Pharmacokinetics of H002, a novel S1PR1 modulator, and its metabolites in rat blood using liquid chromatography–tandem mass spectrometry

• Main Authors: Jiaqi Mi, Manman Zhao, Shu Yang, Shuang Yang, Jing Jin, Xiaojian Wang, Qiong Xiao, Jinping Hu, Yan Li
• Format: Article
• Language: English
• Published: Elsevier 2016-10-01
• Series: Acta Pharmaceutica Sinica B
• Subjects: S1P receptor; S1PR1 modulator; Sphingosine-1-phosphate; S1P analogue; Metabolite; LC–MS/MS; Pharmacokinetics; Periphery blood lymphocyte
• Online Access: http://www.sciencedirect.com/science/article/pii/S2211383515300435
• ISSN: 2211-3835; 2211-3843

A rapid and sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated for the simultaneous determination of H002 and its phosphorylated metabolite, H002-P and hydroxylated metabolite H002-M, in rat blood. H001, an analogue of H002, was used as the internal standard. Blood samples were prepared by simple protein precipitation. The analytes and internal standard were separated on a Zorbax SB-C18 column with a gradient mobile phase consisting of methanol and water containing 0.1% formic acid at a flow rate of 0.2 mL/min with an operating temperature of 20 °C. The detection was performed on a triple quadrupole tandem mass spectrometer with positive electrospray ionization in multiple-reaction monitoring mode. Linear detection responses were obtained from 0.2–100 ng/mL for H002 and H002-M, while 0.5–100 ng/mL for H002-P. The intra- and inter-day precision (RSD%) was within 11.76%, with the accuracy (RE%) ranging from –9.84% to 9.12%. The analytes were shown to be stable during sample storage, preparation and analytic procedures. The method was applied to determine the pharmacokinetics of H002 in rats, and a preliminary study showed that the pharmacokinetics of H002 correlated with its biological effect on peripheral blood lymphocytes.