Development and validation of an HPLC method for stability evaluation of nystatin

Abstract

A liquid chromatography method for evaluating the stability of Nystatin (Nys) in an ointment was developed and validated, since the traditional pharmacopeial microbiological methods are unable to indicate stability. The stress experiments showed that Nys was found to significantly degrade in alkaline and acidic conditions and also under oxidative stress. Lower levels of degradation were detected under heat and with the sample exposed to Xenon light. Resolutions higher than 2 for Nys and degradation products (DP) chromatographic peaks were achieved by using an Inerstil ODS-3 column, isocratic elution with methanol:water and UV detection at 305 nm. The system was found to be linear over a range of 102 to 310 IU mL-1 and proved precise, since the RSD(%) was 0.24% for the six replicates tested. The method also exhibited good levels of recovery (from 98.24% to 100.74%). Therefore, the validation fulfilled pharmacopeial requirements and the procedure was found to be reliable, precise, accurate and selective for determination of Nys and its degradation products.