Determination of obeticholic acid in rat plasma by liquid chromatography-tandem mass spectrometry (LC-MS/MS)
10.16438/j.0513-4870.2017-0841
- VernacularTitle:LC-MS/MS法测定大鼠血浆中奥贝胆酸的浓度
- Author:
Ru-yue ZHANG
1
;
Yuan GU
2
;
Ai-jie ZHANG
2
;
Shi-qi DONG
3
;
Quan-sheng LI
2
;
Guang-li WEI
2
;
Duan-yun SI
2
Author Information
1. Tianjin Medical University Graduate School, Tianjin 300070, China
2. State Key Laboratory of Drug Delivery Technology and Pharmacokinetics, Tianjin Institute of Pharmaceutical Research, Tianjin 300193, China
3. Tianjin University, Tianjin 300191, China
- Publication Type:ORIGINAL ARTICLES
- Keywords:
obeticholic acid;
LC-MS/MS;
plasma concentration;
pharmacokinetics;
primary biliary cholangitis
- From:
Acta Pharmaceutica Sinica
2018;53(2):271-277
- CountryChina
- Language:Chinese
-
Abstract:
A simple and sensitive method was developed for quantitation of obeticholic acid in rat plasma with liquid chromatography-tandem mass spectrometry (LC-MS/MS). After liquid-liquid extraction by methyl tert-butyl ether, the chromatographic separation was carried out on an ACE Excel 2 Super C18 column (50 mm×2.1 mm ID, 1.7 μm) with a gradient mobile phase consisting of acetonitrile and 2 mmol·L-1 ammonium formate at a flow rate of 0.2 mL·min-1. The quantitation analysis was performed using multiple reaction monitoring (MRM) at the specific ion transitions of m/z 418.9[M-H]-→401.2 for obeticholic acid and m/z 469.0[M-H]-→ 425.2 for glycyrrhetinic acid (internal standard) in the negative ion mode with electrospray ionization (ESI) source. This validated LC-MS/MS method yielded a good linearity over the range of 5 -5 000 ng·mL-1 with the lower limit of quantitation (LLOQ) of 5 ng·mL-1. The intra and inter-assay precisions (RSD) were all less than 9.82% and the accuracy (RE) was within ±6.90%. The extraction recovery of obeticholic acid was from 85.4% to 88.5%, and the matrix effect of obeticholic acid ranged from 78.9% to 82.5%. Stability test suggest that obeticholic acid in rat plasma was stable for 24 h on workbench, up to 1 month at -70℃, and after three cycles of freeze-thaw. Extracted samples were stable for more than 24 h in an auto-sampler at 6℃. The precision was less than 7.25%, and the accuracy was within ±11.2%, after being diluted 10 times by blank rat plasma. The method has been successfully applied to a pharmacokinetic study of obeticholic acid in rats following oral administration at the dose of 2.5 mg·kg-1.
