DEVELOPMENT AND VALIDATION OF SENSITIVE LC-ESI-MS/MS METHOD FOR THE SIMULTANEOUS ESTIMATION OF DAPAGLIFLOZIN AND SAXAGLIPTIN IN HUMAN PLASMA
Keywords:Dapagliflozin, Saxagliptin, LC-ESI-MSMS, Validation, Precision and Accuracy
Objective: To develop and validate a sensitive liquid chromatography-electrospray ionization-tandem mass spectrometric (LC-ESI-MS/MS) technique for the quantification of dapagliflozin and saxagliptin in plasma by linagliptin as internal standard.
Methods: Chromatography was achieved on hypersil C18 (50 mmx4 mm) 5 µ analytical column with 0.1% formic acid and acetonitrile (25:75 V/V) as mobile phase at 0.7 ml/min flow rate. Dapagliflozin, saxagliptin, and linagliptin were detected at m/z 409.14/135.0, m/z 316.2/180.13 and m/z 472.54/456.21 respectively. Drugs and internal standard were extracted by LLE (liquid-liquid extraction).
Results: Developed technique was validated over 0.5-1500.0 ng/ml linear concentration range for dapagliflozin and 2.00-2000.0 ng/ml for saxagliptin. This method established with intra-batch and inter-batch precision within 2.44-8.12% and 1.25-7.14 % for dapagliflozin and 1.84-7.5 % and 1.02–6.00 % for saxagliptin. This method established with intra-batch and inter-batch accuracy for dapagliflozin within 98.86-103% and 96.98-102 % respectively and for saxagliptin within 98.05-109.06 % and 97.00-104.00 % respectively.
Conclusion: Both dapagliflozin and saxagliptin were stable during three freeze-thaw cycles, long term and bench-top stability studies. The developed method was useful for the routine analysis of dapagliflozin and saxagliptin simultaneously in plasma samples.
Obermeier M, Yao M, Khanna A, Koplowitz B. In vitro characterization and pharmacokinetics of dapagliflozin (BMS-512148), a potent sodium-glucose cotransporter type II inhibitor, in animals and humans. Drug Metab Dispos 2010;38:405-14.
Kasichayanula S, Liu X, Lacreta F, Griffen SC, Boulton DW. Clinical pharmacokinetics and pharmacodynamics of dapagliflozin, a selective inhibitor of sodium-glucose cotransporter type 2. Clin Pharmacokinet 2014;53:17-27.
Scirica B, Bhatt DL, Braunwald E. Saxagliptin and cardiovascular outcomes in patients with type 2 diabetes mellitus. New England J Med 2013;369:1317-26.
Ali S, Fonseca V. Saxagliptin overview: special focus on safety and adverse effects. Expert Opin Drug Safety 2013;12:103–9.
Dhillon S, Weber J. Saxagliptin. Drugs 2009;69:2103–14.
Raveendra BG, Kumar RA, Shaheen SD, Greeshma A, Satyanarayana M. A novel stability-indicating method for the simultaneous estimation of saxagliptin and dapagliflozin in rat serum by using UV spectroscopy. Pharm Anal Acta 2018;9:2-5.
Gajanan Vithoba Mante, Krishna Radheshyam Gupta, Atul Tryambakrao Hemke. Estimation of dapagliflozin from its tablet formulation by UV-spectrophotometry. Pharm Methods 2017;8:102-7.
Patel PD, Pandya SS. Validated RP–HPLC method for simultaneous estimation of dapagliflozin and saxagliptin hydrochloride in the tablet dosage form. Int J Pharm Res Scholars 2018;7:32-41.
Deepan Thiyagarajan, Dasaratha Dhanaraju. Stability indicating HPLC method for the simultaneous determination of dapagliflozin and saxagliptin in bulk and tablet dosage form. Curr Issues Pharm Med Sci 2018;31:39-43.
Sarath Nalla, Seshagiri Rao JVLN. A stability indicating RP-HPLC method for simultaneous estimation of dapagliflozin and saxagliptin in combined tablet dosage forms. Inventi Rapid: Pharm Anal Quality Assurance 2017;4:1-11.
Sharmila Donepudi. Simultaneous estimation of saxagliptin and dapagliflozin in human plasma by validated HPLC-UV method. Turkish J Pharm Sci 2018;1-23. Doi:10.4274/tjps.46547.
Ghadir A Khalil, Salama Ismail, Gomaa Mohammed, Helal Mohammed. Validated RP-HPLC method for simultaneous determination of canagliflozin, dapagliflozin, empagliflozin and metformin. Int J Pharm Chem Biol Sci 2018;8:1-13.
Batta N, Pilli NR, Derangula VR. A rapid and sensitive LC-MS/MS assay for the determination of saxagliptin and its active metabolite 5-hydroxy saxagliptin in human plasma and its application to a pharmacokinetic study. Drug Res 2015;65:133-40.
Shuchi Dave Mehta, Sarvesh Paliwal. Phytochemical analysis, liquid chromatography, and mass spectroscopy and in vitro anticancer activity of annona squamosa seeds linn. Asian J Pharm Clin Res 2018;11:101-3.
US FDA, Guidance for Industry Bioanalytical Method Validation, Food and Drug Administration, Center for Drug Evaluation and Research (CDER), Rockville, Maryland, USA; 2001.
Marlina Ika, Rizka Andalusia, Supandi Supandi, Yahdiana Harahap. Simultaneous analytical method development of 6-mercaptopurine and 6-methylmercaptopurine in dried blood spot using ultra performance liquid chromatography-tandem mass spectrometry. Int J Appl Pharm 2017;9:168-71.
Swathi P, Vidyadhara S, Sasidhar RLC, Kalyan Chakravarthi K. Method development and validation for the estimation of entecavir in bulk and pharmaceutical dosage forms by RP-HPLC. Int J Curr Pharm Res 2017;9:107-11.
International Conference on Harmonization (ICH) of Technical Requirements for the Registration of Pharmaceuticals for Human Use, Validation of analytical procedures: Methodology, ICH-Q2B, Geneva; 1996.
Revathi Naga Lakshmi Ponnuri, Prahlad Pragallapati, Ravindra N, Venkata Basaveswara Rao Mandava. A rapid and sensitive liquid chromatography-mass spectrometry/mass spectrometry method for estimation of pioglitazone, keto pioglitazone and hydroxy pioglitazone in human plasma. Asian J Pharm Clin Res 2017;10:120-8.
Validation of Analytical Procedures: Methodology. ICH Harmonised Tripartite Guidelines; 1995.
Rezk MR, Basalious EB, Karim IA. Development of a sensitive UPLC-ESI-MS/MS method for quantification of abacavir and its metabolite, GS-331007, in human plasma: application to a bioequivalence study. J Pharm Biomed Anal 2015;114:97-104.
Rezk MR, Basalious EB, Amin ME. Novel and sensitive UPLC-MS/MS method for quantification of sofobuvir in human plasma: application to a bioequivalence study. Biomed Chromatogr 2016;30:1354-62.
Shi X, Zhu D, Lou J, Gan D. Evaluation of a rapid method for the simultaneous quantification of ribavirin, sofobuvir and its metabolites in rat plasma by UPLC-MS/MS method. J Chromatogr B: Anal Technol Biomed Life Sci 2015;1002:353-7.
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.
Yahdiana Harahap, Norma Andriyani, Harmita. Method development and validation of lercanidipine in human plasma by liquid chromatography tandem mass spectrometry. Int J Appl Pharm 2018;10:87-91.