title: Stability-indicating validation of acitretin and isoacitretin in human plasma by LC-ESI-MS/MS bioanalytical method and its application to pharmacokinetic analysis.
abstract: LC- ESI- MS/MS simultaneous bioanalytical method was developed to determine acitretin and its metabolite isoacitretin in human plasma using acitretin-d3 used as the internal standard for both analytes. The compounds were extracted using protein precipitation coupled with liquid-liquid extraction with flash freezing technique. Negative mass transitions (m/z) of acitretin, isoacitretin and acitretin-d3 were detected in multiple reactions monitoring (MRM) mode at 325.4 → 266.3, 325.2 → 266.1 and 328.3 → 266.3, respectively, with a turbo ion spray interface. The chromatographic separation was achieved on an Ascentis-RP amide column (4.6 × 150 mm, 5 µm) with mobile phase delivered in isocratic mode. The method was validated over a concentration range of 1.025-753.217 ng/mL for acitretin and 0.394-289.234 ng/mL for isoacitretin with a limit of quantification of 1.025 and 0.394 ng/mL. The intra-day and inter-day precisions were below 8.1% for acitretin and below 13.8% for isoacitretin, while accuracy was within ±7.0 and ±10.6% respectively. For the first time, the best possible conditions for plasma stability of acitretin and isoacitretin are presented and discussed with application to clinical samples.
authors.affiliation: Ranbaxy Research Laboratories Ltd, HSIDC, Gurgaon, India.
authors.forename: Ajay
authors.initials: A
authors.lastname: Kumar
authors.forename: Tausif
authors.initials: T
authors.lastname: Monif
authors.forename: Arshad
authors.initials: A
authors.lastname: Khuroo
authors.forename: Dinakar
authors.initials: D
authors.lastname: Sasmal
authors.forename: Dipanjan
authors.initials: D
authors.lastname: Goswami
authors.forename: Vijay Kumar
authors.initials: VK
authors.lastname: Lahkar
mesh_terms: D017255:Acitretin / Q000097:blood* / Q000493:pharmacokinetics; D019540:Area Under Curve; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D016018:Least-Squares Analysis; D008297:Male; D010782:Photolysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D017255:Acitretin