The present work demonstrates a simple, rapid, precise, specific, and sensitive reverse-phase high-performance liquid
chromatography (RP-HPLC) method for analyzing glimepiride in pure and tablet forms. The present method was developed
using a C18 column 150 × 4.6 mm, with 5 μm, and packing L1 maintained at a temperature of 30°C. The mobile phase was
prepared by dissolving 0.5 gram of monobasic sodium phosphate in 500 mL of distilled water, pH of the solution adjusted
to 2.1 to 2.7 with 10% phosphoric acid, and added 500 mL of acetonitrile. The mobile phase was pumped in the highperformance
liquid chromatography (HPLC) system at a flow rate of 1 mL/min, and separation was carried out at 228 nm,
using an ultraviolet (UV) detector. The chromatographic separation was achieved with peak retention time (RT) at about
9.30 minutes, and the method was found to be linear over a concentration range of 40 to 140 μg/mL. The specificity of the
method represented no interference of the excipients during the analysis, and stability testing after 24 hours also showed that
the method is suitable and specific.
The accuracy was between 99.93 to 99.96%, with limit of detection (LOD) and limit of quantitation (LOQ) being 0.354 μg/mL,
1.18 μg/mL, respectively. Satisfactory results were found for precision and robustness parameters during the development
and validation stage for the analytical method. The proposed method was also adopted for the analysis of glimepiride tablets
to improve the overall quality control.
Using this method, symmetric peak shape was obtained with reasonable retention time. The retention time of glimepiride for
six repetitions is 9.3 ± 0.1 minutes; the run time is 21 minutes.
The proposed RP-HPLC method is a modification of the United States Pharmacopeia (USP) method, and it was found to be
valid for glimepiride within concentration ranges 40 to 140 μg/mL, using C18 analytical columns, and isocratic elution with
UV detection, and at 1 mL/min of flow rate.
Analytical method development and validation of simultaneous estimation of rabeprazole, pantoprazole, and itopride by reverse-phase high-performance liquid chromatography
