title: Development and validation of a LC-MS/MS method for quantification of hetrombopag for pharmacokinetics study.
abstract: Hetrombopag as the derivative of ethylidene hydrazine carboxamide was recently developed into a novel patented non-peptide thrombopoietin mimetic and thrombopoietin receptor agonist to treat idiopathic thrombocytopenic purpura. To study the pharmacokinetics of hetrombopag, a highly sensitive, rapid and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for determination of hetrombopag in rat plasma. After protein precipitation extraction, the chromatography separation of analyte and internal standard named eltrombopag as an marketed analog of hetrombopag was performed on an Synergi Polar-RP column at the flow rate of 0.5 mL/min, and the determination was conducted on an API4000 triple quadrupole mass spectrometry in the multiple reaction monitoring mode using the respective [M+H](+) ions m/z 459.2 → 200.9 for hetrombopag and m/z 443.2 → 229.0 for IS. The lower limit of quantification was established to be 1 ng/mL, and the linear scope of standard curve was 1-1000 ng/mL. Both the precision (RSD%) and accuracy (RE%) were within the acceptable criterion of below 15 %. The validated method was successfully applied to quantify hetrombopag in the rat plasma and investigate the pharmacokinetics.
authors.affiliation: Guangdong Metabolic Diseases Research Center of Integrated Chinese and Western Medicine, Guangdong TCM Key Laboratory against Metabolic Diseases, Institute of Chinese Medical Sciences, Guangdong Pharmaceutical University, Guangzhou Higher Education Mega Centre, Guangzhou, 510006 People's Republic of China.
authors.forename: Tao
authors.initials: T
authors.lastname: Chen
authors.affiliation: Department of Pharmacology, West China School of Pharmacy, Sichuan University, Chengdu, 610041 People's Republic of China.
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Chen
authors.affiliation: CarysBio Holdings Limited, Foshan, 528000 People's Republic of China.
authors.forename: Suxing
authors.initials: S
authors.lastname: Zhang
authors.affiliation: CarysBio Holdings Limited, Foshan, 528000 People's Republic of China.
authors.forename: Kezhi
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Guangdong Metabolic Diseases Research Center of Integrated Chinese and Western Medicine, Guangdong TCM Key Laboratory against Metabolic Diseases, Institute of Chinese Medical Sciences, Guangdong Pharmaceutical University, Guangzhou Higher Education Mega Centre, Guangzhou, 510006 People's Republic of China.
authors.forename: Laiyou
authors.initials: L
authors.lastname: Wang
keywords: Hetrombopag; Idiopathic thrombocytopenic purpura; LC–MS/MS; Pharmacokinetics