The present invention relates to the field of animal extraction engineering, specifically a method of extracting livestock bile to prepare bile acid products.
Background technology:
Bile acid is an important component of bile and plays an important role in fat metabolism. Bile acids mainly exist in the enterohepatic circulatory system and play a certain protective role through recycling. Only a small proportion of bile acids enter the peripheral circulation. The driving force that promotes enterohepatic circulation of bile acids is the transport system of hepatocytes---absorption and secretion of bile acids into bile, cholecystokinin-induced gallbladder contraction, propulsive peristalsis of the small intestine, active transport of ileal mucosa, and blood flow to the portal vein. inflow.
At present, there are two main types of bile acid preparation: chemical synthesis and natural bile extraction. The currently disclosed methods for extracting bile acids from natural bile have shortcomings such as complex operations, low purity, low yield, large consumption of reagents, high cost, and difficulty in industrial application; the chemical synthesis method is complex to operate and uses a large amount of methanol. When extracting cholesterol with chloroform and methanol, the stratification is not clear, it is difficult to master, errors are prone to occur, it is difficult to achieve the purpose of separating cholesterol and lipids, and the cost is too high.
Technical implementation elements:
The purpose of the present invention is to provide a method for extracting livestock bile to prepare bile acid products, so as to solve the problems raised in the above background technology.
To achieve the above objectives, the present invention provides the following technical solutions:
A method of extracting livestock bile to prepare bile acid products, the process steps are as follows:
1 ) Take fresh livestock bile, soak and clean it with 65% alcohol for 20 to 30 minutes, cut the livestock bile into pieces, and take out the livestock bile;
2) Add 1 to 3 times the volume of sodium hydroxide solution to the collected bile. , stir and heat for 30 to 50 minutes, the heating temperature is 50 to 60°C, and when it is left to cool to 40°C, use hydrochloric acid with a volume concentration of 30 to 35 to adjust the pH value to 2.2 to 4.6. After standing, the precipitation and transformation are complete;
3) Filter and collect the precipitate, add 2 to 5 times the mass of water, add hydrogen peroxide solution to make the concentration reach 2.2 to 4.5 of the volume of the mixed suspension, then add sodium carbonate solid powder according to 15 to 20 of the mass of bile, so that Precipitate transformation, collect the filtrate by filtration, adjust the pH of the filtrate to 2.2-4.6 with hydrochloric acid with a volume concentration of 30-35, let it stand, filter to obtain the precipitate, wash with water until the pH is 6.5-7.5, and dry to obtain total bile acids Crude product;
4) Add 95 alcohol solution to the crude product, adjust the solution concentration to 50-60, stir for 30-50 minutes, filter and collect the filtrate, add styrene-divinylbenzene at a volume of 1:1 for extraction and degreasing. Take the alcohol layer and adjust the pH value to 7.5-8.5 with ammonia water, recover the alcohol solution, add water to the original volume, add calcium chloride aqueous solution with a mass-volume ratio of 12-15, stir for 30-50 minutes until the precipitation is complete, filter and collect the precipitate;
5) Wash the precipitate with water until it is light yellow, heat to dissolve it, and filter while it is hot. Use hydrochloric acid solution to adjust the pH of the filtrate to 3 to 5. Filter and collect the precipitate. Wash the precipitate with water until the pH is 6.5 to 7.5. , elute with 65% alcohol, and continuously detect with thin layer chromatography during this period. When the appearance of the precipitate is pure, collect the eluent until it is complete;
6) Add a mass ratio of 1.2 to 1.5 to the eluate After the activated carbon is decolorized, the alcohol is evaporated under vacuum to obtain the bile acid product crystals, which are heated in a water bath at 50 to 60°C for 12 hours. The product crystals are taken out and dried under vacuum at -10 to -5°C for 3 to 5 hours to obtain the bile acid product. .
As a further solution of the present invention: the resting time in step 2) is 1.5 to 3 hours.
As a further solution of the present invention: the resting time in step 3) is 3 to 5 hours.
As a further solution of the present invention: the mass-to-volume ratio of the sodium hydroxide solution in step 1) is 50 to 90.
Compared with the existing technology, the beneficial effects of the present invention are: soaking and cleaning with alcohol before preparation, effectively ensuring the quality of livestock bile; using livestock bile extract to prepare health-care drugs, the raw materials are rich and low-priced; The preparation method is safe, non-toxic, simple, efficient and low-cost; the entire preparation process has mild conditions and low reagent consumption, making it easy to apply to industrial production.
Detailed Embodiments
The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention; All examples. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the scope of protection of the present invention.
In the embodiment of the present invention, a method of extracting bile of livestock to prepare bile acid products, and a method of extracting bile of livestock to prepare bile acid products, the process steps are as follows:
1) Take fresh livestock bile, soak it with 65% alcohol and clean it for 20 to 30 minutes, cut the livestock bile into pieces, and take out the livestock bile;
2) Add 1 to 3 times the volume of sodium hydroxide solution to the collected bile. Stir and heat for 30 to 50 minutes. The heating temperature is 50 to 60°C. The mass-to-volume ratio of the sodium hydroxide solution is 50 to 90. When it is left to cool to 40°C, use hydrochloric acid with a volume concentration of 30 to 35 to adjust the pH value to 2.2~4.6, let stand for 1.5~3h, and the precipitation will be transformed completely;
3) Filter to collect the precipitate, add 2 to 5 times the mass of water, and add hydrogen peroxide solution to make the concentration reach 2.2 to 4.5 of the volume of the mixed suspension. , then add sodium carbonate solid powder according to 15 to 20% of the bile mass to convert the precipitate, filter and collect the filtrate, adjust the pH value of the filtrate to 2.2 to 4.6 with hydrochloric acid with a volume concentration of 30 to 35, let it stand for 3 to 5 hours, and filter to obtain the precipitate , wash with water until the pH is 6.5-7.5, and dry to obtain the crude total bile acid product;
4) Add 95 alcohol solution to the crude product, adjust the solution concentration to 50-60, stir for 30-50 minutes, and filter Collect the filtrate, add styrene-divinylbenzene at a volume of 1:1 for extraction and degreasing, take the alcohol layer and adjust the pH value to 7.5-8.5 with ammonia water, recover the alcohol solution, add water to the original volume, and add chlorine with a mass-to-volume ratio concentration of 12-15 Calcium aqueous solution, stir for 30 to 50 minutes until the precipitation is complete, filter and collect the precipitate;
5) Wash the precipitate with water until it is light yellow, heat to dissolve, filter while hot, and adjust the pH value of the filtrate with hydrochloric acid solution to 3 to 5, collect the precipitate by filtration, wash the precipitate with water until the pH is 6.5 to 7.5, and elute with 65 alcohol. During this period, continue to use thin layer chromatography to detect. When the appearance of the precipitate is pure, collect the eluent until it is complete;
6) Add activated carbon with a mass ratio of 1.2 to 1.5 to the eluate for decolorization, evaporate under vacuum to recover the alcohol, and obtain bile acid product crystals. Place them in a 50 to 60°C water bath and heat for 12 hours to take out the product crystals. Vacuum dry at -10~-5°C for 3~5 hours to obtain fine bile acid.
Using livestock bile extract to prepare health-care drugs, the raw materials are abundant and low-priced; the preparation method is safe, non-toxic, simple, efficient, and low-cost; the entire preparation process has mild conditions, low reagent consumption, and is easy to be applied to industrial production.