APPLICATION OF DOE AND STATISTICAL ANALYSIS FOR DEVELOPMENT AND VALIDATION OF ANALYTICAL METHOD FOR CHLROHEXIDINE GLUCONATE AND CETRIMIDE IN ITS BULK AND PHARMACEUTICAL DOSAGE FORMS

The present work includes development of analytical method for simultaneous estimation of Chlorhexidine gluconate and Cetrimide by RP-HPLC method. The chromatographic conditions were successfully optimised for the separation of Chlorhexidine gluconate and Cetrimide by using Hypersil BDS C18 column (4.6 × 150 mm) 5µ, flow rate of 0.8 ml/min, mobile phase ratio of (30:55:15 v/v/v) ACN: methanol: phosphate buffer (KH₂PO₄ and K₂HPO₄) phosphate pH 3 (pH was adjusted with orthophosphoric acid) and detection wavelength used was 210 nm. The retention times were found to be 3.10 min and 3.9 min for Cetrimide and Chlorhexidine gluconate respectively. The % purity of Chlorhexidine gluconate and Cetrimide was found to be 99.92% and 100.45% respectively. The analytical method was validated according to ICH guidelines (ICH, Q2 (R1). The linearity study of Chlorhexidine gluconate and Cetrimide was found in concentration range of 3 µg – 18 µg and 30 µg – 180 µg. DOE was applied in validation for checking robustness of the developed method using Box-Behnken design and also normal distribution of data was verified by Anderson-Darling normality test. The method developed was found to be specific, selective, and robust and can be applied for routine analysis of marketed formulation in laboratory premises.