ABSTRACT
As a precious traditional Chinese medicine(TCM), snake bile has been widely used in numerous Chinese medicine prescriptions. Bile acid(BA) derivatives have been demonstrated as the primary chemical family in snake bile. In-depth chemical characterization of BAs is of great importance towards the establishment of quality standards and clarification of the effective material basis for snake bile. This study firstly employed ~1H-NMR to preliminarily analyze the chemical profiles of snake bile, an automated fraction collector was subsequently implemented to obtain the fractions-of-interest. The fraction was then concentrated and re-analyzed by LC-MS. Based on ~1H-NMR, BAs were found to be the main components of snake bile, and six BAs including CDCA, CA, TCDCA, TCA, TDCA and GCA were tentatively identified from the representative spectrum with the assistance of literature and reference compounds. Whereas the content of TCA in snake bile was too great, resulting in a great obstacle for the detection of trace components, the automated fraction collector was subsequently implemented to obtain the fractions-of-interest for LC-MS analysis. According to matching MS/MS information and retention time with reference compounds as well as database retrieval, a total of 57 BAs were detected and annotated. Because of the combination of ~1H-NMR and LC-MS platforms, the findings are beneficial for the in-depth characterization of BAs in snake bile, which provides references for the establishment of quality control and evaluation methods of snake bile.
Subject(s)
Animals , Bile , Bile Acids and Salts , Chromatography, Liquid , Snakes , Tandem Mass SpectrometryABSTRACT
OBJECTIVE： To study the bile acid components in snake bile and other animal bile, and establish the quality control method of the snake bile and its preparations. METHODS： Acetonitrile-methanol and 10 mmol•L-1 ammonium acetate was used as mobile phase system and 18 alkyl silane bonded silica gel was used as filling agent. Liquid chromatography-mass spectrometry was used to qualitatively identify and quantitatively analyze the bile acid components in snake bile and other animal bile, and the adulteration control and content determination method of bile acid in snake bile and its preparations were optimized to establish a suitable quality control method of snake bile and its preparations. RESULTS： The examination method was highly specialized and suitable for the quality control of snake bile and its preparations.The linear relationship of sodium taurocholate between 0.302-9.053 μg is good, r=0.999 3, the repeatability test and accuracy test results are good, and the RSD is smaller than 3.0%. CONCLUSION： The established quality control method can guarantee the quality and safety of snake bile and its preparations.
ABSTRACT
Objective: To establish the method for determination of taurocholic acid in the Traditional Chinese Medicine Shedanchuanbei Oral Liquid and snake bile. Methods: The sample was prepared as mixed solution containing methanol and KH 2PO 4. The mixed solution was injected into Sep Pak C 18 cartridge for the purpose of sample purity. In this processing, the substances which having strong retain action and could harm analytic column were hold in the Sep Pak C 18 cartridge. The eluting solution that the Sep Pak C 18 cartridge had be over loading for taurocholic acid was used as the test solution. The test solution was measured by RP HPLC. The chromatographic conditions were as followed: Supelcosil LC 8 column(150mm?4.6nm,5?m) as analytic column, detect wavelength at 203nm, and MeOH 0.4%KH 2PO 4 mixed solution(56∶44, V/V ) as mobile phase. The inject volume was 50?L. Results: The linear response range of sodium taurocholate was from 0.0253mg?mL -1 to 0.253mg?mL -1 , and the correlation coefficient was 0.9999. The average recovery rate was 101.3%, RSD was 0.40%( n =6). Conclusion: This method was simple, efficient and suitable to the quality control for Shedanchuanbei Oral Liquid and snake bile.