DEVELOPMENT AND VALIDATION OF NEW HPLC-METHODS FOR ESTIMATION OF FLUCONAZOLE IN DIFFERENT SIMULATED BIOLOGICAL FLUIDS: A COMPARATIVE STUDY

In the present work, a new reverse phase-high performance liquid chromatography (RP-HPLC) method was developed, validated, and compared for estimation of the bis-triazole antifungal fluconazole (FLZ), in different simulated biological fluids, that is, gastric fluid simulant (GFS, pH=1.5), vaginal fluid simulant (VFS, pH=4.2), topical simulated media-phosphate buffer (PB, pH=6.8), blood plasma simulant-phosphate buffer saline (PBS, pH=7.4) and in methanol. For RP-HPLC the chromatography was carried out on a octadecyl silane (ODS-3) Hypersil C-18 column (250mmx4.6mmx5 mu m) using water (pH 5.2, adjusted with orthophosphoric acid) and acetonitrile (80:20, v/v) as the mobile phase at a flow rate of 2.5mL/min with detection at 260nm. The calibration curve of standard FLZ was linear in the range 0.2-40 mu g/mL, 0.1-40 mu g/L, 0.2-40 mu g/mL, 0.1-40 mu g/mL, and 0.2-50 mu g/mL, in GFS, VFS, PB, PBS, and in methanol, respectively. All the methods were validated with respect to linearity, precision, accuracy, and specificity for intra- and inter-day and satisfied the International Conference on Harmonisation (ICH) and United States Food and Drug Administration (USFDA'S) guidelines.