Influences Of Different Factors On Cholic Acid Extraction From Ovine, Bovine Bile And Their Mixed Bile

Cholic acid (CA) is one of the main effective components from the bile of sheep, cattle and other animals, which has the actions of antipyretic, antibacterial, anti-inflammatory. In addition, CA has stronger effect of hemolysis. CA as one of the main component of artificial bezoar had used to synthesis of chenodeoxycholic acid and ursodeoxycholic acid. Modern Chinese medicine preparations add extracted CA to medicine, such as compound bezoar scattered and Xiaozhiling capsules were used to the treatment of fever, stroke, throat in swelling and pain, and sore in mouth. The aim of the present study was to investigate the effects of CA extraction from bovine, sheep and mixed bile by crystallization in ethanol, and choose the optimum technology of CA extraction.The influences on the extraction process, the time of alkali resolved, acidulated condition, ethanol concentration, the mixture ratio of sheep and bovine bile to the extractive quantity and purity of CA was analysed by orthogonal experimental design which used the orthogonal table L4 (23), screening out the best extraction process of CA. Qualitative and quantitative analysis of CA were carried out by using thin layer chromatography and high performance liquid chromatography.Results of the orthogonal experiment showed that main factors of the amount of cholic acid(CA) extraction were ethanol concentration and alkaline hydrolysis time from sheep bile, and factors of the purity CA extracted were alkaline hydrolysis time and acidulated condition. The optimal conditions of CA extraction from sheep bile were ethanol concentration (75%), alkaline hydrolysis time (15h) and acidification conditions (20% hydrochloric acid). Main factors of the amount of CA extraction were ethanol concentration and acidulated condition from cattle, the optimal conditions of CA extraction from cattle bile were ethanol concentration (75%), alkaline hydrolysis time (20h) and acidification conditions (10% hydrochloric acid). Main factors of the amount of CA extraction mixed bile from sheep and cattle were alkaline hydrolysis time and acidulated condition, The optimal conditions of CA extraction from mixed bile were the proportion of mixed bile from sheep and cattle (7:3), alkaline hydrolysis time (20h) and acidification conditions (20% hydrochloric acid)The orthogonal test was employed for selecting the optimal extraction process. Compared with the traditional process, it had characteristics such as large amount of acid extraction, high purity, simple operation, low cost of raw materials and short production cycle.