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.
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 BEH(TM) C18 column (50 × 2.1 mm, 1.7 µm) under an isocratic mobile phase, consisting of acetonitrile: 10 mM 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 (r(2)  ≥ 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.
Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, PO Box 2457, Riyadh, 11451, Saudi Arabia.|Bioavailability Laboratory, College of Pharmacy, King Saud University, Riyadh, 11451, Saudi Arabia.
Muzaffar
M
Iqbal
Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, PO Box 2457, Riyadh, 11451, Saudi Arabia.
Mashooq A
MA
Bhat
Center of Excellence in Biotechnology Research (CEBR), King Saud University, PO Box 2460, Riyadh, 11451, Saudi Arabia.
Faiyaz
F
Shakeel
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000081005:Cyclohexane Monoterpenes; D004355:Drug Stability; D007538:Isoniazid / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D039821:Monoterpenes / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012545:Schiff Bases; D053719:Tandem Mass Spectrometry / Q000379:methods*
D000081005:Cyclohexane Monoterpenes; D039821:Monoterpenes; D012545:Schiff Bases; C006923:carvone; D007538:Isoniazid
CSB-INH; UHPLC-MS/MS; pharmacokinetics; rat plasma