RAPID, METHOD FOR DETERMINATION OF METFORMIN AND CANAGLIFLOZIN IN PLASMA BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETER, APPLICATION TO BIOEQUIVALENCE STUDY

SAMBASIVA RAO PURAM; NITHYA G; SUBBA RAO NB; RAMAN BATHEJA

doi:10.22159/ajpcr.2019.v12i2.29027

Authors

SAMBASIVA RAO PURAM Department of Bioanalytical, Vergo Clinicals, Corlim, Goa, India.
NITHYA G Department of Chemistry, School of basic sciences, Vels University, Chennai, Tamil Nadu, India.
SUBBA RAO NB Department of Bioanalytical, Vergo Clinicals, Corlim, Goa, India.
RAMAN BATHEJA Department of Bioanalytical, Vergo Clinicals, Corlim, Goa, India.

DOI:

https://doi.org/10.22159/ajpcr.2019.v12i2.29027

Keywords:

Metformin canagliflozin, Metformin and canagliflozin in plasma, Bioequivalence of metformin and canagliflozin, Canagliflozin metformin by liquid chromatography-tandem mass spectrometer

Abstract

Objective: The objective of the study was to develop and validate a rapid selective bioanalytical method for the simultaneous determination of metformin and canagliflozin in plasma by liquid chromatography-tandem mass spectrometer (LC-MS/MS) to facilitate bioequivalence study sample analysis. Stock solutions and spiking solutions were prepared accurately in methanol and 80% methanol in water, respectively.

Methods: Chromatography monitored using Analyst 1.6.2 software. Method was found selective, no matrix effect, reproducible and consistent recovery, accuracy, precision, and stable in aqueous as well as extracted/matrix samples. Method found linear over the range 10–2000 ng/ml for metformin and 30–6000 ng/ml for canagliflozin. The method is successfully applied to analyze samples collected in a bioequivalence study after administration of metformin/canagliflozin 1000/150 mg to 24 healthy male volunteers.

Results: Extraction was carried out using a simple solid phase extraction using mobile phase elution. 5 μl sample delivered as injection volume in 1.000 ml/min isocratic mobile phase flow on turbo ion electron spray technique for positive mode on API 4000 MS.

Discussion: Chromatography achieved within 3 min using a mobile phase containing acetonitrile and 10 mM ammonium formate buffer in a ratio of 70:30 on chromolith C18 analytical column. Q1/Q3 are 130.1/70.1, 136.3/77.1, 462.2/267.2 (with ammonium adduct), and 466.4/267.1 for metformin, metformin D6, canagliflozin, and canagliflozin D4, respectively.

Conclusion: Rapid, sensitive method for the simultaneous estimation of metformin and canagliflozin in plasma by LC-MS/MS is successfully developed, validated and applied to analyze 960 unknown samples of a bioequivalence study. Incurred sample reanalysis was revealed great reproducibility.

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