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DEVELOPMENT AND VALIDATION OF A HIGHLY SENSITIVE METHOD FOR THE DETERMINATION OF VORICONAZOLE AND POSACONAZOLE IN HUMAN SERUM BY LC-MS/MS-ESI: APPLICATION TO A PATIENT
Prem Pal*, Amrendra Thakur and Dhanuja
ABSTRACT A highly sensitive, rapid assay method has been developed and validated for the estimation of Voriconazole (VOR) and Posaconazole (POS) in human serum with liquid chromatography coupled to tandem mass spectrometry with electro spray ionization in the positive-ion mode. The assay procedure involves extraction of VOR, POS and Voriconazole d3 and Posaconazole d5 (internal standards, IS) from human serum with simple protein precipitation extraction process. Chromatographic separation was achieved using an gradient mobile (2mM ammonium acetate in water (0.1formic Acid) : 2mM ammonium acetate in methanol (0.1formic Acid) at a flow rate of 0.60 mL/min on an Acquity UPLC@BEH C18 column maintained at 40 °C with a total run time of 3.5 min. The MS/MS ion transitions monitored were 350.15 127.05 for VOR AND POS and 1353.23.2 127.05 for IS. Method validation was performed as per CLSI guidelines and the results met the acceptance criteria. The lower limit of quantitation achieved was 122.27 ng/mL for VOR and 122.54 ng/mL, the linearity range extended from 120.3 to 6002.3 ng/mL for VOR and 120.3ng/mL to 6003.6 ng/mL for POS. The intra and inter-day precisions were in the range of 1.80-8.90 for VOR and 0.5-6.2 for POS & 3.90-9.40 for VOR and 2.50-9.70 for POS % in human serum. Keywords: Voriconazole and posaconazole; LC-MS/MS; method validation; human serum. [Download Article] [Download Certifiate] |