Quantitative Determination of Fluconazole by Ultra-Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS) in Human Plasma and its Application to a Pharmacokinetic Study

Abstract

In this study, a simple, rapid and sensitive ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method is described for determination of fluconazole (FLA) in human plasma samples using phenacetin as the internal standard (IS). Sample preparation was accomplished through one-step protein precipitation by methanol, and chromatographic separation was performed on an Acquity BEH C18 column (2.1 mm×50 mm, 1.7 μm) with mobile phase consisted of acetonitrile and water containing 0.1% formic acid (40:60, v/v) at a flow of 0.45 mL/min. Mass spectrometric analysis was performed using a QTrap 5500 mass spectrometer coupled with an electro-spray ionization (ESI) source in the positive ion mode. The MRM transition of m/z 307.2→238.2 was used to quantify for FLA. The linearity of this method was found to be within the concentration range of 10-6 000 ng/mL for FLA in human plasma. Only 1.0 min was needed for an analytical run. The method herein described was superior to previous methods and was successfully applied to the pharmacokinetic study of FLA in healthy Chinese volunteers after oral administration.

Development and Validation of Simple RP-HPLC Method for Intracellular Determination of Fluconazole Concentration and Its Application to the Study ofCandida albicansAzole Resistance

“…A literature survey reveals some HPLC methods that are reported for the determination of FLC in pharmaceutical dosage formulations as anticipated with the variation of mobile phase, column, and detector. Different HPLC methods [ 11 , 12 ] for individual assay are available for FLC in official pharmacopoeia and several LC-MS/MS methods were used for determination of FLC in human plasma [ 13 – 15 ]. Hence, an attempt has been made to develop a simple, efficient, and selective RP-HPLC method for intracellular determination of FLC concentration and its application to azole-resistant C. albicans .…”