title: Rapid and specific approach for direct measurement of glimepiride in human plasma by LC-ESI-MS-MS employing automated 96 well format: application to a bioequivalence study.
abstract: A rapid liquid chromatographic method with electrospray ionization tandem mass spectrometric (LC-MS-MS) detection is developed and validated for quantification of glimepiride in heparinized human plasma. Plasma samples, without a drying and reconstitution step, are extracted by solid-phase extraction (SPE) and eluted with 0.9 mL of acetonitrile-methanol (1:1, v/v) containing 0.05% formic acid. The analyte and glimepiride d8 (internal standard, IS) are chromatographed on a C(18) column; the mobile phase is acetonitrile-2 mm ammonium formate (88:12, v/v), with the pH adjusted to 3.5 with formic acid, at a flow rate of 0.5 mL/min. The retention times of glimepiride and the IS are 0.93 min, and the runtime is 1.6 min per sample. Selected reaction monitoring of MH(+) at m/z 491.20 and 499.26 result in stable fragment ions with m/z 351.80 and 359.96 for glimepiride and the IS, respectively. The response was a linear function of the concentration in the range of 2.0-650.0 ng/mL, with r ≥ 0.9994. The recovery of glimepiride and the IS ranged from 81.91 to 83.36%. The assay has excellent characteristics and has been successfully used for the analysis of glimepiride in healthy human subjects in a bioequivalence study. It was well suited to clinical studies of the drug involving large numbers of samples.
authors.affiliation: Padmashri Vikhe Patil College, Pravaranagar, Loni Kurd, Pin-413713, Dis-Ahmednagar, Maharastra State, India.
authors.forename: M L
authors.initials: ML
authors.lastname: Kundlik
authors.forename: B H
authors.initials: BH
authors.lastname: Zaware
authors.forename: S R
authors.initials: SR
authors.lastname: Kuchekar
mesh_terms: D000328:Adult; D001331:Automation; D002853:Chromatography, Liquid / Q000379:methods*; D018592:Cross-Over Studies; D004355:Drug Stability; D006801:Humans; D016014:Linear Models; D008297:Male; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000295:instrumentation / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods; D013453:Sulfonylurea Compounds / Q000097:blood* / Q000493:pharmacokinetics*; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D013810:Therapeutic Equivalency
chemical_list: D013453:Sulfonylurea Compounds; C057619:glimepiride