Abstract
A validated ultra-performance liquid chromatography mass spectrometric method (UPLC–MS/MS) was used for the simultaneous quantitation of candesartan (CN) and hydrochlorothiazide (HCT) in human plasma. The analysis was performed on UPLC–MS/MS system using turbo ion spray interface. Negative ions were measured in multiple reaction monitoring (MRM) mode. The analytes were extracted using a liquid–liquid extraction (LLE) method by using 0.1mL of plasma volume. The lower limit of quantitation for CN and HCT was 1.00ng/mL whereas the upper limit of quantitation was 499.15ng/mL and 601.61ng/mL for CN and HCT respectively. CN d4 and HCT-13Cd2 were used as the internal standards for CN and HCT respectively. The chromatography was achieved within 2.0min run time using a C18 Phenomenex, Gemini NX (100 mm×4.6 mm, 5µm) column with organic mixture:buffer solution (80:20, v/v) at a flow rate of 0.800mL/min. The method has been successfully applied to establish the bioequivalence of candesartan cilexetil (CNC) and HCT immediate release tablets with reference product in human subjects.
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