A Rapid and Sensitive Method for the Pharmacokinetic Study of Janumet (Sitagliptin and Metformin) Tablets by LC-MS/MS Coupled with Ion-Pair Solid Phase Extraction

Author(s): Xiaonian Han*, Jing Wang, Jing Huang, Lirong Peng.

Journal Name: Current Pharmaceutical Analysis

Volume 15 , Issue 7 , 2019

Become EABM
Become Reviewer

Graphical Abstract:


Background: As first-line treatments for diabetes, sitagliptin and metformin have been widely prescribed as a combination to enhance the therapeutic effect.

Objective: To establish a methodology to simultaneously monitor the two drugs in vivo by a reversedphase Liquid Chromatography-Tandem Mass Spectrometric (LC-MS/MS) method.

Methods: The two drugs were extracted from 50 μl human plasma by ion-pair solid phase extraction. The separation of the plasma samples was implemented on an Agilent Zorbax SB-CN column (150×4.6 mm, 5.0 µm). The mobile phase was the mixture (80:20, v/v) of methanol and 5.0 mM ammonium formate in water (pH 4.5). An ion trap spectrometer equipped with an electrospray ionization source was utilized to detect the elution in positive mode. Quantification of the analytes was achieved by Multiple Reaction Monitoring (MRM) using the transitions of m/z 408.3→235.1 for sitagliptin and m/z 130.1→ 60.2 for metformin.

Results: Sitagliptin and metformin demonstrated good linearity among the range of 1.00-1000 ng/mL and 5.00-4000 ng/mL. The intra-day and inter-day investigations displayed precisions of ≤ 3.6% and an accuracy range of -7.5% to 6.0% for the two drugs. The mean recovery of the two drugs was 96.0% and 98.5%. Under mandatory storage conditions, both the drugs gave an acceptable stability. The throughput of the assay was found to be more than 100 plasma samples per day ascribed to the run time of 3.0 min for each sample.

Conclusion: The developed method was successfully applied to a pharmacokinetic study for a fixeddose tablet formulation containing 50 mg sitagliptin and 500 mg metformin in 12 healthy volunteers.

Keywords: Type 2 diabetes mellitus, pharmacokinetics, liquid chromatography, tandem mass spectrometry, sitagliptin, metformin.

Tadic, M.; Cuspidi, C. Type 2 diabetes mellitus and atrial fibrillation: From mechanisms to clinical practice. Arch. Cardiovasc. Dis., 2015, 108(4), 269-276.
Trierweiler, H.; Kisielewicz, G.; Jonasson, T.H.; Petterle, R.R.; Moreira, C.A. Victória Borba ZC Sarcopenia: A chronic complication of type 2 diabetes mellitus. Diabetol. Metab. Syndr., 2018, 10(25), 1-9.
Arnold, S.V.; Lipska, K.J.; Wang, J.Y.; Seman, L.; Mehta, S.N.; Kosiborod, M. Use of intensive glycemic management in older adults with diabetes mellitus. J. Am. Geriatr, 2018, 66(6), 1190-1194.
Vijayakumar, T.M.; Jayram, J.; Cheekireddy, V.M.; Himaja, D.; Dharma, T.Y.; Narayanasamy, D. Safety, efficacy, and bioavailability of fixed-dose combinationsin Type 2 diabetes mellitus: A systematic updated review. Curr. Ther. Res. , 2017, 84(C), 4-9.
Scott, L.J. Sitagliptin: A review in type 2 diabetes. Drugs, 2017, 77(2), 209-224.
Shah, P.A.; Shah, J.V.; Sanyal, M.; Shrivastav, P.S. LC-tandem mass spectrometry method for the simultaneous determination of metformin and sitagliptin in human plasma after ion-pair solid phase extraction. J. Pharmaceut. Biomed., 2016, 131, 64-70.
Siddiqui, M.R.; AlOthman, Z.A.; Rahman, N. Analytical techniques in pharmaceutical analysis: A review. Arab. J. Chem., 2017, 1, S1409-S1421.
AlOthman, Z.A.; Rahman, N.; Siddiqui, M.R. Review on pharmaceutical impurities, stability studies and degradation products. Rev. Adv. Sci. Eng., 2013, 2, 155-166.
Rahman, N.; Azmi, S.N.H.; Wu, H.F. The importance of impurity analysis in pharmaceutical products: An integrated approach. Accredit. Qual. Assur., 2006, 11, 69-74.
Ramalingam, P.; Bhaskar, V.U.; Reddy, Y.P.; Kumar, K.V. Stability-indicating RP-HPLC method for the simultaneous determination of sitagliptin and simvastatin in tablets. Indian J. Pharm. Sci., 2014, 76(5), 407-414.
Salim, M.; Elenany, N.; Belal, F.; Walash, M.; Patonay, G. Simultaneous determination of sitagliptin and metformin in pharmaceutical preparations by capillary zone electrophoresis and its application to human plasma analysis. Anal. Chem. Insights, 2012, 7(1), 31-46.
Suresh, P.S.; Srinivas, N.R.; Mullangi, R. A concise review of the bioanalytical methods for the quantitation of sitagliptin, an important Dipeptidyl Peptidase-4 (DPP4) inhibitor, utilized for the characterization of the drug. Biomed. Chromatogr., 2016, 30(5), 749-771.
Georgita, C.; Albu, F.; David, V.; Medvedovici, A. Simultaneous assay of metformin and glibenclamide in human plasma based on extraction-less sample preparation procedure and LC/(APCI)MS. J. Chromatogr. B , 2007, 854(1-2), 211-218.
Mistri, H.N.; Jangid, A.G.; Shrivastav, P.S. Liquid chromatography tandem mass spectrometry method for simultaneous determination of antidiabetic drugs metformin and glyburide in human plasma. J. Pharmaceut. Biomed, 2007, 45(1), 97-106.
Swales, J.G.; Gallagher, R.T.; Denn, M.; Peter, R.M. Simultaneous quantitation of metformin and sitagliptin from mouse and human dried blood spots using laser diode thermal desorption tandem mass spectrometry. J. Pharmaceut. Biomed, 2011, 55(3), 544-551.
Scherf-Clavel, M.; Hogger, P. Analysis of metformin, sitagliptin and creatininein human dried blood spots. J. Chromatogr. B , 2015, 997, 218-228.
Reddy, S.; Ahmed, I.; Mukhopadhyay, A.; Thangam, S. Development and validation of a method for simultaneous estimation of metformin andsitagliptin in human plasma by LC-MS-MS and its application in a bioequivalence study. J. Chromatogr. Sci., 2015, 53(9), 1549-1556.
Bonde, S.L.; Bhadane, R.P.; Gaikwad, A.; Katale, D.; Gavali, S.; Narendiran, A.S. A simple and sensitive method for determination of metformin and sitagliptinin human plasma using liquid chromatography and tandem massspectrometry. Int. J. Pharm. Pharm. Sci., 2013, 5, 463-470.
El-Bagary, R.I.; Elkady, E.F.; Ayoub, B.M. Liquid chromatographic determination of sitagliptin either alone or in ternary mixture with metformin and sitagliptin degradation product. Talanta, 2011, 85(1), 673-680.
Nirogi, R.; Kandikere, V.; Mudigonda, K.; Komarneni, P.; Aleti, R.; Boggavarapu, R. Sensitive liquid chromatography tandem mass spectrometry method for thequantification of sitagliptin, a DPP-4 inhibitor, in human plasma using liquid-liquid extraction. Biomed. Chromatogr., 2008, 22(2), 214-222.
AbuRuz, S.; Millership, J.; McElnay, J. The development and validation of liquid chromatography method for the simultaneous determination of metformin and glipizide gliclazide, glibenclamide or glimperide in plasma. J. Chromatogr. , 2005, 817(2), 277-286.
Zeng, W.; Musson, D.G.; Fisher, A.L.; Chen, L.; Schwartz, M.S.; Woolf, E.J.; Wang, A.Q. Determination of sitagliptin in human urine and hemodialysate usingturbulent flow online extraction and tandem mass spectrometry. J. Pharm. Biomed. Anal., 2008, 46(3), 534-542.
Zeng, W.; Xu, Y.; Constanzer, M.; Woolf, E.J. Determination of sitagliptinin human plasma using protein precipitation and tandem mass spectrometry. J. Chromatogr. B , 2010, 878(21), 1817-1823.
Hess, C.; Musshoff, F.; Madea, B. Simultaneous identification and validated quantification of 11 oral hypoglycaemic drugs in plasma by electrospray ionisation liquid chromatography-mass spectrometry. Anal. Bioanal. Chem., 2010, 400(1), 33-41.
Burugula, L.; Mullangi, R.; Pilli, N.R.; Makula, A.; Lodagala, D.S.; Kandhagatla, R. Simultaneous determination of sitagliptin and simvastatin in human plasma by LC-MS/MS and its application to a human pharmacokinetic study. Biomed. Chromatogr., 2013, 27(1), 80-87.
Herman, G.A.; Mistry, G.C.; Yi, B.; Bergman, A.J.; Wang, A.Q.; Zeng, W.; Chen, L.; Snyder, K.; Ruckle, J.L.; Larson, P.J.; Davies, M.J.; Langdon, R.B.; Gottesdiener, K.M.; Wagner, J.A. Evaluation of pharmacokinetic parameters and dipeptidyl peptidase-4 inhibition following single doses of sitagliptin in healthy, young Japanese males. Br. J. Pharmacol., 2011, 71(3), 429-436.

Rights & PermissionsPrintExport Cite as

Article Details

Year: 2019
Page: [776 - 784]
Pages: 9
DOI: 10.2174/1573412914666181011141714
Price: $65

Article Metrics

PDF: 27
PRC: 1