Simple and Sensitive RP-HPLC and UV Spectroscopic Methods for the Determination of Remogliflozin Etabonate in Pure and Pharmaceutical Formulations

Abstract

Simple, novel and selective reverse phase-high performance liquid chromatography (RP-HPLC) and ultraviolet (UV) spectroscopic methods have been developed and optimized for the determination of remogliflozin etabonate (RMZ) in bulk and dosage forms. In the HPLC method, the principal peak and internal standard peak were eluted separately at different retention times (RT) with the chromatographic conditions such as, mobile phase consisting of 0.02 M ammonium acetate buffer (pH was adjusted to 4.0 by 1.0 M ortho phosphoric acid), acetonitrile and tetrahydrofuran in the ratio 50:45:05, respectively (v/v) and the stationary phase used was C18, 5 μm, 4.6 mm x 250 mm kromasil column. The flow rate was 2.0 mL min-1, sample injection volume was 10 μL, and the wavelength of detection was fixed at 228 nm. In case UV spectroscopic method, the RMZ was diluted with pure ethanol. The RMZ showed a maximum absorbance at 228 nm. Hence throughout analysis 228 nm was used for the determination of RMZ. The RT of RMZ and internal standard, atorvastatin (ATST) were 6.2 min and 7.0 min, respectively. The resolution between the peaks was found to be more than 2.0. The total run time was fixed at 10 min. The linearity range for RP-HPLC method was found to be 10 μg mL-1 to 50 μg mL-1, at a fixed concentration of ATST. The linearity range for the UV spectroscopic method was found to be in the range of 100 to 250 μg mL-1. Regression coefficients (R2) were found above 0.999 for both of the techniques. The limit of detection and quantification for RMZ were found to be 1.0 μg mL-1 and 3.5 μg mL-1 respectively, in RP-HPLC method and 10.0 μg mL-1 and 40 μg mL-1, respectively, in UV spectroscopic method. The developed methods were found to be simple, accurate, reproducible, and precise. The RMZ can be analyzed in dual techniques, i.e., chromatographic and UV spectroscopic methods for its routine analysis.