Development and Validation of Rapid RP-HPLC and Green Second-Derivative UV Spectroscopic Methods for Simultaneous Quantification of Metformin and Remogliflozin in Formulation Using Experimental Design

Mahesh Attimarad,Abdulmalek Ahmed Balgoname,Katharigatta N Venugopala,Sreeharsha Nagaraja,Rafea Elamin Elgack Elgorashe,Mohammed Monirul Islam,Rajasekaran Subramaniam

doi:10.3390/separations7040059

Abstract

Recently, a new formulation containing metformin HCl (MFH) and remogliflozin etabonate (RGE) has been approved for the management of diabetes mellitus. However, only one analytical method has been reported for the simultaneous determination of both the analytes. Therefore, the current study was designed to develop simple UV derivative spectroscopic and rapid RP-HPLC methods for simultaneous determination of MFH and RGE. The chromatographic separation of MFH and RGE was performed using a monolithic C18 column with an optimized chromatographic conditions carried out by full factorial Box–Behnken design model. The spectroscopic technique was based on the determination of peak amplitude of second-order derivative UV spectra at zero crossings. Further, both the methods were validated and compared statistically using Student’s-t-test and F-test, and employed for the concurrent estimation of MFH and RGE in laboratory mixed solutions and formulations. Perturbation plots and response surface models showed the effect of chromatographic parameters and the final chromatographic condition was selected from 47 solutions suggested by the desirability function. Further, UV spectroscopic and HPLC procedures showed good linearity in the range of 1–24 µg/mL and 2–150 µg/mL for RGE and 2–30 µg/mL and 5–200 µg/mL for MFH, respectively. The average percent assay was found to be 99.51% and 99.80% for MFH and 99.60% and 100.07% for RGE by spectroscopic and HPLC methods, respectively. The proposed methods were simple, accurate, precise, and rapid. Therefore, they can be used for regular quality control of MFH and RGE formulations and dissolution studies as well.

Highlights

Diabetes mellitus is a leading cause of death all over the world [1,2,3]
The second-order derivative UV spectrophotometric technique was developed for the simultaneous quantification of remogliflozin etabonate (RGE) and metformin HCl (MFH) because zero-order UV spectra of RGE and MFH showed complete overlap (Figure 2A)
Two simple analytical procedures were established for the concurrent determination of RGE and MFH from laboratory mixed solutions and tablets without prior separation

Summary

Introduction

Diabetes mellitus is a leading cause of death all over the world [1,2,3]. Better glycemic control is essential to reduce diabetic complications such as kidney failure, retinopathy, neuropathy, cardiovascular complications, etc. Combination therapy using oral hypoglycemic agents having Separations. Metformin HCl (MFH, Figure 1A) is extensively used as an oral hypoglycemic drug, which has Diabetes mellitus is a leading of death all over the world [1,2,3]. Bybetter glycemic multiple modes of action, namely,cause reducing the intestine absorption of glucose increasing the control is essential to reduce diabetic complications such as kidney failure, retinopathy, neuropathy, anaerobic metabolism of glucose in enterocytes, decreasing the rate of hepatic gluconeogenesis, and cardiovascular complications, etc. Combination therapy using hypoglycemic agents having a insulin sensitivity, thereby increasing the utilization of glucose byoral peripheral cells [6,7,8]

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Conclusion