Abstract

DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF FLUCYTOSINE AND POSACONAZOLE

D. N. Madhusudana*, K. P. Channabasavaraj and A. Namratha

ABSTRACT

A simple, sensitive, precise, rapid and accurate reverse phase high performance liquid chromatography (RP- HPLC) method was developed and validated for simultaneous estimation of Flucytosine and Posaconazole High performance Liquid Chromatograph SHIMADZU-SPD-20AT prominence auto sampler. The Chromatographic separation was achieved by using C18 LiChrospher RP-HPLC (250 mm×4.6 mm, 5 μm) column as stationary phase and mobile phase consists of Acetonitrile:15 mM of Potassium dihydrogen orthophosphate anhydrous buffer (pH 6.0 adjusted with 0.1N NaOH) in the ratio of (75:25%, V/V) with a flow rate of 1 mL/min. The analysis was performed at ambient temperature and the eluent was monitored at 246 nm using UV detector. The retention time of Flucytosine and Posaconazole were found to be 2.6 min and 4.2 min, respectively. The calibration curves were linear (r2 = 0.9999 and 0.9998) over a concentration range of 25-200 μg/mL for Flucytosine and Posaconazole, respectively. The limit of detection (LOD) was observed to be 0.4 μg/mL and 0.3 μg/mL for Flucytosine and Posaconazole, respectively. The limit of quantification (LOQ) was observed to be 1.4 μg/mL and 1 μg/mL for Flucytosine and Posaconazole, respectively.The developed method was validated as per ICH guidelines using parameters like linearity, specificity, system suitability, precision, ruggedness, robustness, accuracy, sensitivity. All the validation parameters were found to be well within the acceptance criteria. Hence the proposed method is sensitive and specific without any interference from the excipients for the routine analysis of Flucytosine and Posaconazole in bulk and tablet dosage forms.

Keywords: Flucytosine (FLU), Posaconazole (PSZ), RP-HPLC, and ICH guidelines.