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LC–QToF–MS method for quantification of ethambutol, isoniazid, pyrazinamide and rifampicin in human plasma and its application
Author(s) -
Fachi Mariana Millan,
Vilhena Raquel Oliveira,
Boger Beatriz,
Domingos Eric Luiz,
Santos Josiane Marlei Müller Fernandes,
Junkert Allan Michael,
Fátima Cobre Alexandre,
Momade Danilo Raul Ossufo,
BeraldiMagalhães Francisco,
Liz Marcus Vinicius,
CordeiroSantos Marcelo,
Pontarolo Roberto
Publication year - 2020
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.4812
Subject(s) - pyrazinamide , chemistry , ethambutol , protein precipitation , chromatography , formic acid , ammonium formate , isoniazid , rifampicin , bioanalysis , therapeutic drug monitoring , pharmacokinetics , high performance liquid chromatography , pharmacology , tuberculosis , medicine , biochemistry , pathology , antibiotics
In this research, we developed and validated a liquid chromatography coupled to mass spectrometry (LC–QToF–MS) method for simultaneous quantification of the anti‐tuberculosis drugs ethambutol, isoniazid, pyrazinamide and rifampicin in human plasma. Plasma samples spiked with cimetidine (internal standard) were extracted using protein precipitation with acetonitrile containing 1% formic acid. Separation was performed using a C 18 column under flow gradient conditions with water and acetonitrile, both containing 5 m m ammonium formate and 0.1% formic acid. The method was validated according to the ANVISA and US Food and Drug Administration guidelines for bioanalytical method validation. The calibration curve was linear over a concentration range of 0.2–5 μg ml −1 for ethambutol, 0.2–7.5 μg ml −1 for isoniazid, 1–40 μg ml −1 for pyrazinamide and 0.25–2 μg ml −1 for rifampicin, all with adequate precision and accuracy. The method was reproducible, selective and free of carryover and matrix effects. The validated LC–QToF–MS method was successfully applied to real samples and shown to be applicable to future therapeutic and pharmacokinetic monitoring studies.