Method Development Validation and In Vivo Bioequivalence Studies of Anti Diabetic Drugs

Abstract

Diabetes mellitus (DM) is one of the most common chronic diseases occurring all over newlinethe world. Type II diabetes which is non-insulin-dependent diabetes mellitus is the most newlinecommon form of diabetes. A recent meta-analysis assessed the benefit of early metforminbased newlinecombinations versus metformin alone and demonstrated superior results of the newlinecombination therapy with better HbA1c reduction. SGLT2 inhibitors are one such novel class newlineof drugs which have shown valuable efficacy profile when used as an add-on with other oral newlineanti-diabetic drugs. Initial combination therapy of empagliflozin and metformin has been shown to be more effective in HbA1c reduction versus empagliflozin monotherapy or newlinemetformin monotherapy. newlineA novel, selective and sensitive methods are developed for simultaneous estimation newlineof SGLT2 inhibitors in presence of metformin and are successfully applied to bioequivalence newlinestudies in healthy Indian volunteers. The current study reports the development, newlineoptimization, and validation of liquid chromatography-mass spectrometry (LC-MS/MS) newlinemethod for simultaneous quantification of SGLT2 inhibitors in combination with metformin newlinein human plasma. newlineFor CN and ME, the solid-phase extraction technique was employed where strata X newlinepolymeric reverse phase (30 mg-1 cc) SPE cartridges were used for the extraction of analytes newlineand IS from plasma. The ACE 5 C18 column (50 x 4.6 mm, 5and#956;) was used to chromatograph newlinethe prepared samples. The mobile phase consisted of methanol and 5 mM ammonium newlinetrifluoroacetate in water, pH 5 (50:50, v/v) at a flow rate of 0.8 mL/min. Detection was newlineperformed by positive ion Turbo ion spray in Multiple reaction monitoring (MRM) mode, newlinemonitoring the transitions m/z 461.9and#8594;m/z 191.1 and m/z 461.9and#8594;m/z 267.2, for newlinequantification of canagliflozin. The response of canagliflozin fragments m/z 461.9and#8594;m/z newline191.1 and m/z 461.9and#8594;m/z 267.2 was combined. Also, for metformin transitions were newlinemonitored at m/z 130.0and#8594;m/z 71.1.