阅读说明：本技术 沙棘提取物和多步萃取的提取方法及其用途 (Sea-buckthorn extract and multi-step extraction method and application thereof ) 是由 王洪伦 苏晨玉 胡娜 董琦 于 2020-08-05 设计创作，主要内容包括：本发明公开了沙棘提取物和多步萃取的提取方法及其用途,将沙棘与乙醇混合超声提取后,移除乙醇得预提取物,用水和石油醚对预提取物进行萃取,再将石油醚萃取后剩余的部位用乙酸乙酯萃取；再将乙酸乙酯萃取后剩余的部位用正丁醇萃取,剩余部分为水部位,本发明通过不同的溶剂萃取得到的沙棘提取物均对α-葡萄糖苷酶的活性具有很好的抑制作用,实现了沙棘有效物质的充分提取和利用,抑制能力优于现有的α-葡萄糖苷酶抑制剂阿卡波糖,可用于制备α-葡萄糖苷酶抑制剂以及预防和/或治疗以α-葡萄糖苷酶为靶点的疾病的产品,安全高效。(The invention discloses a sea-buckthorn extract and a multi-step extraction method and application thereof, wherein sea-buckthorn and ethanol are mixed and ultrasonically extracted, the ethanol is removed to obtain a pre-extract, the pre-extract is extracted by water and petroleum ether, and the rest part after the petroleum ether extraction is extracted by ethyl acetate; the method has the advantages that the sea buckthorn extracts obtained by extracting different solvents have good inhibition effect on the activity of alpha-glucosidase, the full extraction and utilization of active substances of sea buckthorn are realized, the inhibition capability is superior to that of the existing alpha-glucosidase inhibitor acarbose, and the sea buckthorn extracts can be used for preparing alpha-glucosidase inhibitors and products for preventing and/or treating diseases with alpha-glucosidase as a target point, and are safe and efficient.)

1. The extraction method of the seabuckthorn extract A is characterized by comprising the following steps: mixing fructus Hippophae pomace with ethanol, extracting, removing ethanol to obtain pre-extract, extracting the pre-extract with water and petroleum ether, mixing petroleum ether phases, and removing solvent to obtain extract A.

2. A method for extracting a seabuckthorn extract B is characterized by comprising the following steps: the aqueous phase remaining after extraction by the process of claim 1 is A1, aqueous phase A1 is extracted with ethyl acetate, and the ethyl acetate phases are combined to remove the solvent to give extract B.

3. A method for extracting seabuckthorn fruit extract C is characterized by comprising the following steps: the aqueous phase A1 was extracted with ethyl acetate according to the method of claim 2, the remaining aqueous phase B1 was extracted with n-butanol, and the n-butanol phases were combined to remove the solvent to give extract C.

4. The extraction method according to any one of claims 1 to 3, wherein the ratio of sea buckthorn pomace: the feed-liquid ratio of ethanol is 1 g: 1-15 mL, preferably 1 g: 10 mL;

further, the temperature during ultrasonic treatment is 30-80 ℃, and preferably 45 ℃;

further, the ultrasonic time is 1-5 h, preferably 1-3 h, and more preferably 2 h;

further, the ethanol concentration is 95%;

further, the ultrasonic power is 200W, and the frequency is 40 KHz.

5. The extraction process according to any one of claims 1 to 3, characterized in that the pre-extract: the proportion of water is 1 g: 2-15 mL; further 1 g: 3-10 mL; further 1 g: 9.5-10.5 mL;

pre-extraction: the proportion of petroleum ether is 1 g: 2-15 mL; further 1 g: 3-10 mL; further 1 g: 9.5-10.5 mL;

further, water: volume of petroleum ether is 1: 0.5 to 1.5, preferably 1: 1;

further, the pre-extract is dissolved in water and then mixed with petroleum ether for extraction.

6. Extraction process according to claim 2 or 3, characterized in that the pre-extract: the proportion of ethyl acetate was 1 g: 2-10 mL; further 1 g: 3-9 mL; further 1 g: 8-9 mL;

further, the pre-extract: the volume ratio of n-butanol is 1 g: 1-10 mL, further 1 g: 2-8 mL; further 1 g: 7-8 mL.

7. A sea buckthorn extract characterized by being obtained by extraction according to the method of any one of claims 1 to 6.

8. The use of the hippophae rhamnoides extract of claim 7 in the preparation of a product for preventing and/or treating one or more of impaired glucose tolerance, diabetes, obesity, chronic hepatitis b, aids, and tumors.

9. Use of the hippophae rhamnoides extract of claim 7 for the preparation of alpha-glucosidase inhibition.

10. A product comprising one or more of the hippophae rhamnoides extracts of claim 7.

Technical Field

The invention relates to the field of extraction of effective substances in sea-buckthorn, in particular to a sea-buckthorn extract, a multi-step extraction method and application thereof.

Background

Seabuckthorn (the name of Latin, Hippophae rhamnoides Linn.) is a deciduous shrub of the genus Hippophae of the family Elaeagnaceae, and is drought-resistant and sand-resistant, and can survive in saline-alkali lands, thus being widely used for water and soil conservation. Sea buckthorn is planted in great amount in northwest China for desert greening.

The sea buckthorn fruit is rich in nutrition, and contains active substances such as various vitamins, flavonoids, triterpenoids, oil, fatty acid, phenols, volatile oils, trace elements, phospholipids, 5-hydroxytryptamine and the like, and various amino acids and proteins required by a human body according to measurement.

In addition, the current sea-buckthorn is widely applied to the production of sea-buckthorn fruit juice and sea-buckthorn raw pulp, and in the production process, a large amount of sea-buckthorn pomace is produced after the sea-buckthorn is squeezed, and the sea-buckthorn pomace is usually discarded as waste or produced into dry powder to be used as feed, so that the price is very low. If the utilization value of the seabuckthorn fruit residues can be developed more, the method has great economic significance.

Disclosure of Invention

The invention mainly solves the technical problem of providing a seabuckthorn extract and an extraction method thereof, which can extract an extract with active value from seabuckthorn pomace.

Alpha-glucosidase plays an important role in the absorption process of food, and must be bound thereto for the food to be digested and absorbed. The sugar-reducing mechanism of the alpha-glucosidase inhibitor is that the speed of decomposing starch into glucose is slowed down by inhibiting the alpha-glucosidase on the intestinal mucosa, and the absorption of glucose by the small intestine is reduced and delayed, so that the blood sugar is reduced, and the effect on postprandial hyperglycemia is obvious.

Experiments show that the sea buckthorn extracts obtained by extracting with different solvents have a good inhibition effect on the activity of alpha-glucosidase, the inhibition capability of the sea buckthorn extracts is superior to that of acarbose serving as the existing alpha-glucosidase inhibitor, and the sea buckthorn extracts can be used for preparing the alpha-glucosidase inhibitor, wherein the inhibition effect of the sea buckthorn extracts obtained by extracting with ethyl acetate is optimal.

The alpha-glucosidase inhibitor does not stimulate the secretion of insulin, and the single use of the medicine does not usually cause hypoglycemia, so that the alpha-glucosidase inhibitor can help reduce the fluctuation of blood sugar, can obviously reduce the probability of cardiovascular diseases of diabetics, and has the most obvious effect of improving myocardial infarction. Is one of the few oral hypoglycemic drugs which can interfere with impaired glucose tolerance. The alpha-glucosidase inhibitor has been reported to have definite curative effect on diabetes, and also has certain curative effect on obesity, chronic hepatitis B, AIDS and tumor.

Based on the experimental results of the invention, the invention can provide the following technical scheme:

a method for extracting seabuckthorn fruit extract A comprises the following steps: mixing fructus Hippophae pomace with ethanol, extracting, removing ethanol to obtain pre-extract, extracting the pre-extract with water and petroleum ether, mixing petroleum ether phases, and removing solvent to obtain extract A.

The extraction method of the sea-buckthorn pomace and ethanol mixed extraction is not limited, and any extraction method available in the field can be applied to the method, such as ultrasonic extraction, reflux extraction, ultrahigh pressure extraction, microwave extraction and the like.

In a specific embodiment of the invention, the sea buckthorn pomace and ethanol are mixed and extracted by ultrasonic extraction.

In the present invention, the "extraction of the pre-extract with water and petroleum ether" means that the extraction is performed using a solvent system of water and petroleum ether. If the material to be extracted contains a proper amount of water, only petroleum ether can be added to form a water and petroleum ether solvent system for extraction; if the material to be extracted contains no water or has extremely low water content, petroleum ether and proper amount of water can be added to form a water and petroleum ether solvent system for extraction; and so on.

The invention also provides an extraction method of the seabuckthorn extract B, and by adopting the method, after the pre-extract is extracted by using water and petroleum ether, the residual water phase is A1, the water phase A1 is extracted by ethyl acetate, and the ethyl acetate phase is combined to remove the solvent to obtain the extract B.

The invention also provides an extraction method of the seabuckthorn extract C, the method is adopted, after ethyl acetate is used for extracting the water phase A1, the residual water phase is B1, the water phase B1 is extracted by n-butanol, and the n-butanol phase is combined to remove the solvent to obtain the extract C.

And concentrating the residual water phase after n-butanol extraction under reduced pressure to obtain water fraction D.

In a specific embodiment of the present invention, the sea buckthorn pomace: the feed-liquid ratio of ethanol is 1 g: 1-15 mL, preferably 1 g: 10 mL;

further, the temperature during ultrasonic treatment is 30-80 ℃, and preferably 45 ℃;

further, the ultrasonic time is 1-5 h, preferably 1-3 h, and more preferably 2 h;

further, the ethanol concentration was 95%.

Further, the ultrasonic power is 200W, and the frequency is 40 KHz.

In a particular embodiment of the invention, the pre-extract: the proportion of water is 1 g: 2-15 mL; further 1 g: 3-10 mL; further 1 g: 9.5-10.5 mL;

pre-extraction: the proportion of petroleum ether is 1 g: 2-15 mL; further 1 g: 3-10 mL; further 1 g: 9.5-10.5 mL;

further, water: volume of petroleum ether is 1: 0.5 to 1.5, preferably 1: 1;

further, the pre-extract is dissolved in water and then mixed with petroleum ether for extraction.

In a particular embodiment of the invention, the pre-extract: the proportion of ethyl acetate was 1 g: 2-10 mL; further 1 g: 3-9 mL; further 1 g: 8-9 mL.

Further, the pre-extract: the volume ratio of n-butanol is 1 g: 1-10 mL, further 1 g: 2-8 mL; further 1 g: 7-8 mL.

The invention also provides the seabuckthorn extract obtained by the method.

The invention also provides the application of the seabuckthorn extract in preparing products for preventing and/or treating one or more of impaired glucose tolerance, diabetes, obesity, chronic hepatitis B, AIDS and tumors.

Impaired glucose tolerance, i.e., pre-diabetes, mainly includes Impaired Fasting Glucose (IFG) and Impaired Glucose Tolerance (IGT). Can effectively improve impaired glucose regulation, prevent and/or treat impaired glucose tolerance and/or diabetes, and simultaneously prevent or improve diabetic complications, and can be used for preparing related products.

The invention also provides application of the seabuckthorn extract in preparation of alpha-glycosidase inhibition.

The invention also provides a product which comprises the seabuckthorn extract.

In the present invention, the products include, but are not limited to, pharmaceuticals, nutraceuticals, foods, and the like.

The invention has the beneficial effects that:

(1) the extraction method has simple steps, is simple, convenient and quick, can efficiently obtain the seabuckthorn extract, and realizes the full extraction and utilization of effective substances of the seabuckthorn by a multi-step extraction mode.

(2) The different sea buckthorn extracts obtained by extracting with different solvents are proved by experiments to be capable of inhibiting the activity of the alpha-glucosidase, have better effect than acarbose, can be used for preparing alpha-glucosidase inhibitor products, and can also be used for preparing products for preventing and/or treating diseases taking the alpha-glucosidase as a target point, such as products for treating diseases of impaired glucose tolerance, diabetes, hypertension, obesity, chronic hepatitis B, AIDS, tumors and the like, thereby realizing waste utilization and changing waste into valuable.

Detailed Description

The technical solutions of the present invention are described clearly and completely below, and it is obvious that the described embodiments are some, not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.