Fluconazole Bioequivalence Study: Quantification by Tandem Mass Spectrometry

Abstract

To develop a new method for quantifying fluoconazole in human plasma and to compare the bioavailability of two fluconazole capsule formulations, an open, randomized, two-period crossover study with a one-week washout interval was conducted in 24 healthy volunteers. Plasma samples were obtained up to 168 hours after drug administration and the serum fluconazole concentrations were analyzed using electrospray tandem mass spectrometry coupled to liquid chromatography using multiple reaction monitoring mode. The pharmacokinetic parameters obtained for fluconazole after the administration of each formulation included the Area under the curve (AUC)(0-168h), AUC(0-infinity), Cmax, Cmax/AUC(0-168h), Tmax, elimination rate constant (Ke), and half-life (T1/2). Within- and between-run imprecision was less than 2.3% and 8.2%, respectively. Inaccuracy within and between runs was -1.5% and -9.7%, respectively. The pharmacokinetic parameters for bioequivalence showed a normal distribution, and the variance of AUC(0-168h), AUC(0-infinity), and Cmax were homoscedastic. The geometric mean for the Fluconal/Zoltec (Fluconal; Libbs Farmacêutica Ltda, São Paulo, Brazil; Zoltec; Laboratórios Pfizer Ltda., São Paulo, Brazil) individual percent ratio was 94.9% for AUC(0-168h), 94.7% for AUC(0-infinity), 80.1% for Cmax, 102.6% for Ke, 97.5% for T1/2, and 0.93 for Tmax (arithmetic mean of individual differences). We have developed a method in which liquid chromatography is coupled with electrospray tandem mass spectrometry to improve the pharmacokinetic analysis of fluconazole. Because the 90% CI AUC is within the interval proposed for the Food and Drug Administration, we concluded that Fluconal is bioequivalent to Zoltec in terms of absorption. The CV was 27.5% for the Cmax parameter, indicating that fluconazole's absorption rate is highly variable. The European Union Regulatory Agency accepts an interval of 70-143%, and because the 90% CI for Cmax is within the interval proposed for the European Union agency, we conclude that Fluconal is bioequivalent to Zoltec for the rate of absorption.

Bioequivalence evaluation of two formulations of fluconazole 150 mg capsule in healthy Arab men

“…1 of 0.108 to 0.023 h À1 [1,6]. These values are similar to the corresponding values obtained in the current study, suggesting that ethnicities may have little or no effect on the elimination process, half-life or extent of absorption of fluconazole.…”

“…Previous studies from Brazil [1,6] and Thailand [7] reported a K el of 0.021 to 0.023 h À1 [1,7], T max of 1.18 to 1.59 h [7], C max of 2.61 to 3.84 mg/ml [1,6], AUC 0!1 of 152 to 181 mg.h/ml [1,6] and C max /AUC 0 ! 1 of 0.108 to 0.023 h À1 [1,6].…”

Rapid and sensitive method for the determination of sertraline in human plasma using liquid chromatography–tandem mass spectrometry (LC–MS/MS)

“…The analytical techniques employed in recent years for the estimation of sertraline in human plasma include HPLC, GC or in combination with MS i.e. liquid chromatography and tandem mass spectrometry (LC-MS/MS) and GC-MS/MS [7][8][9][10][11][12][13][14][15][16][17][18]. All these reported methods require either a lengthy extraction and/or derivatization procedure, yet the desired sensitivity is not achieved.…”

“…Kim et al [13] have reported a GC-MS/MS method with sensitivity of 0.2 ng/ml, but the extraction procedure involved deproteination, liquid-liquid extraction followed by derivatization. The upper limit of quantification (10.0 ng/ml) of their linearity range (0.2-10 ng/ml) for sertraline is inadequate to quantify the achieved C max for 50 mg [7] as well as for 100 mg dose [8]. Also, these assay methods do not meet modern drug development needs, which require a rapid and accurate feedback of analytical information of pre-clinical and clinical pharmacokinetic studies.…”

Multiplex Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry Method for Simultaneous Quantification in Human Plasma of Fluconazole, Itraconazole, Hydroxyitraconazole, Posaconazole, Voriconazole, Voriconazole- N -Oxide, Anidulafungin, and Caspofungin

“…Analytical methods using liquid chromatography coupled with triple-quadrupole mass spectrometry (LC-MS/MS) have been reported for the quantification of single antifungals in biological fluids, including fluconazole (FLC) (14,34,49), itraconazole (ITZ) (39,62) and its active metabolite (hydroxyitraconazole [OH-ITZ]) (4,6,27,53), voriconazole (VRC) (2,15,26,54,63), posaconazole (PSC) (10, 40a, 44, 52), caspofungin (CASPO) (7,16,40), and anidulafungin (ANI) (13). LC-MS/MS assays were recently reported for the simultaneous quantification of posaconazole, voriconazole, isavuconazole, caspofungin, anidulafungin, and micafungin in different peripheral blood compartments (19) and of itraconazole, voriconazole, and posaconazole in blood (3).…”