title: Development, optimization and validation of a highly sensitive UPLC-ESI-MS/MS method for simultaneous quantification of amlodipine, benazeprile and benazeprilat in human plasma: application to a bioequivalence study.
abstract: A rapid, simple, sensitive and specific LC-MS/MS method has been developed and validated for the simultaneous estimation of amlodipine (AML), benazepril (BEN) and benazeprilat (BNT) using eplerenone and torsemide as internal standards (IS). The Xevo TQD LC-MS/MS was operated under the multiple-reaction monitoring mode using electrospray ionization. Sample preparation involves both extraction and precipitation techniques. The reconstituted samples were chromatographed on Acquity UPLC BEH C18 (50mm×2.1mm, 1.7μm) column by pumping 0.1% formic acid and acetonitrile in a gradient mode at a flow rate of 0.45ml/min. A detailed validation of the method was performed as per the FDA guidelines and the standard curves were found to be linear in the range of 0.1-5ng/ml for AML; 5-1200ng/ml for both BEN and BNT. The intra-day and inter-day precision and accuracy results were within the acceptable limits. A run time of 2.5min for each sample made it possible to analyze more than 300 human plasma samples per day. The developed assay method was successfully applied to a bioequivalence study in human volunteers.
authors.affiliation: Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, 11562, Cairo, Egypt. Electronic address: mamdouhreda75@yahoo.com.
authors.forename: Mamdouh R
authors.initials: MR
authors.lastname: Rezk
authors.affiliation: Pharmaceutics Department, Faculty of Pharmacy, Delta University for Science and Technology, Egypt; Advanced Research Center (ARC), Nasr City, Cairo, Egypt.
authors.forename: Kamal A
authors.initials: KA
authors.lastname: Badr
mesh_terms: D017311:Amlodipine / Q000097:blood*; D001552:Benzazepines / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D010949:Plasma / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods; D013810:Therapeutic Equivalency
chemical_list: D001552:Benzazepines; D017311:Amlodipine; C052133:benazeprilat; C044946:benazepril
keywords: Amlodipine; Benazepril; Benazeprilat; Bioequivalence; UPLC–MS/MS