DESIGN OF EXPERIMENT UTILIZATION TO DEVELOP AND VALIDATE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY TECHNIQUE FOR ESTIMATION OF SERTACONAZOLE NITRATE

A novel method of estimation and validation of Sertaconazole nitrate (SER) by Reverse Phase-High Performance Liquid Chromatography coupled with Ultra-violet detection was developed which had high potential in determining drug concentration with more precision and high accuracy. The process of elution was conducted using Phenomenex C₁₈ (250 mm × 4.6 mm i.d., 5.0 μm) using 0.01M monobasic sodium phosphate and acetonitrile in a ratio of 28:72% v/v as mobile phase at 4.5 pH and a flow rate of 1.0 mL/min. Detection was carried out using a UV detector at 260 nm. This precise method was linear between a range of 10 to 500 μg/ml with R² close to one (0.999). The limit of detection (LOD) and limit of quantification (LOQ) of SER was found to be 0.1 μg/ml and 0.15 μg/ml respectively. The method was validated for accuracy, precision, linearity, LOD, LOQ, and robustness. Validation studies demonstrated that this HPLC method is simple, specific, rapid, reliable and reproducible. The 3-level 2-factor face-centred central composite design was employed using Design Expert Software ver. 7.0.0 to examine the effect of independent chromatographic factors like pH of the mobile phase and the ratio between 0.01M monobasic sodium phosphate and acetonitrile on the dependent factors like peak area, theoretical plates and tailing factor. The ANOVA studies proved that the model employed for this study was significant. This method can be used as a more convenient and efficient option for the analysis of SER to establish the quality of the drug substance during routine analysis with consistent and reproducible results.