An Effective and Time-Saving HPTLC Densitometry Technique for the Measuring of Antidiabetic Triple-Drug Combination: From TLC Spot Formation to Method Validation

A precise, rapid, linear & robust High-Performance Thin-Layer Chromatography (HPTLC)-Densitometric was successfully developed and validated method for the simultaneous estimation of a triple antidiabetic drug combination comprising Metformin Hydrochloride (MET), Teneligliptin Hydrobromide Hydrate (TEN), and Pioglitazone Hydrochloride (PIO) in bulk and ph'ceutical formulation. Chromatographic separation was achieved using precoated silica gel 60 F254 TLC plates with a mobile phase made up of n-butanol, 1,4-dioxane, glacial acetic acid, and n-hexane in a ratio of 4:2:2:2 (%v/v/v/v). Densitometry was used for detection at 242 nanometers. Spots were obtained using distinct and well-resolved Rf values 0.20 ± 0.01, 0.55 ± 0.02, and 0.90 ± 0.01 for MET, TEN, and PIO, respectively. The ICH Q2 (R1) guidelines were followed in the validation of the approach, demonstrating excellent linearity over the ranges of concentrations of 1000-3500 nanogram/band for MET, 40-140 nanogram/band for TEN & 30-115 nanogram/band for PIO, with correlation coefficients (r2) of 0.9983, 0.9961, and 0.9965, respectively. The recovery studies confirmed the method's accuracy, yielding mean recoveries of 99.92% (MET), 100.03% (TEN) & 100.97% (PIO). The LOD and LOQ were determined to be 154.22 nanogram/band and 467.34 nanogram/band for MET, 8.66 nanogram/band and 26.25 nanogram/band for TEN, and 10.95 ng/spot and 33.18 ng/spot for PIO, respectivelyThe assay results further confirmed the reliability of the developed method, showing percentage recoveries of 99.03% (MET), 99.34% (TEN), and 99.03% (PIO). Overall, the proposed HPTLC-densitometric method provides a simple, economical and effective analytical tool for the routine quality control and simultaneous quantification of Metformin hydrochloride, Teneligliptin hydrobromide hydrate, and Pioglitazone