Romanian Society of Pharmaceutical Sciences
 

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LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY DETERMINATION OF FLUCONAZOLE LEVELS IN HUMAN PLASMA FOR BIOAVAILABILITY STUDIES

LÉNÁRD FARCZÁDI1,2, ORSOLYA MELLES1, LAURIAN VLASE2*, BRINDUSA TILEA3

1.Vim Spectrum SRL, 409 Sighisoarei St., RO-547367, Corunca, Tirgu-Mures, Romania
2.Department of Pharmaceutical Technology and Bio-pharmaceutics, Faculty of Pharmacy, “Iuliu Hatieganu” University of Medicine and Pharmacy, 8 Victor Babes St., RO-400012, Cluj-Napoca, Romania
3.Infectious Diseases, Department M4, Clinical Sciences, Faculty of Medicine, University of Medicine and Pharmacy, 38 Gheorghe Marinescu St., RO-540139, Tirgu Mures, Romania

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The increase of fungal infections requires active antimycotic agents, fluconazole being widely used. A high-throughput tandem mass spectrometry coupled liquid chromatography method (LC-MS/MS) was developed and validated for the determination of fluconazole. Separation was performed using a Zorbax SB-C18 column with mobile phase consisting of methanol and 0.1% formic acid in water (45:55, v/v) in isocratic elution, with a flow rate of 0.6 mL/min. Detection of fluconazole was performed in multiple reaction monitoring mode (m/z 219.9 from m/z 306.9) with positive electrospray ionization. Plasma samples were processed using a protein precipitation method with methanol. As internal standard, prednisone was used. Method validation was carried out for selectivity, linearity (r > 0.9978), intra-day and inter-day precision (CV < 7.3%) and accuracy (bias < 13.5%) over the concentration range of 105 - 5600 ng/mL plasma. Recovery for fluconazol was 100.6 - 108.2%. The analytical method developed is simple, sensitive and selective, requires small volumes of plasma and can be used in clinical and bioequivalence studies.