Development and Validation of a Green UPLC Analytical Procedure for Glibenclamide Determination in Pharmaceutical Product Using Response Surface Methodology

Abstract

Glibenclamide (GB) is one of the drugs that is used in the first line for the management of the diabetes mellitus. The aim of this work is to develop a unique green UPLC analytical procedure to analyze GB in pharmaceutical products by implementing QbD (Quality by Design) approach. 32 full factorial design was applied to study the effect of three levels of formic acid (X1; 0.05-0.5%) and temperatures (X2; 25-55oC), for their effects on retention time (Y1), peak area (Y2), peak height (Y3), and HPLC-EAT; environmental assessment tool (Y4). A green mobile phase, which is composed of Methanol 70% and water 30% containing different concentrations of formic acid, was pumped in Isocratic elution mode at a flow rate of 0.2 ml/min, with a total run time of 3.0 min with UV detection at 225 nm. Subsequently, this method was successfully validated using several validation parameters according to ICH guidelines. The results showed that temperature had a significant indirect effect on retention time (P value = 0.001), while formic acid concentration had a significant indirect impact on Peak Area (P value = 0.0101). The peak height was highly affected with formic acid concentration as well as the temperature with P < 0.005. Furthermore, HPLC-EAT was highly affected by column temperature in an indirect way. The optimized green method was discovered to be acceptable in terms of selectivity, precision, accuracy, robustness, sensitivity, and specificity. Moreover, the method efficaciously separated GB peak in the presence of its degradation products in different harsh conditions, revealing stability-indicating property.