ABSTRACT
OBJECTIVE To establish the method for determination of bile acid active constituents （cholic acid， hyodeoxycholic acid and sodium taurocholate） in Babao jingfeng powder， and to compare the differences of 3 constituents in Babao jingfeng powder from different manufacturers. METHODS UPLC-MS method was adopted to determine the contents of cholic acid， hyodeoxycholic acid and sodium taurocholate in 9 batches of Babao jingfeng powder from 3 manufacturers. The separation was performed on a Phenomenex Luna® C18 column with mobile phase consisted of acetonitrile-water （gradient elution） at a flow rate of 0.5 mL/min. The column temperature was 40 ℃ and the sample size was 2 μL. MS system was operated in a negative ion single quadrupole mode， and the m/z of cholic acid， hyodeoxycholic acid and sodium taurocholate were 407.2， 392.2 and 514.2， respectively. With the contents of three components as the index， cluster analysis and principle component analysis were performed on 9 batches of samples. RESULTS The linear ranges of cholic acid， hyodeoxycholic acid and sodium taurocholate were 5.48- 548.40， 5.38-538.40， 4.74-474.05 ng/mL， respectively （r≥ 0.999 3）. RSDs of precision， repeatability and stability tests（24 h） were all lower than or equal to 3.7% （n＝6）， and the average recoveries were 103.3%， 103.3% and 101.6% with RSDs of 3.3%， 3.4%， 4.2% （n＝6）， respectively. The contentsof cholic acid ranged 0.702-1.711 mg/g， those of deoxycholic acid ranged 0.599-2.049 mg/g， and those of sodium taurocholate ranged 0.664-1.752 mg/g in 9 batches of samples. The average content of 3 components in samples from manufacturer A was high， and the difference between batches was the smallest； the average content of 3 components from manufacturer C was the lowest， and the difference between batches was large. Cluster analysis could basically distinguish samples from different manufacturers； cluster analysis was conducted by using the comprehensive score of principal components as an indicator， the results of which were basically consistent with those of cluster analysis for content. CONCLUSIONS The method is established successfully for the content determination of the 3 bile acid active constituents of artificial bezoar in Babao jingfeng powder. The contents of 3 components in Babao jingfeng powder from 3 manufacturers are significantly different.
ABSTRACT
OBJECTIVE:To study the effects of different extraction methods on the content of cholalic acid from Artificial Bezoar. METHODS: Artificial Bezoar from 3 different habitats was used to extract cholalic acid by hydrolization vs. hydrolization-free method, and the content of cholalic acid was determined by HPLC-ELSD(high performance liquid chromatography with evaporative light scattering detection). RESULTS: The content of cholalic acid in the hydrolyzed sample was higher than that in the non-hydrolyzed sample of the same batch. CONCLUSION: Through hydrolization, the binding cholalic acid in the artificial bezoar was transformed into free cholalic acid so that the content of cholalic acid was increased. Therefore, it is more reasonable to control the quality of Artificial Bezoar by taking the contents of total cholalic acid and free cholalic acid as indexes.
