title: Advantages of automation in plasma sample preparation prior to HPLC/MS/MS quantification: application to the determination of cilazapril and cilazaprilat in a bioequivalence study.
abstract: An HPLC/MS/MS method characterized by complete automation and high throughput was developed for the determination of cilazapril and its active metabolite cilazaprilat in human plasma. All sample preparation and analysis steps were performed by using 2.2 mL 96 deep-well plates, while robotic liquid handling workstations were utilized for all liquid transfer steps, including liquid-liquid extraction. The whole procedure was very fast compared to a manual procedure with vials and no automation. The method also had a very short chromatographic run time of 1.5 min. Sample analysis was performed by RP-HPLC/MS/MS with positive electrospray ionization using multiple reaction monitoring. The calibration curve was linear in the range of 0.500-300 and 0.250-150 ng/mL for cilazapril and cilazaprilat, respectively. The proposed method was fully validated and proved to be selective, accurate, precise, reproducible, and suitable for the determination of cilazapril and cilazaprilat in human plasma. Therefore, it was applied to a bioequivalence study after per os administration of 2.5 mg tablet formulations of cilazapril.
authors.affiliation: University of Athens, Division of Pharmaceutical Chemistry, Department of Pharmacy, Panepistimioupoli Zografou GR-157 71, Athens, Greece.
authors.forename: Filomila
authors.initials: F
authors.lastname: Kolocouri
authors.forename: Yannis
authors.initials: Y
authors.lastname: Dotsikas
authors.forename: Constantinos
authors.initials: C
authors.lastname: Apostolou
authors.forename: Constantinos
authors.initials: C
authors.lastname: Kousoulos
authors.forename: Georgios-Stefanos
authors.initials: GS
authors.lastname: Soumelas
authors.forename: Yannis L
authors.initials: YL
authors.lastname: Loukas
mesh_terms: D000806:Angiotensin-Converting Enzyme Inhibitors / Q000737:chemistry / Q000493:pharmacokinetics; D001331:Automation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D017315:Cilazapril / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000493:pharmacokinetics; D004355:Drug Stability; D006801:Humans; D015394:Molecular Structure; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013810:Therapeutic Equivalency
chemical_list: D000806:Angiotensin-Converting Enzyme Inhibitors; D017315:Cilazapril; C045980:cilazaprilat