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

  • 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.

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.

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

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RAO PURAM, S., N. G, S. RAO NB, and R. BATHEJA. “RAPID, METHOD FOR DETERMINATION OF METFORMIN AND CANAGLIFLOZIN IN PLASMA BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETER, APPLICATION TO BIOEQUIVALENCE STUDY”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 12, no. 2, Jan. 2019, pp. 93-97, doi:10.22159/ajpcr.2019.v12i2.29027.
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