A Comparative Validation Study of Fluconazole by HPLC and UPLC with Forced Degradation Study

The simplest stability indicating reversed phase Isocratic HPLC and UPLC methods has been developed and validated for the determination of fluconazole in bulk and solid pharmaceutical dosage form. A SunFire C18 (250 × 4.5？mm, 5？μm particle size) column has been used for HPLC and BEH C18 (100 × 2.1？mm, 1.7？μm particle size) column used for UPLC. The Mobile phase consisted of Methanol？:？Water (70？:？30) for HPLC and Methanol？:？Water (55？:？45？v/v) for UPLC. Isocratic flow was set at 1？mL/min and 0.30？mL/min, respectively, for HPLC and UPLC. For both HPLC and UPLC system detection has been performed at 211？nm with 30°C column oven temperature (good elution was obtained at 30°C) and injection volume, respectively, 2？μL and 20？μL for HPLC and UPLC. 1. Introduction Fluconazole [2-(2,4-difluorophenyl)-1,3-bis-(1H-1,2,4-triazole-1-yl)-2-propanol] [1] is mainly used as an antifungal antibiotic [2–4] (Figure 1) is and also used to prevent yeast infections in patients who are likely to become infected, because they are being treated with chemotherapy or radiation therapy before a bone marrow transplant. Several HPLC methods were reported for the analysis of fluconazole in pharmaceutical preparations and in body fluids as plasma, serum, and human urine sample [1, 5–11]. Few of GC and HPLC methods have been reported for the determination of fluconazole in biological fluids, eye drops, and creams [12]. The UV method was also reported for syrups, capsules and intravenous solution, and microbiological assay for capsules [13]. Few of HPLC and UPLC methods have also been reported for the simultaneous determination of fluconazole and its combination dosage tablet [14, 15]. LC-MS/MS method was also reported for the determination of fluconazole in human plasma [16]. Review literature suggested that there is no single UPLC method reported for the determination of fluconazole in pharmaceutical dosage forms or in API with acidic, alkali (Figure 5), oxidative (Figure 6), thermal (Figure 7), and photo degradation (Figure 8) study. Ultraperformance liquid chromatography (UPLC) (Figure 2) is well known and widely used analytical technique for the analysis of drug products and drug substance [17]. Current work has advantages over the earlier in terms of speed and simplicity. This work also deals with the stability indicating study and forced degradation of fluconazole under conditions such as oxidation, acid hydrolysis, base hydrolysis, and thermal and photolytic stress [18]. This method was validated using both the instruments HPLC and UPLC by different parameters such as linearity,