The Study On Separation And Identification Of The Sturgeon Bile By Chromatography

Sturgeon bile which was collected from Tianxia Sturgeon industry limited company,the biggest resource pool with the largest scale in nation and overseas t and he most variety Sturgeon was chosen as the object in this study. Firstly the solvent was screened and optimized by Thin Layer Chromatography , then the valid components of Sturgeon bile was seperated by Silica-Gel Column Chromatography , followed by the influence of different solvent systems,different filling height and different particle size of Silica-Gel on the separation effect,finally the analysis and identification of what was separated were carried out in this paper.In the former experiment gradient elution was carried out at the condition that the silica-gel with 200~300 mesh , chloroform and methanol mixed at the ratio of 5:1 by volume were used.And then used Petroleum ether,acetone and ethanol mixed at the ratio of 3:1:1 by volume to elute other components.However,the separation effect was not perfectly ideal in that component 2 and component 3,eluting from most of the liquid at the same time,were not completely separated,and only single component(component 1)was separated.Following,chloroform,chloroform and ethanol mixed at the ratio of 15:1 by volume, Petroleum ether,acetone and ethanol mixed at the ratio of 3:1:1 by volume,ethanol,acetone and water mixed at the ratio of 4:2:1 by volume ,water were chosen as solvent systems,sequently,six main components of Sturgeon bile were separated. In two separation experiments,silica-gel with 100~200 mesh and 200~300 mesh,and filling with the height of 290mm and 390mm were respectively used,the results showed that the seperation effect was greatly improved by increasing the height of silica-gel filling and reducing particle size of silica-gel.It could be preliminarily inferred via the combination of ultraviolet - visible spectrogram and infra-red spectrogram that component 1 was free lithocholic acid , component 2 was possibly free cholic acid or chenodeoxycholic acid , which both microscaly existed in Sturgeon bile.component 3 was still uncertain and needed the further study.component 4 was sodium salt or potassium salt of taurocholic acid or taurochenodeoxycholic acid which existed in Sturgeon bile.component 5 was glycochenodeoxycholic acid which existed in Sturgeon bile as an acid.component 6 was sodium salt or potassium salt of biliverdin;Extraction technology of Bile acid from the farmed sturgeon was studied by dilute alkali,extraction rate of total Bile acid was as a study object,single factor experiment and orthogonal experiment was carried out.The results show that the optimal condition of extraction was 9% of NaOH, 120oC of temperatureand 17h of hydrolysis time.