阅读说明：本技术 一种牛骨源柠檬酸钙的提取方法 (Method for extracting bovine bone source calcium citrate ) 是由 谭春萍 于 2019-09-29 设计创作，主要内容包括：本发明涉及牛骨处理技术领域,具体涉及一种牛骨源柠檬酸钙的提取方法,包括如下步骤：S1破碎；S2脱脂；S3渗透；S4速冻；S5解冻；S6微波处理；采用本发明的方法较好地保留了骨中的氨基酸、多糖等成分的完整性,使得蛋白质不易变性,同时,转化了钙离子形成柠檬酸钙,提高了钙的吸收利用率和溶出率,由于未使用强酸强碱进而避免了生产设备的腐蚀。(The invention relates to the technical field of bovine bone treatment, in particular to a method for extracting bovine bone-derived calcium citrate, which comprises the following steps: s1, crushing; s2 degreasing; s3 infiltration; s4, quick freezing; s5, unfreezing; s6 microwave processing; the method of the invention well keeps the integrity of amino acid, polysaccharide and other components in the bone, so that the protein is not easy to denature, simultaneously, calcium ions are converted to form calcium citrate, the absorption utilization rate and the dissolution rate of calcium are improved, and the corrosion of production equipment is avoided because strong acid and strong alkali are not used.)

1. The method for extracting the bovine bone-derived calcium citrate is characterized by comprising the following steps of:

s1 crushing: cleaning fresh ox bone, crushing to 50-80 mesh;

s2 degreasing: placing Os bovis Seu Bubali in Lactobacillus rhamnosus activating solution, performing shake fermentation culture at 20-35 deg.C and 100 rpm and 200 rpm for 10-20h, and filtering to obtain Os bovis Seu Bubali powder;

s3 penetration: placing the bovine bone meal in a container, and spraying water vapor containing citric acid for 120-240 s; the mass fraction of the citric acid in the water vapor containing the citric acid is 10-20%; the spraying rate of each 100g of the bovine bone meal is 300-400 mL/s;

s4 quick freezing: freezing the bovine bone powder subjected to the permeation treatment at the temperature of-10 ℃ to-30 ℃ for 1-3 h;

s5, unfreezing: placing the quick-frozen Os bovis Seu Bubali powder under 2 atmospheric pressure, treating at 121 deg.C under high temperature and high pressure for 5min, and increasing to 4 atmospheric pressure for 10 min;

s6 microwave treatment: transferring the unfrozen bovine bone powder into a microwave instrument, adding a citric acid solution to adjust the pH value to 3-6, carrying out microwave treatment and filtration, and heating and concentrating the filtrate to obtain the calcium citrate.

2. The method for extracting bovine bone-derived calcium citrate as claimed in claim 1, wherein the preparation method of the lactobacillus rhamnosus activating solution comprises the following steps: mixing rhamnose milkActivating the lyophilized bacillus powder with MRS liquid culture medium at 30-35 deg.C for 1-4 hr to obtain suspension; pouring the suspension into potassium-containing solution, standing for 10-20h, and treating for 10-25min under heat stress at 45-55 deg.C; the total viable count of the lactobacillus rhamnosus activating solution is (1.7-3.7) multiplied by 10⁷cfu/mL; the volume ratio of the suspension to the potassium-containing solution is 1: (0.2-0.4).

3. The method for extracting bovine bone-derived calcium citrate according to claim 2, wherein the formula of the MRS liquid culture medium is as follows: 6-8g of peptone, 2-4g of yeast extract, 22-27g of glucose, 1-3g of sodium acetate, 3-4g of diammonium citrate, 801-2mL of tween and MgSO₄·7H₂O 0.1-0.3g，MnSO₄·4H₂O 0.03-0.04g，K₂HPO₄2-4g, 12-16g of agar powder and 1300mL of water 1100-.

4. The method for extracting bovine bone-derived calcium citrate according to claim 2, wherein the yeast extract is prepared by the following steps:

(1) mixing yeast milk and hot water with the temperature of 75-84 ℃ according to the proportion of 1: (2-5) mixing and stirring for 20-25 min;

(2) cooling the product obtained in the step (1) to 25-35 ℃, adjusting the pH value to 6.5-7.0, and then adding complex enzyme to perform hydrolysis reaction for 8-12 h;

(3) and (3) heating the hydrolysate obtained in the step (2) to 75-85 ℃, inactivating for 0.5-2h, separating, and concentrating or spray drying the separated liquid to obtain the yeast extract.

5. The method for extracting bovine bone-derived calcium citrate as claimed in claim 4, wherein the adding amount of the complex enzyme is calculated by thousands of the total weight of the yeast in the yeast milk, the adding amount of the trypsin is 1-3 per mill, the adding amount of the nuclease is 1-3 per mill, and the adding amount of the glutaminase is 10-15 per mill.

6. The method for extracting bovine bone-derived calcium citrate as claimed in claim 1, wherein the amount of the lactobacillus rhamnosus activating solution is 3-7 times of the bovine bone powder by mass.

7. The method as claimed in claim 1, wherein the microwave treatment is performed under the conditions of 600-800MHz for 200-300s, increasing the frequency to 1800-2000MHz for 200-300s, and decreasing the frequency to 250-450MHz for 100-160 s.

8. The method for extracting bovine bone-derived calcium citrate as claimed in claim 7, wherein the rate of raising the frequency is 1-2 MHz/s.

9. The method for extracting bovine bone-derived calcium citrate according to claim 7, wherein the rate of frequency reduction is 2.5 to 3.5 MHz/s.

10. The method for extracting bovine bone-derived calcium citrate according to claim 1, wherein the citric acid solution is 50 to 60 mass%.

Technical Field

The invention relates to the technical field of bovine bone treatment, and particularly relates to a method for extracting bovine bone-derived calcium citrate.

Background

Calcium is an indispensable important nutrient for human bodies, and a plurality of calcium preparations on the market exist at present, but all have certain limitations. The calcium citrate is odorless and tasteless white powder, has absorption efficiency 2.6 times of common calcium carbonate, has small irritation to intestines and stomach, and can be used as calcium supplement for human and animals in pharmaceutical industry for diagnosing and treating osteoporosis; in the food industry, calcium citrate is also a safe food additive, buffering agent and acidity regulator; calcium citrate is also considered a potentially market value feed additive for use in aquaculture. In addition, the calcium citrate can ensure that the feed does not absorb moisture, effectively improve the flavor of the feed, improve the food intake of animals, relieve the reaction of the animals to violent phenomena, improve the body shapes and meat colors of the animals and improve the quality of meat.

The composition of the hard bones of the cattle is stable, 70 percent of the hard bones are mineral substances mainly containing calcium salts, more than 98 percent of the calcium salts are calcium phosphate and calcium carbonate, but the hydroxyapatite of the bone calcium has extremely poor water solubility, and the bones and the collagen fiber structures generally cause low calcium extraction rate, and if the bone powder is directly eaten, the absorption and utilization rate of the calcium is low, so that the effective utilization of the calcium in the hard bones of the cattle is very important. The existing acidolysis method and alkaline hydrolysis method can cause protein irreversible deformation, thereby being not beneficial to the transformation of free calcium. For example, patent No. CN201810507693.1 discloses a method for extracting calcium from bovine bone, but still remains in the dissolution rate.

Disclosure of Invention

The invention provides a method for extracting bovine bone-derived calcium citrate to solve the technical problems.

The method is realized by the following technical scheme:

the extraction method of bovine bone-derived calcium citrate comprises the following steps:

s1 crushing: cleaning fresh ox bone, crushing to 50-80 mesh;

s2 degreasing: placing Os bovis Seu Bubali in Lactobacillus rhamnosus activating solution, performing shake fermentation culture at 20-35 deg.C and 100 rpm and 200 rpm for 10-20h, and filtering to obtain Os bovis Seu Bubali powder; (ii) a

S3 penetration: placing the bovine bone meal in a container, and spraying water vapor containing citric acid for 120-240 s; the mass fraction of the citric acid in the water vapor containing the citric acid is 10-20%; the spraying rate of each 100g of the bovine bone meal is 300-400 mL/s;

s4 quick freezing: freezing the bovine bone powder subjected to the permeation treatment at the temperature of-10 ℃ to-30 ℃ for 1-3 h;

s5, unfreezing: placing the quick-frozen Os bovis Seu Bubali powder under 2 atmospheric pressure, treating at 121 deg.C under high temperature and high pressure for 5min, and increasing to 4 atmospheric pressure for 10 min;

s6 microwave treatment: transferring the unfrozen bovine bone powder into a microwave instrument, adding a citric acid solution to adjust the pH value to 3-6, carrying out microwave treatment and filtration, and heating and concentrating the filtrate to obtain the calcium citrate.

The preparation method of the lactobacillus rhamnosus activation liquid comprises the following steps: collecting freeze-dried powder of Lactobacillus rhamnosusActivating with MRS liquid culture medium at 30-35 deg.C for 1-4 hr to obtain suspension; pouring the suspension into potassium-containing solution, standing for 10-20h, and treating for 10-25min under heat stress at 45-55 deg.C; the total viable count of the lactobacillus rhamnosus activating solution is (1.7-3.7) multiplied by 10⁷cfu/mL; the volume ratio of the suspension to the potassium-containing solution is 1: (0.2-0.4).

The MRS liquid culture medium comprises the following components in percentage by weight: 6-8g of peptone, 2-4g of yeast extract, 22-27g of glucose, 1-3g of sodium acetate, 3-4g of diammonium citrate, 801-2mL of tween and MgSO₄·7H₂O 0.1-0.3g，MnSO₄·4H₂O 0.03-0.04g，K₂HPO₄2-4g, 12-16g of agar powder and 1300mL of water 1100-.

The preparation method of the yeast extract comprises the following steps:

(1) mixing yeast milk and hot water with the temperature of 75-84 ℃ according to the proportion of 1: (2-5) mixing and stirring for 20-25 min;

(2) cooling the product obtained in the step (1) to 25-35 ℃, adjusting the pH value to 6.5-7.0, and then adding complex enzyme to perform hydrolysis reaction for 8-12 h;

(3) and (3) heating the hydrolysate obtained in the step (2) to 75-85 ℃, inactivating for 0.5-2h, separating, and concentrating or spray drying the separated liquid to obtain the yeast extract.

The compound enzyme is composed of trypsin, nuclease and glutaminase.

The adding amount of the complex enzyme is calculated by thousandth of the total weight of yeast in the yeast milk, the adding amount of the trypsin is 1-3 thousandth, the adding amount of the nuclease is 1-3 thousandth, and the adding amount of the glutaminase is 10-15 thousandth.

The dosage of the lactobacillus rhamnosus activating solution is 3-7 times of the mass of the bovine bone meal.

The microwave treatment is carried out for 200-300s under the conditions of 600-800MHz, 200-300s under the conditions of 1800-2000MHz and 100-160s under the conditions of 250-450 MHz.

The rate of raising the frequency is 1-2 MHz/s.

The rate of frequency reduction is 2.5-3.5 MHz/s.

The citric acid solution is 50-60% by mass.

The potassium-containing solution is any one of KCl, potassium nitrate and potassium phosphate; the mass fraction of potassium in the potassium-containing solution is 5-9%.

Has the advantages that:

the method of the invention well keeps the integrity of amino acid, polysaccharide and other components in the bone, so that the protein is not easy to denature, simultaneously, calcium ions are converted to form calcium citrate, the absorption utilization rate and the dissolution rate of calcium are improved, and the corrosion of production equipment is avoided because strong acid and strong alkali are not used.

The invention takes ox bone as raw material, through the water vapor treatment containing citric acid, not only utilizes water to dissolve soluble calcium, but also utilizes the action of heat energy and citric acid to enable the citric acid to deeply penetrate into bones, and then utilizes the combination of quick freezing and unfreezing treatment to decompose crystal tissues and release beneficial elements such as soluble substances, bone calcium and the like, thereby enabling the penetrated citric acid to be mutually combined with calcium. The invention uses multi-frequency microwave treatment to destroy the combination of ossein and hydroxyapatite, accelerate the penetration and diffusion of citric acid and promote the dissolution and conversion of calcium.

In addition, the effect of the lactobacillus rhamnosus is utilized, a large amount of insoluble calcium is converted into calcium elements in free forms such as calcium lactate and calcium amino acid, and the calcium elements are activated by using a potassium-containing solution, so that the activation efficiency is improved, and the adverse effect of components such as glucose on calcium extraction is weakened; the yeast extract is used for activating the lactobacillus rhamnosus, so that the activity of the lactobacillus rhamnosus is enhanced, the dissociation and dissolution of nutritional ingredients in bone meal can be improved, proteins are decomposed into polypeptides, the dissolution rate of calcium is improved, the physiological activity of the proteins is improved, and meanwhile, a potassium-containing solution can inhibit Maillard reaction.

Detailed Description

The following is a detailed description of the embodiments of the present invention, but the present invention is not limited to these embodiments, and any modifications or substitutions in the basic spirit of the embodiments are included in the scope of the present invention as claimed in the claims.