Citation: LIN Chu-Hui, CHAI Rui-Ping, ZHANG Lu, FU Zhi-Bo, ZHANG Hong-Yang, ZHANG Min, HU Ping. Determination of Six Kinds of Bile Acids in NiuhuangJiedu Tablets Using High Performance Liquid Chromatography Coupled with Charged Aerosol Detector[J]. Chinese Journal of Analytical Chemistry, ;2022, 50(5): 764-771. doi: 10.19756/j.issn.0253-3820.210910 shu

Determination of Six Kinds of Bile Acids in NiuhuangJiedu Tablets Using High Performance Liquid Chromatography Coupled with Charged Aerosol Detector

1.
Shanghai Key Laboratory of Functional Materials Chemistry, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai 200237, China;
2.
Thermo Fisher Scientific China Co. Ltd., Shanghai 200000, China;
3.
Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China

Corresponding author: HU Ping, huping@ecust.edu.cn
Received Date: 31 December 2021
Revised Date: 27 February 2022

Fund Project: Supported by the National Natural Science Foundation of China (No.81973285).

A high performance liquid chromatography-charge aerosol detector (HPLC-CAD) method for determination of six kinds of bile acids(Glycocholicacid, hyodeoxycholic acid, cholic acid, glycodeoxycholic acid, chenodeoxycholic acid and deoxycholic acid) in Niuhuang Jiedu tablets was developed. The tablet samples were extracted by ultrasonication in 70% methanol and concentrated under nitrogen flow. The separation of bile acids was achieved on an Agilent Poroshell 120 EC C₁₈ column (4.6 mm × 100 mm, 2.7 μm) with gradient elution using methanol and aqueous solution containing 0.1% formic acid and 5 mmol/L ammonium formate. CAD evaporation temperature was set at 55℃ and the power function value (PFV) was 1.35. The linearity ranges were 2.10-972 μg/mL for glycocholic acid, 2.09-1046 μg/mL for hyodeoxycholic acid, 2.05-978 μg/mL for cholic acid, 2.56-1022 μg/mL for glycodeoxycholic acid, 1.25-658 μg/mL for chenodeoxycholic acid and 2.05-1025 μg/mL for deoxycholic acid, respectively. All relative coefficients (R²) were higher than 0.9991. The limits of detection and the limits of quantitation ranged from 0.86 to 1.53 μg/mL and 1.20 to 2.56 μg/mL, respectively. The average recoveries of six kinds of bile acids were in the range of 94.2%-105.3% with RSD ≤ 1.63%. The method was used to determine the real tablet samples, and the results indicated that the contents of six bile acids had an obvious difference among the Niuhuang Jiedu tablets of different manufacturers. The HPLC-CAD method for the determination of six kinds of bile acids was sensitive, accurate and reliable, which could be applied to evaluate the quality of the Niuhuang Jiedu tablets.
- Niuhuang Jiedu tablet,
- High performance liquid chromatography,
- Charged aerosol detector,
- Bile acid
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