Co-production process of bilirubin and bile acid

Abstract: Both bile acid and bilirubin products are present in the bile of animals. These two products have great potential for development as feed additives. This article describes the production process of bile acids and bilirubin.

Key words: bile acid; bilirubin; production process

Bile acid is the main active ingredient of bile. It is a series of sterols produced during cholesterol metabolism in animals. It is a complex mixture of components with multiple biological activities [1]. At present, the frequent stress in the intensive farming process, the toxic and harmful substances in the feed ingredients, and the one-sided pursuit of premature farming mode have intensified the burden on the animal liver [2], so that the liver synthesizes bile acids and secretes bile. The function is inhibited, causing nutritional diarrhea, fatty liver, hepatitis and other diseases, resulting in increased breeding costs in the aquaculture industry, resulting in unnecessary waste [3]. As a feed additive, bile acid can effectively alleviate the above problems. Shandong Longchang Animal Health Products Co., Ltd. has been committed to the research of bile acids, and has achieved relatively good results.

The study found that bilirubin is not only a metabolite in the body, it is also an endogenous strong antioxidant at a certain concentration, with antioxidant, anti-lipid peroxidation, protecting cells from damage and enhancing vitamin C and vitamins. The antioxidant capacity of E and other effects [4].

Both bile acids and bilirubin are present in the bile of animals. In order to make better use of resources, Shandong Longchang Animal Health Products Co., Ltd. co-produces these two products, saving production costs without wasting resources.

1 bilirubin production process

1.1 Test reagent 1‰NaHSO3 aqueous solution, NaOH, chloroform, 3N hydrochloric acid aqueous solution, 95% ethanol, ether

1.2 Test material Shrimp gallbladder provided by Shuanghui Biochemical Factory

1.3 test instrument stainless steel knife, stainless steel reaction tank, distillation bottle, suction filter bottle

1.4 Test methods

1.4.1 Heating bile: Place the cut bitter bile in a stainless steel reaction tank with a stainless steel knife or scissors. Add 1 ‰NaHSO3 aqueous solution when heated to 50 °C.

1.4.2 Saponification reaction: When the bile is heated to 50-57 ° C, slowly add NaOH, adjust the pH to 11-12, continue heating without stirring, stop heating after 3 minutes of boiling, and quickly cool down to below 20 °C. .

1.4.3 Acidification treatment: add 0.5-1‰NaHSO3 aqueous solution after cooling, add about 50% of the total weight of bile chloroform, stir and add 45N/kg of 3N hydrochloric acid solution to adjust the pH to 4.5, the bile is divided into three layers. .

1.4.4 Distillation: The stratified chloroform is slowly discharged and placed in a distillation flask for distillation. The bile in the reaction tank was further added with 40% chloroform while stirring, and after 30 minutes, it was discharged and combined and distilled in a distillation flask.

1.4.5 Filtering: When chloroform solution is distilled until the bottom of the distillation flask is bilirubin, add the same amount of 95% ethanol to evaporate. When the chloroform smell is not heard, the remaining liquid is filtered and then ether. Wash and dry, dry, and get the finished product.

2 Production process of bile acid

2.1 Test reagent 10% NaOH solution, decolorizer, 3N hydrochloric acid aqueous solution

2.2 Test material Pig gallbladder provided by Shuanghui Biochemical Factory

2.3 test equipment reaction tank, oxidation tank, acidification tank, suction filter bottle, drying oven

2.4 Test methods

2.4.1 Saponification process: The bilirubin-producing liquid is added to the bile 10 times volume of 10% NaOH solution, placed in a reaction tank for heating, boiled for about 10 hours, and the heating is stopped. After stratification, the liquid is discharged. .

2.4.2 Decolorization process: After dissolving the sodium salt of bile acid, the temperature is raised to 80 ° C and decolorization agent is added to decolorize. When the liquid color is close to yellow, it is placed in the tank after filtration.

2.4.3 Acidification treatment: The liquid after decolorization is lowered to room temperature, and after stirring, a 3N hydrochloric acid aqueous solution is added to adjust the pH to about 4.5. Thus, the solid is substantially completely precipitated.

2.4.4 Filtration: The solid after acid adjustment is subjected to suction filtration. Then, it is washed with water, and when the color of the solid is close to white, it is drained and dried to obtain a finished product.

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3 Conclusion

The content of bilirubin in bile is relatively low. The raw material of 1000 kg of bitter bile is fed in batches. The yield of bilirubin is almost 0.25 ‰, and the average purity of bilirubin is above 90%.

After the extraction of bilirubin, the content of bile acid is relatively high, the yield is about 50%, and the average purity is about 97%.

3 Precautions for the production process

3.1 Bitter gallbladder is best to use fresh, frozen or stored bitterness for too long to affect the yield.

3.2 Bitter gall and bile avoid contact with iron.

3.3 Brush the container and utensils preferably with distilled water or deionized water.

references:

[1] Zeng Rui, Li Li, Wang Qunchu, Liu Wanhan Feed Additives, 2002, (5): 19-20

[2] Magni, Ambrogio; Picol Oreste; Ascheri, Antonio. Preparation of 7-hydroxy2substituted bile acids by stereoseletive hydrogenation of the 7-Keto group in bile acids using nickel catalysts. EP 230,085, Jul 1987 (CA 1988, 108: 221466c) .

[3] Guo Ying, Xie Jianping, Mo Zhengji. New progress in pharmacological research and clinical application of penehyclidine hydrochloride [J]. West China Medical Journal, 2004, 19 (4): 702 - 703.

[4] Xu Shuyun. Pharmacological Experimental Methodology [M]. 3rd ed. Beijing: People's Medical Publishing House, 2001. 1376.