• Adluri Phanindra Research Scholar, Mewar University, Chittorgarh, Rajasthan, India
  • Y. Shravan Kumar Research Supervisor, Mewar University, Chittorgarh, Rajasthan, India



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.


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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.



How to Cite

Phanindra, A., and Y. Shravan Kumar. “DEVELOPMENT AND VALIDATION OF SENSITIVE LC-ESI-MS/MS METHOD FOR THE SIMULTANEOUS ESTIMATION OF DAPAGLIFLOZIN AND SAXAGLIPTIN IN HUMAN PLASMA”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 11, no. 4, Apr. 2019, pp. 55-59, doi:10.22159/ijpps.2019v11i4.31249.



Original Article(s)