Research Journal of Pharmacy and Technology

Open Access Open Access  Restricted Access Subscription Access
Open Access Open Access Open Access  Restricted Access Restricted Access Subscription Access

Development and Validation of an Ultra Performance Liquid Chromatography-tandem Mass Spectrometry Method with Liquid-liquid Extraction for the Quantification of Sitagliptin in Human Plasma Using Nebivolol as Internal Standard


Affiliations
1 Department of Analytical Chemistry Politeknik AKA Bogor, Bogor 16158, Indonesia
2 PT. Equilab International, Jakarta 12430,, Indonesia
     

   Subscribe/Renew Journal


We aimed to investigate the performance of the development method for sitagliptin quantification analysis in human plasma by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS) using liquid-liquid extraction (LLE) and Nebivolol as internal standard (IS). The parameters of this presented validation method are selectivity, the lower limit of quantification (LLOQ), linearity, accuracy, precision with five different concentration (LLOQ, Low QC, Medium QC, High QC, Upper Limit Of Quantification (ULOQ)), Integrity of dilution, matrix effect, and test for stability. Based on this study, the multiple reaction monitoring (MRM) transitions were m/z 408,23 → 127,02 for Sitagliptin and m/z 406,25 → 151,06 for IS Nebivolol. The selectivity test obtained % interference of sitagliptin and IS nebivolol by (0.00 - 0.38) % and (0.04 - 0.24) %, respectively. LLOQ test obtained a concentration value of 10.32 ng/mL and % RSD (n = 5) of 5.27 %. When the Sitagliptin concentration ranged from 1 to 1000 ng/mL, the method showed strong linearity with a coefficient of correlation of 0.9991. Accuracy test obtained % differentiation of (-12.58 - 7.77) % and precision test obtained % RSD of (1.62 - 5.32) %. The dilution integrity test obtained % differentiation at 4 and 2 times dilution was -9.20 % and -9.16 %. Matrix effect and stability data were in line with the stipulated European Medicines Agency (EMA) guidelines for validating the bioanalytical method.

Keywords

Sitagliptin, Human Plasma, Nebivolol, Liquid-liquid Extraction, UPLC-MS, Validation.
Subscription Login to verify subscription
User
Notifications
Font Size


  • Soujanya Kumari. Ravi Kumar. Yamunappa. Pooja Shetty. Prathibha Suvarna. Narayana Swamy VB. Formulation and Evaluation of Gastro Retentive Matrix Tablets of Sitagliptin. Asian J. Res. Pharm. Sci. 2016; 6(1):1-14. doi: 10.5958/2231-5659.2016.00001.1

  • Aschner P. Kipnes MS. Lunceford JK. Sanchez M. Mickel C. Williams-Herman DE. Effect of the dipeptidyl peptidase-4 inhibitor sitagliptin as monotherapy on glycemic control in patients with type 2 diabetes. Diabetes Care. 2006; 29:2632–7. doi: 10.2337/dc06-0703

  • Arnolds S. Dellweg S. Clair J. Dain MP. Nauck MA. Rave K. et al. Further improvement in postprandial glucose control with addition of exenatide or Sitagliptin to combination therapy with insulin glargine and metformin: a proof-of-concept study. Diabetes Care. 2010; 33:1509–15. doi: 10.2337/dc09-2191

  • Iwamoto Y. Taniguchi T. Nonaka K. Okamoto T. Okuyama K. Arjona Ferreira JC, et al. Dose-ranging efficacy of Sitagliptin, a dipeptidyl peptidase-4 inhibitor, in Japanese patients with type 2 diabetes mellitus. Endocr J. 2010; 57:383–94. doi: 10.1507/endocrj.k09e-272

  • Takahara M. Shiraiwa T. Kaneto H. Katakami N. Matsuoka TA. Shimomura I. Efficacy of Sitagliptin on blood glucose fluctuation in Japanese type 2 diabetic patients with basal-supported oral therapy. Endocr J. 2012; 59:1131–6. doi: 10.1507/endocrj.ej12-0220

  • Sachin Patil. B Ramesh. AR Hareesh. Kiran Patil. Validated UV Spectrophotometric Method for Estimation of Sitagliptin Phosphate in Tablet Dosage Form. Research J. Pharm. and Tech. 2010; 3(3):798-800. Available on: https://rjptonline.org/AbstractView.aspx?PID=2010-3-3-28

  • Lobhe Gayatri A. Shah Amol. Singhvi Indrajeet. Development and Validation of A Stability-Indicating RP-HPLC Method for the Determination of Sitagliptin Phosphate and Simvastatin in the Presence of their Degradation Products in Bulk and Binary Mixture. Asian J. Res. Pharm. Sci. 2016; 6(3):191-197. doi: 10.5958/2231-5659.2016.00026.6

  • Sathish Kumar Konidala. Pampana V V Suresh Babu. Ranj. K S D Ranjitha. RP-HPLC Method Development and Validation for the Simultaneous Estimation of Sitagliptin and Simvastatin in Pharmaceutical Formulation. Asian J. Pharm. Ana. 2016; 6(2):68-76. doi: 10.5958/2231-5675.2016.00011.9

  • W Zeng. DG Musson. AL Fisher. AQ Wang. Determination of MK-0431 in human plasma using high turbulence liquid chromatography online extraction and tandem mass spectrometry. Rapid Commun. Mass Spectrom. 2006; 20.1169. doi: 10.1002/rcm.2426.

  • W Zeng. DG Musson. AL Fisher. L Chen. MS Schwartz. EJ Woolf. AQ Wang. Determination of Sitagliptin in human urine and hemodialysate using turbulent flow online extraction and tandem mass spectrometry. J. Pharm. Biomed. Anal. 2008; 48.534. doi: 10.1016/j.jpba.2007.11.003.

  • Oliver SC. M Kinzig. MS Stoffel. U Fuhr. F Sorgel. A HILIC-MS/MS assay for the quantification of metformin and Sitagliptin in human plasma and urine: A tool for studying drug transporter perturbation. Journal of Pharmacetical and Biomedical Analysis. 2019; 175.112754. doi: 10.1016/j.jpba.2019.07.002

  • Deepak Prashar. Process Validation: An Overview. Research J. Pharma. Dosage Forms and Tech. 2011; 3(6):247-250. Available on: https://rjpdft.com/AbstractView.aspx?PID=2011-3-6-2

  • R Nirogi. V Kandikere. K Mudigonda. P Komarneni. R Aleti and R Boggavarapu. Sensitive liquid chromatography tandem mass spectrometry method for the quantification of Sitagliptin, a DPP-4 inhibitor, in human plasma using liquid–liquid extraction. Biomed. Chromatogr. 2018; 22: 214–222. doi: 10.1002/bmc.926.

  • S Madhavi. A Prameela Rani. Development and Validation of Bioanalytical Method for the Determination of Cobicistat from Human Plasma. Asian J. Pharm. Ana. 2016; 6(4): 216-226. http://dx.doi.org/10.5958/2231-5675.2016.00033.8

  • Shashikala P. Sireesha D. Vasudha B. Development and Validation of Bioanalytical Method for the Estimation of Carisoprodol in Human Plasma using LC-MS/MS. Asian J. Pharm. Ana. 2015; 5(4):181-186. doi: 10.5958/2231-5675.2015.00029.0

  • Sachin R Shinde. Suvarna I Bhoir. Namdev S Pawar. Suman B Yadav. Ajay S Ghumatkar. Ashok M Bhagwat. Simultaneous Determination of Valsartan and Nebivolol HCl in Tablet Dosage Form by RP-HPLC. Asian J. Research Chem. 2009; 2(4):519-522. Available on: https://ajrconline.org/AbstractView.aspx?PID=2009-2-4-40

  • Europe Medicine Agency: Guideline on bioanalytical method validation. 2011. Available at: https://www.ema.europa.eu/en/bioanalytical-method-validation, accessed May 2019.

  • Oskar González. Rosa María Alonso. Validation of bioanalytical chromatographic methods for the quantification of drugs in biological fluids. Methods of Therapeutic Drug Monitoring including Pharmacogenetics. 2020; doi: 10.1016/B978-0-444-64066-6.00006-X

  • B Booth. ME Arnold. B DeSilva. L Amaravadi. S Dudal. E Fluhler. B Gorovits. SH Haidar. J Kadavil. S Lowes. R Nicholson. M Rock. M Skelly. L Stevenson. S Subramaniam. R Weiner. E Woolf. Workshop Report: Crystal City V-Quantitative Bioanalytical Method Validation and Implementation: The 2013 Revised FDA Guidance. AAPS J. 2015; 17(2):277-88. doi: 10.1208/s12248-014-9696-2.

  • Amruta S. Kadam, Nayana V. Pimpodkar, Puja S.Gaikwad, Sushila D. Chavan. Bioanalytical Method Validation. Asian J. Pharm. Ana. 2015; 5(4):219-225. Available on: https://ajpaonline.com/AbstractView.aspx?PID=2015-5-4-9

  • RB Saudagar. PG Thete. Bioanalytical Method Validation: A Concise Review. Asian J. Res. Pharm. Sci. 2018; 8(2):107-114. doi: 10.5958/2231-5659.2018.00019.X


Abstract Views: 143

PDF Views: 0




  • Development and Validation of an Ultra Performance Liquid Chromatography-tandem Mass Spectrometry Method with Liquid-liquid Extraction for the Quantification of Sitagliptin in Human Plasma Using Nebivolol as Internal Standard

Abstract Views: 143  |  PDF Views: 0

Authors

Henny Rochaeny
Department of Analytical Chemistry Politeknik AKA Bogor, Bogor 16158, Indonesia
Candra Irawan
Department of Analytical Chemistry Politeknik AKA Bogor, Bogor 16158, Indonesia
Lintannisa Rahmatia
Department of Analytical Chemistry Politeknik AKA Bogor, Bogor 16158, Indonesia
Nilna Izzatul Mawaddah
Department of Analytical Chemistry Politeknik AKA Bogor, Bogor 16158, Indonesia
Ismail Dwi Saputr
PT. Equilab International, Jakarta 12430,, Indonesia

Abstract


We aimed to investigate the performance of the development method for sitagliptin quantification analysis in human plasma by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS) using liquid-liquid extraction (LLE) and Nebivolol as internal standard (IS). The parameters of this presented validation method are selectivity, the lower limit of quantification (LLOQ), linearity, accuracy, precision with five different concentration (LLOQ, Low QC, Medium QC, High QC, Upper Limit Of Quantification (ULOQ)), Integrity of dilution, matrix effect, and test for stability. Based on this study, the multiple reaction monitoring (MRM) transitions were m/z 408,23 → 127,02 for Sitagliptin and m/z 406,25 → 151,06 for IS Nebivolol. The selectivity test obtained % interference of sitagliptin and IS nebivolol by (0.00 - 0.38) % and (0.04 - 0.24) %, respectively. LLOQ test obtained a concentration value of 10.32 ng/mL and % RSD (n = 5) of 5.27 %. When the Sitagliptin concentration ranged from 1 to 1000 ng/mL, the method showed strong linearity with a coefficient of correlation of 0.9991. Accuracy test obtained % differentiation of (-12.58 - 7.77) % and precision test obtained % RSD of (1.62 - 5.32) %. The dilution integrity test obtained % differentiation at 4 and 2 times dilution was -9.20 % and -9.16 %. Matrix effect and stability data were in line with the stipulated European Medicines Agency (EMA) guidelines for validating the bioanalytical method.

Keywords


Sitagliptin, Human Plasma, Nebivolol, Liquid-liquid Extraction, UPLC-MS, Validation.

References