Objective: To establish a method for simultaneous determination of 22 cholic acids(glycoursodeoxycholic acid, glycohyocholic acid, taurocholic acid, 7-oxodeoxycholic acid, 12-oxocholic acid, glycocholic acid, 3-oxocholic acid, taurochenodeoxycholic acid, ursodeoxycholic acid, 3α-hydroxy-7-oxo-5β-cholanic acid, hyocholic acid, taurodeoxycholic acid sodium salt hydrate, 12-ketodeoxycholic acid, hyodeoxycholic acid, cholic acid, glycochenodeoxycholic acid, glycodehydrocholic, taurolithocholic acid sodium salt, glycolithocholic acid, chenodeoxycholic acid, deoxycholic acid and lithocholic acid) in Niuhuang Shedan Chuanbei liquid by ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Methods: The samples were extracted by methanol ultrasonic extraction. Using Thermo Fisher Scientfic Bremen HYPERSI GOLD (100 mm×2.1 mm, 1.9 μm) as chromatographic column, 0.1% formic acid aqueous solution (A)-methanol (B) as mobile phase with gradient elution. The flow rate was 0.2 mL·min-1, the column temperature was 40 ℃, the injection volume was 2 μL. The mass spectrometry adopted heating electrospray ion source (HESI) with negative ion mode and parallel reaction monitoring (PRM) mode. The spray voltage was 3.0 kV (-). Results: The established method showed a good linear relationship in a certain range (r≥0.999 4). The precision, repeatability and stability of the tested samples were good within 48 h. The recovery rate were 94.9%-105.2% and the RSDs were 0.50%-5.2%. The results of content determination showed that the contents of taurocholic acid, cholic acid, porcine cholic acid, glycine cholic acid and porcine deoxycholic acid in Niuhuang Shedan Chuanbei liquid were higher, and there were great differences among different manufacturers. Cluster analysis could divided the samples into three categories. It was found that the contents of taurocholic acid, 3-oxocholic acid, 3α-hydroxy-7-oxo-5β-cholanic acid, 12-oxocholic acid and cholic acid in class Ⅱ were significantly higher than those in class Ⅱ and Ⅲ, and the content of porcine cholic acid in class Ⅲ was significantly higher than that in class Ⅰ and Ⅱ. Conclusion: This method is convenient, efficient, accurate and sensitive, and can be used for the determination of 22 cholic acids in Niuhuang Shedan Chuanbei liquid.
YOU Jing-jing, LIN Chen, LIN Shi-ling, LIN Yi-han, CHI Wen-jie, HUANG Ming-qing
. Determination of 22 cholic acid components in Niuhuang Shedan Chuanbei liquid by UHPLC-MS/MS*[J]. Chinese Journal of Pharmaceutical Analysis, 2023
, 43(11)
: 1847
-1857
.
DOI: 10.16155/j.0254-1793.2023.11.06
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