Analytical Evaluation of the Mobile Phase Variables by Factorial Design for Efavirenz Separation Using High Performance Liquid Chromatography

Abstract

A statistical optimization strategy was effectively used to investigate the effect of system factors on the chromatographic analysis of Efavirenz. To improve the procedure for excellent chromatographic responses, the effect of altering the flow rate, temperature, and concentration of % Acetonitrile in buffer (phosphate buffer 25mM, pH 7.5) was investigated. Response surface methodology (RSM) was used with Plackett-Burman designs to discover the optimum circumstances. The optimal regions for flow rate (0.8 ml/min) and temperature (25◦C) were determined from the response surface graphs as -1, 0, and +1. According to the Pareto ranking, temperature was the most influential variable influencing the selected responses. Linearity was seen in the 30-70 ug/ml range, with a considerably high correlation coefficient (r2 = 0.999). LOD and LOQ values The concentrations were 0.30 and 0.81 µg/ml, respectively. The devised approach underwent validation for accuracy, precision, linearity, range, and specificity. The approach was successfully employed to examine the chromatographic performance of a tablet formulation, and it was determined to be 100.88% with a standard deviation of ±1.36.