, 39.5%, 39.75%, 40%, 40.25%, 40.5%, 40.75%, 41%, 41.25%, 41.5%, 41.75%, 42%, 42.25%, 42.5%, 42.75%, 43%, 43.25%, 43.5%, 43.75%, 44%, 44.25%, 44.5%, 44.75%, 45%, 45.25%, 45.5%, 45.75%, 46%, 46.25%, 46.5%, 46.75%, 47%, 47.25%, 47.5%, 47.75%, 48%, 48.25%, 48.5%, 48.75%, 49%, 49.25%, 49.5%, 49.75%, or 50%.

The MDG in the aqueous phase may be about 0.001% to about 30% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. The MDG in the aqueous phase may be about 0.1% to about 20% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. The MDG in the aqueous phase may be about 1% to about 5% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. For example, the MDG in the aqueous phase may be about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.25%, 1.5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%, 4.25%, 4.5%, 4.75%, 5%, 5.25%, 5.5%, 5.75%, 6%, 6.25%, 6.5%, 6.75%, 7%, 7.25%, 7.5%, 7.75%, 8%, 8.25%, 8.5%, 8.75%, 9.75%, 10.75%, 11.75%, 10.75%, 14.75%, 10%, 10.75%, 10%, 10.75%, 10%, 11.75%, 10.75%, 10%, 10.75%, 10%, 10.75%, 10%, 10.75%, 1.75%, 1% of the nutritional composition after centrifugation at 20%, 14.75%, 15%, 15.25%, 15.5%, 15.75%, 16%, 16.25%, 16.5%, 16.75%, 17%, 17.25%, 17.5%, 17.75%, 18%, 18.25%, 18.5%, 18.75%, 19%, 19.25%, 19.5%, 19.75%, 20%, 20.25%, 20.5%, 20.75%, 21%, 21.25%, 21.5%, 21.75%, 22%, 22.25%, 22.5%, 22.75%, 23%, 23.25%, 23.5%, 23.75%, 24%, 24.25%, 24.5%, 24.75%, 25%, 25.25%, 25.5%, 25.75%, 26%, 26.25%, 26.5%, 26.75%, 27%, 27.25%, 27.5%, 27.75%, 28%, 28.25%, 28.5%, 28.75%, 29%, 29.25%, 29.75%, or 30%.

The MDG in the aqueous phase may be about 0.001% to about 20% of the nutritional composition after centrifugation at 100,000x g for 1 hour at 20 ℃. The MDG in the aqueous phase may be about 0.01% to about 15% of the nutritional composition after centrifugation at 100,000x g for 1 hour at 20 ℃. The MDG in the aqueous phase may be about 0.1% to about 10% of the nutritional composition after centrifugation at 100,000x g for 1 hour at 20 ℃. The MDG in the aqueous phase may be about 1% to about 5% of the nutritional composition after centrifugation at 100,000x g for 1 hour at 20 ℃. For example, the MDG in the aqueous phase may be about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.6%, 3.8%, 4.5%, 4.6%, 4.5%, 4%, 4.6%, 4%, 6%, 4.5%, 6%, 1.5%, 1%, 1.6%, 1%, 6%, 1.6%, 5%, 4.6%, 1%, 5%, 4.6%, 1.6%, 4.6%, 1%, 1.6%, 1%, 1.6%, 4% of the nutritional composition after centrifugation at 20 ℃ for 1.5% at 100,25% of a period of a 1.2, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11%, 11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%, 11.7%, 11.8%, 11.9%, 12.1%, 12.12%, 12.5%, 12.12.12%, 12.5%, 14.6%, 14.7%, 14.6%, 14.9%, 14.7%, 14.6%, 14.4%, 14.6%, 14.7%, 14.6%, 14.2%, 14.12.12%, 14.12%, 14.12.12.12%, 14.12.12%, 14.2%, 14.6%, 14.7%, 14.6%, 14.2%, 14.7%, 14.9%, 14.6%, 14.2%, 14.6%, 14.7%, 14.2%, 14.7%, 14.6%, 14.9%, 14.6%, 14.2%, 14.6%, 14.7%, 14.2%, 14.9%, 14.2%, 14.7%, 14.9%, 14.9.9%, 14.9%, 14.6%, 14.9%, 14.2%, 14.6%, 14.9%, 14.6%, 14.2%, 14.9%, 14.6%, 14.2%, 14.9%, 14.6%, 14.9%, 14.7%, 14.2%, 14.9.9.2%, 14.9%, 9%, 14.9%, 9%, 14.9%, 9%, 14.9%, 9.9.9%, 14.9.9.2%, 14.1%, 14.9%, 14.2%, 14.9%, 9%, 14.2%, 14.9%, 14.6%, 14.2%, 9%, 9.6%, 9%, 9.7%, 14.6%, 9%, 14.6%, 14.9%, 14.1%, 14.6%, 14.2%, 9%, 14.9, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16%, 16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%, 16.7%, 16.8%, 16.9%, 17%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%, 17.9%, 18%, 18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8%, 19.9%, or 20%.

The MDG in the assembly may be about 0.001% to about 20% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. The MDG in the assembly may be about 0.01% to about 15% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. The MDG in the assembly may be about 0.1% to about 10% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. The MDG in the assembly may be about 1% to about 5% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. For example, the MDG in the assembly may be about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.5%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.5%, 3.7%, 4.5%, 4%, 6%, 4.5%, 4%, 3.5%, 4%, 4.5%, 6%, 1%, 1.5%, 1%, 1.5%, 1.6%, 1%, 1.6%, 1%, 1.6%, 6%, 1.6%, 1%, 1.6%, 4% of the nutritional composition after centrifugation at 20% at 20 ℃ for 1.5%, 1%, 1.6%, 1% of a 1.6% of a period of a nutritional composition after 1.6% at 20, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11%, 11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%, 11.7%, 11.8%, 11.9%, 12.12%, 12.7%, 12.12.12%, 12.5%, 12.12.12%, 14.7%, 14.12.12.12%, 14.6%, 14.7%, 14.9.9.9%, 14.7%, 14.6%, 14.7%, 14.9%, 14.6%, 14.7%, 14.6%, 14.7%, 14.6%, 14.1%, 14.6%, 12.6%, 14.6%, 12.6%, 14.6%, 14.7%, 14.6%, 14.1%, 14.6%, 12.6%, 14.6%, 14.1%, 14.6%, 12.6%, 14.6%, 12.6%, 14.6%, 14.1%, 14.6%, 14.1%, 14.6%, 12.6%, 12, 15%, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16%, 16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%, 16.7%, 16.8%, 16.9%, 17%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%, 17.9%, 18%, 18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8%, 19.9%, or 20%.

The lutein (lipophilic compound discussed below) in the aqueous phase may be about 0.001% to about 50% of the nutritional composition after centrifugation at 1,000x g for 1 hour at 20 ℃. The lutein in the aqueous phase may be about 0.5% to about 40% of the nutritional composition after centrifugation at 1,000x g for 1 hour at 20 ℃. The lutein in the aqueous phase may be about 5% to about 30% of the nutritional composition after centrifugation at 1,000x g for 1 hour at 20 ℃. The lutein in the aqueous phase may be about 15% to about 25% of the nutritional composition after centrifugation at 1,000x g for 1 hour at 20 ℃. For example, the lutein in the aqueous phase may be about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.25%, 1.5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%, 4.25%, 4.5%, 4.75%, 5%, 5.25%, 5.5%, 5.75%, 6%, 6.25%, 6.5%, 6.75%, 7%, 7.25%, 7.5%, 7.75%, 8%, 8.25%, 8.5%, 8.9%, 9.75%, 10.75%, 11.75%, 10.75%, 11.75%, 13%, 10%, 10.75%, 10%, 1.75%, 1%, 1.75%, 1%, 1.75%, 1% of the nutritional composition, 14.5%, 14.75%, 15%, 15.25%, 15.5%, 15.75%, 16%, 16.25%, 16.5%, 16.75%, 17%, 17.25%, 17.5%, 17.75%, 18%, 18.25%, 18.5%, 18.75%, 19%, 19.25%, 19.5%, 19.75%, 20%, 20.25%, 20.5%, 20.75%, 21%, 21.25%, 21.5%, 21.75%, 22%, 22.25%, 22.5%, 22.75%, 23%, 23.25%, 23.5%, 23.75%, 24%, 24.25%, 24.5%, 24.75%, 25%, 25.25%, 25.5%, 25.75%, 26%, 26.25%, 26.5%, 26.75%, 27%, 27.25%, 27.5%, 27.75%, 28%, 28.25%, 28.5%, 28.75%, 29%, 29.25%, 29.5%, 29.30.25%, 30.25%, 30.75%, 31.75%, 33.75%, 31.25%, 31.75%, 33.75%, 33.25%, 31.25%, 31.75%, 31.25%, 31.75%, 33.25%, 31.25%, 31.75%, 31.25%, 35%, 31.75%, 31.25%, 35%, 31.25%, 35%, 31%, 35%, 31.25%, 31%, 35%, 31.25%, 35%, 31%, 35%, 31.25%, 35%, 31.25%, 35%, 31.25%, 35%, 31.25%, 35%, 31.25%, 35%, 31.25, 36%, 36.25%, 36.5%, 36.75%, 37%, 37.25%, 37.5%, 37.75%, 38%, 38.25%, 38.5%, 38.75%, 39%, 39.25%, 39.5%, 39.75%, 40%, 40.25%, 40.5%, 40.75%, 41%, 41.25%, 41.5%, 41.75%, 42%, 42.25%, 42.5%, 42.75%, 43%, 43.25%, 43.5%, 43.75%, 44%, 44.25%, 44.5%, 44.75%, 45%, 45.25%, 45.5%, 45.75%, 46%, 46.25%, 46.5%, 46.75%, 47%, 47.25%, 47.5%, 47.75%, 48%, 48.25%, 48.5%, 48.75%, 49%, 49.25%, 49.5%, 49.75%, or 50%.

The lutein in the aqueous phase may be about 0.001% to about 30% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. The lutein in the aqueous phase may be about 0.1% to about 20% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. The lutein in the aqueous phase may be about 1% to about 5% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. For example, lutein in the assembly can be about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.25%, 1.5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%, 4.25%, 4.5%, 4.75%, 5%, 5.25%, 5.5%, 5.75%, 6%, 6.25%, 6.5%, 6.75%, 7%, 7.25%, 7.5%, 7.75%, 8%, 8.25%, 8.5%, 9.25%, 9.75%, 10.75%, 11.75%, 10.75%, 10%, 10.75%, 10%, 1%, 1.75%, 1%, 1.75%, 1% of the nutritional composition at 20%, 1% of the composition, 14.5%, 14.75%, 15%, 15.25%, 15.5%, 15.75%, 16%, 16.25%, 16.5%, 16.75%, 17%, 17.25%, 17.5%, 17.75%, 18%, 18.25%, 18.5%, 18.75%, 19%, 19.25%, 19.5%, 19.75%, 20%, 20.25%, 20.5%, 20.75%, 21%, 21.25%, 21.5%, 21.75%, 22%, 22.25%, 22.5%, 22.75%, 23%, 23.25%, 23.5%, 23.75%, 24%, 24.25%, 24.5%, 24.75%, 25%, 25.25%, 25.5%, 25.75%, 26%, 26.25%, 26.5%, 26.75%, 27%, 27.25%, 27.5%, 27.75%, 28%, 28.25%, 28.5%, 28.75%, 29%, 29.25%, 29.5%, 29.75%, or 30%.

The lutein in the aqueous phase may be about 0.001% to about 20% of the nutritional composition after centrifugation at 100,000x g for 1 hour at 20 ℃. The lutein in the aqueous phase may be about 0.01% to about 15% of the nutritional composition after centrifugation at 100,000x g for 1 hour at 20 ℃. The lutein in the aqueous phase may be about 0.1% to about 10% of the nutritional composition after centrifugation at 100,000x g for 1 hour at 20 ℃. The lutein in the aqueous phase may be about 1% to about 5% of the nutritional composition after centrifugation at 100,000x g for 1 hour at 20 ℃. For example, the lutein in the aqueous phase may be about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.4%, 3.6%, 3.7%, 3.8%, 4.5%, 4%, 6%, 4.5%, 6%, 4.6%, 1%, 1.6%, 1%, 5%, 1%, 5%, 6%, 1.6%, 5%, 1.5%, 4.6%, 1%, 4.6%, 1.6%, 1%, 1.5%, 4%, 6%, 4.6%, 4% of the nutritional composition after centrifugation at 20 ℃ for 1% at 20% of a period, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11%, 11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%, 11.7%, 11.8%, 11.9%, 12.12%, 12.7%, 12.12.12%, 12.5%, 12.12.12%, 14.7%, 14.12.12.12%, 14.6%, 14.7%, 14.9.9.9%, 14.7%, 14.6%, 14.7%, 14.9%, 14.6%, 14.7%, 14.6%, 14.7%, 14.6%, 14.1%, 14.6%, 12.6%, 14.6%, 12.6%, 14.6%, 14.7%, 14.6%, 14.1%, 14.6%, 12.6%, 14.6%, 14.1%, 14.6%, 12.6%, 14.6%, 12.6%, 14.6%, 14.1%, 14.6%, 14.1%, 14.6%, 12.6%, 12, 15%, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16%, 16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%, 16.7%, 16.8%, 16.9%, 17%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%, 17.9%, 18%, 18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8%, 19.9%, or 20%.

The lutein in the assembly may be about 0.001% to about 20% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. The lutein in the assembly may be about 0.01% to about 15% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. The lutein in the assembly may be about 0.1% to about 10% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. The lutein in the assembly may be about 1% to about 5% of the nutritional composition after centrifugation at 31,000x g for 1 hour at 20 ℃. For example, lutein in the assembly may be about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 4.5%, 4.6%, 4.5%, 4.6%, 4.5%, 4%, 6%, 4.5%, 4%, 6%, 4.5%, 6%, 1.5%, 1%, 3.6%, 3.5%, 4%, 4.5%, 4% of the nutritional composition after centrifugation at 20 ℃ for 1.5% at 31.6% for 1.6% for 1.5% for 1% of a period of a time, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11%, 11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%, 11.7%, 11.8%, 11.9%, 12.12%, 12.7%, 12.12.12%, 12.5%, 12.12.12%, 14.7%, 14.12.12.12%, 14.6%, 14.7%, 14.9.9.9%, 14.7%, 14.6%, 14.7%, 14.9%, 14.6%, 14.7%, 14.6%, 14.7%, 14.6%, 14.1%, 14.6%, 12.6%, 14.6%, 12.6%, 14.6%, 14.7%, 14.6%, 14.1%, 14.6%, 12.6%, 14.6%, 14.1%, 14.6%, 12.6%, 14.6%, 12.6%, 14.6%, 14.1%, 14.6%, 14.1%, 14.6%, 12.6%, 12, 15%, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16%, 16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%, 16.7%, 16.8%, 16.9%, 17%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%, 17.9%, 18%, 18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8%, 19.9%, or 20%.

MDG plays a key role in the formation and stability of compounds associated with the assembly, where MDG increases the concentration of proteins and bound lipophilic material remaining in the aqueous phase after centrifugation relative to a control without MDG. MDG in the nutritional composition may increase the concentration of protein in the aqueous phase by about 0.001% to about 20% after centrifugation at 31,000x g for 1 hour at 20 ℃ compared to a control without MDG. MDG in the nutritional composition may increase the concentration of protein in the aqueous phase by about 0.01% to about 15% after centrifugation at 31,000x g for 1 hour at 20 ℃ compared to a control without MDG. MDG in the nutritional composition may increase the concentration of protein in the aqueous phase by about 0.1% to about 10% after centrifugation at 31,000x g for 1 hour at 20 ℃ compared to a control without MDG. MDG in the nutritional composition may increase the concentration of protein in the aqueous phase by about 1% to about 5% after centrifugation at 31,000x g for 1 hour at 20 ℃ compared to a control without MDG. For example, after centrifugation at 31,000x g for 1 hour at 20 ℃, the protein in the aqueous phase can be increased by about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 4.5%, 3.5%, 4.5%, 4%, 4.5%, 4%, 4.5%, 4%, 4.5%, 4%, 6%, 4%, 4.5%, 4% or 5%, 4% of a, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%, 11.7%, 11.6%, 11.12%, 12.5%, 12.12%, 12.7%, 13.12.12%, 13.6%, 14.7%, 14.6%, 12.6%, 14.7%, 12.12%, 14.12.12%, 14.7%, 14.12%, 13.12%, 14.7%, 14.6%, 9.6%, 9%, 10.6%, 12.6%, 12%, 12.6%, 12.1%, 12.6%, 12.1%, 12.6%, 12.1%, 12.1.1%, 12%, 12.1%, 12, 14.8%, 14.9%, 15%, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16%, 16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%, 16.7%, 16.8%, 16.9%, 17%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%, 17.9%, 18%, 18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8%, 19.9%, or 20%.

MDG in the nutritional composition may increase bound lipophilic compounds in the aqueous phase by about 0.001% to about 20% after centrifugation at 31,000x g for 1 hour at 20 ℃ compared to a control without MDG. MDG in the nutritional composition may increase bound lipophilic compounds in the aqueous phase by about 0.01% to about 15% after centrifugation at 31,000x g for 1 hour at 20 ℃ compared to a control without MDG. MDG in the nutritional composition may increase bound lipophilic compounds in the aqueous phase by about 0.1% to about 10% after centrifugation at 31,000x g for 1 hour at 20 ℃ compared to a control without MDG. MDG in the nutritional composition may increase bound lipophilic compounds in the aqueous phase by about 1% to about 5% after centrifugation at 31,000x g for 1 hour at 20 ℃ compared to a control without MDG. For example, after centrifugation at 31,000x g for 1 hour at 20 ℃, bound lipophilic compounds in aqueous phase and assemblies can be increased by about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.5%, 4.5%, 4%, 4.5%, 4%, 5%, 4.5%, 4%, 5%, 4%, 4.5%, 4%, 5%, 4% of a, 5.9%, 6%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11%, 11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%, 11.2%, 11.5%, 11.6%, 7.4%, 12.12.12%, 12.12%, 13.12.12%, 13.12.6%, 13.12%, 13.6%, 13.7%, 13.9.6%, 7%, 9.8%, 9.6%, 9.7%, 9.6%, 9.7%, 9.8%, 9.7%, 12.8%, 12.6%, 12.12.12.12.12.12%, 13%, 12.12.12.12.12.12.12.12%, 13%, 13.2%, 13%, 12.6%, 13.6%, 12.6%, 13.6%, 12.1%, 13.6%, 12.6%, 13.6%, 12.6%, 13.6%, 12.6%, 12.12.12.6%, 13.6%, 12.6%, 13.6%, 9%, 13.6%, 12.6%, 12.12.6%, 12.6%, 13.12.12.6%, 9%, 13.6%, 12.6%, 13.6%, 13%, 13.6%, 9%, 12.12.12.12.12.12.6%, 9%, 13%, 12.12, 14.5%, 14.6%, 14.7%, 14.8%, 14.9%, 15%, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16%, 16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%, 16.7%, 16.8%, 16.9%, 17%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%, 17.9%, 18%, 18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8%, 19.9%, or 20%.

The size of the assembly depends on the components that make up the nutritional powder, such as MDG, lipophilic compounds, proteins, and the like. The size of the assembly may be critical to the bioavailability of the lipophilic compound. For example, lutein uptake will increase with increasing assembly size. However, assembly sizes larger than outside the scope of the present invention may lead to complications related to solubility and digestibility after consumption. The assembly size can be measured via size exclusion chromatography techniques known in the art and then compared to known calibration curves. The assembly size may also be measured by size analysis techniques known in the art, including, but not limited to, laser diffraction, dynamic light scattering, sieve separation analysis, and image analysis (e.g., using microscopic methods such as optical microscopy or scanning electron microscopy). The assembly size can be measured by Malvern Zetasizer Nano NS. In some embodiments, the nutritional composition may be centrifuged as described above, and the aqueous portion may then be analyzed by a dynamic light scattering machine. In addition, the size of the assembly can be measured via Size Exclusion Chromatography (SEC) after the centrifugation step as described above. This is done by using the reference protein and its SEC elution time and molecular weight. This can then be used to estimate the size of the assembly. By extrapolation from the reference protein plot. The size of the assembly may be from about 15kD to about 3000 kD. The size of the assembly may be from about 100kD to 1000 kD. The size of the assembly may be from about 100kD to about 500 kD. The size of the assembly may be from about 60kD to about 400 kD. For example, the size of the assembly may be about 15kD, 20kD, 25kD, 30kD, 35kD, 40kD, 45kD, 50kD, 55kD, 60kD, 65kD, 70kD, 75kD, 80kD, 85kD, 90kD, 95kD, 100kD, 105kD, 110kD, 115kD, 120kD, 125kD, 130kD, 135kD, 140kD, 145kD, 150kD, 155kD, 160kD, 165kD, 170kD, 175kD, 180kD, 185kD, 190kD, 195kD, 200kD, 205kD, 210kD, 215kD, 220kD, 225kD, 230kD, 235kD, 240kD, 245kD, 250kD, 255kD, 260kD, 265kD, 270kD, 275kD, 280kD, 285kD, 290kD, 295kD, 300kD, 305kD, 310kD, 315kD, 320kD, 325kD, 330kD, 335kD, 340kD, 375kD, 420kD, 380kD, 400kD, 1 kD, 400kD, 1 kD, 150kD, 1 kD, 150kD, 1 kD, 150kD, 1 kD, 150kD, 1 kD, 150kD, 1 kD, 150kD, 1 kD, 1 kD, 150kD, 1 k, 425kD, 430kD, 435kD, 440kD, 445kD, 450kD, 455kD, 460kD, 465kD, 470kD, 475kD, 480kD, 485kD, 490kD, 495kD, 500kD, 505kD, 510kD, 515kD, 520kD, 525kD, 530kD, 535kD, 540kD, 545kD, 550kD, 555kD, 560kD, 565kD, 570kD, 575, 580kD, 585kD, 590kD, 595kD, 600kD, 605kD, 610kD, 615kD, 620kD, 625kD, 630kD, 635kD, 640kD, 645kD, 650kD, 655kD, 660kD, 665kD, 670kD, 675kD, 680kD, 685kD, 690kD, 695kD, 705kD, 710kD, 720kD, 725kD, 730kD, 735kD, 835kD, 840kD, 750kD, 755, 780kD, 760kD, 775, 785kD, 800kD, 300kD, 800kD, 300kD, 800kD, 300kD, 800kD, 300kD, 800kD, 900kD, 300kD, 800kD, 855kD, 860kD, 870kD, 875kD, 880kD, 885kD, 890kD, 895kD, 900kD, 905kD, 910kD, 915kD, 920kD, 925kD, 930kD, 935kD, 940kD, 945kD, 950kD, 955kD, 960kD, 965kD, 970kD, 975kD, 980kD, 985kD, 990kD, 995kD, 1000kD, 1010kD, 1020kD, 1030kD, 1040kD, 1050kD, 1070kD, 1080kD, 1090kD, 1100kD, 1110kD, 1130kD, 1140kD, 1150kD, 1160kD, 1170kD, 1180kD, 1350kD, 1200kD, 1210kD, 1230kD, 1240kD, 1260kD, 1270kD, 128kD, 1310kD, 1300kD, 1330kD, 1420kD, 20kD, 1250kD, 1450kD, 20kD, 300kD, 20kD, 300kD, 150kD, 300kD, 1450kD, 150kD, 300kD, 1450kD, 20kD, 150kD, 20kD, 300kD, 150kD, 300kD, 150kD, 300kD, 150kD, 300kD, 20kD, 300kD, 150kD, 300kD, 150kD, 300kD, 150kD, 300kD, 150kD, 300kD, 1570kD, 1580kD, 1590kD, 1600kD, 1610kD, 1620kD, 1630kD, 1640kD, 1650kD, 1660 kD, 1670 kD, 1680 kD, 1690 kD, 1700 kD, 1710 kD, 1720 kD, 1730 kD, 1740 kD, 1750 kD, 1760 kD, 1770 kD, 1780 kD, 1790 kD, 1800 kD, 1810 kD, 1820 kD, 1830 kD, 1840 kD, 1850 kD, 1870kD 1880 kD, 1890kD, 1900kD, 1920kD, 1930kD, 1940kD, 1950kD, 1960kD, 1970kD, 1980kD, 1990kD, 2000kD, 2010kD, kD, 2030kD, 2040kD, 2050kD, 2060kD, 190kD, 220kD, 190kD, 220kD, 224 kD, 190kD, 220kD, 2340, 220kD, 220kD, 2250kD, 3000kD, 2290kD, 220kD, 2290kD, 190kD, 220kD, 190kD, 220kD, 190kD, 220kD, 190kD, 220kD, etc, 2360 kD, 2370 kD, 2380kD, 2390kD, 2400kD, 2410kD, 2420kD, 2430kD, 2440kD, 2450kD, 2460kD, 2470kD, 2480kD, 2490 kD, 2500 kD, 2510 kD, 2520 kD, 2530 kD, 2540 kD, 2550 kD, 2560 kD, 2570kD, 2590kD, 2600kD, 2610kD, 2620kD, 2630kD, 2640kD, 2650kD, 2660kD, 2670kD, 2680kD, 2690kD, 2700kD, 2710kD, 2720kD, 2730kD, 2750kD, 2760kD, 280kD, 2810kD, 20kD, 30kD, 2840kD, 2850kD, 2860kD, 2880kD, 2790kD, 290kD, 2980kD, or 2980 kD.

The nutritional composition may include an assembly that increases the bioavailability of the lipophilic compound by about 5% to about 80% relative to a control that does not contain MDG and hydrophobic proteins. The nutritional composition may include an assembly that increases the bioavailability of the lipophilic compound in a range from about 40% to about 55% relative to a control that does not contain MDG and hydrophobic proteins. The nutritional composition may include an assembly that increases the bioavailability of the lipophilic compound in a range from about 70% to about 75% relative to a control that does not contain MDG and hydrophobic proteins. The bioavailability of lipophilic compounds depends on the formation of an assembly of nutritional compositions, wherein MDG and hydrophobic proteins are contained within the assembly. The bioavailability of the assembly can be assessed via lymphatic analysis in rats. In particular, rats may be fasted overnight prior to surgery and may be subjected to laparotomy under anesthesia and may be treated according to Tso et al, "The Absorption of Lipid and Lipid Synthesis", Lipid Research Methodology, chapter 5: 191-216(1984) Alan r. loss, inc., NY (incorporated herein by reference to the extent consistent herewith) intubated the intestinal lymphatic vessels. The superior mesenteric artery was isolated, but not occluded. A silicon infusion tube (1.6mm OD) was placed in the stomach for future infusion of the test sample. The fundoplication is closed by purse string sutures. Rats were allowed to recover for 24 hours before infusion was initiated. At 24 hours post-surgery, the animals were intragastrically infused with 3ml of a lipophilic compound-delivering nutritional composition. Lymph was collected in pre-cooling tubes 1 hour before infusion of the nutritional composition (fasting) and then every other hour after infusion started (for 8 hours). At the end of the 8 hour infusion, the rats were sacrificed by exsanguination.

Lymph lipids were extracted and analyzed for lutein concentration using high performance liquid chromatography with programmed wavelength ultraviolet detection (Craft Technologies, Wilson, NC). For example, the assembly can increase the bioavailability of a lipophilic compound by about 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, 16.5%, 17%, 17.5%, 18%, 18.5%, 19%, 19.5%, 20%, 20.5%, 21%, 21.5%, 22%, 22.5%, 23%, 23.5%, 24%, 24.5%, 25%, 25.5%, 26%, 26.5%, 27%, 27.5%, 28%, 28.5%, 29%, 29.5%, 30%, 30.5%, 31.5%, 32%, 32.5%, 33%, 33.5%, 34%, 34.5%, 35%, 35.5%, 36%, 36.5%, 37%, 37.5%, 38%, 38.5%, 39.5%, 40.5%, 40%, 43.5%, 42%, 42.5%, 42%, 40%, 5%, 25.5%, 25%, 22.5%, 25%, 26.5%, 27%, 27.5%, 25%, 30.5%, 31%, 25%, 31.5%, 31%, 25%, or more, 44%, 44.5%, 45%, 45.5%, 46%, 46.5%, 47%, 47.5%, 48%, 48.5%, 49%, 49.5%, 50%, 50.5%, 51%, 51.5%, 52%, 52.5%, 53%, 53.5%, 54%, 54.5%, 55%, 55.5%, 56%, 56.5%, 57%, 57.5%, 58%, 58.5%, 59%, 59.5%, 60%, 60.5%, 61%, 61.5%, 62%, 62.5%, 63%, 63.5%, 64%, 64.5%, 65%, 65.5%, 66%, 66.5%, 67%, 67.5%, 68%, 68.5%, 69%, 69.5%, 70%, 70.5%, 71%, 71.5%, 72%, 72.5%, 73%, 73.5%, 74%, 74.5%, 75%, 75.5%, 76%, 76.5%, 77%, 77.5%, 78%, 78.5%, 79%, 79.5%, or 80%.

(1) Activating assembly

The assembly may be an activated assembly. The activating assembly may be contained in a powder, suspension, solution, emulsion, or combination thereof. The activation assembly may comprise MDG, at least one lipophilic compound and at least one hydrophobic protein. The activation assembly may be in, but is not limited to, a premix, a protected premix, and an activation premix. The activation assembly may contact the aqueous liquid as part of the activation premix.

(2) Premix/protected premix

The premix may or may not contain an assembly. The protected premix may or may not contain an assembly.

(3) Activating premix assemblies

The activation premix may comprise the assembly. The assembly may be in a protected activation premix. The assembly in the activation premix may comprise MDG, at least one lipophilic compound, and at least one hydrophobic protein. The assemblies in the protected activation premix may comprise MDG, at least one lipophilic compound, and at least one hydrophobic protein. The activation premix comprising the assembly may be added to other components of the nutritional composition. The protected activated premix comprising the assembly may be added to other components of the nutritional composition.

In some embodiments, the premix is contacted with a protein-in-water slurry to form an activated premix. The activation premix is then added to the other components of the nutritional composition. In some embodiments, a premix comprising the assembly is contacted with a protein-in-oil slurry to form an activated premix. The activation premix is then added to the other components of the nutritional composition. In some embodiments, a premix comprising the assembly is contacted with a carbohydrate-mineral slurry and a protein-in-oil slurry to form an activated premix. The activation premix is then added to the other components of the nutritional composition. In some embodiments, a premix comprising the assembly is contacted with the slurry oil to form an activated premix. The activation premix is then added to the other components of the nutritional composition. In some embodiments, a premix comprising the assembly is contacted with the final mixed slurry to form an activated premix. The activation premix is then added to the other components of the nutritional composition.

(4) Shelf life

The nutritional composition may include an assembly having an enhanced shelf life. The assembly may have enhanced shelf life in, but is not limited to, powders, premixes, activation premixes, protected activation premixes, suspensions, liquids, emulsion mixtures, or combinations thereof.

The nutritional composition may include an assembly that is stable at 23 ℃ for at least about 1 day to at least about 36 months. The nutritional composition may include an assembly that is stable at 23 ℃ for at least about 10 days to at least about 36 months. The nutritional composition may include an assembly that is stable at 23 ℃ for at least about 6 months to at least about 12 months. The nutritional composition may include an assembly that is stable at 23 ℃ for at least about 1 day to at least about 24 months. The stability of the assembly depends on the combination of MDG and the ability of the hydrophobic protein to interact with the lipophilic compound, which keeps the lipophilic compound solubilized, thereby enhancing its biocompatibility. The stability of the assembly may be investigated at different time points after formulation of the nutritional composition by size exclusion chromatography techniques known in the art. The assembly can be stable at 23 ℃ for at least about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 24 months, 25 months, 26 months, 27 months, 28 months, 29 months, 30 months, 31 months, 32 months, 33 months, 34 months, 35 months, or 36 months.

Lipophilic compounds

The nutritional composition may comprise at least one lipophilic compound. The lipophilic compound may be present within the premix, the activation premix, and/or the protected premix. Lipophilic compounds may be incorporated into the assembly. Lipophilic compounds have poor water solubility, which results in reduced bioavailability when incorporated into nutritional compositions. However, by incorporating lipophilic compounds together with MDG and at least one hydrophobic protein within the above-described assembly, the solubility and stability of the lipophilic compounds is higher. The lipophilic compound may be a carotenoid, zeaxanthin, alpha-or beta-cryptoxanthin, retinol, gangliosides, phosphoinositides, lipoproteins, phospholipids, glycolipids, glycophospholipids, oil soluble vitamins (such as the synthetic and natural forms of vitamin A, E, D or K), lycopene or mixtures thereof. The lipophilic compound may be a source of long chain polyunsaturated fatty acids (LCPUFAs). Some examples of LCPUFAs include, but are not limited to, eicosapentaenoic acid ("EPA"), arachidonic acid (ARA), linoleic acid in the n-6 pathway (18:2n-6), gamma-linolenic acid (18:3n-6), dihomo-gamma-linolenic acid (20:3n-6), alpha-linolenic acid (18:3n-3), stearidonic acid (18:4n-3), arachidonic acid (20:4n-3), eicosapentaenoic acid (20:5n-3), and DHA (22:6 n-3). The lipophilic compound may be lutein. In addition, the lipophilic compound may be a mixture of different lipophilic compounds.

Suitable non-limiting examples of lipophilic agents that may be included in the compositions of the present invention may be selected from: oil soluble drugs (e.g., immunosuppressants such as cyclosporine)^TMProtease inhibitors such as Ritonavir^TMMacrolide antibiotics and oil-soluble anesthetics such as Propofol^TM) (ii) a Synthetic and natural forms of steroid hormones such as estrogen, estradiol, progesterone, testosterone, cortisone, phytoestrogens, Dehydroepiandrosterone (DHEA), and growth hormone; oil-soluble acids and alcohols (e.g., tartaric acid, acyl lactic acid, butyl hydroxy anisole, butylated hydroxytoluene, lignin, phytosterols, flavonoids such as quercetin and resveratrol, and diallyl disulfide); and combinations thereof. Other lipophilic agents may be found in U.S. patent application No. 13/452,033 filed 4/20/2012, and which is hereby incorporated by reference in a manner consistent with the present disclosure.

Lipophilic compounds are valuable nutritional sources for infants, toddlers, children, and adults. Lipophilic compounds may be used to treat or maintain appropriate health, such as but not limited to bone, eye, cardiovascular and brain health.

(1) Xanthophyll

The lipophilic compound of the nutritional composition may be lutein. Lutein can be incorporated into the assembly. Incorporation of lutein as part of the assembly enhances its solubility, thereby enhancing its bioavailability for treating a health condition relative to controls that do not contain MDG or hydrophobic proteins. The term "lutein" as used herein, unless otherwise specified, refers to one or more of the following: free lutein, lutein esters, lutein salts, or other lutein derivatives or related structures as described or otherwise suggested herein. Xanthophylls or xanthophyll sources suitable for use in the nutritional compositions of the present disclosure include free lutein as well as esters, salts or other derivatives thereof or related structures, including those that conform to the following formula:

The above formula includes the general structure of lutein and related derivatives or structures. For example, the free lutein corresponds to wherein R₁And R₂Are all of the formula hydrogen and include the cis and trans isomers thereof and salts thereof, e.g., sodium, potassium salts.

Lutein esters suitable for use in the nutritional compositions of the present disclosure include any of the lutein esters of the above formula, wherein R is₁And R₂Are identical or different and are a nutritionally acceptable monovalent salt, hydrogen or the acyl residue of a carboxylic acid, with the proviso that R₁And R₂Is an acyl residue of a carboxylic acid. Suitable lutein esters also include cis and trans isomers. R₁And R₂Partially saturated or unsaturated C₁To C₂₂Non-limiting examples of residues of fatty carboxylic acids include formic acid, acetic acid, propionic acid, butyric acid, valeric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, and oleic acid.

Any natural or synthetic source of lutein is suitable for use herein, provided that such source is also known for use in or otherwise suitable for use in nutritional compositions and is compatible with other selected ingredients within the composition. The source of lutein may be provided as an individual ingredient or in combination with other materials or sources, including sources such as multivitamin premixes, mixed carotenoid premixes, and pure lutein sources. In some embodiments, the components of the nutritional composition may inherently contain lutein (such as lutein-rich oils, surfactants, and the like).

Non-limiting examples of sources of lutein suitable for use herein include those available from Kemin Foods, Des Moines, Iowa, USACrystalline lutein; and LUTIMAX lutein esters supplied by OmniActive, Mumbai, India.

In some embodiments, it may be desirable to mix the lutein with the MDG oil to form an MDG premix prior to mixing the MDG oil or lutein with other ingredients of the composition. To prepare the MDG premix, lutein can be mixed with MDG oil in the presence of heat or at ambient temperature and in some embodiments with agitation to allow the lutein to dissolve, disperse or suspend in the MDG oil.

In some embodiments, it may be desirable to form an MDG protected premix comprising lutein, surfactant, and MDG oil prior to mixing the MDG oil or lutein with other ingredients of the composition. In these embodiments, the lutein and surfactant may be mixed with the MDG oil in the presence of heat or at ambient temperature and in some embodiments with agitation to allow the lutein to dissolve, disperse or suspend in the MDG oil.

Lutein is a good source of nutrition for infants, young children, children and adults. Lutein can be used in nutritional compositions to maintain and supplement healthy vision and eye health. Lutein can be used in nutritional compositions to maintain and supplement brain health and development.

The lutein in the nutritional composition may be about 0.01mg/kg to about 50 mg/kg. The lutein in the nutritional composition may be about 0.02mg/kg to about 20 mg/kg. The lutein in the nutritional composition may be about 0.1mg/kg to about 20 mg/kg. The lutein in the nutritional composition may be about 1mg/kg to about 15 mg/kg. For example, the xanthophyll in the nutritional composition may be about 0.01mg/kg, 0.02mg/kg, 0.03mg/kg, 0.04mg/kg, 0.05mg/kg, 0.1mg/kg, 0.25mg/kg, 0.5mg/kg, 0.75mg/kg, 1mg/kg, 1.25mg/kg, 1.5mg/kg, 1.75mg/kg, 2mg/kg, 2.25mg/kg, 2.5mg/kg, 2.75mg/kg, 3mg/kg, 3.25mg/kg, 3.5mg/kg, 3.75mg/kg, 4mg/kg, 4.25mg/kg, 4.5mg/kg, 4.75mg/kg, 5mg/kg, 5.25mg/kg, 5.5mg/kg, 5.75mg/kg, 6mg/kg, 6.25mg/kg, 6.5mg/kg, 6.75mg/kg, 7.75mg/kg, 7mg/kg, 7.25mg/kg, 7mg/kg, 7.75mg/kg, 7.5mg/kg, 7.75mg/kg, 8mg/kg, 8.25mg/kg, 8.5mg/kg, 8.75mg/kg, 9mg/kg, 9.25mg/kg, 9.5mg/kg, 9.75mg/kg, 10mg/kg, 10.25mg/kg, 10.5mg/kg, 10.75mg/kg, 11mg/kg, 11.25mg/kg, 11.5mg/kg, 11.75mg/kg, 12mg/kg, 12.25mg/kg, 12.5mg/kg, 12.75mg/kg, 13mg/kg, 13.25mg/kg, 13.5mg/kg, 13.75mg/kg, 14mg/kg, 14.25mg/kg, 14.5mg/kg, 14.75mg/kg, 15mg/kg, 15.25mg/kg, 15.5mg/kg, 15.75mg/kg, 16.75mg/kg, 16.25mg/kg, 16.5mg/kg, 16mg/kg, 16.5mg/kg, 16.75mg/kg, 17mg/kg, 17.25mg/kg, 17.5mg/kg, 17.75mg/kg, 18mg/kg, 18.25mg/kg, 18.5mg/kg, 18.75mg/kg, 19mg/kg, 19.25mg/kg, 19.5mg/kg, 19.75mg/kg, 20mg/kg, 20.25mg/kg, 20.5mg/kg, 20.75mg/kg, 21mg/kg, 21.25mg/kg, 21.5mg/kg, 21.75mg/kg, 22mg/kg, 22.25mg/kg, 22.5mg/kg, 22.75mg/kg, 23mg/kg, 23.25mg/kg, 23.5mg/kg, 23.75mg/kg, 24mg/kg, 24.25mg/kg, 24.5mg/kg, 24.75mg/kg, 25mg/kg, 25.25mg/kg, 25mg/kg, 25.5mg/kg, 25mg/kg, 25.75mg/kg, 25mg/kg, 19.75mg/kg, 20.75mg/kg, 20.25mg/kg, 21.75mg/kg, 21.25mg/kg, 20.75mg/kg, 20.25mg/kg, 21.25 mg/20.25 mg/kg, 2mg/kg, 2.75 mg/5 mg/20.25 mg/kg, 2.25mg/kg, 2mg/kg, 2.25mg/kg, 2.75mg/kg, 2.25 mg/g, 2mg/kg, 2.25mg/kg, 2mg/kg, 2mg/kg, 2mg/kg, 2.75mg/kg, 2mg/kg, 2.75mg/kg, 2mg/kg, 2.75mg/kg, 2mg/kg, 2.75mg/kg, 2mg/kg, 2, 26mg/kg, 26.25mg/kg, 26.5mg/kg, 26.75mg/kg, 27mg/kg, 27.25mg/kg, 27.5mg/kg, 27.75mg/kg, 28mg/kg, 28.25mg/kg, 28.5mg/kg, 28.75mg/kg, 29mg/kg, 29.25mg/kg, 29.5mg/kg, 29.75mg/kg, 30mg/kg, 30.25mg/kg, 30.5mg/kg, 30.75mg/kg, 31mg/kg, 31.25mg/kg, 31.5mg/kg, 31.75mg/kg, 32mg/kg, 32.25mg/kg, 32.5mg/kg, 32.75mg/kg, 33mg/kg, 33.25mg/kg, 33.5mg/kg, 33.75mg/kg, 34mg/kg, 34.25mg/kg, 34.5mg/kg, 34.75mg/kg, 34.25mg/kg, 34.5mg/kg, 34.75mg/kg, 35.75mg/kg, 28.75mg/kg, 35.25mg/kg, 35.5mg/kg, 35.75mg/kg, 36mg/kg, 36.25mg/kg, 36.5mg/kg, 36.75mg/kg, 37mg/kg, 37.25mg/kg, 37.5mg/kg, 37.75mg/kg, 38mg/kg, 38.25mg/kg, 38.5mg/kg, 38.75mg/kg, 39mg/kg, 39.25mg/kg, 39.5mg/kg, 39.75mg/kg, 40mg/kg, 40.25mg/kg, 40.5mg/kg, 40.75mg/kg, 41mg/kg, 41.25mg/kg, 41.5mg/kg, 41.75mg/kg, 42mg/kg, 42.25mg/kg, 42.5mg/kg, 42.75mg/kg, 43mg/kg, 43.25mg/kg, 43.5mg/kg, 43.75mg/kg, 44mg/kg, 44.25mg/kg, 44mg/kg, 44.75mg/kg, 44mg/kg, 44.5mg/kg, 44.75mg/kg, 45mg/kg, 45.25mg/kg, 45.5mg/kg, 45.75mg/kg, 46mg/kg, 46.25mg/kg, 46.5mg/kg, 46.75mg/kg, 47mg/kg, 47.25mg/kg, 47.5mg/kg, 47.75mg/kg, 48mg/kg, 48.25mg/kg, 48.5mg/kg, 48.75mg/kg, 49mg/kg, 49.25mg/kg, 49.5mg/kg, 49.75mg/kg or 50 mg/kg.

(2) Vitamin D

The lipophilic compound of the nutritional composition may be vitamin D. Vitamin D can be incorporated into the assembly. Incorporation of vitamin D as part of the assembly enhances its solubility, thereby enhancing its bioavailability for treating a health condition relative to controls that do not contain MDG or hydrophobic proteins. "vitamin D" refers to a group of lipophilic compounds or "isovitamins" associated with steroids. The term "vitamin D" covers several isovitamins, but of which important is a calciferol, also known as vitamin D₂：

And cholecalciferol, also known as vitamin D₃：

For the purposes of this disclosure, unless otherwise specified, the term "vitamin D" as used herein encompasses all forms of vitamin D, whether as individual isovitamins such as vitamin D2 or vitamin D3, or as a combination of two or more isovitamins.

Vitamin D can be ingested from the diet and vitamin D3 is also synthesized in mammalian skin by the reaction of cholesterol with UV radiation from sunlight. Once inside the body, vitamin D isovitamins are metabolized to other chemical forms that regulate the concentration of calcium and phosphate in the bloodstream and promote healthy growth and maintenance of bone. Vitamin D can be used in nutritional compositions to maintain and supplement healthy bone health and maintenance. Vitamin D can be used in nutritional compositions for activating the innate immune system and suppressing the adaptive immune system as well as assisting cognitive development.

Any natural or synthetic source of vitamin D is suitable for use in the compositions disclosed herein, provided that such source is suitable for use in an ingestible composition and is compatible with the other ingredients within the composition. Vitamin D can be provided as an individual ingredient or in combination with other materials or sources, including sources such as multivitamin premixes. For example, vitamin D may be mixed with other oil soluble vitamins such as vitamin a, vitamin E, or vitamin K.

Non-limiting examples of vitamin D sources suitable for use herein include vitamin D3 supplied by BASF Corporation (Florham Park, New Jersey, USA), vitamin D3 supplied by Fermenta Biotech Ltd (Kullu, Himachal Pradesh, India), or DSM Nutritional Products AG (Kaiserarst, Switzerland)

Vitamin D in the nutritional composition may be from about 1 μ g/kg to about 30 μ g/kg. Vitamin D in the nutritional composition may be from about 10 μ g/kg to about 30 μ g/kg. Vitamin D in the nutritional composition may be from about 5 μ g/kg to about 25 μ g/kg. Vitamin D in the nutritional composition may be from about 10 μ g/kg to about 20 μ g/kg. For example, vitamin D in the nutritional composition may be about 1. mu.g/kg, 1.2. mu.g/kg, 1.4. mu.g/kg, 1.6. mu.g/kg, 1.8. mu.g/kg, 2. mu.g/kg, 2.2. mu.g/kg, 2.4. mu.g/kg, 2.6. mu.g/kg, 2.8. mu.g/kg, 3. mu.g/kg, 3.2. mu.g/kg, 3.4. mu.g/kg, 3.6. mu.g/kg, 3.8. mu.g/kg, 4. mu.g/kg, 4.2. mu.g/kg, 4.6. mu.g/kg, 4.8. mu.g/kg, 5. mu.g/kg, 5.2. mu.g/kg, 5.4. mu.g/kg, 5.6. mu.g/kg, 5.8. mu.g/kg, 6.g/kg, 6.2. mu.g/kg, 6.6.g/kg, 6.8. mu.g/kg, 6.g/kg, 6.8. mu.8. mu.g/kg, 6.g/kg, 6.6.6.6.g/kg, 6.6.6.6.6.g/kg, 6.g/kg, 6.8. mu.g/kg, 6.g/kg, 2 g/kg, 2, 7. mu.g/kg, 7.2. mu.g/kg, 7.4. mu.g/kg, 7.6. mu.g/kg, 7.8. mu.g/kg, 8. mu.g/kg, 8.2. mu.g/kg, 8.4. mu.g/kg, 8.6. mu.g/kg, 8.8. mu.g/kg, 9. mu.g/kg, 9.2. mu.g/kg, 9.4. mu.g/kg, 9.6. mu.g/kg, 9.8. mu.g/kg, 10. mu.g/kg, 10.2. mu.g/kg, 10.4. mu.g/kg, 10.6. mu.g/kg, 10.8. mu.g/kg, 11. mu.g/kg, 11.2. mu.g/kg, 11.4. mu.g/kg, 116. mu.g/kg, 118. mu.g/kg, 12. mu.g/kg, 12.2. mu.2. mu.g/kg, 12.4. mu.g/kg, 12.6. mu.g/kg, 12.8.g/kg, 13.4. mu.g/kg, 13.g/kg, 13.4. mu.g/kg, 13.g/kg, 13.8. mu.g/kg, 13.6. mu.g/kg, 13.8. mu.g/kg, 14. mu.g/kg, 14.2. mu.g/kg, 14.4. mu.g/kg, 14.6. mu.g/kg, 14.8. mu.g/kg, 15. mu.g/kg, 15.2. mu.g/kg, 15.4. mu.g/kg, 15.6. mu.g/kg, 15.8. mu.g/kg, 16. mu.g/kg, 16.2. mu.g/kg, 16.4. mu.g/kg, 16.6. mu.g/kg, 16.8. mu.g/kg, 17. mu.g/kg, 17.2. mu.g/kg, 17.6. mu.g/kg, 17.8. mu.g/kg, 18. mu.g/kg, 18.2. mu.g/kg, 18.4. mu.g/kg, 18.6. mu.8. mu.g/kg, 19.2. mu.g/kg, 19.6. mu.g/kg, 19.g/kg, 19.8. mu.g/kg, 20.8. mu.g/kg, 20.2. mu.g/kg, 20.4. mu.g/kg, 20.6. mu.g/kg, 20.8. mu.g/kg, 21. mu.g/kg, 21.2. mu.g/kg, 21.4. mu.g/kg, 21.6. mu.g/kg, 21.8. mu.g/kg, 22. mu.g/kg, 22.2. mu.g/kg, 22.4. mu.g/kg, 22.6. mu.g/kg, 22.8. mu.g/kg, 23. mu.g/kg, 23.2. mu.g/kg, 23.6. mu.g/kg, 23.8. mu.g/kg, 24. mu.g/kg, 24.2. mu.g/kg, 24.4. mu.g/kg, 24.6. mu.g/kg, 24.8. mu.g/kg, 25.2. mu.g/kg, 25.4. mu.g/kg, 25.6. mu.8. mu.g/kg, 26.g/kg, 26.8. mu.8. mu.g/kg, 26.g/kg, 26.8. mu.g/kg, 22.g/kg, 22.8. mu.g/kg, 22.g/kg, 23.g/kg, 23.6. mu.g/kg, 23.g/kg, 23.6. mu.g/kg, 23.6. mu.6. g/kg, 2. mu.g/kg, 23.g/kg, 23.6. g/kg, 23.g/kg, 2. g/kg, 23.g/kg, 2. g/kg, 23.g/g/kg, 23.6. mu.g/kg, 23.g/kg, 23.6. g/kg, 23.g/kg, 23.6. g/kg, 23.6. g/kg, 23.g/g/kg, 23.6. g/kg, 24.g/kg, 23.6. mu.6. g/kg, 23.6. g/kg, 24.6. mu.8 g/kg, 23.6. mu.6. g/kg, 24.g/kg, 23.g/g/kg, 23.g/kg, 23.6 g/kg, 23.g/g/kg, 23.g/g/kg, 23.g/g/kg, 26.8. mu.g/kg, 27. mu.g/kg, 27.2. mu.g/kg, 27.4. mu.g/kg, 27.6. mu.g/kg, 27.8. mu.g/kg, 28. mu.g/kg, 28.2. mu.g/kg, 28.4. mu.g/kg, 28.6. mu.g/kg, 28.8. mu.g/kg, 29. mu.g/kg, 29.2. mu.g/kg, 29.4. mu.g/kg, 29.6. mu.g/kg, 29.8. mu.g/kg or 30. mu.g/kg.

(3) Vitamin E

The lipophilic compound of the nutritional composition may be vitamin E. Vitamin E can be incorporated into the assembly. Incorporation of vitamin E as part of the assembly enhances its solubility, thereby enhancing its bioavailability for treating a health condition relative to controls that do not contain MDG or hydrophobic proteins. Vitamin E can be used in nutritional compositions to provide antioxidants and maintain and supplement cardiovascular health. Vitamin E may also be referred to as tocopherol. Tocopherols can exist in four forms, α, β, γ and δ, which differ in the number and position of methyl groups on the chroman ring (see table 1 below). Additionally, tocopherols can exist in many stereoisomeric forms depending on the chirality of the phytyl tail. Among the alpha tocopherols, RRR-alpha tocopherol (also known as "natural vitamin E") has the greatest biological activity and is reported to be the predominant form of alpha tocopherol in the brain. In one aspect, the composition may comprise RRR-alpha tocopherol, meaning that the composition is formulated or otherwise prepared to contain RRR-alpha tocopherol by the addition of RRR-alpha tocopherol. As used herein, the term "RRR-alpha tocopherol" refers to free RRR-alpha tocopherol from both external and intrinsic sources as well as RRR-alpha tocopherol esters such as RRR-o-alpha tocopherol acetate present in a composition. Intrinsic sources include RRR-alpha tocopherol, which is itself present in the components present in the nutritional composition and may include, for example, various oils and fats. The external source of RRR-alpha tocopherol includes RRR-alpha tocopherol acetate added to the nutritional composition but not as part of another component. Any source of RRR-alpha tocopherol is suitable for use in the present invention, provided that the finished product contains RRR-alpha tocopherol.

RRR-alpha tocopherol is a single stereoisomer, while synthetic vitamin E (all-rac-alpha tocopherol or tocopherol acetate) is an equimolar mixture of eight isomers, only one of which is RRR-alpha tocopherol. The fact that the main form of alpha tocopherol is RRR-alpha tocopherol (based on animal studies) strongly suggests: the other seven chiral isomers must be absorbed by the brain at a slower rate or oxidized at a faster rate. Cholesterol is the major component of myelin and it is likely that stimulated cholesterol synthesis may stimulate neonatal neuronal myelination.

Table 1: tocopheryl type

Compound (I)	R1	R2	R3
				Alpha-tocopherol	Me	Me	Me
Beta-tocopherol	Me	H	Me
				Gamma-tocopherol	H	Me	Me
Delta-tocopherol	H	H	Me

The vitamin E in the nutritional composition may be from about 5mg/kg to about 75 mg/kg. The vitamin E in the nutritional composition may be from about 10mg/kg to about 60 mg/kg. The vitamin E in the nutritional composition may be from about 20mg/kg to about 40 mg/kg. For example, vitamin E in the nutritional composition may be about 5mg/kg, 5.5mg/kg, 6mg/kg, 6.5mg/kg, 7mg/kg, 7.5mg/kg, 8mg/kg, 8.5mg/kg, 9mg/kg, 9.5mg/kg, 10mg/kg, 10.5mg/kg, 11mg/kg, 11.5mg/kg, 12mg/kg, 12.5mg/kg, 13mg/kg, 13.5mg/kg, 14mg/kg, 14.5mg/kg, 15mg/kg, 15.5mg/kg, 16mg/kg, 16.5mg/kg, 17mg/kg, 17.5mg/kg, 18mg/kg, 18.5mg/kg, 19mg/kg, 19.5mg/kg, 20mg/kg, 20.5mg/kg, 21.5mg/kg, 22.5mg/kg, 22mg/kg, 22.5mg/kg, 23mg/kg, 23.5mg/kg, 24mg/kg, 24.5mg/kg, 25mg/kg, 25.5mg/kg, 26mg/kg, 26.5mg/kg, 27mg/kg, 27.5mg/kg, 28mg/kg, 28.5mg/kg, 29mg/kg, 29.5mg/kg, 30mg/kg, 30.5mg/kg, 31mg/kg, 31.5mg/kg, 32mg/kg, 32.5mg/kg, 33mg/kg, 33.5mg/kg, 34mg/kg, 34.5mg/kg, 35mg/kg, 35.5mg/kg, 36mg/kg, 36.5mg/kg, 37mg/kg, 37.5mg/kg, 38mg/kg, 38.5mg/kg, 39mg/kg, 39.5mg/kg, 40mg/kg, 40.5mg/kg, 41mg/kg, 41.5mg/kg, 42mg/kg, 42.5mg/kg, 43mg/kg, 43.5mg/kg, 44mg/kg, 44.5mg/kg, 45mg/kg, 45.5mg/kg, 46mg/kg, 46.5mg/kg, 47mg/kg, 47.5mg/kg, 48mg/kg, 48.5mg/kg, 49mg/kg, 49.5mg/kg, 50mg/kg, 50.5mg/kg, 51mg/kg, 51.5mg/kg, 52mg/kg, 52.5mg/kg, 53mg/kg, 53.5mg/kg, 54mg/kg, 54.5mg/kg, 55mg/kg, 55.5mg/kg, 56mg/kg, 56.5mg/kg, 57mg/kg, 57.5mg/kg, 58mg/kg, 58.5mg/kg, 59mg/kg, 59.5mg/kg, 59.5mg/kg, 60mg/kg, 60.5mg/kg, 61mg/kg, 61.5mg/kg, 62mg/kg, 62.5mg/kg, 63mg/kg, 63.5mg/kg, 64mg/kg, 64.5mg/kg, 65mg/kg, 65.5mg/kg, 66mg/kg, 66.5mg/kg, 67mg/kg, 67.5mg/kg, 68mg/kg, 68.5mg/kg, 69mg/kg, 69.5mg/kg, 70mg/kg, 70.5mg/kg, 71mg/kg, 71.5mg/kg, 72mg/kg, 72.5mg/kg, 73mg/kg, 73.5mg/kg, 74mg/kg, 74.5mg/kg or 75 mg/kg.

(4) Docosahexaenoic acid (DHA)

The lipophilic compound of the nutritional composition may be DHA. DHA may be incorporated into the assembly. The incorporation of DHA as part of the assembly enhances its solubility, thereby enhancing its bioavailability for treating a health condition relative to a control that does not contain MDG or hydrophobic proteins. DHA may be used in nutritional compositions to maintain and supplement conditions such as healthy brain function. DHA is an omega-3 fatty acid with 22 carbons in the lipophilic chain, which is commonly found in, for example, cold water fish and algae:

DHA is a major structural component of the human brain, cerebral cortex and retina, and is an essential fatty acid for proper development of the eye, brain and nervous system of infants and young children. Breast milk contains DHA, and many infant formulas are rich in DHA because it plays a key role in the growth and development of these systems in infants. There is increasing interest in providing supplemental DHA to pregnant women. DHA is believed to reduce the risk of developing lacrimal keratoconjunctivitis, glaucoma, and macular degeneration. DHA is also required for healthy brain function in adults. DHA deficiency can lead to a decline in mental function in healthy elderly, and initial studies have shown that early intervention with DHA supplementation can improve memory and learning in adults over 50 years of age.

Non-limiting examples of sources of DHA suitable for use herein include Life's available from DSM diagnostic Products, Kaiser ugst, SwitzerlandAnd Maxomega available from BASF Pharma ltd, Callanish, Scotland^TMDHA。

The DHA in the nutritional composition may be from about 10mg/kg to about 10 g/kg. The DHA in the nutritional composition may be from about 100mg/kg to about 5 g/kg. The DHA in the nutritional composition may be from about 500mg/kg to about 5 g/kg. The DHA in the nutritional composition may be from about 50mg/kg to about 1 g/kg. For example, DHA in the nutritional composition may be about 10mg/kg, 20mg/kg, 30mg/kg, 40mg/kg, 50mg/kg, 60mg/kg, 70mg/kg, 80mg/kg, 90mg/kg, 100mg/kg, 110mg/kg, 120mg/kg, 130mg/kg, 140mg/kg, 150mg/kg, 160mg/kg, 170mg/kg, 180mg/kg, 190mg/kg, 200mg/kg, 210mg/kg, 220mg/kg, 230mg/kg, 240mg/kg, 250mg/kg, 260mg/kg, 270mg/kg, 280mg/kg, 290mg/kg, 300mg/kg, 310mg/kg, 320mg/kg, 330mg/kg, 340mg/kg, 350mg/kg, 360mg/kg, 370mg/kg, 380mg/kg, 390mg/kg, 400mg/kg, 410mg/kg, 420mg/kg, 430mg/kg, 440mg/kg, 450mg/kg, 460mg/kg, 470mg/kg, 480mg/kg, 490mg/kg, 500mg/kg, 510mg/kg, 520mg/kg, 530mg/kg, 540mg/kg, 550mg/kg, 560mg/kg, 570mg/kg, 580mg/kg, 590mg/kg, 600mg/kg, 610mg/kg, 620mg/kg, 630mg/kg, 640mg/kg, 650mg/kg, 660mg/kg, 670mg/kg, 680mg/kg, 690mg/kg, 700mg/kg, 710mg/kg, 720mg/kg, 730mg/kg, 740mg/kg, 750mg/kg, 760mg/kg, 770mg/kg, 780mg/kg, 790mg/kg, 800mg/kg, 810mg/kg, 820mg/kg, 830mg/kg, 840mg/kg, 850mg/kg, 860mg/kg, 870mg/kg, 880mg/kg, 890mg/kg, 900mg/kg, 910mg/kg, 920mg/kg, 930mg/kg, 940mg/kg, 950mg/kg, 960mg/kg, 970mg/kg, 980mg/kg, 990mg/kg, 1g/kg, 1.2g/kg, 1.4g/kg, 1.6g/kg, 1.8g/kg, 2g/kg, 2.2g/kg, 2.4g/kg, 2.6g/kg, 2.8g/kg, 3g/kg, 3.2g/kg, 3.4g/kg, 3.6g/kg, 3.8g/kg, 4g/kg, 4.2g/kg, 4.4g/kg, 4.6g/kg, 4.8g/kg, 5g/kg, 5.2g/kg, 5.4g/kg, 5.6g/kg, 5.8g/kg, 6g/kg, 6.2g/kg, 6.4g/kg, 6.6g/kg, 6.8g/kg, 7g/kg, 7.2g/kg, 7.4g/kg, 7.6g/kg, 7.8g/kg, 8g/kg, 8.2g/kg, 8.4g/kg, 8.6g/kg, 8.8g/kg, 9g/kg, 9.2g/kg, 9.4g/kg, 9.6g/kg, 10.8 g/kg or 10 g/kg.

(5) Combination of DHA and lutein

In one embodiment, it has been unexpectedly found that when DHA and lutein are present in a composition according to the present disclosure, the bioavailability of DHA and lutein after ingestion is improved over nutritional compositions that do not contain a combination of DHA and lutein. The improved bioavailability allows DHA and lutein to be more easily absorbed into the body of the subject. Because the bioavailability of DHA and lutein can be increased using the presently described methods, the level of DHA and lutein fortification required in the composition to provide the desired nutritional benefits can be reduced. DHA and lutein may be incorporated into the nutritional composition at the values listed above.

c. Fat

The nutritional composition may comprise at least one fat. The fat may be incorporated into the assembly. Fats may be present in the premix, the activation premix, and/or the protected premix. The fat may be a monoglyceride, a diglyceride, or a combination thereof (hereinafter, both referred to as "MDG"). Fats allow unique interactions with lipophilic compounds and proteins to form water-soluble stable assemblies. Thus, the presence of fat contributes to the formation of an assembly and the subsequent stability of the assembly of lipophilic substances contained within the composition relative to a control without MDG and hydrophobic proteins. Fats also enhance the bioavailability of lipophilic compounds relative to controls that do not contain MDG or hydrophobic proteins. Fat may also increase the protection of the nutritional composition from dissolved oxygen. In addition to the mixtures of mono-and diglycerides discussed above, non-limiting examples of fats or sources thereof suitable for use in the nutritional compositions described herein may be selected from (but are not limited to) the following group: coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, sunflower oil, high oleic sunflower oil, palm kernel oil, palm olein, rapeseed oil, marine oil, cottonseed oil, linseed oil, hemp seed oil, peanut oil, borage oil, algal oil, fungal oil, MCT oil (medium chain triglycerides), and combinations thereof.

(1) Monoglycerides and Diglycerides (MDG)

The fat of the nutritional composition may be MDG. MDG is a combination of mono-and diglycerides. The MDG may be incorporated into the assembly. Incorporation of MDG as part of the assembly enhances the solubility of lipophilic compounds, thereby enhancing their bioavailability for treating a health condition relative to controls that do not contain MDG or hydrophobic proteins. It is hypothesized, but not bound by a particular theory, that after consumption, MDG in the activation premix or MDG protected premix binds to lipophilic compounds and hydrophobic proteins, making it easier to incorporate into micelles formed in the gut. Thus, lipophilic compounds can be more efficiently delivered to the brush border of the intestine and absorbed into the body. In addition, it is hypothesized, but not bound by a particular theory, that MDG can interact with hydrophobic proteins and increase access to hydrophobic domains within the proteins. Thus, lipophilic compounds can form more stable interactions with the newly accessible hydrophobic domains of proteins.

MDG is a component with limited water solubility. When MDG is combined with at least one hydrophobic protein and at least one lipophilic compound (as in an assembly), its water solubility is significantly increased. It is hypothesized, but not bound by any particular theory, that MDG remaining in the aqueous phase after centrifugation may bind to the assembly due to its limited water solubility. The increased water solubility can be investigated by centrifugation as listed above, wherein the centrifuged aqueous phase exhibits the presence of more MDG than without MDG, lipophilic compound and at least one hydrophobic protein.

Diglycerides, commonly known as Diacylglycerols (DAG), are glycerolipids consisting of two fatty acid chains covalently bonded to a glycerol molecule through ester linkages. Monoglycerides are normal metabolites in the body formed during the breakdown of triglycerides and diglycerides. Non-limiting examples of commercial sources of mono-and diglycerides include natural sources (e.g., animal (bovine or porcine derived) or vegetable, those that can be derived from partially hydrogenated soybean oil, sunflower oil, safflower oil, and coconut oil) or synthetic sources. In some embodiments, the mixture of mono-and diglycerides can be derived from the hydrolysis of triglycerides in safflower oil. The monoglycerides in the MDG of the nutritional composition may be from about 0.1 wt% to about 99.9 wt%, wherein the remainder of the MDG is diglycerides. The monoglycerides in the MDG of the nutritional composition may be from about 10% to about 80% by weight, wherein the remainder of the MDG is diglycerides. The monoglycerides in the MDG of the nutritional composition may be from about 25% to about 50% by weight, wherein the remainder of the MDG is diglycerides. For example, the monoglyceride in the MDG of the nutritional composition may be about 0.1 wt%, 0.5 wt%, 1 wt%, 1.5 wt%, 2 wt%, 2.5 wt%, 3 wt%, 3.5 wt%, 4 wt%, 4.5 wt%, 5 wt%, 5.5 wt%, 6 wt%, 6.5 wt%, 7 wt%, 7.5 wt%, 8 wt%, 8.5 wt%, 9 wt%, 9.5 wt%, 10 wt%, 10.5 wt%, 11 wt%, 11.5 wt%, 12 wt%, 12.5 wt%, 13 wt%, 13.5 wt%, 14 wt%, 14.5 wt%, 15 wt%, 15.5 wt%, 16 wt%, 16.5 wt%, 17 wt%, 17.5 wt%, 18 wt%, 18.5 wt%, 19 wt%, 19.5 wt%, 20 wt%, 20.5 wt%, 21 wt%, 21.5 wt%, 22 wt%, 22.5 wt%, 23.5 wt%, 5 wt%, 6.5 wt%, 11 wt%, 11.5 wt%, 12.5 wt%, 13.5 wt%, 13 wt%, 13.5 wt%, 13 wt%, 13.5 wt%, 19 wt%, 14 wt%, 22.5 wt%, 23 wt%, 23.5 wt%, 23 wt%, or more, 24 wt%, 24.5 wt%, 25 wt%, 25.5 wt%, 26 wt%, 26.5 wt%, 27 wt%, 27.5 wt%, 28 wt%, 28.5 wt%, 29 wt%, 29.5 wt%, 30 wt%, 30.5 wt%, 31 wt%, 31.5 wt%, 32 wt%, 32.5 wt%, 33 wt%, 33.5 wt%, 34 wt%, 34.5 wt%, 35 wt%, 35.5 wt%, 36 wt%, 36.5 wt%, 37 wt%, 37.5 wt%, 38 wt%, 38.5 wt%, 39 wt%, 39.5 wt%, 40 wt%, 40.5 wt%, 41 wt%, 41.5 wt%, 42 wt%, 42.5 wt%, 43 wt%, 43.5 wt%, 44 wt%, 44.5 wt%, 45 wt%, 45.5 wt%, 46 wt%, 46.5 wt%, 47 wt%, 47.5 wt%, 48 wt%, 48.5 wt%, 49.5 wt%, 49 wt%, 49.5 wt%, 38.5 wt%, 39 wt%, 39.5 wt%, 39 wt%, 39.5 wt%, 40 wt%, 46.5 wt%, 49 wt%, 49.5 wt%, 49 wt%, 40 wt%, 49.5 wt%, 25 wt%, 25.5 wt%, 25 wt%, 40 wt%, 25 wt%, 25.5 wt%, 25 wt%, 25.5 wt%, 25 wt%, 25.5 wt%, 35 wt%, 25 wt%, 35 wt%, 35 wt%, 35, 50 wt%, 50.5 wt%, 51 wt%, 51.5 wt%, 52 wt%, 52.5 wt%, 53 wt%, 53.5 wt%, 54 wt%, 54.5 wt%, 55 wt%, 55.5 wt%, 56 wt%, 56.5 wt%, 57 wt%, 57.5 wt%, 58 wt%, 58.5 wt%, 59 wt%, 59.5 wt%, 60 wt%, 60.5 wt%, 61 wt%, 61.5 wt%, 62 wt%, 62.5 wt%, 63 wt%, 63.5 wt%, 64 wt%, 64.5 wt%, 65 wt%, 65.5 wt%, 66 wt%, 66.5 wt%, 67 wt%, 67.5 wt%, 68 wt%, 68.5 wt%, 69 wt%, 69.5 wt%, 70 wt%, 70.5 wt%, 71 wt%, 71.5 wt%, 72 wt%, 72.5 wt%, 73 wt%, 73.5 wt%, 74 wt%, 74.5 wt%, 75 wt%, 75.5 wt%, 57.5 wt%, 57 wt%, 57.5 wt%, 60 wt%, 60.5 wt%, 60 wt%, and 70.5 wt% of the like, 76%, 76.5%, 77%, 77.5%, 78%, 78.5%, 79%, 79.5%, 80%, 80.5%, 81%, 81.5%, 82%, 82.5%, 83%, 83.5%, 84%, 84.5%, 85%, 85.5%, 86%, 86.5%, 87%, 87.5%, 88 wt.%, 88.5 wt.%, 89 wt.%, 89.5 wt.%, 90 wt.%, 90.5 wt.%, 91 wt.%, 91.5 wt.%, 92 wt.%, 92.5 wt.%, 93 wt.%, 93.5 wt.%, 94 wt.%, 94.5 wt.%, 95 wt.%, 95.5 wt.%, 96 wt.%, 96.5 wt.%, 97 wt.%, 97.5 wt.%, 98 wt.%, 98.5 wt.%, 99 wt.%, 99.5 wt.%, or 99.9 wt.%, wherein the remainder of the MDG is diglycerides.

Non-limiting examples of mixtures of mono-and diglycerides suitable for use in the composition include Capmul GMO-40 (MDG derived from high oleic safflower oil, available from Abitec Corporation, Columbus, Ohio), and Kimol CE1089 (MDG derived from corn oil, available from BASF, Ludwigshafen, Gernmann), RADIAMULS MG F038 (MDG derived from high oleic sunflower oil, available from Oleon, Ertveld, Belgium).

The MDG in the nutritional composition may be from about 140mg/kg to about 900 mg/kg. The MDG in the nutritional composition may be from about 145mg/kg to about 890 mg/kg. The nutritional composition may comprise about 140mg/kg to about 700mg/kg MDG. The MDG in the nutritional composition may be from about 145mg/kg to about 600 mg/kg. The MDG in the nutritional composition may be from about 140mg/kg to about 400 mg/kg. For example, the MDG in the nutritional composition may be about 140mg/kg, 145mg/kg, 150mg/kg, 155mg/kg, 160mg/kg, 165mg/kg, 170mg/kg, 175mg/kg, 180mg/kg, 185mg/kg, 190mg/kg, 195mg/kg, 200mg/kg, 205mg/kg, 210mg/kg, 215mg/kg, 220mg/kg, 225mg/kg, 230mg/kg, 235mg/kg, 240mg/kg, 245mg/kg, 250mg/kg, 255mg/kg, 260mg/kg, 265mg/kg, 270mg/kg, 275mg/kg, 280mg/kg, 285mg/kg, 290mg/kg, 295mg/kg, 300mg/kg, 305mg/kg, 310mg/kg, 315mg/kg, 320mg/kg, 325mg/kg, 330mg/kg, 335mg/kg, 340mg/kg, 345mg/kg, 350mg/kg, 355mg/kg, 360mg/kg, 365mg/kg, 370mg/kg, 375mg/kg, 380mg/kg, 385mg/kg, 390mg/kg, 395mg/kg, 400mg/kg, 405mg/kg, 410mg/kg, 415mg/kg, 420mg/kg, 425mg/kg, 430mg/kg, 435mg/kg, 440mg/kg, 445mg/kg, 450mg/kg, 455mg/kg, 460mg/kg, 465mg/kg, 470mg/kg, 475mg/kg, 480mg/kg, 485mg/kg, 490mg/kg, 495mg/kg, 500mg/kg, 505mg/kg, 510mg/kg, 515mg/kg, 520mg/kg, 525mg/kg, 530mg/kg, 535mg/kg, 540mg/kg, 545mg/kg, 550mg/kg, 555mg/kg, 560mg/kg, 565mg/kg, 570mg/kg, 575mg/kg, 580mg/kg, 585mg/kg, 590mg/kg, 595mg/kg, 600mg/kg, 605mg/kg, 610mg/kg, 615mg/kg, 620mg/kg, 625mg/kg, 630mg/kg, 635mg/kg, 640mg/kg, 645mg/kg, 650mg/kg, 655mg/kg, 660mg/kg, 665mg/kg, 670mg/kg, 675mg/kg, 680mg/kg, 685mg/kg, 690mg/kg, 695mg/kg, 700mg/kg, 705mg/kg, 710mg/kg, 715mg/kg, 720mg/kg, 725mg/kg, 730mg/kg, 735mg/kg, 740mg/kg, 745mg/kg, 750mg/kg, 755mg/kg, 760mg/kg, 765mg/kg, 770mg/kg, 775mg/kg, 780mg/kg, 785mg/kg, 790mg/kg, 795mg/kg, 800mg/kg, 805mg/kg, 810mg/kg, 815mg/kg, 820mg/kg, 825mg/kg, 830mg/kg, 835mg/kg, 840mg/kg, 845mg/kg, 850mg/kg, 855mg/kg, 860mg/kg, 865mg/kg, 845mg/kg, 710mg/kg, 855mg/kg, and, 870mg/kg, 875mg/kg, 880mg/kg, 885mg/kg, 890mg/kg, 895mg/kg, or 900 mg/kg.

In some embodiments, the nutritional composition may comprise about 0.1mg/kg to about 20mg/kg MDG and vitamin D. In some embodiments, the nutritional composition may comprise about 0.1mg/kg to about 10mg/kg MDG and vitamin D.

(2) Palmitic acid

The fat of the nutritional composition of the invention may be palmitic acid. Palmitic acid may be incorporated into the assembly. The incorporation of palmitic acid as part of the assembly enhances its solubility, thereby enhancing its bioavailability for treating a health condition relative to a control that does not contain MDG or hydrophobic proteins. Palmitic acid may be used in nutritional compositions to maintain and supplement healthy nutrition, especially in infants.

Palmitic acid is a saturated fatty acid having 16 carbons in the lipophilic chain. Palmitic acid is an important energy source for infant growth. It is also a precursor of long chain fatty acids synthesized in vivo. These long chain fatty acids, such as docosahexaenoic acid, are important structural components in the brain, eye and central nervous system. Thus, palmitic acid is an important nutrient in the healthy development of infants.

The palmitic acid may be provided by HPAV oil comprising greater than about 22% palmitic acid as a percentage of total fatty acids in the HPAV oil. Examples of HPAV oils include, but are not limited to, palm oil and palm olein.

Palmitic acid is found in palm oil and palm olein and constitutes more than 30% of the total fatty acids in each oil. Palmitic acid is also found in cocoa butter, cottonseed oil, and some other oil of the seed plant.

Palm oil is found in the flesh of the oil palm fruit. Palmitic acid constitutes from about 43% to about 45% of the fatty acids found in palm oil. Palm oil also contains from about 37% to about 40% oleic acid, which makes it a good source of monounsaturated fats. Palm oil also contains the essential fatty acid linoleic acid, which constitutes from about 5% to about 11% of the fatty acids in palm oil. Palm oil should not be confused with palm kernel oil, which is derived from the kernel (seed or kernel) of the oil palm fruit. Palm kernel oil has a very different fatty acid composition, containing less than about 10% palmitic acid and mainly saturated fatty acids. Thus, for the purposes of this description, palm kernel oil is not considered HPAV oil.

During refining, the raw material palm oil is fractionated by crystallisation and separation of the liquid fraction from the solid fraction palm oil at a controlled temperature. The liquid fraction is called palm olein. Palm olein contains slightly less palmitic acid (about 33% to about 40%) but more oleic acid (about 42% to about 48%) than palm oil.

The palmitic acid may be provided by HPAV oil comprising greater than about 22% palmitic acid as a percentage of total fatty acids in the HPAV oil. Examples of HPAV oils include, but are not limited to, palm oil and palm olein.

Suppliers of suitable HPAV oils such as palm oil or palm olein include: archer Daniels Midland, Decatur, Illinois, USA; fuji Vegetable Oil, Inc., Savannah, Georgia, USA and California Oils Corporation, Richmond, California, USA.

HPAV oil may be provided in the nutritional composition as an individual ingredient or in combination with other materials or sources. For example, HPAV oil may be provided as a mixture with other oils such as rapeseed oil or corn oil.

In some embodiments, it may be desirable to mix the protected premix comprising HPAV oil, surfactant, and MDG prior to mixing the MDG oil or HPAV oil with the other ingredients of the nutritional composition. In these embodiments, the HPAV oil and surfactant are mixed with the MDG oil in the presence of heat or at ambient temperature and in some embodiments with agitation to allow the HPAV oil to dissolve or disperse in the MDG oil.

In the protected premix containing HPAV oil, HPAV oil is present in an amount relative to the amount of MDG oil, which will provide effective amounts of HPAV oil and MDG oil in the infant formula. For example, the HPAV oil can be present in an amount of 15g, about 20g, about 30g, about 50g, about 75g, about 100g, about 125g, about 150g, about 175g, or about 200g HPAV/1 g MDG oil in the premix. Other ratios of HPAV oil to MDG oil are within the scope of the present disclosure, as long as HPAV oil is completely or substantially dissolved or dispersed in the resulting premix. Based on the disclosure herein, one skilled in the art can calculate a suitable ratio that will allow for effective amounts of HPAV oil and MDG oil to be incorporated into the nutritional composition.

In human breast milk, about 70% of palmitic acid is found in the middle or SN-2 position of triglycerides. The SN-2 position is relatively protected and palmitic acid remains bound to the glycerol backbone until later digestion processes. Thus, palmitic acid in the SN-2 position is in a form that is readily absorbed by the infant's intestine. However, for vegetable oils such as palm oil or palm olein, only about 9% of the palmitic acid is found at the SN-2 position. The remainder of the palmitic acid in these oils is found in the SN-1 or SN-3 position on the glycerol backbone. The fatty acids at the SN-1 or SN-3 positions are less protected and are often cleaved from the glycerol backbone early in the digestion process. These free fatty acids then react with calcium in the gastric fluid to form insoluble calcium-fatty acid soap complexes. These insoluble soap complexes, which bind both fatty acids and calcium, are not absorbed into the intestinal tract, but rather are excreted from the body. Studies have shown that infants fed formulas containing palm oil or palm olein do not absorb much of the fat or calcium as do breastfed infants.

Palmitic acid can be synthetically prepared from fats that more closely mimic the fat in breast milk. These fats are synthesized to have significantly more palmitic acid at the SN-2 position than the SN-2 position found in natural vegetable oils. The nutritional composition may have palmitic acid at the SN-2 position of about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the total palmitic acid content of the nutritional composition.

Palmitic acid in the nutritional composition may be from about 0.5g/kg to about 35 g/kg. Palmitic acid in the nutritional composition may be from about 1g/kg to about 30 g/kg. Palmitic acid in the nutritional composition may be from about 1g/kg to about 20 g/kg. Palmitic acid in the nutritional composition may be from about 1g/kg to about 10 g/kg. For example, palmitic acid in the nutritional composition may be about 0.5g/kg, 0.75g/kg, 1g/kg, 1.25g/kg, 1.5g/kg, 1.75g/kg, 2g/kg, 2.25g/kg, 2.5g/kg, 2.75g/kg, 3g/kg, 3.25g/kg, 3.5g/kg, 3.75g/kg, 4g/kg, 4.25g/kg, 4.5g/kg, 4.75g/kg, 5g/kg, 5.25g/kg, 5.5g/kg, 5.75g/kg, 6g/kg, 6.25g/kg, 6.5g/kg, 6.75g/kg, 7g/kg, 7.25g/kg, 7.5g/kg, 7.75g/kg, 8g/kg, 8.25g/kg, 8.75g/kg, 9.75g/kg, 9g/kg, 9.5g/kg, 9.25g/kg, 9.5g/kg, 9.75g/kg, 10g/kg, 10.25g/kg, 10.5g/kg, 10.75g/kg, 11g/kg, 11.25g/kg, 115g/kg, 1175g/kg, 12g/kg, 12.25g/kg, 12.5g/kg, 12.75g/kg, 13g/kg, 13.25g/kg, 13.5g/kg, 13.75g/kg, 14g/kg, 14.25g/kg, 14.5g/kg, 14.75g/kg, 15g/kg, 15.25g/kg, 15.5g/kg, 15.75g/kg, 16g/kg, 16.25g/kg, 16.5g/kg, 16.75g/kg, 17g/kg, 17.25g/kg, 17.5g/kg, 17.75g/kg, 18.75g/kg, 18g/kg, 18.25g/kg, 18.5g/kg, 18.75g/kg, 19g/kg, 19.25g/kg, 19.5g/kg, 19.75g/kg, 20g/kg, 20.25g/kg, 20.5g/kg, 20.75g/kg, 21g/kg, 21.25g/kg, 21.5g/kg, 21.75g/kg, 22g/kg, 22.25g/kg, 22.5g/kg, 22.75g/kg, 23g/kg, 23.25g/kg, 23.5g/kg, 23.75g/kg, 24g/kg, 24.25g/kg, 24.5g/kg, 24.75g/kg, 25g/kg, 25.25g/kg, 25.5g/kg, 25.75g/kg, 26g/kg, 26.25g/kg, 26.5g/kg, 26.75g/kg, 27.75g/kg, 27.25g/kg, 27.5g/kg, 27.75g/kg, 28g/kg, 28.25g/kg, 28.5g/kg, 28.75g/kg, 29g/kg, 29.25g/kg, 29.5g/kg, 29.75g/kg, 30g/kg, 30.25g/kg, 30.5g/kg, 30.75g/kg, 31g/kg, 31.25g/kg, 31.5g/kg, 31.75g/kg, 32g/kg, 32.25g/kg, 32.5g/kg, 32.75g/kg, 33g/kg, 33.25g/kg, 33.5g/kg, 33.75g/kg, 34g/kg, 34.25g/kg, 34.5g/kg, 34.75g/kg or 35 g/kg.

d. Protein

The nutritional composition may comprise at least one protein. Such a protein may be incorporated into the assembly. The protein may be present within the activation premix and/or the protected premix. The proteins interact uniquely with the lipophilic compounds and fats in the nutritional composition to form the water-soluble stable assemblies discussed above. Thus, the presence of the protein contributes to the formation of the assembly and the subsequent stability of the assembly of lipophilic substances contained within the composition. The protein also enhances the bioavailability of lipophilic compounds relative to controls that do not contain fat (e.g., MDG) or hydrophobic protein.

The protein may be an intact protein, a hydrolyzed protein, or a combination thereof. The protein may be a hydrophobic protein. Non-limiting examples of proteins or sources thereof suitable for use in the nutritional composition include partially hydrolyzed (less than 25% hydrolysis) or non-hydrolyzed proteins (e.g., intact proteins) or protein sources, which may be derived from any known or otherwise suitable source, such as milk (e.g., casein, whey), animals (e.g., meat, fish), grains (e.g., rice, corn), or combinations thereof. Non-limiting examples of such proteins include milk protein isolates, milk protein concentrates as described herein (such as whey protein concentrates), casein isolates, whey proteins, caseinates, whole milk, partially or completely skimmed milk, soy protein isolates, soy protein concentrates, and the like.

In some embodiments, the nutritional composition includes a protein component consisting only of intact and/or partially hydrolyzed proteins; that is, the protein component does not contain any protein having a degree of hydrolysis of about 25% or more. In this regard, the term "partially hydrolyzed protein" is intended to mean a protein having a degree of hydrolysis of less than about 25% (including less than about 20%, including less than about 15%, including less than about 10%), and includes a protein having a degree of hydrolysis of less than about 5%. The degree of hydrolysis is the extent to which peptide bonds are cleaved by the hydrolysis process. The degree of proteolysis of the partially hydrolyzed protein component used to characterize these embodiments is readily determined by one of ordinary skill in the composition art by quantifying the amino nitrogen to total nitrogen ratio (AN/TN) of the protein component of the selected composition. The amino nitrogen component is quantified by the USP titration method for determining amino nitrogen content, while the total nitrogen component is determined by the Tecator Kjeldahl method, all of which are well known to those of ordinary skill in the analytical chemistry art.

(1) Hydrophobic proteins

The protein of the nutritional composition may comprise at least one hydrophobic protein. Hydrophobic proteins may be incorporated into the assembly. Incorporation of a hydrophobic protein as part of the assembly enhances the solubility of the lipophilic compound, thereby enhancing the bioavailability of the lipophilic compound for treating a health condition relative to a control that does not contain MDG or the hydrophobic protein. It is hypothesized, but not bound by any particular theory, that the water solubility of lipophilic compounds is enhanced by binding within the hydrophobic domains of proteins. The hydrophobic protein may be, but is not limited to, beta-lactoglobulin, bovine lactoferrin, gamma ₂-casein, β -casein, α -lactalbumin or a combination thereof. The hydrophobic protein in the nutritional composition may be from about 1g/L to about 100 g/L. The hydrophobic protein in the nutritional composition may be from about 10g/L to about 80 g/L. The hydrophobic protein in the nutritional composition may be from about 20g/L to about 60 g/L. The hydrophobic protein in the nutritional composition may be from about 30g/L to about 50 g/L. For example, the hydrophobin in the nutritional composition can be about 1g/L, 1.5g/L, 2g/L, 2.5g/L, 3g/L, 3.5g/L, 4g/L, 4.5g/L, 5g/L, 5.5g/L, 6g/L, 6.5g/L, 7g/L, 7.5g/L, 8g/L, 8.5g/L, 9g/L, 9.5g/L, 10g/L, 10.5g/L, 11g/L, 11.5g/L, 12g/L, 12.5g/L, 13g/L, 13.5g/L, 14g/L, 14.5g/L, 15g/L, 15.5g/L, 16g/L, 16.5g/L, 17g/L, 17.5g/L, 18g/L, 18.5g/L, 19g/L, 19.5g/L, 20g/L, 20.5g/L, 21g/L, 21.5g/L, 22g/L, 22.5g/L, 23g/L, 23.5g/L, 24g/L, 24.5g/L, 25g/L, 25.5g/L, 26g/L, 26.5g/L, 27g/L, 27.5g/L, 28g/L, 28.5g/L, 29g/L, 29.5g/L, 30g/L, 30.5g/L, 31g/L, 31.5g/L, 32g/L, 32.5g/L, 33g/L, 33.5g/L, 34g/L, 34.5g/L, 35g/L, 35.5g/L, 36g/L, 36.5g/L, 23.5g/L, 23g/L, 23.5g/L, 36.5g/L, 37g/L, 37.5g/L, 38g/L, 38.5g/L, 39g/L, 39.5g/L, 40g/L, 40.5g/L, 41g/L, 41.5g/L, 42g/L, 42.5g/L, 43g/L, 43.5g/L, 44g/L, 44.5g/L, 45g/L, 45.5g/L, 46g/L, 46.5g/L, 47g/L, 47.5g/L, 48g/L, 48.5 g/L 49g/L, 49.5g/L, 50g/L, 50.5g/L, 51g/L, 51.5g/L, 52g/L, 52.5g/L, 53g/L, 53.5g/L, 54g/L, 54.5g/L, 55g/L, 55.5g/L, 56g/L, 56.5g/L, 57g/L, 57.5g/L, 58g/L, 58.5g/L, 59g/L, 59.5g/L, 60g/L, 60.5g/L, 61g/L, 61.5g/L, 62g/L, 62.5g/L, 63g/L, 63.5g/L, 64g/L, 64.5g/L, 65g/L, 65.5g/L, 66g/L, 66.5g/L, 67g/L, 67.5g/L, 68g/L, 68.5g/L, 69g/L, 69.5g/L, 70g/L, 70.5g/L, 71g/L, 71.5g/L, 72g/L, 72.5g/L, 73g/L, 73.5g/L, 74g/L, 74.5g/L, 75g/L, 75.5g/L, 76g/L, 76.5g/L, 77g/L, 77.5g/L, 78g/L, 78.5g/L, 79g/L, 79.5g/L, 80g/L, 80.5g/L, 81g/L, 81.5g/L, 82g/L, 82.5g/L, 83g/L, 83.5g/L, 84g/L, 84.5g/L, 85g/L, 85.5g/L, 86g/L, 86.5g/L, 87g/L, 87.5g/L, 88g/L, 88.5g/L, 89g/L, 89.5g/L, 90g/L, 90.5g/L, 91g/L, 91.5g/L, 92g/L, 92.5g/L, 93g/L, 93.5g/L, 94g/L, 94.5g/L, 95g/L, 95.5g/L, 96g/L, 96.5g/L, 97g/L, 97.5g/L, 98g/L, 98.5g/L, 99g/L, 99.5g/L, or 100 g/L.

Hydrophobic proteins can be characterized by their overall average hydrophilicity index (GRAVY) value. Hydrophilicity is, for example, Creighton TE, Proteins: structures and molecular properties, 2 nd edition, 1993, w.h.freeman and Company, NY, page 160. The hydrophobic protein may have a GRAVY value of about-0.5 to about 0. The hydrophobic protein may have a GRAVY value of about-0.4 to about 0. The hydrophobic protein may have a GRAVY value of about-0.2 to about 0. The GRAVY value of a hydrophobic protein is a measure of hydrophobicity and water solubility and can predict the ability of a protein to bind to a lipophilic molecule such as lutein. The GRAVY values can be obtained from protein databases known in the art, and are referenced to Kyte et al, J.mol.biol. (1982)157, 105-132, which is incorporated herein by reference in its entirety. The GRAVY value for the hydrophobic protein can be about-0.5, -0.498, -0.496, -0.494, -0.492, -0.49, -0.488, -0.486, -0.484, -0.482, -0.48, -0.478, -0.476, -0.474, -0.472, -0.47, -0.468, -0.466, -0.464, -0.462, -0.46, -0.458, -0.456, -0.454, -0.452, -0.45, -0.448, -0.446, -0.444, -0.442, -0.44, -0.438, -0.436, -0.434, -0.432, -0.43, -0.428, -0.426, -0.424, -0.422, -0.42, -0.418, -0.416, -0.414, -0.412, -0.41, -0.408, -0.406, -0.404, -0.402, -0.4, -0.398, -0.396, -0.394, -0.392, -0.39, -0.388, -0.386, -0.384, -0.382, -0.38, -0.378, -0.376, -0.374, -0.372, -0.37, -0.368, -0.366, -0.364, -0.362, -0.36, -0.358, -0.356, -0.354, -0.352, -0.35, -0.348, -0.346, -0.344, -0.342, -0.34, -0.338, -0.336, -0.334, -0.332, -0.33, -0.328, -0.326, -0.324, -0.322, -0.32, -0.318, -0.316, -0.314, -0.312, -0.31, -0.308, -0.306, -0.304, -0.302, -0.3, -0.298, -0.296, -0.294, -0.292, -0.29, -0.288, -0.286, -0.284, -0.282, -0.28, -0.278, -0.276, -0.274, -0.272, -0.27, -0.268, -0.266, -0.264, -0.262, -0.26, -0.258, -0.256, -0.254, -0.252, -0.25, -0.248, -0.246, -0.244, -0.242, -0.24, -0.238, -0.236, -0.234, -0.232, -0.23, -0.228, -0.226, -0.224, -0.222, -0.22, -0.218, -0.216, -0.214, -0.212, -0.21, -0.208, -0.206, -0.204, -0.202, -0.2, -0.198, -0.196, -0.194, -0.192, -0.19, -0.188, -0.186, -0.184, -0.182, -0.18, -0.178, -0.176, -0.174, -0.172, -0.17, -0.168, -0.166, -0.164, -0.162, -0.16, -0.158, -0.156, -0.154, -0.152, -0.15, -0.148, -0.146, -0.144, -0.142, -0.14, -0.138, -0.136, -0.134, -0.132, -0.13, -0.128, -0.126, -0.124, -0.122, -0.12, -0.118, -0.116, -0.114, -0.112, -0.11, -0.108, -0.106, -0.104, -0.102, -0.1, -0.098, -0.096, -0.094, -0.092, -0.09, -0.088, -0.086, -0.084, -0.082, -0.08, -0.078, -0.076, -0.074, -0.072, -0.07, -0.068, -0.066, -0.064, -0.062, -0.06, -0.058, -0.056, -0.054, -0.052, -0.05, -0.048, -0.046, -0.044, -0.042, -0.04, -0.038, -0.036, -0.034, -0.032, -0.03, -0.028, -0.026, -0.024, -0.022, -0.02, -0.018, -0.016, -0.014, -0.012, -0.01, -0.008, -0.006, -0.004, -0.002 or 0.004.

In addition, it has been surprisingly found that hydrophobic proteins bound to the assembly, wherein the hydrophobic proteins constitute from about 10 wt% to about 95 wt% of the assembly, lead to a corresponding increase of lipophilic compounds within the assembly. For example, the hydrophobic protein in the assembly can be about 10 wt%, 10.5 wt%, 11 wt%, 11.5 wt%, 12 wt%, 12.5 wt%, 13 wt%, 13.5 wt%, 14 wt%, 14.5 wt%, 15 wt%, 15.5 wt%, 16 wt%, 16.5 wt%, 17 wt%, 17.5 wt%, 18 wt%, 18.5 wt%, 19 wt%, 19.5 wt%, 20 wt%, 20.5 wt%, 21 wt%, 21.5 wt%, 22 wt%, 22.5 wt%, 23 wt%, 23.5 wt%, 24 wt%, 24.5 wt%, 25 wt%, 25.5 wt%, 26 wt%, 26.5 wt%, 27 wt%, 27.5 wt%, 28 wt%, 28.5 wt%, 29 wt%, 29.5 wt%, 30 wt%, 30.5 wt%, 31 wt%, 31.5 wt%, 32 wt%, 32.5 wt%, 33.5 wt%, 15.5 wt%, 15 wt%, 15.5 wt%, 15 wt%, 15.5 wt%, 16.5 wt%, 16 wt%, 16.5 wt%, 17 wt%, 22.5 wt%, 22 wt%, 22.5 wt%, 22 wt%, 22.5 wt%, 23 wt%, and/wt%, and 23 wt%, and/wt%, 25 wt%, respectively, 34 wt%, 34.5 wt%, 35 wt%, 35.5 wt%, 36 wt%, 36.5 wt%, 37 wt%, 37.5 wt%, 38 wt%, 38.5 wt%, 39 wt%, 39.5 wt%, 40 wt%, 40.5 wt%, 41 wt%, 41.5 wt%, 42 wt%, 42.5 wt%, 43 wt%, 43.5 wt%, 44 wt%, 44.5 wt%, 45 wt%, 45.5 wt%, 46 wt%, 46.5 wt%, 47 wt%, 47.5 wt%, 48 wt%, 48.5 wt%, 49 wt%, 49.5 wt%, 50 wt%, 50.5 wt%, 51 wt%, 51.5 wt%, 52 wt%, 52.5 wt%, 53 wt%, 53.5 wt%, 54 wt%, 54.5 wt%, 55 wt%, 55.5 wt%, 56 wt%, 56.5 wt%, 57 wt%, 57.5 wt%, 58 wt%, 58.5 wt%, 59.5 wt%, 40 wt%, 40.5 wt%, 41.5 wt%, 42.5 wt%, 43.5 wt%, 45 wt%, 46.5 wt%, 47.5 wt%, 49 wt%, 47.5 wt%, 49 wt%, 50 wt%, 56 wt%, 50 wt%, 5 wt%, 50 wt%, 56 wt%, 5 wt%, and 58 wt%, 56 wt%, 5 wt%, 56 wt%, and 5 wt%, and 58.5 wt%, 25.5 wt%, 5 wt%, etc, 60 wt%, 60.5 wt%, 61 wt%, 61.5 wt%, 62 wt%, 62.5 wt%, 63 wt%, 63.5 wt%, 64 wt%, 64.5 wt%, 65 wt%, 65.5 wt%, 66 wt%, 66.5 wt%, 67 wt%, 67.5 wt%, 68 wt%, 68.5 wt%, 69 wt%, 69.5 wt%, 70 wt%, 70.5 wt%, 71 wt%, 71.5 wt%, 72 wt%, 72.5 wt%, 73 wt%, 73.5 wt%, 74 wt%, 74.5 wt%, 75 wt%, 75.5 wt%, 76 wt%, 76.5 wt%, 77 wt%, 77.5 wt%, 78 wt%, 78.5 wt%, 79 wt%, 79.5 wt%, 80 wt%, 80.5 wt%, 81 wt%, 81.5 wt%, 82 wt%, 82.5 wt%, 83 wt%, 83.5 wt%, 84 wt%, 84.5 wt%, 85 wt%, 85.5 wt%, 67 wt%, 67.5 wt%, 67 wt%, and/wt% 86 wt%, 86.5 wt%, 87 wt%, 87.5 wt%, 88 wt%, 88.5 wt%, 89 wt%, 89.5 wt%, 90 wt%, 90.5 wt%, 91 wt%, 91.5 wt%, 92 wt%, 92.5 wt%, 93 wt%, 93.5 wt%, 94 wt%, 94.5 wt%, or 95 wt%. In addition, the higher molecular weight of the hydrophobic protein associated with the assembly is associated with an increase in lipophilic compounds within the assembly, wherein the hydrophobic protein associated with the assembly can have a minimum molecular weight of about 5kD to about 15 kD. For example, the minimum molecular weight of the protein bound to the assembly may be about 5kD, 5.2kD, 5.4kD, 5.6kD, 5.8kD, 6kD, 6.2kD, 6.4kD, 6.6kD, 6.8kD, 7kD, 7.2kD, 7.4kD, 7.6kD, 7.8kD, 8kD, 8.2kD, 8.4kD, 8.6kD, 8.8kD, 9kD, 9.2kD, 9.4kD, 9.6kD, 9.8kD, 10kD, 10.2kD, 10.4kD, 10.6kD, 10.8kD, 11.2kD, 11.4kD, 11.6kD, 11.8kD, 12kD, 12.2kD, 12.4kD, 12.6kD, 12.8kD, 13kD, 13.2kD, 13.4kD, 13.6kD, 13.8kD, 13.14 kD, 14.14 kD, 14kD, 14.14 kD, 14.8kD, 14kD, 14.2kD, 14kD, 14.6kD, 14kD, or 14kD, 13.6kD, 13.8kD, 13kD, 13.2kD, 13kD, 13.4kD, 13kD, 13.4kD, 13kD, or 14kD, 13kD, 8kD, 13kD, or 14kD, 13kD, 8kD, 13kD, or 14kD, 8kD, 13kD, 8kD, 13kD, or 14kD, 13kD, 8kD, 13kD, 8kD, 13kD, or 14kD, 13kD, 8kD, or 14kD, 13 kD. In one embodiment, the minimum weight of protein bound to the assembly is about 11.8 kD.

(a) Beta-casein

The hydrophobin of the nutritional composition may comprise at least beta-casein. Beta-casein may be incorporated into the assembly. The incorporation of beta-casein as part of the assembly enhances the solubility of the lipophilic compound, thereby enhancing the bioavailability of the lipophilic compound for treating a health condition relative to a control that does not contain MDG or hydrophobic proteins. It has been unexpectedly shown that beta-casein is involved in the delivery of certain lipophilic nutrients for absorption, digestion, or both. A2 β -casein is a relatively more hydrophobic form of β -casein (compared to a1 β -casein). Nutritional compositions that replace a portion of the intrinsic beta-casein with genetic variant a2 exhibit enhanced bioavailability of certain lipophilic nutrients.

In the united states, most of the milk proteins used in milk protein containing products are from the Holstein cow (Holstein) breed. The term "Holstein cattle" as used herein should be understood to include Holstein cattle varieties, Friesian cattle varieties and hybrid varieties of both (so-called black-white cattle). Milk from holstein cattle included genetic variant a1 as the major genetic variant. In contrast, milk from the Bos indicus breed has genetic variant a2 as the major genetic variant. Similarly, milk from the gramicifu breed cattle (Bos taurus) has been shown to express high levels of beta-casein variant a2 and low levels of other beta-casein variants.

In certain embodiments, the nutritional composition may comprise protein, including 10% to 100% bovine beta-casein. Bovine beta-casein comprises from about 50% to about 100% of genetic variant a 2. Thus, as a non-limiting example, in a nutritional composition containing 10 grams of protein per serving, about 1-10 grams of the protein will be bovine beta-casein, and about 0.5-10 grams of the protein will be genetic variant a 2. In certain exemplary embodiments, the bovine beta-casein comprises from about 60% to about 100%, including from about 70% to about 100%, including from about 80% to about 100%, and including from about 90% to about 100% by weight bovine beta-casein.

The bovine beta-casein used in certain exemplary embodiments may be from a single source or may otherwise be provided by a combination of sources. Bovine beta-casein according to certain exemplary embodiments will generally be found in milk protein isolates and milk protein concentrates, but may also be found in other milk protein sources, such as whole milk, skim milk powder, milk protein concentrates, total milk protein, milk protein isolates, acid casein, calcium caseinate, sodium caseinate, magnesium caseinate, purified beta-casein, and combinations thereof. It may be possible to purify milk protein isolates (or another milk protein source) that contain too high levels of less desirable genetic variants (e.g., those other than a 2). Non-limiting examples of purification methods that can be used to reduce unacceptably high levels of the beta-casein genetic variant include: preparative chromatographic methods (e.g., affinity chromatography, ion exchange chromatography, reverse phase chromatography) or by selective salt precipitation (e.g., ammonium sulfate). Alternatively, milk protein sources (such as milk protein isolates or milk protein concentrates) containing milk derived primarily from non-holstein cows and thus reducing the number of genetic variants other than a2 may also be used in the nutritional compositions and methods disclosed herein.

The term "milk protein concentrate" is generally used to refer to a milk protein-containing product from which a substantial amount of inherent water has been removed from regular milk and from which inherent fat has also been removed. The term "milk protein isolate" is generally used to refer to a milk protein-containing product from which not only a significant amount of inherent water and inherent fat has been removed from regular milk, but also a certain amount of inherent lactose has been removed. In most cases, the milk protein isolate can be considered a further purified milk protein concentrate. Certain manufacturers may use the term milk protein concentrate to refer to milk-based protein products even though they contain at least 85% by weight protein.

The protein in the nutritional composition according to certain embodiments may be provided by a single protein source or a combination of protein sources. As previously discussed, about 10 to about 100 weight percent of the protein present in the nutritional composition comprises bovine beta-casein. The remainder of the protein (e.g., 0-90% by weight of the total protein present in the nutritional composition) may be selected from one or more other sources. As discussed in more detail below, these additional protein sources are not particularly limited and may include one or more of soy protein, whey protein, or any other protein source, including but not limited to those discussed herein. Furthermore, it will be appreciated that the source of the remainder of the protein may also be selected from milk protein isolate, milk protein concentrate, caseinate or skim milk powder which do not meet the requirements discussed previously, provided that these components are present in amounts which are not so high as to violate other restrictions on the amount of genetic modification discussed previously.

The beta-casein in the nutritional composition may be from about 0.2g/L to about 20 g/L. The beta-casein in the nutritional composition may be from about 0.5g/L to about 15 g/L. The beta-casein in the nutritional composition may be from about 1g/L to about 15 g/L. The beta-casein in the nutritional composition may be from about 5g/L to about 15 g/L. For example, the beta-casein in the nutritional composition may be about 0.2g/L, 0.4g/L, 0.6g/L, 0.8g/L, 1g/L, 1.2g/L, 1.4g/L, 1.6g/L, 1.8g/L, 2g/L, 2.2g/L, 2.4g/L, 2.6g/L, 2.8g/L, 3g/L, 3.2g/L, 3.4g/L, 3.6g/L, 3.8g/L, 4g/L, 4.2g/L, 4.4g/L, 4.6g/L, 4.8g/L, 5g/L, 5.2g/L, 5.4g/L, 5.6g/L, 5.8g/L, 6g/L, 6.2g/L, 6.6g/L, 6g/L, 6.6g/L, 6g/L, 6.6g/L, 8g/L, 6.8g/L, 6g/L, 6.8g/L, 6g/L, 6.8g/L, 6, 7g/L, 7.2g/L, 7.4g/L, 7.6g/L, 7.8g/L, 8g/L, 8.2g/L, 8.4g/L, 8.6g/L, 8.8g/L, 9g/L, 9.2g/L, 9.4g/L, 9.6g/L, 9.8g/L, 10g/L, 10.2g/L, 10.4g/L, 10.6g/L, 10.8g/L, 11g/L, 11.2g/L, 11.4g/L, 11.6g/L, 11.8g/L, 12g/L, 12.2g/L, 12.4g/L, 12.6g/L, 12.8g/L, 13g/L, 13.2g/L, 13.4g/L, 13.6g/L, 13.14 g/L, 14.8g/L, 14.2g/L, 14g/L, 14.6g/L, 14.2g/L, 14g/L, 14.4g/L, 14.6g/L, 14.8g/L, 15g/L, 15.2g/L, 15.4g/L, 15.6g/L, 15.8g/L, 16g/L, 16.2g/L, 16.4g/L, 16.6g/L, 16.8g/L, 17g/L, 17.2g/L, 17.4g/L, 17.6g/L, 17.8g/L, 18g/L, 18.2g/L, 18.4g/L, 18.6g/L, 18.8g/L, 19g/L, 19.2g/L, 19.4g/L, 19.6g/L, 19.8g/L, or 20 g/L.

Table 2: ratio of beta-casein to lipophilic nutrient

e. Optional ingredients

The nutritional compositions may further comprise other optional ingredients that may alter the physical, chemical, aesthetic or processing characteristics of the product or serve as pharmaceutical or additional nutritional components when used in the target population. Many such optional ingredients are known or otherwise suitable for use in other nutritional or pharmaceutical products, and may also be used in the nutritional compositions described herein, provided that such optional ingredients are safe and effective for oral administration and are compatible with the essential and other ingredients in the selected product form.

Non-limiting examples of such optional ingredients include carbohydrates, preservatives, antioxidants, emulsifiers, buffers, pharmaceutically active agents, other nutrients as described herein, colorants, flavorants, thickeners, stabilizers, and the like.

Non-limiting examples of carbohydrates or sources thereof suitable for use in the nutritional compositions described herein may be selected from the group consisting of: maltodextrin, hydrolyzed or modified starch or corn starch, glucose polymers, corn syrup solids, rice-derived carbohydrates, glucose, fructose, lactose, high fructose corn syrup, honey, sugar alcohols (e.g., maltitol, erythritol, sorbitol), and combinations thereof.

The nutritional composition may further comprise vitamins or related nutrients, non-limiting examples of which may be selected from the group consisting of: vitamin a, vitamin K, thiamine, riboflavin, pyridoxine, vitamin B12, carotenoids (other than lutein discussed above), niacin, folic acid, pantothenic acid, biotin, vitamin C, choline, inositol, salts and derivatives thereof, and combinations thereof.

The nutritional composition may further comprise minerals, non-limiting examples of which may be selected from the group consisting of: phosphorus, magnesium, iron, zinc, manganese, copper, sodium, potassium, molybdenum, chromium, selenium, chloride, and combinations thereof.

In some embodiments, the nutritional composition may comprise a compound selected from the group consisting of: hydroxymethylbutyrate, leucine, beta-alanine, epigallocatechin gallate, human milk oligosaccharides, prebiotics, probiotics, and combinations thereof.

The nutritional composition may also include one or more masking agents to reduce or otherwise mask bitter and aftertaste. Suitable masking agents may be selected from the group consisting of: natural and artificial sweeteners, sodium sources such as sodium chloride, and hydrocolloids such as guar gum, xanthan gum, carrageenan, gellan gum, and combinations thereof. The amount of masking in the composition can vary depending on the particular masking agent selected, the other ingredients in the composition, and other composition or product target variables. However, such amounts most often range from at least about 0.1%, including from about 0.15% to about 3.0%, and also including from about 0.18% to about 2.5%, by weight of the composition.

The nutritional composition may also optionally include one or more stabilizers. Suitable stabilizers for formulating the nutritional products include, but are not limited to, acacia, ghatti gum, karaya gum, tragacanth, agar, furcellaran, locust bean gum, pectin, low methoxyl pectin, gelatin, microcrystalline cellulose, CMC (sodium carboxymethylcellulose), methylcellulose, hydroxypropyl cellulose, DATEM (diacetyl tartaric acid esters of mono-and diglycerides), dextran, CITREM (citric acid esters of mono-and diglycerides), and mixtures thereof.

(1) Choline

An optional ingredient of the nutritional composition may comprise choline. Choline can act as a nutrient within the composition as well as an agent that enhances assembly stability and formation. Choline can also increase the lipophilic concentration within the assembly. It is hypothesized, but not bound by any particular theory, that the positive charge on choline interacts with the negative side chain of the hydrophobic protein (e.g., an aspartic acid residue) and changes the 3D configuration of the protein. This may allow more access of lipophilic compounds into the hydrophobic domains of the protein. In addition, the use of choline may be lutein specific, as choline selectively increases the amount of lutein within the assembly relative to other lipophilic compounds.

Choline can be added at different stages of the composition. At what stage choline is added to the composition is critical to the formation of a stable water-soluble assembly. In particular, the addition of choline to the protein slurry is more advantageous than the direct addition to the premix/activation premix of MDG and lipophilic compound.

Choline serves as a source of methyl groups for biosynthesis of other methylated products. It is a precursor of the neurotransmitter acetylcholine. Administration of choline has been shown to be beneficial to patients suffering from any condition associated with defective cholinergic neurotransmission.

Choline, along with lecithin, is also a major component of phospholipids and sphingomyelins. Due to its essential function in the membrane structure, choline deficiency leads to various phospholipid abnormalities, which are clinically manifested as fatty liver, kidney injury (hemorrhagic kidney necrosis), and lipoprotein metabolism disorder. When the diet is deficient in choline, cholesterol esters and fat accumulate in the liver.

Choline chloride and choline bitartrate are mentioned as nutritional/dietary supplements in the US Code of Federal Regulations (US Code of Federal Regulations), which have been granted GRAS status (generally recognized as safe).

The choline can be in the nutritional composition from about 5mg/kg to about 1 g/kg. The choline can be in the nutritional composition from about 50mg/kg to about 1 g/kg. The choline can be in the nutritional composition from about 100mg/kg to about 1 g/kg. The choline can be in the nutritional composition from about 500mg/kg to about 1 g/kg. For example, choline in the nutritional composition may be about 5mg/kg, 10mg/kg, 15mg/kg, 20mg/kg, 25mg/kg, 30mg/kg, 35mg/kg, 40mg/kg, 45mg/kg, 50mg/kg, 55mg/kg, 60mg/kg, 65mg/kg, 70mg/kg, 75mg/kg, 80mg/kg, 85mg/kg, 90mg/kg, 95mg/kg, 100mg/kg, 105mg/kg, 110mg/kg, 115mg/kg, 120mg/kg, 125mg/kg, 130mg/kg, 135mg/kg, 140mg/kg, 145mg/kg, 150mg/kg, 155mg/kg, 160mg/kg, 165mg/kg, 170mg/kg, 175mg/kg, or, 180mg/kg, 185mg/kg, 190mg/kg, 195mg/kg, 200mg/kg, 205mg/kg, 210mg/kg, 215mg/kg, 220mg/kg, 225mg/kg, 230mg/kg, 235mg/kg, 240mg/kg, 245mg/kg, 250mg/kg, 255mg/kg, 260mg/kg, 265mg/kg, 270mg/kg, 275mg/kg, 280mg/kg, 285mg/kg, 290mg/kg, 295mg/kg, 300mg/kg, 305mg/kg, 310mg/kg, 315mg/kg, 320mg/kg, 325mg/kg, 330mg/kg, 335mg/kg, 340mg/kg, 345mg/kg, 350mg/kg, 355mg/kg, 360mg/kg, 365mg/kg, 370mg/kg, 375mg/kg, 380mg/kg, 385mg/kg, 390mg/kg, 395mg/kg, 400mg/kg, 405mg/kg, 410mg/kg, 415mg/kg, 420mg/kg, 425mg/kg, 430mg/kg, 435mg/kg, 440mg/kg, 445mg/kg, 450mg/kg, 455mg/kg, 460mg/kg, 465mg/kg, 470mg/kg, 475mg/kg, 480mg/kg, 485mg/kg, 490mg/kg, 495mg/kg, 500mg/kg, 505mg/kg, 510mg/kg, 515mg/kg, 520mg/kg, 525mg/kg, 530mg/kg, 535mg/kg, 540mg/kg, 545mg/kg, 550mg/kg, 555mg/kg, 560mg/kg, 565mg/kg, 570mg/kg, 575mg/kg, 580mg/kg, 585mg/kg, 590mg/kg, 595mg/kg, 600mg/kg, 605mg/kg, 610mg/kg, 615mg/kg, 620mg/kg, 625mg/kg, 630mg/kg, 635mg/kg, 640mg/kg, 645mg/kg, 650mg/kg, 655mg/kg, 660mg/kg, 665mg/kg, 670mg/kg, 675mg/kg, 680mg/kg, 685mg/kg, 690mg/kg, 695mg/kg, 700mg/kg, 705mg/kg, 710mg/kg, 715mg/kg, 720mg/kg, 725mg/kg, 730mg/kg, 735mg/kg, 740mg/kg, 745mg/kg, 750mg/kg, 755mg/kg, 760mg/kg, 765mg/kg, 770mg/kg, 775mg/kg, 780mg/kg, 785mg/kg, 790mg/kg, 795mg/kg, 800mg/kg, 805mg/kg, 810mg/kg, 815mg/kg, 820mg/kg, 825mg/kg, 830mg/kg, 835mg/kg, 840mg/kg, 845mg/kg, 850mg/kg, 855mg/kg, 860mg/kg, 865mg/kg, 870mg/kg, 875mg/kg, 880mg/kg, 885mg/kg, 890mg/kg, 895mg/kg, 900mg/kg, 905mg/kg, 910mg/kg, 915mg/kg, 920mg/kg, 925mg/kg, 930mg/kg, 935mg/kg, 940mg/kg, 945mg/kg, 950mg/kg, 955mg/kg, 960mg/kg, 965mg/kg, 970mg/kg, 975mg/kg, 980mg/kg, 985mg/kg, 990mg/kg, 995mg/kg or 1 g/kg.

(2) Lecithin

An optional ingredient of the nutritional composition may comprise lecithin. Lecithin can serve as a nutritional source within the composition as well as an agent that enhances assembly stability and formation. Lecithin may also increase the lipophilic concentration within the assembly. Lecithin is primarily a mixture of glycerophospholipids (e.g., phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol). Phosphatidylcholine is generally the major glycerophospholipid component. Lecithin may also contain other compounds such as free fatty acids, monoglycerides, diglycerides, triglycerides, glycolipids and other lipid/fatty acid containing compounds. Lecithins are sometimes classified as glycerophospholipids or phospholipids. This class of compounds has amphiphilic properties and therefore an emulsifying function.

Lecithin is typically added to liquid food products, including nutritional compositions, so that the liquid product remains homogeneous and does not separate. Lecithin is approved for human consumption by the United States Food and Drug Administration and is in a state of "recognized safety". Non-limiting examples of lecithins suitable for use herein include egg yolk lecithin, wheat lecithin, corn lecithin, soy lecithin, modified lecithins, and combinations thereof. Lecithin may be provided in a defatted or liquid form or in a phosphatidylcholine-enriched form. In addition, lecithin may be derived from sources including, but not limited to, organic soy, canola, skim milk powder, or whey protein.

In one embodiment, the nutritional powder may comprise de-oiled lecithin, wherein the de-oiled lecithin comprises: 1% of water; 3% triglyceride; 24% phosphatidylcholine; 20% phosphatidylethanolamine; 14% phosphatidylinositol; 7% phosphatidic acid; 8% of a small amount of phospholipid; 8% of complex sugar; and 15% glycolipid.

In another embodiment, the nutritional powder may comprise liquid lecithin, wherein the liquid lecithin comprises: 1% of water; 37% triglycerides; 16% phosphatidylcholine; 13% phosphatidylethanolamine; 10% phosphatidylinositol; 5% phosphatidic acid; 2% of a small amount of phospholipid; 5% of complex sugar; and 11% glycolipid.

Lecithin suitable for use herein may be obtained from any known or otherwise suitable nutritional source. Non-limiting examples include soy lecithin from ADM Specialty Food Ingredients, Decatur, il., USA; soy lecithin from Solae, LLC, st.louis, mo., USA; and soy Lecithin from American Lecithin Company, Oxford, Conn., USA.

The lecithin in the nutritional composition may be from about 10mg/kg to about 5 g/kg. The lecithin in the nutritional composition may be from about 50mg/kg to about 5 g/kg. The lecithin in the nutritional composition may be from about 100mg/kg to about 1 g/kg. The lecithin in the nutritional composition may be from about 500mg/kg to about 1 g/kg. For example, the lecithin in the nutritional composition may be about 10mg/kg, 15mg/kg, 20mg/kg, 25mg/kg, 30mg/kg, 35mg/kg, 40mg/kg, 45mg/kg, 50mg/kg, 55mg/kg, 60mg/kg, 65mg/kg, 70mg/kg, 75mg/kg, 80mg/kg, 85mg/kg, 90mg/kg, 95mg/kg, 100mg/kg, 105mg/kg, 110mg/kg, 115mg/kg, 120mg/kg, 125mg/kg, 130mg/kg, 135mg/kg, 140mg/kg, 145mg/kg, 150mg/kg, 155mg/kg, 160mg/kg, 165mg/kg, 170mg/kg, 175mg/kg, 180mg/kg, 175mg/kg, 185mg/kg, 190mg/kg, 195mg/kg, 200mg/kg, 205mg/kg, 210mg/kg, 215mg/kg, 220mg/kg, 225mg/kg, 230mg/kg, 235mg/kg, 240mg/kg, 245mg/kg, 250mg/kg, 255mg/kg, 260mg/kg, 265mg/kg, 270mg/kg, 275mg/kg, 280mg/kg, 285mg/kg, 290mg/kg, 295mg/kg, 300mg/kg, 305mg/kg, 310mg/kg, 315mg/kg, 320mg/kg, 325mg/kg, 330mg/kg, 335mg/kg, 340mg/kg, 345mg/kg, 350mg/kg, 355mg/kg, 360mg/kg, 365mg/kg, 370mg/kg, 375mg/kg, 380mg/kg, 385mg/kg, 390mg/kg, 395mg/kg, 400mg/kg, 405mg/kg, 410mg/kg, 415mg/kg, 420mg/kg, 425mg/kg, 430mg/kg, 435mg/kg, 440mg/kg, 445mg/kg, 450mg/kg, 455mg/kg, 460mg/kg, 465mg/kg, 470mg/kg, 475mg/kg, 480mg/kg, 485mg/kg, 490mg/kg, 495mg/kg, 500mg/kg, 505mg/kg, 510mg/kg, 515mg/kg, 520mg/kg, 525mg/kg, 530mg/kg, 535mg/kg, 540mg/kg, 545mg/kg, 550mg/kg, 555mg/kg, 560mg/kg, 565mg/kg, 570mg/kg, 575mg/kg, 580mg/kg, 585mg/kg, 590mg/kg, 595mg/kg, 600mg/kg, 605mg/kg, 610mg/kg, 615mg/kg, 620mg/kg, 625mg/kg, 630mg/kg, 635mg/kg, 640mg/kg, 645mg/kg, 650mg/kg, 655mg/kg, 660mg/kg, 665mg/kg, 670mg/kg, 675mg/kg, 680mg/kg, 685mg/kg, 690mg/kg, 695mg/kg, 700mg/kg, 705mg/kg, 710mg/kg, 715mg/kg, 720mg/kg, 725mg/kg, 730mg/kg, 735mg/kg, 220mg/kg, 725mg/kg, 220mg/kg, 260mg/kg, 220mg/kg, 160mg/kg, 740mg/kg, 745mg/kg, 750mg/kg, 755mg/kg, 760mg/kg, 765mg/kg, 770mg/kg, 775mg/kg, 780mg/kg, 785mg/kg, 790mg/kg, 795mg/kg, 800mg/kg, 805mg/kg, 810mg/kg, 815mg/kg, 820mg/kg, 825mg/kg, 830mg/kg, 835mg/kg, 840mg/kg, 845mg/kg, 850mg/kg, 855mg/kg, 860mg/kg, 865mg/kg, 870mg/kg, 875mg/kg, 880mg/kg, 885mg/kg, 890mg/kg, 895mg/kg, 900mg/kg, 905mg/kg, 910mg/kg, 915mg/kg, 920mg/kg, 150mg/kg, 925mg/kg, 930mg/kg, 935mg/kg, 940mg/kg, 945mg/kg, 950mg/kg, 955mg/kg, 960mg/kg, 965mg/kg, 970mg/kg, 975mg/kg, 980mg/kg, 985mg/kg, 990mg/kg, 995mg/kg, 1g/kg, 1.2g/kg, 1.4g/kg, 1.6g/kg, 1.8g/kg, 2g/kg, 2.2g/kg, 2.4g/kg, 2.6g/kg, 2.8g/kg, 3g/kg, 3.2g/kg, 3.4g/kg, 3.6g/kg, 3.8g/kg, 4g/kg, 4.2g/kg, 4.4g/kg, 4.6g/kg, 4.8g/kg, or 5 g/kg.

4. Manufacturing method

To prepare a nutritional composition having enhanced bioavailability of the lipophilic compounds contained therein, methods according to the present disclosure may be utilized. The method may comprise the steps of: forming a premix; adding the premix to a solution to form an activating premix or a protected premix, and adding the activating premix to a composition, wherein the assembly can be formed at least in the activating premix.

a. Premix compound

In some embodiments, a premix comprising MDG is provided. The premix may comprise only MGD, MGD and at least one lipophilic compound, as well as other optional ingredients as described above. The premix may be activated. The premix may be heated prior to activation. In some embodiments, the premix is heated to about 85 ° F for 30 minutes. In some embodiments, the premix is heated to 120 ° F for about 10 minutes. The skilled artisan will appreciate that the premix composition can affect the temperature and time required to heat the premix. The premix may be protected, for example, by a surfactant.

(1) Activating premix

The premix may be added to an aqueous solution, such as a protein-in-water slurry, to form an "activation premix". The activation premix is then added to a composition that may comprise at least one lipophilic compound.

Notably, to form an "activating premix," the premix may not be added to the oil in an amount sufficient to cause separation of the MDG from the assembly in the oil prior to adding the premix to the aqueous solution. In this case, the activation premix may be protected to limit the separation of the MDG and lipophilic compounds from each other.

When the "activation premix" is subsequently added to the fat-containing solution, at least a portion of the MDG is not separated from the assembly by the fat in the fat-containing solution. To determine the amount of mono-and diglycerides that are not separated in fat, the "separation MDG (mono-and diglycerides) test" may be utilized.

The isolated MDG test is as follows: the fat-containing solution was subjected to high speed centrifugation (31,000x g; 20 ℃; 4h) as described above. HPLC analysis of the resulting aqueous fraction was performed as described in "Determination of food emulsions in general additives and food products by liquid Chromatography/apparatus-compression chemical Chromatography", M.Sum an et al, Journal of Chromatography A, 1216(2009)3758-3766, to obtain the mono-and diglyceride content. The levels of mono-and diglycerides detected in the aqueous fraction are compared to the levels added to the fat containing solution to yield the percentage of the mixture of mono-and diglycerides that is not separated by fat.

Using the split MDG test, the effect of using an activated premix in a composition versus using a premix that has not been activated can be determined. This can be achieved by comparing the amount of mono-and diglycerides present in the aqueous portion of the composition comprising the activated premix with the amount of mono-and diglycerides present in the aqueous portion of the same composition wherein the premix has not been activated. The aqueous portion of the composition comprising the activated premix will typically contain higher amounts of mono-and diglycerides than the same composition wherein the premix is not activated.

The difference between the content of mono-and diglycerides in the aqueous portion of a composition comprising an activated premix and the content of mono-and diglycerides in the aqueous portion of the same composition comprising an unactivated premix can be quantified. In some embodiments, the content of monoglycerides and diglycerides in the aqueous portion of the composition comprising the activated premix may be at least about 5%, 10%, or 15% higher than in the same composition comprising the unactivated premix.

(b) Variants of manufacture

The premixes, activation premixes, and compositions may comprise different embodiments, some of which are listed below. It should be noted that the nutritional composition is not limited to the representative embodiments listed below.

In some embodiments, a premix comprising a lipophilic compound and MDG is provided. To prepare the premix, the lipophilic compound and MDG may be mixed in the presence of heat or at ambient temperature and in some embodiments with agitation to allow the lipophilic compound to dissolve in the monoglycerides and diglycerides. The premix is added to an aqueous solution, such as a protein-in-water slurry, to form an activated premix containing the assembly. The "activation premix" is then added to the composition.

In some embodiments, a premix comprising a lipophilic compound, a surfactant, MDG is provided. To prepare the premix, the lipophilic compound and MDG can be mixed in the presence of heat or at ambient temperature and in some embodiments with agitation to allow the lipophilic compound to dissolve in the mixture of mono-and diglycerides. The premix is added to a fat-containing solution, such as a fat-in-protein slurry, to form a "protected premix" containing the assembly. The "protected premix" is then added to the composition.

Without wishing to be bound by theory, it is believed that when the premix is added to the fat-containing solution, the surfactant combines with the lipophilic compound, the mono-and diglycerides to form a microemulsion. Thus, in a "protected premix", the monoglycerides and diglycerides can be protected from separation in the fat-containing solution.

The activation premix or protected premix may be added to any other ingredient of the nutritional or pharmaceutical composition at any useful point in the manufacturing process. In some embodiments, the nutritional or pharmaceutical composition may then be dried using any method known in the art to form a powdered composition. For example, a nutritional powder may be prepared by preparing at least two slurries, later mixing the two slurries together (and may be further mixed with an activation premix), heat treating, standardizing, secondary heat treating, evaporating to remove water, and spray drying to form a powdered nutritional composition.

Slurries to which the activation premix or protected premix may be added include carbohydrate-mineral (CHO-MIN) slurries and protein-in-oil (PIO) slurries. First, a CHO-MIN slurry may be formed by dissolving selected carbohydrates (e.g., lactose, galactooligosaccharides, etc.) in hot water with agitation, followed by the addition of minerals (e.g., potassium citrate, magnesium chloride, potassium chloride, sodium chloride, choline chloride, etc.). The resulting CHO-MIN slurry may then be maintained under constant heating and gentle agitation until it is later mixed with other prepared slurries.

Protein-in-oil (PIO) slurries can be formed by heating and mixing oils (e.g., high oleic safflower oil, soybean oil, coconut oil, monoglycerides) and emulsifiers (e.g., soybean lecithin), and then adding oil soluble vitamins, proteins (e.g., milk protein concentrate, milk protein hydrolysate, etc.), carrageenan (if any), calcium carbonate or tricalcium phosphate (if any), and arachidonic acid (ARA) oil and docosahexaenoic acid (DHA) oil (if any) with continued heating and stirring. The resulting PIO slurry can be maintained under constant heating and gentle agitation until it is later mixed with other prepared slurries.

The water was heated and then combined with the CHO-MIN slurry, skim milk (if any), and PIO slurry under sufficient agitation. The pH of the resulting mixture was adjusted to 6.6-7.0 and the mixture was kept under gentle heating agitation. In some embodiments, ARA oil and DHA oil are added at this stage. In some embodiments, the activation premix or protected premix is mixed with one or more of hot water, CHO-MIN slurry, skim milk (if any), and PIO slurry. Alternatively, the activation premix or protected premix may be mixed into the composition after further processing and immediately prior to drying the composition.

The composition may then be subjected to High Temperature Short Time (HTST) processing during which the composition is heat treated, emulsified and homogenized, and then cooled. Water soluble vitamins and ascorbic acid are added, the pH adjusted to the desired range if necessary, flavors (if any) are added, and water is added to achieve the desired total solids level. In some embodiments, the emulsion may then be further diluted and subsequently processed and packaged into a ready-to-drink or concentrated liquid. In some embodiments, the emulsion is evaporated, heat treated and then processed and packaged into a reconstitutable powder (e.g., spray-dried, dry-blended, agglomerated).

The spray-dried or dry-blended powdered nutritional compositions may be prepared by any collection of known or otherwise effective techniques suitable for preparing and formulating nutritional powders. For example, the spray-drying step may likewise include any spray-drying technique known for or otherwise suitable for use in the production of nutritional powders. Many different spray-drying processes and techniques are known for use in the nutritional field, all of which are suitable for the manufacture of spray-dried powdered nutritional compositions. After drying, the finished powder may be packaged in suitable containers.

5. Application method

The nutritional compositions may be packaged and sealed in single-use or multi-use containers and then stored under ambient conditions or under refrigeration for up to 36 months or more, more typically from about 6 months to about 24 months. For multiple use containers, these packages may be opened and then capped for reuse by the end user. For liquid embodiments, the package opened and then reclosed is typically stored under refrigerated conditions, and the contents are used within about 7 days. For powder embodiments, the package opened and then reclosed can typically be stored under ambient conditions (e.g., to avoid extreme temperatures) and the contents used within about a month. Non-limiting examples of ways in which the present formulations can be utilized include use as the following products: beverages, such as coffee beverages, cocoa or chocolate beverages, malt beverages, fruit or juice beverages, carbonated beverages, soft drinks, or milk-based beverages; performance nutritional products, such as performance nutritional bars, powders, or ready-to-drink beverages; a medical nutrition product; dairy products, such as milk drinks, yogurt or other fermented dairy products; an ice cream product; confectionery products, such as chocolate products; functional foods or drinks such as weight-loss products, fat-burning products, products for improving mental performance or preventing deterioration of mental ability, or products for improving skin. The beverage according to the invention may be in the form of a powder or liquid concentrate to be mixed with a suitable liquid (e.g. water or milk) before consumption; or in the form of a ready-to-drink beverage.

In embodiments directed to enhancing the absorption of a lipophilic nutrient, the lipophilic nutrient in a nutritional composition comprising the lipophilic nutrient and the lipophilic MDG carrier oil of the nutritional composition is administered to a subject. The absorption of lipophilic nutrients may be enhanced in one or more measurable ways. Absorption of lipophilic nutrients can be enhanced by increasing the maximum plasma concentration of the nutrient in the blood of a subject ingesting the nutrient. Absorption of lipophilic nutrients can be enhanced by prolonged plasma absorption of the nutrients in the blood of a fed subject over a period of days or weeks. Absorption of lipophilic nutrients can be enhanced by increasing the total plasma concentration of the nutrient in the blood of a fed subject over a period of days or weeks.

In embodiments where the lipophilic compound is lutein, a powdered nutritional composition including lutein may be administered to ameliorate age-related macular degeneration as well as other retinal diseases and conditions. Although in some embodiments, the methods of the present disclosure may be directed to individuals having age-related macular degeneration or other retinal diseases and disorders, the methods of the present disclosure as described herein are also intended in some embodiments to include the use of such methods in "at-risk" individuals, including individuals who are not affected by or otherwise have such diseases or disorders, for preventing, minimizing, or delaying the progression of such diseases and conditions over time. For such prophylactic purposes, the methods of the present disclosure preferably comprise continuous daily administration of a composition as described herein. Such prevention methods may be directed to adults or others, especially the elderly, who are susceptible to the development of age-related macular degeneration or other retinal diseases and disorders due to genetic factors, environmental factors, and the like.

A composition comprising vitamin D can be administered to a subject to promote healthy skeletal development, reduce vitamin D deficiency, increase bone strength, maintain or increase bone mineralization, and maintain or increase bone mineral density. Compositions comprising vitamin D may also be administered to subjects suffering from or at risk of diseases and conditions associated with insufficient bone mineralization, such as rickets, osteomalacia, osteoporosis, osteopenia and bone fractures. Subjects receiving compositions containing vitamin D can include, but are not limited to, pregnant and postpartum women, infants, children, adolescents, adults, postmenopausal women, and the elderly. In some embodiments, the methods of the present disclosure are directed to a pregnant or postpartum woman for promoting healthy skeletal development of a developing fetus or infant. In some embodiments, the methods of the present disclosure are directed to infants and children for promoting healthy skeletal development and preventing rickets or bone fractures in growing children. In some embodiments, the methods of the present disclosure are directed to adolescents and adults for promoting increased bone mineralization and bone mineral density. In some embodiments, the methods of the present disclosure are directed to a postpartum woman for preventing or slowing the onset of osteoporosis. In some embodiments, the methods of the present disclosure are directed to adults with osteomalacia, osteoporosis, osteopenia, or other bone diseases and disorders. In some embodiments, the methods of the present disclosure are directed to "at-risk" individuals, including individuals who are not affected by or otherwise have cartilage disease, osteoporosis, osteopenia, loss of balance and fall, bone fracture, or other bone disease or disorder, for preventing, minimizing, or delaying the progression of such diseases and conditions over time. For such development, maintenance and prevention purposes, the methods of the present disclosure preferably comprise continuous daily administration of a composition as described herein. Such development, maintenance and prevention methods are directed to subjects such as pregnant and postpartum women, infants, children, adolescents, adults, postmenopausal women and the elderly. The method of prevention is particularly directed to elderly persons who are susceptible to the development of age-related osteoporosis, osteopenia, loss of balance or falls, bone fractures or other diseases and disorders associated with inadequate vitamin D intake or inadequate bone mineralization.

In these embodiments, the subject desirably consumes a portion of the composition that provides an effective amount of vitamin D per day. Effective amounts of vitamin D range from about 50IU to about 7,500 IU/day, including from about 100 IU/day to about 5,000 IU/day, including from about 200 IU/day to about 2,500 IU/day, including from about 250 IU/day to about 1,500 IU/day, including from about 400 IU/day to about 1,000 IU/day, and including from about 500 IU/day to about 800 IU/day.

In embodiments where the lipophilic nutrient is vitamin E, a nutritional composition including vitamin E may be administered to a subject to provide antioxidants, promote cardiovascular health, and prevent or reduce the risk of certain types of cancer. Subjects receiving a nutritional composition containing vitamin E may include, but are not limited to, pregnant and postpartum women, infants, children, and adults. In some embodiments, the methods of the present disclosure may be directed to a pregnant or postpartum woman for promoting healthy development of a developing fetus or infant. In some embodiments, the methods of the present disclosure may be directed to infants and children for promoting healthy development in a growing child. In some embodiments, the methods of the present disclosure may be directed to adults with cardiovascular disease. In some embodiments, the methods of the present disclosure are also directed to "at-risk" individuals, including individuals who are not affected by or otherwise suffering from cancer, cardiovascular disease, or other diseases or disorders, for preventing, minimizing, or delaying the progression of such diseases and conditions over time. For such development, maintenance and prevention purposes, the methods of the present disclosure preferably comprise continuous daily administration of the nutritional compositions as described herein. Such development, maintenance and prevention methods may be directed to subjects such as pregnant and postpartum women, infants, children and adults (especially the elderly) who are susceptible to developing cancer, cardiovascular disease or other diseases and conditions due to genetic factors, environmental factors, and the like.

In embodiments where the lipophilic nutrient is DHA, the nutritional composition including DHA can be administered to a subject to promote healthy eye, brain, and central nervous system development, maintain overall brain health, prevent or reduce age-related mental function decline, and reduce cardiovascular disease, among other diseases and disorders. Subjects receiving a nutritional composition containing DHA may include, but are not limited to, pregnant and postpartum women, infants, children, and adults. In some embodiments, the methods of the present disclosure may be directed to a pregnant or postpartum woman for promoting development of the eye, brain and central nervous system of a developing fetus or infant. In some embodiments, the methods of the present disclosure may be directed to infants and children for promoting eye, brain, and central nervous system development in a growing child. In some embodiments, the methods of the present disclosure may be directed to adults for maintaining overall brain health, preventing or reducing age-related mental function decline, and reducing cardiovascular disease, as well as other diseases and disorders. In some embodiments, the methods of the present disclosure are also directed to "at-risk" individuals, including individuals who are not affected by age-related mental decline or cardiovascular disease or who do not otherwise have such disease, for preventing, minimizing, or delaying the progression of such diseases and conditions over time. For such development, maintenance and prevention purposes, the methods of the present disclosure preferably comprise continuous daily administration of the nutritional compositions as described herein. Such development, maintenance and prevention methods may be directed to subjects such as pregnant and postpartum women, infants, children and adults (especially the elderly) who are predisposed to age-related decline in mental function, cardiovascular disease or other diseases and conditions due to genetic factors, environmental factors, and the like.

The nutritional composition including HPAV oil and activated MDG oil components or protected premixes can be administered to infants to enhance palmitic acid absorption, promote healthy skeletal development, and maintain healthy bone mineral density. Subjects receiving compositions with HPAV oil and activated MDG oil components or protected premixes include, but are not limited to, preterm and term infants. In some embodiments, the methods of the present disclosure may be directed to infants for promoting healthy skeletal development and maintaining healthy bone mineral density in a growing infant. For such development and maintenance purposes, the methods of the present disclosure preferably comprise continuous daily administration of the nutritional compositions as described herein. Such development and maintenance methods may be directed to subjects such as preterm and term infants.

In these embodiments, the infant ideally consumes a portion of the nutritional composition that provides an effective amount of HPAV oil per day. An effective amount of HPAV oil ranges from about 0.6g to about 18 g/day, including from about 1 g/day to about 15 g/day, including from about 2.5 g/day to about 12.5 g/day, including from about 5 g/day to about 10 g/day, and including from about 6 g/day to about 8 g/day.

A nutritional composition including DHA and lutein may be administered to a subject to promote healthy eye and retinal development, maintain overall eye health, and prevent or reduce age-related macular degeneration as well as other diseases and disorders of the eye. Subjects receiving a nutritional composition containing DHA and lutein may include, but are not limited to, pregnant and postpartum women, infants, children, and adults. In some embodiments, the methods of the present disclosure may be directed to pregnant or postpartum women for promoting eye and retina development and healthy vision in a developing fetus or infant. In some embodiments, the methods of the present disclosure may be directed to infants for promoting eye and retina development and visual acuity in growing children. In some embodiments, the methods of the present disclosure may be directed to children and adults for improving visual acuity, improving light stress recovery, or reducing glare sensitivity. In some embodiments, the methods of the present disclosure may be directed to adults with age-related macular degeneration, glaucoma, cataracts, lacrimal keratoconjunctivitis, or other diseases and disorders of the eye. In some embodiments, the methods of the present disclosure are also directed to "at-risk" individuals, including individuals who are not affected by or otherwise have age-related macular degeneration, glaucoma, cataracts, lacrimal keratoconjunctivitis, diabetic retinopathy, or other eye diseases or disorders, for preventing, minimizing, or delaying the progression of such diseases and conditions over time. For such development, maintenance and prevention purposes, the methods of the present disclosure preferably comprise continuous daily administration of the nutritional compositions as described herein. Such development, maintenance and prevention methods may be directed to subjects such as pregnant and postpartum women, infants, children and adults (especially the elderly) who are susceptible to the development of age-related macular degeneration or other eye diseases and disorders due to genetic factors, environmental factors, and the like.

In these embodiments, the subject ideally consumes a portion of the composition that provides an effective amount of DHA and lutein per day. An effective amount of DHA is in the range of about 5mg to about 10 g/day, including about 10mg to about 1 g/day, including about 20mg to about 500 mg/day, including about 40mg to about 200 mg/day, and including about 80mg to about 150 mg/day. An effective amount of lutein is in the range of about 5 μ g to about 10 mg/day, including about 10 μ g to about 5 mg/day, including about 25 μ g to about 1 mg/day, including about 50 μ g to about 500 μ g/day, and including about 100 μ g to about 250 μ g/day.

In these embodiments, the individual desirably consumes at least one serving of the nutritional composition per day, and in some embodiments may consume two, three, or even more servings per day. Each serving is desirably administered as a single undivided dose, however the serving may also be divided into two or more portions or divided serving amounts for ingestion in two or more portions during the day. The methods of the present disclosure include continuous daily administration as well as regular or limited administration, although continuous daily administration is generally desirable. The nutritional composition may be for infants, children and adults.

The nutritional compositions have a number of aspects illustrated by the following non-limiting examples.

5. Examples of the embodiments

It should be understood that the foregoing detailed description and accompanying examples are illustrative only and should not be taken as limiting the scope of the invention, which is defined solely by the appended claims and equivalents thereof.

Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications, including but not limited to those relating to the chemical structures, substituents, derivatives, intermediates, syntheses, compositions or methods of use of the invention, may be made without departing from the spirit and scope thereof.

The following examples illustrate specific embodiments and/or features of nutritional compositions comprising an activating premix and compositions comprising a protected premix. The examples given are for illustrative purposes only and should not be taken as limitations of the present disclosure, as many variations thereof are possible without departing from the spirit and scope of the present disclosure. All exemplified amounts are weight percentages based on the total weight of the composition, unless otherwise specified.

Exemplary compositions can be prepared according to the manufacturing methods described herein such that each exemplary nutritional or pharmaceutical composition has an increased bioavailability of the lipophilic compound.

The following details may be applied to the embodiments listed below where appropriate. Variations other than those listed below are set forth in the specific embodiments.

The Size Exclusion Chromatography (SEC) system used in the following examples was as follows: column: superdex Peptide 10/300GL, GE Healthcare P/N17-5176-01; mobile phase: 700mL Milli-Q Plus water, 300mL acetonitrile; 1.0mL of trifluoroacetic acid; flow rate: 0.4 mL/min; temperature: ambient temperature (about 21 ℃); and (3) detection: UV light at 214nm, 280 nm; visible light at 476 nm; and (3) injection: 10 mu L of the solution; operating time: 70 minutes; sample preparation: diluting 0.6-1.1g to 10mL with mobile phase; calibration: the protein was purified by 6, 14.2-160kD reference proteins.

The high molecular weight xanthophyll assemblies were determined as follows: SEC peak elution ≤ 20.0 min, detected with visible light at 476nm (maximum absorbance of lutein). The high MW lutein assembly is a protein-based water dispersible or water soluble macromolecular aggregate with lutein incorporated therein. The size of the high MW xanthophyll assemblies is approximately in the 60-380kD range (approximately ). The concentration of the high MW lutein assembly was quantified as peak area (mAU-min at 476 nm)/mg injected sample. The value reported as high MW lutein assembly is not the absolute concentration of bound lutein but rather a relative measure of bound lutein in these high MW but water dispersible/soluble macromolecular aggregates.

The protein > 137kD was determined as follows: SEC peak elution ≤ 18.0 min, using UV detection at 214nm (peptide bond and amino acid side chain signals). The peak consists mainly of aggregated proteins; for example, it is larger in size than all major milk proteins (about 14 to 25kD), so the proteins in this peak (> 137kD) are considered aggregates but are water dispersible/soluble and capable of encapsulating lutein and other oil soluble vitamins/nutrients. It is quantified as g protein/kg sample. The peak area of the protein was measured relative to known concentrations to provide g/kg protein.

The bound lipophilic substance index was determined as follows: non-specific indicator of lipophilic nutrients (including lutein in combination with other oil-soluble vitamins) in the high MW protein peak (as described above). The values reported are the peak area ratio (280nm/214nm) for SEC peak elution ≦ 18.0 min, which is a relative indicator of the lipophilic nutrient concentration in the high MW aggregates. The validity of this index is based on the fact that: the ratio of 280nm/214nm of the lipophilic nutrient is greater than the ratio of 280nm/214nm of the protein.

Examples 1 to 5

Examples 1-5 illustrate powdered infant formulas of the present disclosure, the ingredients of which are listed in the following table. Unless otherwise specified, all ingredient amounts are listed in kg/1000kg batch.

Table 3: examples 1-5 and other exemplary compositions.

Examples 6 to 10

Examples 6-10 illustrate powdered infant formulas of the present disclosure, the ingredients of which are listed in the following table. Unless otherwise specified, all ingredient amounts are listed in kg/1000kg batch.

Table 4: nutritional composition ingredients for examples 6-10 and other exemplary compositions.

Examples 11 to 15

Examples 11-15 illustrate powdered adult formulations of the present disclosure, the ingredients of which are listed in the table below. Unless otherwise specified, all ingredient amounts are listed in kg/1000kg batch.

Table 5: examples 11-15 and other exemplary compositions of nutritional composition ingredients.

Examples 16 to 18

Examples 16-18 illustrate liquid adult formulations of the present disclosure, the ingredients of which are listed in the following table. Unless otherwise specified, all ingredient amounts are listed in kg/1000kg batch.

Table 6: examples 16-18 and other exemplary compositions.

Example 19

In this example, the effect of mixing lutein (a polar lipid nutrient) into a combination of mono-and diglycerides on the bioavailability of lutein was evaluated and compared to the bioavailability of lutein mixed in a typical triglyceride base oil.

Male Sprague Dawley rats weighing between 280 and 330 grams were all fed for one week with standard commercial Purina (Purina) rat chowder. The rats were then fasted overnight before surgery and subjected to laparotomy under anesthesia and a laparotomy, and a surgery performed according to Tso et al, "The Absorption of Lipid and Lipid Synthesis", Lipid Research Methodology, chapter 5: 191-216(1984) Alan r. loss, inc., NY (incorporated herein by reference to the extent consistent herewith) intubated the intestinal lymphatic vessels. The superior mesenteric artery was isolated, but not occluded. A silicon infusion tube (1.6mm OD) was placed in the stomach for future infusion of the test sample. The fundoplication is closed by purse string sutures. Rats were allowed to recover for 24 hours before infusion was initiated.

The test samples included the following solutions: (1) safflower oil solution of lutein (control); (2) a solution of lutein in mono-and diglycerides derived from corn oil; and (3) a solution of lutein in mono-and diglycerides derived from high oleic sunflower oil.

Rats were randomly divided into three groups. At 24 hours post-surgery, the animals were infused intragastrically with 20mg/kg lutein from the corresponding lutein-containing solution.

Lymph was collected in pre-cooling tubes 1 hour prior to lutein infusion (fasting) and then every other hour after infusion started (for 6 hours). At the end of the 6 hour infusion, the rats were sacrificed by exsanguination.

Lymph lipids were extracted and analyzed for lutein concentration using high performance liquid chromatography with programmed wavelength ultraviolet detection (Craft Technologies, Wilson, NC).

Figure 1 shows the lymph flow rate per hour over 6 hours after lutein administration. Figure 2 shows the hourly lutein output in lymph within 6 hours after lutein administration. Figure 3 shows the cumulative lymph lutein absorption over 6 hours after lutein administration. Figure 4 shows the percent change in lymphatic lutein absorption over a 6 hour period with mono-and diglycerides versus control.

As shown in figure 1, the mean fasting lymphatic flow varied between 2.3 and 2.6mL/h for all three rat groups. In all groups, lymphatic flow increased significantly after lutein infusion and reached maximum output between 2.7 and 3.9mL/h during 2-3 hours after lipid infusion. There was no difference in lymphatic flow rates between groups, indicating that the use of lutein mono-and di-glyceride solutions did not significantly affect lymphatic flow rates relative to lutein triglyceride control solutions. Figure 2 shows the lymphatic lutein output (in μ g/h) during the first 6 hours after feeding 3 different lutein samples. In all groups, the lymphatic output of lutein increased during the first 3 hours and reached steady state 3 to 5 hours after lutein administration. However, the output of lymphatic lutein 2-6 hours after lutein administration along with mono-and diglycerides (corn and HOSO) was significantly increased compared to lutein in triglyceride control. Figure 3 shows the above-described improvement in lutein absorption obtained using mono-and diglycerides by demonstrating a significantly higher cumulative lutein absorption (area under the curve (AUC)) over a 6 hour absorption period relative to a triglyceride control solution of lutein. When this data is expressed as a percentage change in lympholutein uptake, as shown in figure 4, the lympholutein uptake of lutein plus mono-and diglycerides increased by 78% -88% over the 6 hour infusion period relative to the control. This data indicates that the polar lipid nutrient lutein has increased bioavailability in a mixture of mono-and diglycerides compared to triglyceride oil.

Example 20

In this example, the effect of the addition of the activating premix comprising lutein and the addition of the protected premix on the bioavailability of lutein in the nutritional composition was evaluated and compared to the bioavailability of lutein typically added to the triglyceride based oil prior to addition to the nutritional composition.

Male Sprague Dawley rats weighing between 280 and 330 grams were all fed for one week with standard commercial Purina (Purina) rat chowder. The rats were then fasted overnight before surgery and subjected to laparotomy under anesthesia and a laparotomy, and a surgery performed according to Tso et al, "The Absorption of Lipid and Lipid Synthesis", Lipid Research Methodology, chapter 5: 191-216(1984) Alan r. loss, inc., NY (incorporated herein by reference to the extent consistent herewith) intubated the intestinal lymphatic vessels. The superior mesenteric artery was isolated, but not occluded. A silicon infusion tube (1.6mm OD) was placed in the stomach for future infusion of the test sample. The fundoplication is closed by purse string sutures. Rats were allowed to recover for 24 hours before infusion was initiated.

The test samples included the following nutritional compositions: (AET-C) lutein was added to high oleic safflower oil for 10 minutes at 120 ℃ F. to the lipo-coated protein slurry (control); (AET-1) lutein was premixed with mono-and diglycerides at 120 ° F for 10 minutes, lecithin was added to the heated premix and allowed to mix for 5 minutes, added to the fat-coated protein slurry to form the MDG protected premix, then added to the rest of the composition; (AET-2) lutein is premixed with mono-and diglycerides at 120 ° F for 10 minutes, added to a protein-in-water slurry to form an activated premix, then added to the remainder of the composition; (AET-3) lutein was premixed with mono-and diglycerides at 120 ° F for 10 minutes, added to the lipo-coated protein slurry, then added to the remainder of the composition; and (AET-4) lutein was premixed with mono-and diglycerides at 120 ° F for 10 minutes, choline was added as choline chloride to the heated premix and allowed to mix for 5 minutes, added to the protein-in-water slurry to form an activated premix, and then added to the remainder of the composition. The composition of each test sample is provided in detail in the table below.

Table 7: example 20 and other exemplary compositions of nutritional composition ingredients.

The rats were randomly divided into five groups. After 24 hours of surgery, the animals were infused intragastrically with 3ml of the nutritional formula delivering 5.8 μ g lutein/animal from the corresponding lutein-containing solution.

Lymph was collected in pre-cooling tubes 1 hour before infusion of the nutritional composition (fasting) and then every other hour after infusion started (for 8 hours). At the end of the 8 hour infusion, the rats were sacrificed by exsanguination.

Lymph lipids were extracted and analyzed for lutein concentration using high performance liquid chromatography with programmed wavelength ultraviolet detection (Craft Technologies, Wilson, NC).

Fig. 5 shows lymph flow rate per hour over a period of 8 hours after administration of liquid infant composition containing lutein. Fig. 6 shows the hourly lutein output in lymph over 8 hours after administration of a liquid infant composition containing lutein. Fig. 7 shows the cumulative lymph lutein absorption over 6 and 8 hours after administration of liquid infant composition containing lutein. Figure 8 shows the percentage change in lymphatic lutein absorption after administration of liquid infant composition containing lutein versus 6 and 8 hours after administration of control.

As shown in fig. 5, the mean fasting lymph flow for all three rat groups varied between 2.4 and 2.7 mL/h. In all groups, lymphatic flow increased significantly after infusion of the nutritional composition and reached a maximum output between 2.8 and 3.1mL/h during 2-3 hours after infusion of the nutritional composition. There was no difference in lymph flow rates between groups, indicating that the lymph flow rate was not significantly affected using a nutritional composition including an activated premix with lutein or an MDG protected premix with lutein relative to a control nutritional composition including a lutein triglyceride solution. Figure 6 shows the lympholutein output (in μ g/h) during the first 8 hours after feeding 5 different nutritional composition samples. In all groups, the lymphatic output of lutein increased during the first 3 hours and reached steady state 2 to 3 hours after administration of the nutritional formula. However, when lutein is given as an activated premix (AET-2, AET-4) or lutein is given as an MDG protected premix (AET-1), there is a significant increase in lymphatic lutein output 2-8 hours after lutein administration compared to lutein or non-activated premixes containing lutein, monoglycerides and diglycerides (AET-C and AET-3, respectively) mixed directly with triglyceride controls. Figure 7 shows the improvement in lutein absorption obtained using either the activated premix with lutein or the MDG protected premix with lutein by demonstrating significantly higher cumulative lutein absorption (area under the curve (AUC)) over the 6 and 8 hour absorption periods relative to the lutein control solution and the non-activated sample. When this data is expressed as a percentage change in lympholutein uptake, as shown in figure 8, the lympholutein uptake of the composition comprising the activated premix with lutein or the MDG protected premix with lutein increased by 106% -170% over the 6 hour infusion period relative to the control and non-activated samples. The lympholutein uptake of the compositions comprising the activated premix with lutein or the MDG protected premix with lutein increased by 121% -165% over the 8 hour infusion period relative to the control and non-activated samples. These data indicate that the lipophilic compound lutein has increased bioavailability when in the activation premix or MDG protected premix compared to when in triglyceride oil.

Example 21

In this example, the test samples evaluated in example 20 were further evaluated for lutein that was not solubilized by the fat in the sample nutritional composition. Five samples, AET-C, AET-1, AET-2, AET-3, and AET-4, were each subjected to high speed centrifugation (31,000x g; 20 ℃; 4 h). The visible absorbance of the resulting aqueous fraction (after dilution with water 2: 8 followed by syringe filtration through a 0.45 μm PTFE membrane) was measured at 476nm (maximum absorbance of lutein). The absorbance readings, expressed as milliabsorbance units per gram of aqueous fraction (mAU/g), and the in vivo lutein reaction grade of the same variant tested in example 20 are reported in the table below.

Table 8: relationship between lutein in aqueous fraction and in vivo reactions

As can be seen from the table, the absorbance values correlate with in vivo study results, where each of the sample nutritional compositions comprising either the activated premix (AET-2 and AET-4) or the MDG protected premix (AET-1) had higher lutein absorbance values than those without the activated premix (AET-C and AET-3). This means that more lutein is present in the aqueous portion of the sample nutritional composition comprising the activated premix or MDG protected premix. Thus, the bioavailability of lutein is accurately predicted in vitro using this method, at least with respect to the sample nutritional composition.

In this example, the effect of adding an activation premix or an MDG protected premix to a nutritional composition on the bioavailability of other lipophilic compounds, such as total triglycerides, phospholipids, arachidonic acid (ARA) and docosahexaenoic acid (DHA), was evaluated and compared to the bioavailability of lipophilic compounds consumed without the activation premix or MDG protected premix.

A similar protocol was followed for animal infusion and lymph collection as described in example 20, including the use of the same test sample. Lymph lipids were extracted and analyzed for triglyceride, phospholipid, ARA and DHA concentrations using high performance liquid chromatography.

Fig. 9 shows the lymphatic triglyceride output per hour over 6 hours after administration of the nutritional composition. Fig. 10 shows the hourly lymphophospholipid output over 6 hours after administration of the nutritional composition.

Fig. 11 and 12 show the lymphatic absorption changes of ARA and DHA, respectively, from fasting within 6 hours after administration of the nutritional composition.

As shown in figure 9, lymphatic output of triglycerides increased during the 6 hour period after administration of the nutritional composition samples in all groups. However, the output of lymphatic triglycerides 2-6 hours after the sample administration with the activated premix (AET-2, AET-4) or the MDG protected premix (AET-1) was significantly increased compared to the samples with mono-and diglycerides mixed directly with triglycerides in the non-activated premix or the non-MDG protected premix (AET-C and AET-3). Similarly, figure 10 shows that in all groups, the output of lymphophospholipids increased over a 6 hour period after sample administration. The output of lymphophospholipids 2-6 hours after administration of the samples given with the activated premix (AET-2, AET-4) or the MDG protected premix (AET-1) was significantly increased compared to administration of samples (AET-C and AET-3) with mono-and diglycerides mixed directly with triglycerides in the non-activated premix or the non-MDG protected premix.

As shown in figure 11, lymph levels of ARA increased significantly from fasting during the 6 hour period after administration of the nutritional compositions containing either the activated premix (AET-2) or the MDG protected premix (AET-1) compared to the non-activated or non-MDG protected controls. Figure 12 shows similar results by showing that during 6 hours after administration of the nutritional formula containing the activated premix (AET-2) or the MDG protected premix, the lymphatic levels of DHA increased significantly from fasting compared to the non-activated or non-MDG protected controls.

Example 22

A study was conducted to compare commercially representative nutritional powders with and without MDG (details of the powder components are listed below). The nutritional meal batches (control and MDG) were reconstituted (152g/L), centrifuged (31,000x g/20 ℃/4h), and analyzed as outlined below. The metric used to analyze bound lutein is the early elution peak of the SEC chromatogram, which corresponds to the high molecular weight peak. The value of this peak is the peak area at 476nm per microliter of injected sample. In addition, the cream layer (upper layer after high speed centrifugation) was used to examine the mechanism of action of the enhanced solubility of lutein and to determine which proteins are responsible for binding the lutein assembly.

The index for bound lutein (high MW Size Exclusion Chromatography (SEC) peak area at 476 nm) for the MDG batch was significantly higher than the control. The cream layer protein comparison and accompanying chromatograms are consistent with the assembly of natural proteins (casein, alpha-lactalbumin, beta-lactoglobulin) and lutein that MDG promotes the formation of high MW but water dispersible (aqueous fraction) and even water soluble (100k ultrafiltrate).

Table 9: example 22 and other exemplary compositions of nutritional composition ingredients.

Example 23

A study was conducted to compare the size estimates of bound lutein in the aqueous fraction. The size estimation of bound lutein was studied by SAXS, SEC and MALDI-TOF.

In general, bound xanthophylls can be divided into two or three classes: (a) large size (about Or > 250kD) water-soluble/water-dispersible aggregates which are not discrete structures and have a broad size range as determined by SAXS and SEC analysis; (b) natural protein/lutein complex (about 10-50kD, about 27 to) They are generally discrete, well-defined structures, supported by MALDI-TOF and SEC data; and (c) smaller (about)Or about 3-4kD) crystals. The compact structure may or may not contain lutein.

Example 24

A study was conducted to examine the overall average hydrophilicity index (GRAVY) of milk proteins. GRAVY values are a measure of hydrophobicity and water solubility and can predict the ability of a protein to bind to lipophilic molecules such as lutein.

The lutein binding capacity of the eight major milk proteins was predicted from the GRAVY values using the graphs listed below. The GRAVY values for milk proteins and their predicted lutein binding capacity are listed in the table below. It has been found that milk protein gamma ₂Casein, beta-lactoglobulin, beta-casein and alpha-lactalbumin have the highest predicted lutein binding capacity of < 2. mu.M, which correlates with MALDI-TOF data, suggesting these proteins as binding partners in the lutein/protein complex. Thus, the GRAVY values can be correlated with the precise lutein binding tendencies of proteins that have been demonstrated to interact effectively with lutein within the nutritional product. This yields GRAVY values with the potential to identify other potential proteins and can be used to optimize the nutritional product system.

Table 10: GRAVY value analysis of different proteins

Example 25

Studies were conducted to explore the size/mass of high molecular weight xanthophyll assemblies. SEC calibration curves were determined to analyze the high molecular weight assemblies described above. These values were also compared to the data provided in example 3 regarding the size of the high molecular weight xanthophyll assemblies.

The reference proteins, their SEC elution times and molecular weights are listed in the table below and used to estimate the size of the high molecular weight lutein assembly. SEC analysis was performed as in the previous examples. By extrapolation from the reference protein curve, the high molecular weight xanthophylls group was foundThe size of the contents is in the range of 60-380kD, with a peak at 160 kD. These values correspond to Particle size diameter of (1), wherein the peak is atThe following steps. The estimates provided below appear to be consistent with the SAXS data provided in example 23, where it is consistent withThe intermediate size category of (1). It should be noted that particles in the large class provided in example 23 would be excluded from the ultrafiltrate and the aqueous fraction may have larger xanthophyll assemblies present prior to ultrafiltration. Furthermore, it should be noted that the 214nm/280nm ratio of the high molecular weight xanthophyll assemblies is in the range of the reference protein, indicating the presence of protein in the high molecular weight assemblies.

Table 11: assembler size calibration using known protein molecular weights

Example 26

A study was conducted to examine the high molecular weight lutein assemblies in commercial nutritional powder batches before and after centrifugation. The nutritional powder batches (control and incorporation into MDG) were reconstituted (152g/L) and analyzed before and after centrifugation (31,000x g/20 ℃/4 h). The components of these powders are listed in the table of example 22. The table below shows the analysis of high molecular weight lutein assemblies by SEC and expressed as mAU-min/mg injections. MDG compositions exhibit a higher presence of initial lutein in both the whole product and the aqueous part, which exhibits even higher differences relative to the control composition.

It was found that the high molecular weight lutein assemblies present in the aqueous fraction were 22% higher than in the control batch. This indicates a significant increase in solubility and/or stability of the MDG composition. In addition, the chromatogram indicates that some fraction (12.3% to 14.5%) of the high molecular weight lutein assemblies are recovered in the supernatant after high speed centrifugation, which indicates that these assemblies are sufficiently soluble and stable that they are not removed or dispersed by the stresses associated with high speed centrifugation.

Table 12: analysis of commercially representative nutritional compositions comprising MDG

Example 27

A study was conducted to evaluate the promotion of high molecular lutein assembly by the presence of MDG versus the presence of High Oleic Safflower Oil (HOSO). Laboratory scale model systems were prepared in skim milk powder (NFDM), where NFDM used was 10% (weight/weight), lipids (HOSO or MDG) and lutein. Lutein was used in variable amounts and the formation of high molecular weight assemblies was then evaluated for these compositions. The different compositions were centrifuged (31,000x g/20 ℃/1h) to obtain an aqueous fraction, which was compared by pH 2.5 SEC as detailed above and the lutein assembly was analyzed by measuring the visible light absorbance at 476nm and measuring the visible light transmittance at 860 nm. In addition, there was a group of MDG batches that were not centrifuged, so the complete product was analyzed for the presence of the xanthophyll assemblies.

The results show that HOSO has a very low ability to promote high molecular weight lutein assemblies at any of the different lutein concentrations. In contrast, MDG compositions exhibited significant formation of high molecular weight xanthophyll assemblies, which can be seen by higher absorbance at 476 nm. Furthermore, this experiment revealed that when the lutein concentration is greater than 1x, lutein assembly formation is reduced. These data indicate that there is an optimal ratio of lutein to MDG for the formation of high molecular weight lutein assemblies. Furthermore, the non-centrifuged sample revealed recovery to about 36% of the total assembly after high speed centrifugation.

Example 28

Studies were conducted to analyze the association of proteins with high molecular weight xanthophyll assemblies. These studies were performed on batches containing MDG or HOSO. Different batch variations were examined using the same techniques as listed above. Specifically, the laboratory scale model system was prepared in skim milk powder where NFDM used was 10% (weight/weight), lipids (HOSO or MDG) and lutein. Variable amounts of lutein were used, and the formation of high molecular weight assemblies was then evaluated for these compositions. The different compositions were centrifuged (31,000x g/20 ℃/1h) to obtain an aqueous fraction, which was compared by pH 2.5 SEC as detailed above. The proteins were analyzed to examine the different molecular weight fractions and their binding to lutein.

The results show that: MDG enhances the solubility and/or stability of both molecular weight moieties (> 57kD and > 137 kD). This situation was particularly enhanced in the 137kD fraction of the MGD batch. It should be noted that the density of the protein assembly may affect its recovery by centrifugation, and as expected, when the lipid content of the assembly is significantly increased, the assembly may be more easily removed by high speed centrifugation. Additionally, as shown in the previous examples, incorporating lutein in the composition at greater than 1x lutein actually reduces assembly formation. This further indicates that an optimal ratio/range of MDG and lutein is required.

Table 13: model system study of assembly formation

Example 29

Studies were conducted to explore the formation of high molecular weight xanthophyll assemblies at different concentrations of MDG, HOSO and lutein. Specifically, the lutein assembly was evaluated by measuring mAU-min/mg at 476nm and by determining the presence of > 137kD protein present in the aqueous fraction after centrifugation. These tests were performed as described above, wherein the different compositions were centrifuged (31,000x g/20 ℃/1h) to obtain an aqueous fraction, which was analyzed by SEC as detailed above.

Overall, the MDG batches outperformed the HOSO batches in terms of the presence of lutein high molecular weight components and higher amount of > 137kD fraction in the aqueous fraction. In addition, it should be noted that although the MDG/lutein ratio remains the same, the size of the assembly still changes. The use of MDG and HOSO in the composition may be advantageous as its use in combination increases the lutein content.

Table 14: model system study of Assembly formation II

Example 30

A study was conducted to evaluate the amino acid profile of proteins involved in the high molecular weight lutein assembly from the sample in example 29. To compare SEC peak areas (independently measured responses to protein concentration), each sample was fractionated by SEC and amino acids were tested in individual fractions. In each case, 10x4-min fractions were collected (manually) over the elution range of the hydrolysate (from about 20min to about 60min) so that the volume of each fraction was 1.6mL (i.e., 0.4mL/min x4 min). Each fraction was digested with acid (6M HCl, 110 ℃, 22h) and the eight amino acids (arginine [ R ] in each digest were tested by reverse phase hplc (rplc) of its 9-Fluorenylmethoxycarbonyl (FMOC) derivative using a modification of methods known in the art]Serine [ S ]]、ASX、[B]Aspartic acid [ D]+ asparagine [ N]、GLX[Z]Glutamic acid [ E ═]+ Glutamine [ Q]Threonine [ T]And glycine [ G ]]). The FMOC derivatives of the six amino acids of the derivatized standards and sample solutions were tested by RPLC using an Agilent model 1100 HPLC system (Agilent Technologies, Wilmington, DE, USA) and a fluorescence detector model Agilent G1321A (FLD, Agilent Technologies, Wilmington, DE, USA). The RPLC system was equipped with a 4.6x250mm i.d., 5 μm, YMC-Pack ODS-AM reverse phase column (Waters, Milford, MA, USA) by passing through a thermostatic column compartment of type G1316A (Agilent Tech)nologies, Wilmington, DE, USA) was maintained at 20 ℃. RPLC mobile phase A was 65% (v/v) 0.05M citric acid (pH 3.0) plus NaOH, and 35% (v/v) ACN; RPLC mobile phase B was 20% (v/v) 0.05M citric acid (pH 3.0) plus NaOH, and 80% (v/v) ACN. The flow rate was 0.5mL/min and the elution procedure was as follows: 0.0-2.0min, 0% B; 2.0-25.0min, 0% B to 50% B (linear); 25.1-48.0min, 100% B; 48.1-63.0min, 0% B. The FLD excitation wavelength was 262nm, the emission wavelength was 310nm, and the gain was set to 10. The injection volume was 4. mu.L. R, S, B (D + N), Z (E + Q), T and G concentrations in the same solution (i.e., prepared from SEC fractions) were calculated from the linear regression of the corresponding separately constructed standard curves.

The following table shows different amino acid ratios for different proteins and experimental groups. For example, NFDM, casein, whey, and proteins found in the aqueous fraction and native proteins. It is noted that native protein refers to monomeric unaggregated protein having a molecular weight in the range of about 14-25 kD. Tube 1 refers to the sample with the highest amount of lutein assembly, while tube 6 refers to the sample with the lowest amount of lutein assembly. They are chosen to best compare the distribution of individual proteins within the assembly.

The data presented show that: more casein is present in the high molecular weight fraction than in the native protein.

Table 15: analysis of proteins bound to assemblies

Example 31

Studies were conducted to evaluate the presence of specific proteins bound to high molecular weight lutein complexes. Commercial representative nutritional powder batches (control and incorporation into MDG) were reconstituted (152g/L) and analyzed before and after centrifugation (31,000x g/20 ℃/2 h). The components of these powders are listed in the table of example 22. Reverse phase HPLC was then used to compare the distribution of individual proteins.

The determination of the proteins bound to the lutein complex was carried out on an Agilent model 1100 HPLC system (Agilent Technologies, Wilmington, DE, USA) with a Diode Array Detection (DAD) system (Agilent Technologies) model G1315A. The system was equipped with 250mm x 4.6mm i.d., 5 μm,YMC-Pack ODS-AQ reverse phase column (Waters, Milford, MA, USA). The column was maintained at 40 ℃ during analysis by a thermostatted column compartment (agilent technologies) type G1316A. A sample prepared with a standard sample of 250mm x 4.6mm i.d., 5 μm,direct determination of the intact protein was carried out on the same system of Jupiter C18 reverse phase column (Phenomenex, Torrance, Calif., USA), wherein the column temperature was also maintained at 40 ℃.

The reverse phase HPLC method used a binary gradient elution with mobile phase A (water, Milli-Q Plus) and mobile phase B (650mL of 0.02M KH2PO4, pH 2.9+175mL acetonitrile +175mL isopropanol) degassed under vacuum. The injection volume was 2 μ L and the elution procedure was: 0% B at 0.3mL/min, from 0.0 to 20.0 min; 0.5mL/min of 16% B, from 20.2 to 40.0 min; 0.5mL/min of 100% B, from 40.1 to 45.0 min; 0% B at 0.5mL/min, from 45.1 to 59.0 min; and 0.3mL/min of 0% B, from 59.5 to 60.0 min. The intact protein method also used a binary gradient elution using vacuum degassed mobile phase A (800mL Milli-Q Plus water +200mL acetonitrile +0.500mL trifluoroacetic acid) and mobile phase B (250mL Milli-Q Plus water +750mL acetonitrile +0.500mL trifluoroacetic acid). The elution procedure with a constant flow rate of 0.6mL/min was: 0% B, from 0.0 to 5.0 min; 0 to 100% B, from 5.0 to 40.0 min; 100% B, from 40.0 to 45.0 min; and 0% B, from 45.0 to 60.0 min. The detection wavelengths were 214nm and 280nm, with the reference at 590 nm. The injection volume was 10 μ L and the run time was 60 min/injection.

The results show that: complete product in MDG batchSecond-order natural with significant reduction_α-casein. This suggests that calcium solubility in MDG compositions may be enhanced. In addition, the aqueous portion of the MDG composition has significantly increased beta-casein, indicating that protein may be included in the high molecular weight lutein assembly. The presence of beta-casein is further evidenced by a reduced presence in the cream layer.

Table 16: analysis of assembly-bound proteins in commercially representative nutritional compositions

Example 32

Studies were conducted to explore the effect of variable concentrations of MDG and HOSO and the presence of different proteins on the formation of high molecular weight xanthophyll assemblies. Model systems for different proteins: sodium caseinate, whole milk protein, whey protein concentrate and whey protein hydrolysate were incorporated into the composition so that the protein in the system was 3.7 wt%. Details of the amounts of MDG, HOSO and lutein are detailed in the table below. The samples were then centrifuged (31,000x g/20 ℃/1h) and the aqueous fraction was tested by SEC. The mobile phase used for SEC was 70% H₂O, 30% acetonitrile and 0.1% trifluoroacetic acid.

And (3) displaying data: in all groups, the MDG composition increased the xanthophyll assemblies and the high molecular protein fraction compared to the HOSO composition. The addition of whey protein hydrolysate exhibited the highest amount of bound lutein. This indicates that it may be advantageous to include whey protein hydrolysates in MDG-type compositions. This finding is interesting in that: it is likely that the aggregation capability of whey protein hydrolysate results in enhanced lutein assembly. In addition, the SEC profiles of sodium caseinate and whole milk protein indicate the presence of casein within the lutein complex.

Table 17: model system study of Assembly formation III

Example 33

A study was conducted to evaluate the effect of Relative Centrifugal Force (RCF) on the analysis of high molecular weight xanthophyll assemblies in the aqueous phase. The components of these powders are listed in the table of example 22. The samples were centrifuged and the aqueous fraction was tested by SEC. The mobile phase used for SEC was 70% H₂O, 30% acetonitrile, 0.1% trifluoroacetic acid. The different RCFs used were: 1x g, 484x g, 1935x g, 7741x g and 31,000x g. These variables and other variables used in the compositions are listed in the table below. The compositions were then evaluated for the presence of high molecular weight xanthophyll assemblies and proteins > 137kD, as well as for the bound lipophilic substance index, as performed above. The bound lipophilic substance index (relative indicator of lipophilic nutrients in high molecular weight assemblies) was determined as the peak ratio (280nm/214nm) for proteins > 137 kD.

The results show that: MDG compositions yielded high molecular weight lutein assemblies at all RCFs that were significantly more water soluble and/or more stable to external stress than the control compositions.

Table 18: analysis of variable centrifugal force of an assembly

Example 34

A study was conducted to evaluate the effect of pH during SEC on the analysis of the presence of high molecular weight xanthophyll assemblies. The components of these powders are listed in the table of example 22. In this experiment, the MDG and control samples were subjected to neutral pH SEC. After rehydration of the samples, they were analyzed via SEC under the conditions listed below. The samples were not centrifuged prior to SEC.

The results show that: under these conditions, MDG-type compositions performed better than controls in terms of the formation of high molecular weight xanthophyll assemblies, the formation of high molecular weight proteins and the index of bound lipophilic substances. In addition, this example demonstrates that the difference between the MDG sample and the control is not a result of the analytical technique used to evaluate the xanthophyll assemblies.

Table 18: effect of pH on the characterization of the Assembly

Example 35

Studies were conducted to explore the effect of choline on the formation of high molecular weight xanthophyll assemblies. Model system of protein content (whey protein concentrate or NFDM, 3.7 wt% protein), MDG (4.4g/L) and lutein (6.7 mg/L). In addition, different experimental groups also had lecithin (949mg/L), choline (45mg/L), or a combination thereof. The samples were then centrifuged (31,000xg/20 ℃/1h) and the aqueous fraction was tested by SEC to analyze the high molecular weight lutein assembly, high molecular weight proteins and bound lipophilic material index. The data are presented in the table below.

In general, choline has a significant positive impact on the formation of high molecular weight lutein assemblies in whey protein concentrate assemblies. It is hypothesized, but not bound by a particular theory, that choline is a basic compound that can interact with acidic aspartate side chains. These interactions can alter the 3-dimensional configuration of the major whey protein beta-lactoglobulin, making the hydrophobic domain more susceptible to binding lutein.

It should also be noted that choline can selectively enhance lutein incorporation, as even though lutein increases in the presence of choline, the bound lipophilic material index actually decreases.

Table 19: model system study of assembly formation IV

Example 36

A study was conducted to examine the variation of commercial representative nutritional composition batches. Samples were prepared as detailed above, with and without MDG in the composition. The samples were centrifuged (7,7741xg/20 ℃/2h) and the aqueous fraction was tested by SEC to analyze the high molecular weight lutein assembly, high molecular weight proteins and bound lipophilic material index. The data are presented in the table below.

Overall, the high molecular weight lutein assembly was 10-30% higher in MGD composition relative to in the control composition. In addition, MDG compositions are associated with high molecular weight protein peaks. The estimated mass of the high molecular peaks of these batches was in the range of 240 kD. There is also evidence that other lipophilic nutrients (e.g., vitamin A, D, E, K, etc.) may be present in the high molecular weight assembly.

Table 20: analysis of commercial representative nutritional compositions in the formation of assemblies

Example 37

Studies were conducted to explore the use of neutral pH as a SEC run condition and its subsequent impact on high molecular weight xanthophyll assemblies during separation. Samples were prepared as detailed above, with and without MDG in the composition. Samples were tested by SEC to analyze high molecular weight lutein assemblies and bound lipophilic substance index. SEC was performed using 0.05M HEPES (pH 7.0) and Superdex Peptide columns. The data are presented in the table below.

The results show that high molecular weight lutein assemblies and proteins are increased in all MDG batches relative to the control batch. This example supports the integrity of the analytical technique for analysis of lutein assemblies.

Table 21: effect of pH on Assembly characterization in commercial representative nutritional compositions

Example 38

The effect of choline on the formation of high molecular weight xanthophyll assemblies was evaluated by conducting a study by providing choline at variable points during the composition. A mixture of water, NFDM, MDG, lutein and choline chloride, wherein choline is added to MDG and lutein or to NFDM slurry. The samples were then centrifuged (31,000x g/20 ℃/1h), ultrafiltered through a 100k filter, and the aqueous fraction was tested by SEC to analyze high molecular weight lutein assemblies, high molecular weight proteins, and native protein-bound lutein. The supernatant was diluted 1: 4 (vol/vol) with SEC mobile phase (0.05M KH2PO4, 0.15M NaCl, pH 7.5) before running on SEC. The different parameters examined were high molecular weight lutein assembly, > 137kD protein and intermediate range protein (which corresponds to native protein bound lutein). Additionally, ultrafiltrate analysis was performed on the xanthophyll assemblies and high molecular weight protein metrics. The data are presented in the table below.

The enhancement of lutein incorporation within high molecular weight assemblies by choline appears to be process dependent. In particular, it was found that the bound lutein was significantly improved when choline was added to NFDM slurry relative to that added to MDG and lutein.

Table 22: model system study of assembly formation V

Example 39

A study was conducted to evaluate the effect of variable components on the formation of high molecular weight xanthophyll assemblies. Different compositions include different protein components (NFDM, β -lactoglobulin or bovine lactoferrin); different amounts of MDG and HOSO, and lutein. The specific details of the above variables are set forth in the following table. The samples were then centrifuged (31,000x g/20 ℃/1h) and bound to lutein via SEC test.

The results show that: when MDG is present in the composition, the measurement of both lutein assembly and proteins > 137kD increases. Interestingly, the combination of MDG and HOSO exhibited the greatest amount of lutein assembly formation and a protein of > 137kD compared to MDG and HOSO alone. In addition, the use of beta-lactoglobulin or bovine lactoferrin alone as part of the composition exhibits the ability to form lutein assemblies and proteins > 137 kD.

Table 23: model system study of assembly formation VI

Example 40

Studies were conducted to explore the effect of variable components on the formation of high molecular weight xanthophyll assemblies. Different compositions included different protein ingredients (NFDM, beta-lactoglobulin or bovine lactoferrin) (37g protein/kg), 6.62g/kg MDG and 10.1mg/kg lutein. Beta-lactoglobulin and bovine lactoferrin were tested with and without centrifugation. Samples were then analyzed by SEC for high molecular weight lutein assemblies, high molecular weight proteins (> 137KD) with or without centrifugation. If the sample is centrifuged, it is performed at 31,000x g/20 ℃/1 h. Details of the ability of the different experimental groups to produce high molecular weight xanthophyll assemblies are shown in the table below.

And (3) displaying data: both beta-lactoglobulin and bovine lactoferrin are capable of forming stable high molecular weight xanthophyll assemblies. Furthermore, these assemblies were still present after centrifugation demonstrating high solubility/stability of the conjugated lutein assemblies.

Table 24: effect of different proteins on formation of assemblies in model systems

EXAMPLE 41

Studies were conducted to examine the effect of variable components on the formation of high molecular weight xanthophyll assemblies. The sample is subjected to an appropriate heat treatment during which the composition is preheated to 165-. After homogenization, each variable was subjected to a High Temperature Short Time (HTST) treatment of about 165-185F for 16 seconds and then cooled to about 40F. All samples were then tested by SEC after centrifugation (31,000x g/20 ℃/1h) or without centrifugation. The data are presented in the table below.

The results show that: the combination of MDG and HOSO resulted in the highest amount of lutein assembly formation and a protein > 137kD prior to centrifugation.

Table 25: model system study of Assembly formation VII

Example 42

A study was conducted using commercially available nutritional powders to examine the effect of variable components on the formation of high molecular weight lutein assemblies. Samples (without dilution or centrifugation) were tested for bound lutein and related attributes by SEC. Comparison of attributes is as follows, and the bound lutein values (peak area at 476 nm/mg injectate) are also compared in the table below.

The results show that: the combination of MDGs provides the highest amount of the following metrics relative to the control. Specifically, MDG/choline outperformed all other groups in all metrics.

Table 26: effect of different variables on the formation of assemblies in commercially representative nutritional compositions

Example 43

Studies were conducted to examine the effect of variable components on the formation of high molecular weight xanthophyll assemblies. Each variable was subjected to an appropriate heat treatment during which the composition was preheated to 165- & 185 ℃ F. and then passed through a homogenizer (first at 1000psig then at 4000 psig). After homogenization, each variable was subjected to a High Temperature Short Time (HTST) treatment of about 165-185F for 16 seconds and then cooled to about 40F. Samples with the following parameters were homogenized and pasteurized: protein ("P" ═ skim milk powder at 10% w/w), lutein ("L"), HOSO ("H"), MDG ("M"), lecithin ("Le"), and/or choline ("C"). These samples were tested for binding to lutein and related attributes by Size Exclusion Chromatography (SEC). Each variable was tested before and after centrifugation (at 31,000x g/20C/1 h).

The results show that: the presence of MDG leads to the formation of lutein assemblies and proteins > 137 kD.

Table 27: model systems study of Assembly formation VIII

Example 44

Studies were conducted to examine the effect of variable components on the formation of high molecular weight xanthophyll assemblies. The sample is subjected to an appropriate heat treatment during which the composition is preheated to 165-. After homogenization, each variable was subjected to a High Temperature Short Time (HTST) treatment of about 165-185F for 16 seconds and then cooled to about 40F. All samples were then tested by SEC after centrifugation (31,000x g/20 ℃/1h) or without centrifugation. The data are presented in the table below.

Samples prepared with lutein, lipids (HOSO or MDG) and different levels of protein (skim milk powder) were tested for binding to lutein and related attributes by Size Exclusion Chromatography (SEC). In particular, the parameters studied were high molecular weight xanthophyll assemblies, > 137kD protein and bound lipophilic substance index. The results are compared below for samples prepared with or without centrifugation (31,000x g; 20C; 1 h).

The results show that: both MDG and HOSO produced higher amounts of xanthophyll assembly formation with increasing amounts of protein, > 137kD protein and bound lipophilic substance index. In addition, the presence of MDG mostly results in higher amounts of the above metric relative to HOSO. Interestingly, MDG was more clearly superior to HOSO, as protein increased via all metrics.

Table 28: model systems study of Assembly formation VIII

Example 45

A study was conducted to examine the presence of tocopherols in high molecular weight assemblies. The components of the test powders are listed in the table of example 22. All samples were prepared and then tested by SEC after centrifugation (31,000x g/20 ℃/3 h). The data are presented in the table below.

Table 29: analysis of tocopherols in an assembly

The results show that: the aqueous portion of the commercial nutritional powder with MDG contained 11% higher levels of tocopherol than the control composition. The aqueous fraction concentrations (about 1.69 and about 1.51mg/kg, respectively) appear to represent an appreciable proportion of total tocopherols (about 20-30%) within a commercial nutritional composition. The solubility of tocopherol provided by MDG in the composition is very different from the reported limit of vitamin E dissolution in water (20.9 mg/kg at 33 ℃). This indicates that the assembly comprising MDG enhances the water solubility of tocopherol.

Example 46

Table 30: analysis of casein bound to assemblies

Table 31 below shows part of the compositional information (protein content) of a commercial liquid nutritional composition designed for individuals with diabetes, and compared to exemplary liquid nutritional compositions according to embodiments disclosed herein. Both indicate liquid nutritional products with different sources of caseinate and with a caseinate SPC combination. The beta-casein utilized in the right column of the embodiment is derived from a boswellia having a2 as the major genetic variant.

Table 31: partial compositional information for commercial nutritional compositions

Nutritional compositions suitable for use in infants and for utilizing the proteins disclosed herein are described in table 32 below. 14 grams of protein per liter of nutritional composition comprises about 18 wt.% bovine beta-casein.

Table 32: example 46 and other exemplary nutritional composition Components

Nutritional compositions suitable for utilizing the proteins disclosed herein are described in table 33 below. 34 grams of protein per liter of nutritional composition comprises about 25% by weight bovine beta-casein.

Table 33: example 46 and other exemplary compositions of nutritional composition ingredients.

In certain exemplary embodiments disclosed herein, the nutritional composition is orally administered (or consumed) as needed to provide the desired nutritional levels. In certain of these embodiments, the nutritional composition is administered (or consumed) in the form of one to two servings per day or in one or two or more divided doses per day. In certain embodiments, when the nutritional composition is a liquid, the serving size may be 150 milliliters to 500 milliliters. In certain other embodiments, when the nutritional composition is a liquid, the serving size is 237 milliliters (about 8 fluid ounces). In other embodiments, when the nutritional composition is a liquid, the serving size is 177 milliliters to 414 milliliters (about 6 fluid ounces to about 14 fluid ounces). In still other embodiments, when the nutritional composition is a liquid, the serving size is 207 milliliters to 266 milliliters (about 7 fluid ounces to about 9 fluid ounces). Each serving of the nutritional composition according to the first, second and third embodiments disclosed herein may have a different caloric content, typically 25 to 500Kcal, including 50 to 400 Kcal; 100 to 350Kcal or 150 to 350 Kcal. In addition, a serving may be understood as any amount intended to be consumed in a meal or within an hour or less.

To the extent that the term "includes" is used in either the detailed description or the claims, it is intended to be inclusive in a manner similar to the term "comprising" as "comprising" is interpreted when employed as a transitional word in a claim. Further, to the extent that the term "or" (e.g., a or B) is used, it is intended to mean "a or B or both. When applicants intend to indicate "only a or B but not both," the term "only a or B but not both" will be used. Thus, use of the term "or" herein is the inclusive, and not the exclusive use. See Bryan A. Garner, A Dictionary of Modern Legal Usage 624 (second edition, 1995). Additionally, to the extent that the term "in …" or "into …" is used in this specification or the claims, it is intended to mean "on …" or "on …" additionally. Furthermore, to the extent that the term "connected" is used in either the specification or the claims, it is intended to mean not only "directly connected," but also "indirectly connected," such as through another component or components.

While the present application has been illustrated by a description of embodiments thereof, and while the embodiments have been described in considerable detail, it is not the intention of the applicants to restrict or in any way limit the scope of the appended claims to such detail. Additional advantages and modifications will readily appear to those skilled in the art. The application, therefore, in its broader aspects, is not limited to the specific details, the representative apparatus, and illustrative examples shown and described. Accordingly, departures may be made from such details without departing from the spirit or scope of the applicant's general inventive concept.

Examples 47 to 53

Examples 47-49 illustrate soy-based powdered infant formulas of the present disclosure having the ingredients listed in Table 34. Unless otherwise specified, all ingredient amounts are listed in kg/1000kg batch.

Table 34: nutritional composition Components of examples 47-49 and other exemplary compositions

Examples 50-52 illustrate milk-based powdered infant formulas of the present disclosure, the ingredients of which are listed in table 35. Unless otherwise specified, all ingredient amounts are listed in kg/1000kg batch.

Table 35: nutritional composition Components of examples 50-52 and other exemplary compositions

Example 53 illustrates a ready-to-eat milk-based liquid infant formula of the present disclosure, the ingredients of which are listed in table 36. Unless otherwise specified, all ingredient amounts are listed in kg/1000kg batch.

Table 36: example 53 and other exemplary nutritional composition Components

Example 54

In this example, the effect of using the activated MDG oil component or the MDG protected premix on the absorbance of triglycerides and fatty acids in the nutritional composition was evaluated and compared to the absorbance of triglycerides and fatty acids in the nutritional composition without the MDG oil. The fatty acids measured in this test include palmitic acid, linoleic acid and alpha-linolenic acid.

The following nutritional compositions were tested:

triglyceride oil control (AET-C): adding high oleic safflower oil to the fat-in-protein slurry and then to the remainder of the composition;

MDG protected premix (AET-1): premixing the MDG oil with lecithin for 5 minutes; adding the premix to the fat-in-protein slurry to form an MDG protected premix and then to the remainder of the composition;

activated MDG oil component (AET-2): adding the MDG oil to the protein-in-water slurry to form an activated MDG oil component, then to the remainder of the composition;

unactivated MDG oil (AET-3): adding the MDG oil to the fat-in-protein slurry and then to the remainder of the composition; and

activated MDG oil component with choline (AET-4): premixing the MDG oil with choline for 5 minutes; this premix was added to the protein-in-water slurry to form the activated MDG oil component and then added to the remainder of the composition.

Table 37: example 54 and other exemplary nutritional composition Components of the compositions

Male Sprague Dawley rats weighing between 280 and 330 grams were all fed for one week with standard commercial Purina (Purina) rat chowder. The rats were then fasted overnight before surgery and subjected to laparotomy under anesthesia and a laparotomy, and a surgery performed according to Tso et al, "The Absorption of Lipid and Lipid Synthesis", Lipid Research Methodology, chapter 5: 191-216(1984) Alan r. loss, inc., NY (incorporated herein by reference to the extent consistent herewith) intubated the intestinal lymphatic vessels. The superior mesenteric artery was isolated, but not occluded. A silicon infusion tube (1.6mm OD) was placed in the stomach for future infusion of the test sample. The fundoplication is closed by purse string sutures. Rats were allowed to recover for 24 hours before infusion was initiated.

Rats were randomly assigned to receive one of the five nutritional compositions tested. (the number of rats in each group, e.g., "(n-7)" is shown in the prompt box of fig. 18.) the animals were intragastrically infused with 3ml of their respective test nutritional compositions 24 hours post-surgery.

Lymph was collected in a pre-cooling tube (for 1 hour) to measure the fasting levels of triglycerides, palmitic acid, linoleic acid and alpha-linolenic acid before the start of the nutritional composition infusion. Lymph was then collected in pre-cooling tubes at one hour intervals (for 6 hours) after infusion began. At the end of the experiment, the rats were sacrificed by exsanguination.

Lymphoid lipids were extracted and analyzed for triglycerides, palmitic acid, linoleic acid and alpha-linolenic acid using high performance liquid chromatography (Craft Technologies, Wilson, NC).

Fig. 18 shows lymph output of triglycerides over 6 hours after infusion of a liquid nutritional composition. Fig. 19 shows lymphatic absorption of palmitic acid within 6 hours after infusion of the liquid nutritional composition. Fig. 20 shows lymphatic absorption of linoleic acid within 6 hours after infusion of the liquid nutritional composition. Figure 21 shows lymphatic uptake of alpha-linolenic acid within 6 hours after infusion of a liquid nutritional composition.

As shown in figure 18, the average fasting lymphatic triglyceride output was varied between about 2.0 and 3.5mg/h for all rat groups. In all groups, lymphatic triglyceride output increased after the start of infusion of each nutritional composition and reached a maximum output of about 8 to about 17.5mg/h two hours after the start of infusion. The control group (AET-C) and the group receiving the unactivated MDG oil (AET-3) exhibited a small increase in lymphatic triglyceride output, and had triglyceride outputs of about 8 to about 9mg/h two hours after the start of infusion. However, the group receiving the activated MDG component (AET-2 and AET-4) or the MDG protected premix (AET-1) exhibited higher triglyceride output and had a triglyceride output of about 13 to about 17.5mg/h two hours after the start of the nutritional composition infusion.

As shown in figure 18, the average fasting lymphatic triglyceride output was varied between about 2.0 and 3.5mg/h for all rat groups. In all groups, lymphatic triglyceride output increased after the start of infusion of each nutritional composition and reached a maximum output of about 8 to about 17.5mg/h two hours after the start of infusion. The control group (AET-C) and the group receiving the unactivated MDG oil (AET-3) exhibited a small increase in lymphatic triglyceride output, and had triglyceride outputs of about 8 to about 9mg/h two hours after the start of infusion. However, the group receiving the activated MDG component (AET-2 and AET-4) or the MDG protected premix (AET-1) exhibited higher triglyceride output and had a triglyceride output of about 13 to about 17.5mg/h two hours after the start of the nutritional composition infusion.

Fig. 20 shows lymphatic absorption of linoleic acid within 6 hours after infusion of the five nutritional compositions. In all groups, lymphatic absorption of linoleic acid increased during the first two hours and decreased to a steady state about 4 to 6 hours after the start of the nutritional composition infusion. However, the nutritional compositions containing some forms of MDG oil (AET-1 to AET-4) had a significant increase in linoleic acid lymphatic absorption two hours after the start of infusion when compared to the nutritional composition containing the triglyceride control without MDG oil (AET-C).

Figure 21 shows lymphatic uptake of alpha-linolenic acid within 6 hours after infusion of five nutritional compositions. In all groups, lymphatic uptake of alpha-linolenic acid increased during the first two hours and decreased to a steady state about 4 to 6 hours after the start of the nutritional composition infusion. However, the nutritional compositions containing some forms of MDG oil (AET-1 to AET-4) had a significant increase in alpha-linolenic acid lymphatic uptake two hours after the start of infusion when compared to the nutritional composition containing the triglyceride control without MDG oil (AET-C).

Example 55

In this example, lutein, which was not solubilized by the fat in the nutritional composition, was further tested against the nutritional composition evaluated in example 1. Five compositions, AET-C, AET-1, AET-2, AET-3, and AET-4, were each subjected to high speed centrifugation (31,000x g; 20 ℃; 4 h). The aqueous portion of each centrifuged nutritional composition was diluted 2: 8 with water and the diluted aqueous portion was filtered through a 0.45 μm PTFE membrane. The visible light absorbance of the diluted aqueous portion was measured at 476nm (maximum absorbance of lutein). The absorbance readings (milliabsorbance units/gram aqueous fraction (mAU/g)) and the in vivo lutein reaction grade of the same composition tested in example 1 are reported in table 3 below.

Table 38: relationship between lutein in aqueous fraction and in vivo reactions

As can be seen from the table, the absorbance values correlate with in vivo results, with nutritional compositions comprising either the activated MDG premix (AET-2 and AET-4) or the MDG protected premix (AET-1) each having a higher lutein absorbance value than either the control nutritional composition (AET-C) or the nutritional composition without the activated premix (AET-3). More lutein is present in the aqueous portion of the nutritional composition comprising the activated MDG premix or the MDG protected premix. Thus, the relative bioavailability of lutein as shown by in vivo testing is accurately predicted by in vitro testing using this method.

Example 56

In this example, the effect on the bioavailability of DHA in a nutritional composition containing an activated MDG premix or an MDG protected premix was evaluated and compared to the bioavailability of DHA in a nutritional composition without an activated MDG premix or an MDG protected premix.

Similar protocols were followed for animal infusion and lymph collection as described in example 1, including infusion of the same nutritional compositions (e.g., compositions AET-C, AET-1, AET-2, AET-3, and AET-4). Lymph lipids were extracted and analyzed for DHA concentration using high performance liquid chromatography. Fig. 22 shows the change in DHA lymphatic output over time. For nutritional compositions containing either the activated premix (AET-2) or the MDG protected premix (AET-1), the lymphatic levels of DHA increase significantly from the initial (fasting) levels to DHA levels within 6 hours after infusion. In contrast, the control nutritional composition (AET-C) and the nutritional composition not comprising the activation premix (AET-3) exhibited only a small increase in the lymphatic level of DHA over time.

Example 57

A study was conducted to compare commercial representative nutritional powders with and without MDG. The nutritional powder batches (control and MDG) were rehydrated (152g/L), centrifuged (31,000x g/20 ℃/4h), and the MDG concentration in the aqueous portion of the sample was estimated by HPLC determination of monolinolein and expressed as mg MDG/kg aqueous portion.

Table 39: MDG presence in the centrifuged aqueous portion of the nutritional composition

Batches of	08/15/13	09/19/13	11/05/13	02/15/14
					AET-C (without MDG)	＜70	＜40	＜20	NT (not tested)
AET-1	144	120	NT	NT
					AET-2	NT	150	NT	NT
AET-3	NT	90	118	NT
					AET-4	178	140	115	NT
AET4-5	NT	NT	127	NT
					A455 (without MDG)	NT	NT	NT	<10
A456	NT	NT	NT	30
					A457	NT	NT	NT	40
A458	NT	NT	NT	40

Measurable MDG was detected in the aqueous portion of all MDG samples and it was confirmed by LC/UV analysis that no MDG was contained in the aqueous portion of all control batches. The significance of the data is: (a) MDG is present in the aqueous fraction (although its water solubility is very low); and (b) MDG concentration is usually related to lutein availability in vivo.

Example 58

Studies were conducted to explore the distribution of lutein and MDG within the nutritional composition. Samples were prepared using homogenization at 150/20bar and then centrifugation at 100,000x g for 1 hour. The different layers (e.g., cream, aqueous and pellet) were then analyzed for the presence of lutein and MDG.

The results show that the nutritional composition comprising MDG provides MDG and lutein in the aqueous phase, where these molecules have very limited water solubility. In particular, it has been found that lutein and MDG are present in the aqueous phase after centrifugation at about 4% and about 8% by weight of the nutritional composition, respectively. This indicates that the lutein found in the aqueous phase is bound to the water soluble complex. It should be noted that the pellets contain about 1% lutein, which suggests that lutein may also be bound to the precipitable assemblies.

Example 59

Studies were conducted to explore the dispersibility/solubility of MDG (mono-and diglycerides) in different aqueous formulations, as assessed by HPLC. MDG was added to water (at 95 ℃, RT) and then the samples were subjected to high speed centrifugation at 20 ℃ and recorded in the table below.

Table 40: MDG alone has limited solubility in the aqueous fraction

The MDG present after centrifugation is very limited and very different from the MDG present in the aqueous phase of the nutritional composition after centrifugation. This indicates that the solubility of MDG together with lipophilic compounds is increased in a nutritional composition comprising MDG, at least one lipophilic compound and at least one hydrophobic protein.

Example 60

Studies were conducted to further explore the distribution of lutein and MDG within the nutritional composition. The sample comprises MDG/lutein/water; MDG/lutein/NFDM/water; MDG/lutein/WPC/water; and MDG/water.

The premix is provided as follows: in the sample containing lutein, heat lutein with shaking at 50 ℃ for 2 h. In a sample containing MDG (GMO-40) and lutein, lutein was added to MDG and heated to 50-60 ℃ and mixed for 10 minutes. In the protein-containing sample, skim milk powder was added to water (50-60 ℃) with agitation and allowed to hydrate for about 10 min.

The samples were prepared as follows: for solution 1, water was heated to 50-60 ℃, a mixture of lutein in GMO-40 and turrax (30sec 10000rpm) were added; for solution 2, NFDM was placed in an aqueous solution heated to 50-60 ℃, a mixture of lutein in GMO-40 and turrax (30sec 10000rpm) were added; for solution 3, water was heated to 50-60 ℃ and GMO-40 mixture and turrax (30sec 10000rpm) were added. The solutions are listed as x.1 in the table below, where 1 is the solution.

Table 41: batch details of example 60

The samples were formulated as detailed above and then characterized by ultrafiltration with different membranes and ultracentrifugation at different times and rates (e.g., 100,000x g, 4,500x g, and 1,000x g). Details of the different centrifugation/filtration methods are listed below.

Centrifugal speed: 1000x g; centrifuging time: 15 min; centrifugation temperature: 20 ℃; centrifuging the tube: 50ml Cellstar tubes from Greiner bio-one; pipe diameter: 23 mm; amount of product in tube: 45ml of the solution; a centrifuge: eppendorf 5810R type; the aqueous phase was separated by syringe through the cream layer.

Centrifugal speed: 4500x g; centrifuging time: 15 min; centrifugation temperature: 20 ℃; centrifuging the tube: 50ml Cellstar tubes from Greiner bio-one; pipe diameter: 23 mm; amount of product in tube: 45ml of the solution; a centrifuge: eppendorf 5810R type; the aqueous phase was separated by syringe through the cream layer.

Centrifugal speed: 100,000x g; centrifuging time: 1 h; centrifugation temperature: 20 ℃; centrifuging the tube: ultra Clear centrifuge tubes from Beckmann Coulter; tube size: 14x89 mm; amount of product in tube: 11.3 g; a centrifuge: beckmann Coulter type L-90K; the aqueous phase was separated by freezing the test tube with the contents in liquid nitrogen and cutting the portion of the tube including the wall of the tube with the slurry phase, followed by thawing the contents of this portion of the test tube.

Film formation: centrifugal filtration units from Amicon Ultra, 100kDa and 10 kDa; centrifugal speed: 4000 g; centrifuging time: 30 min; centrifugation temperature: 20 ℃; centrifuging the tube: 50ml Cellstar tubes from Greiner bio-one; pipe diameter: 23 mm; amount of product in tube: 45ml of the solution; a centrifuge: eppendorf 5810R type; the slurry was separated by removing the filter from the tube and pouring the filtrate.

Centrifugation speed (Lumifuge): 1000 g; centrifuging time: 255 min; light factor: 1; centrifugation temperature: 20 ℃, Lumifuge: a lumiview model 110 centrifuge from LUM Gmbh.

After filtration and/or ultrafiltration, the samples were analyzed for the presence of lutein and MDG in different phases.

Lutein analysis was performed by RP-HPLC-DAD and MDG analysis was performed by RP-HPLC-UV-ELSD. In the MDG analysis, samples were prepared as follows: weighing 0.15g of sample and putting the sample into a 20mL tell bottle; stirring the solution in a fume hood on a magnetic stirrer; 15.0mL or 13.5mL of 80% acetone + 20% chloroform was added with a graduated cylinder.

The solution was stirred on a magnetic stirrer in a water bath (warm water from tap) at ± 50 ℃ for 10 minutes; the solution was sonicated for 10 minutes at room temperature; centrifuge 1.5mL of sample in a 1.5mL centrifuge tube at 20.000x g for 10 min; 20uL of the supernatant was injected onto an HPLC column. UV spectroscopy of the samples was performed at 20 ℃ using a Cary 4000UV-Vis Spectrophotometer (Varian BV) with wavelength scanning (200 and 900nm, 5nm intervals). All measurements were made in a quartz cuvette with a path length of 1 mm.

Qualitatively, the protein-free sample resulted in a more significant amount of precipitated lutein along the sidewall of the sample flask. This suggests that there is an optimal interaction between lutein, hydrophobic proteins and MDG to achieve a water soluble assembly.

In the ultrafiltration assay, none of the lutein or MDG in all samples passed through the 100kD filter (FIG. 23).

These results also indicate that the presence of protein makes the aqueous phase more lutein and MDG. Furthermore, when comparing WPC with NFDM, WPC leads to more lutein and MDG in the aqueous phase. WPC batches may have different particle characteristics (e.g. size) which results in a larger distribution of lutein and MDG in the aqueous phase relative to NFDM.

Centrifugation at 100,000x g: lutein is only found in the aqueous phase of the protein-containing sample; negligible lutein was found in the aqueous phase of the sample containing only lutein and MDG.

Centrifugation at 4,500x g: similar amounts of lutein were found in the aqueous phase of the NFDM and WPC containing samples. More MDG was found in the aqueous phase of the WPC containing sample than in the NFDM containing sample. In addition, the concentration of lutein and MDG in the aqueous phase was much higher compared to the protein free system centrifuged at 1,000x g.

Centrifugation at 1,000x g: significantly higher amounts of lutein were produced in the aqueous phase of the NFDM containing samples compared to the NFDM free samples. Specifically, at this centrifugation speed, about 20% lutein is found in the aqueous phase. Overall, the data indicate that there is an optimal interaction between lutein, hydrophobic proteins and MDG to achieve a water soluble assembly.

Table 42: centrifugation/ultrafiltration analysis of variable MDG batches

Example 61

Studies were conducted to explore the effect of shelf life on MGD batches. Different MDG batches were included: 18-month old batches, 24-month old batches (AET-1) and controls. Different MDG batches were examined in a lymph model as described in example 20.

The results show that: older batches showed similar levels in bioavailability of lutein to newer batches (fig. 24). This indicates that the assembly of the nutritional composition is stable for at least 24 months.