Analytical Quality by Design-Based RP-HPLC Method for Simultaneous Estimation of Lobeglitazone sulfate and Glimeperide in Pharmaceutical Formulations

A simple, precise, accurate, and robust High-Performance Liquid Chromatographic (HPLC) method has been developed for simultaneous estimation of Lobeglitazone sulfate and Glimeperide both in bulk and tablet dosage form. Fishbone diagram was used to identify variables that can affect quality of analytical method. Analytical quality by design (AQbD) approach was implemented for method development process. Box-Behnken Design (BBD) was used to optimise chromatographic conditions. Three factors were selected to design the matrix- percent organic ratio, mobile phase pH and flow rate. Analysis of variance (ANOVA) was applied to confirm if selected factors were significant. The analysis was done on Phenomenex C₁₈(250mm, 4.6mm, 5μm) column, using mobile phase as Acetonitrile: Water [% Orthophosphoric acid (OPA)] (80:20% v/v), (pH: 2.5), flow rate of 1ml/min with 243nm detecting wavelength. ICHQ2 (R2) guidelines were followed for method validation. The method was found to be linear in the range of 5-25μg/ml for Lobeglitazone sulfate and 10-50μg/ml for Glimeperide with correlation coefficients of 0.999. Percent recovery for both drugs was found to be in the range of 98-102%. To estimate precision and repeatability Percent Relative Standard Deviation (%RSD) was calculated. It was found to be below 2% confirming that the proposed method showed good accuracy, robustness.