title: Simultaneous determination of pirfenidone and its metabolite in human plasma by liquid chromatography-tandem mass spectrometry: application to a pharmacokinetic study.
abstract: A simple and rapid analytical method for the simultaneous determination of pirfenidone and its metabolite, 5-carboxy-pirfenidone, in human plasma using liquid chromatography-tandem mass spectrometry has been developed and validated. Aliquots of plasma (0.1 mL) containing pirfenidone and 5-carboxy-pirfenidone, as well as deuterium-labeled internal standards (ISs), were deproteinized using acetonitrile. An Agilent Zorbax Plus C18 column was used for the chromatography, with isocratic elution. The mobile phase was a mixture of acetonitrile and aqueous ammonium formate solution (5 mM) containing 0.1% formic acid (60 : 40, v/v). Using multiple reaction monitoring in positive ionization mode, transitions m/z 186.1 → 65.1, m/z 216.0 → 77.0, m/z 191.1 → 65.1 and m/z 221.0 → 81.0 were chosen to quantify pirfenidone, 5-carboxy-pirfenidone and the two ISs, respectively. The time of analysis was <3 min. The calibration curve was linear over the concentration ranges 0.005-25 μg/mL for pirfenidone, and 0.005-15 μg/mL for 5-carboxy-pirfenidone. The lower limit of quantification for both analytes was 0.005 μg/mL. The intra- and interday precision and relative errors in quality control samples were between -11.7 and 1.3% for pirfenidone and between -5.6 and 2.5% for 5-carboxy-pirfenidone, with mean recoveries ≥90%. The method that has been developed is easy to carry out, sensitive and rapid, and has been successfully used to investigate the pharmacokinetics of pirfenidone in healthy human volunteers.
authors.affiliation: Department of Pharmacy, Guangzhou Brain Hospital, Guangzhou Medical University, 36 Mingxin Road, Guangzhou 510370, China wenyuguandede@163.com.
authors.forename: Yu-Guan
authors.initials: YG
authors.lastname: Wen
authors.affiliation: Department of Pharmacy, Guangzhou Brain Hospital, Guangzhou Medical University, 36 Mingxin Road, Guangzhou 510370, China Department of Pharmacy, Shandong Jiaotong Hospital, Wuyingshan Road, Jinan 250031, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Pharmacy, People's Hospital of Xinhui District, Longshan Road, Jiangmen 529100, China.
authors.forename: Xiu-Ling
authors.initials: XL
authors.lastname: He
authors.affiliation: Department of Pharmacy, Guangzhou Brain Hospital, Guangzhou Medical University, 36 Mingxin Road, Guangzhou 510370, China.
authors.forename: De-Wei
authors.initials: DW
authors.lastname: Shang
authors.affiliation: Department of Pharmacy, Guangzhou Brain Hospital, Guangzhou Medical University, 36 Mingxin Road, Guangzhou 510370, China.
authors.forename: Xiao-Jia
authors.initials: XJ
authors.lastname: Ni
authors.affiliation: Department of Pharmacy, Guangzhou Brain Hospital, Guangzhou Medical University, 36 Mingxin Road, Guangzhou 510370, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of Pharmacy, Guangzhou Brain Hospital, Guangzhou Medical University, 36 Mingxin Road, Guangzhou 510370, China.
authors.forename: Zhan-Zhang
authors.initials: ZZ
authors.lastname: Wang
authors.affiliation: Department of Pharmacy, Guangzhou Brain Hospital, Guangzhou Medical University, 36 Mingxin Road, Guangzhou 510370, China.
authors.forename: Jin-Qing
authors.initials: JQ
authors.lastname: Hu
authors.affiliation: Department of Pharmacy, Guangzhou Brain Hospital, Guangzhou Medical University, 36 Mingxin Road, Guangzhou 510370, China.
authors.forename: Chang
authors.initials: C
authors.lastname: Qiu
mesh_terms: D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D064368:Healthy Volunteers; D006801:Humans; D057230:Limit of Detection; D011728:Pyridones / Q000097:blood* / Q000493:pharmacokinetics; D011786:Quality Control; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D011728:Pyridones; C000599556:5-carboxy-pirfenidone; C093844:pirfenidone