Development and validation of UHPLC‐MS/MS assay for rapid determination of a carvone Schiff base of isoniazid (CSB‐INH) in rat plasma: application to pharmacokinetic study.

Journal Article

Shakeel, Muzaffer Iqbal, Mashooq A. Bhat, Faiyaz . 2015

• Magazine \ Newspaper: Biomedical Chromatography
• Volume Number: 29
• Pages: 876-882

Publication Abstract

In this study, a fast UHPLC-MS/MS method was developed and validated for the determination of a novel potent
carvone Schiff base of isoniazid (CSB-INH) in rat plasma using carbamazepine as an internal standard (IS). After a single-step
protein precipitation by acetonitrile, CSB-INH and IS were separated on an Acquity BEHTM C18 column (50 × 2.1 mm, 1.7 μm)
under an isocratic mobile phase, consisting of acetonitrile: 10mM ammonium acetate (95:5, v/v), at a flow rate of 0.3 mL/
min. Quantification was performed on a triple quadrupole tandem mass spectrometer in multiple reactions monitoring mode
by using positive electrospray ionization source. The precursor to product ion transitions were set at m/z 270.08→79.93 for
CSB-INH and m/z 237.00→178.97 for IS. The proposed method was validated in compliance with US Food and Drug
Administration and European Medicines Agency guidelines for bioanalytical method validation. The method was found to
be linear in the range of 0.35–2500 ng/mL (r2 ≥ 0.997) with a lower limit of quantification of 0.35 ng/mL. The intra- and
inter-day precision values were ≤12.0% whereas accuracy values ranged from 92.3 to 108.7%. In addition, other validation
results were within the acceptance criteria and the method was successfully applied in a pharmacokinetic study of
CSB-INH in rats.