RAPID, METHOD FOR DETERMINATION OF METFORMIN AND CANAGLIFLOZIN IN PLASMA BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETER, APPLICATION TO BIOEQUIVALENCE STUDY
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.
2. Devineni D, Curtin CR, Ariyawansa J, Weiner S, Stieltjes H, Vaccaro N, et al. Bioequivalence of canagliflozin/metformin immediate release fixed dose combination tablets compared with concomitant administration of single components of canagliflozin and metformin in healthy fed participants. J Bioequiv Availab 2014;6:164-73.
3. Attimara MV, Harsha NS, Nair AB, Aldhubaib BE, Alhaider IA. LCMS Method for the Simultaneous Determination of Metformin and Miglitol in Human Plasma: Application to Pharmacokinetic Studies. Available from: http://www.msacl.org/2015.
4. Kobuchi S, Yano K, Ito Y, Sakaeda T. A validated LC-MS/MS method for the determination of canagliflozin, a sodium-glucose co-transporter 2 (SGLT-2) inhibitor, in a lower volume of rat plasma: Application to pharmacokinetic studies in rats. Biomed Chromatogr 2016;30:1549-55.
5. Madhukar A, Prince A, Kumar R, Sanjeeva Y, Jagadeeshwar K, Raghupratap D. Simple and sensitive analytical method development and validation of metformin hydrochloride by RP-HPLC. Int J Pharm Pharm Sci 2011;3:3.
6. Gaware D, Patil RN, Harole M. A validated stability indicating RP-HPLC method for simultaneous determination of metformin and canagliflozin in pharmaceutical formulation. World J Pharm Pharm Sci 2015;4:631-40.
7. Pandya RH, Rathod R, Maheswari DG. Bioanalytical method development and validation for simultaneous determination of linagliptin and metformin drugs in human plasma by rp-hplc method. Pharmacophore 2014;5:202-18.
8. Devineni D, Curtin CR, Polidori D, Gutierrez MJ, Murphy J, Rusch S, et al. Pharmacokinetics and pharmacodynamics of canagliflozin, a sodium glucose co-transporter 2 inhibitor, in subjects with Type 2 diabetes mellitus. J Clin Pharmacol 2013;53:601-10.
9. Gaikwad A, Gavali S, Narendiran AS. A simple and sensitive method for determination of metformin and sitagliptin in human plasma using liquid chromatography and tandem mass spectrometry. Int J Pharm Pharm Sci 2013;5:463-70.
10. Polagani SR, Pilli NR, Gajula R, Gandu V. Simultaneous determination of atorvastatin, metformin and glimepiride in human plasma by LC-MS/MS and its application to a human pharmacokinetic study. J Pharm Anal 2013;3:9-19.
11. Heinig K, Bucheli F. Fast liquid chromatographic-tandem mass spectrometric (LC-MS-MS) determination of metformin in plasma samples. J Pharm Biomed Anal 2004;34:1005-11.
12. Jian W, Ying-Wu WG, Jing-Kai G. Determination of metformin in human plasma by liquid chromatography-tandem mass spectrometric assay. Chem Res Chin Univ 2005;21:246-50.
13. Zhanga W, Hana E, Zhaoa H. Determination of Metformin in Rat Plasma by HILIC-MS/MS Combined with Tecan Automation and Direct Injection. Available from: http://www.wileyonlinelibrary.com/bmc.2673.
14. Iqbal M, Khalil NY, Alanazia AM, Al-Rasod KA. A simple and sensitive high performance liquid chromatography assay with a fluorescence detector for determination of canagliflozin in human plasma. Anal Methods 2015;7:45-7.
15. Chhetri HP, Thapa P, Van Schepdael A. Simple HPLC-UV method for the quantification of metformin in human plasma with one step protein precipitation. Saudi Pharm J 2014;22:483-7.
16. Uçaktürk E. The development and validation of a gas chromatography-mass spectrometry method for the determination of metformin in human plasma. Anal Methods 2013;2013:18.
17. Duttaa P, Rayb S, Nagarajanc K. Docking study of some glutamic acid derivatives as potent antineoplastic agents. Int J Pharm Pharm Sci 2014;6:419-22.
18. Sharma RB, Chetia D. Docking studies on quinine analogs for plasmepsin-II of malaria parasite using bioinformatics tools. Int J Pharm Pharm Sci 2013;5:681-5.
19. Swetha A, Kuber BR. A novel stability-indicating reverse phase liquid chromatographic method for the simultaneous estimation of metformin and teneligliptin in pure and pharmaceutical formulations. Int J Appl Pharm 2018;10:274-80.
20. Das D, Chakraborty J, Dash S. Bioequivalence study of antidiabetic activity between two marketed formulations of metformin on glucocorticoid-induced hyperglycemia in rabbit. Int J Curr Pharm Res 2017;9:47-50.
21. Kulkarni AA, Vaidya IS. Flow injection analysis: An overview. J Crit Rev 2015;2:19-24.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.