Simultaneous determination of the potent anti-tuberculosis regimen—Pyrazinamide, ethambutol, protionamide, clofazimine in beagle dog plasma using LC–MS/MS method coupled with 96-well format plate

Autor: Yuefen Lou, Guorong Fan, Shengyuan Wu, Chih-Ming Ho, Liai Lan, Xianting Ding, Jingjing Jiang
Jazyk: angličtina
Rok vydání: 2019
Předmět:
TB
tuberculosis
Clinical Biochemistry
Antitubercular Agents
Pharmaceutical Science
PZA
pyrazinamide
Tandem mass spectrometry
01 natural sciences
Clofazimine
Beagle dog plasma
Analytical Chemistry
chemistry.chemical_compound
Tandem Mass Spectrometry
Drug Discovery
PTO
protionamide
Spectroscopy
ESI
electrospray ionization
Chemistry
FSC
Feedback System Control
HIV
human immunodeficiency virus
Calibration
Ethambutol
medicine.drug
Formic acid
Electrospray ionization
AIDS
acquired immune deficiency syndrome
Mass spectrometry
Article
Dogs
Pharmacokinetics
LC–MS/MS
medicine
Animals
EMB
ethambutol
MRM
multiple-reaction monitoring
Chromatography
010405 organic chemistry
Elution
Anti-tuberculosis drug
010401 analytical chemistry
Reproducibility of Results
CFZ
clofazimine
Pyrazinamide
0104 chemical sciences
LC–MS/MS
liquid chromatography tandem mass spectrometry
Prothionamide
PRS
Parabolic Response Surface
96-well format plate
Chromatography
Liquid
Zdroj: Journal of Pharmaceutical and Biomedical Analysis
ISSN: 1873-264X
0731-7085
Popis: Highlights • LC–MS/MS method for determination of Pyrazinamide, Ethambutol, Protionamide and Clofazimine in Beagle Dog Plasma. • Method validation was conducted according to FDA and NMPA guidelines. • Hemolysis effect was investigated in detail. • The method is robust and high throughput cooperated with 96-well format plates.
Tuberculosis is one of the top concerns in the world and acutely threatens human health. A new potent candidate regimen containing pyrazinamide (PZA), ethambutol (EMB), protionamide (PTO) and clofazimine (CFZ) was proposed by Parabolic Response Surface/Feedback System Control (FSC/PRS) system and showed excellent outcomes in vitro and vivo studies. Here, a convenient liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) method was developed for the simultaneously determination of four compounds in beagle dog plasma. The plasma samples, 50 μL for each, were pretreated by methanol on 96-well format plates and a further dilution step was designed to reduce predictable matrix effect and lessen the burden of subsequent analysis. The chromatographic separation was achieved on an Agilent SB-Aq column (4.6 mm × 150 mm, 5 μm) at 30 °C by a gradient elution within 6 min. The mobile phase was a mixture of 0.2% formic acid-5 mM ammonium acetate aqueous solution (phase A) and 0.2% formic acid methanol (phase B) with a total flow rate of 1 mL/min. The 30% of post-column eluant was injected into mass spectrometer, equipped with electrospray ionization (ESI) source under positive mode and multiple-reaction monitoring (MRM). This quantification method was proved to be satisfied in selectivity, accuracy, precision, linearity (r2 > 0.998), recovery, matrix effect and stability. Under the specialized conditions, the calibration curves ranged from 20 to 5000 ng/mL for PZA, 1 to 500 ng/mL for EMB, 1 to 500 ng/mL for PTO, and 1 to 200 ng/mL for CFZ. The quantitative accuracy was further assessed under different degrees of hemolyses in detail. This method was proved to be robust and efficient, and successfully applied to the pharmacokinetic study of the new regimen in Beagle dogs.
Databáze: OpenAIRE
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