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

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.

Smart UV Derivative Spectrophotometric Methods for Simultaneous Determination of Metformin and Remogliflozin: Development, Validation and Application to the Formulation

“…The basic principle of ratio derivative spectroscopic method involves division of UV spectra of a mixture of analytes metformin (M) and remogliflozin (R) (M + R) with UV spectra of the appropriate concentration of one of the component (R') to generate a ratio spectra. (Figure 4) 27 The newly generated ratio spectra of components are presented as equation 1…”

“…The mean percentage recovery was found to be from 99.77% to 101.83% for REM and 98.41 % to 99.85 % for MET and it is in agreement with the amount of analytes in the formulations. Further, the suggested techniques were statistically compared to the reported HPLC method 26 concerning accuracy and precision. No significant discrepancy was witnessed in the assay outcomes acquired by the suggested UV derivative techniques and reported HPLC method.…”

“…22 Two stability-indicating liquid chromatographic procedures for the assay of REM from the bulk and formulations were also reported in the literature. 23,24 Recently, Tammisetty et al 25 reported UPLC method and our team has reported RP-HPLC and derivative spectroscopic procedures 26,27 for the concurrent determination of MET and REM from formulations. The reported UV derivative spectroscopic methods were based on first and 2 nd derivative spectroscopic methods, however, the use of higher derivative is preferred for better resolution of interfering analytes, hence in the present work third derivative spectroscopic method was developed.…”

“…a mean of six determinations; b critical vales at p=0.05; SD : standard deviation; c Reference method:26 Chromolith (50 mm × 4.6 mm i.d. 5 µm) C 18 column, Mobile phase acetonitrile and mixture of 25 mM sodium dodecyl sulfate, 10 mM potassium dihydrogen phosphate (pH 3.5) in a ratio of 42%:58% (v/v) respectively, 2 ml /min flow rate, at 230 nm.…”

Development and Validation of Green UV Derivative Spectrophotometric Methods for Simultaneous Determination Metformin and Remogliflozin from Formulation: Evaluation of Greenness

“…There were few reports in the literature for the simultaneous estimation of metformin along with SGLT-2 inhibitors [ 32 , 33 ]. However, UPLC [ 34 ], HPLC and UV second derivative spectroscopic methods [ 35 ] have been reported for the concurrent determination of metformin and remogliflozin. However, UPLC and HPLC methods requires sophisticated expensive instruments and toxic solvents such acetonitrile and methanol as mobile phase.…”