NOVEL RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF SAXAGLIPTIN AND GLIMEPIRIDE
Objective: The objective of the present study was to develop, and validate a novel reverse phase high performance liquid chromatographic (RP-HPLC) method, for simultaneous determination of saxagliptin (SAXA) and glimepiride (GLIM), in bulk mixtures, and in tablets.
Methods: Determination of the drugs, SAXA and GLIM, was carried out employing ODS C18 column (250 mm X 4.6 mm i. d, 5 Âµm particle size), with diode array detector at Î»max of 230 nm. The mobile phase employed for the current study, composed of two solvents, i.e., A (acetonitrile), and B (0.1 % w/v sodium di-hydrogen orthophosphate buffer, pH 3.8 adjusted with orthophosphoric acid). The mobile phase was pumped at a flow rate of 0.75 ml/min in the gradient mode. The validation study with respect to specificity, linearity, precision, accuracy, robustness, limit of detection (LOD), and limit of quantification (LOQ), was carried out employing the ICH Guidelines.
Results: The developed method was selective and linear for both the drugs, i.e., between 15.63 Âµg/ml and 250.00 Âµg/ml for SAXA, and 7.81 Âµg/ml and 125.00 Âµg/ml for GLIM, with a correlation coefficient (R2) 0.9977 and 0.9982, for SAXA, and GLIM, respectively. The % recovery obtained was 102.98Â±0.14% for SAXA, and 101.84Â±1.96% for GLIM. The LOD and LOQ values for SAXA were obtained to be 1.30 Âµg/ml, and 3.94 Âµg/ml, respectively, while for GLIM, LOD was 0.82 Âµg/ml and LOQ was 2.48 Âµg/ml. The method also exhibits good robustness for different chromatographic conditions like wavelength, flow rate, mobile phase and injection volume.
Conclusion: The method was successfully employed, for the quantification of SAXA and GLIM, in the quality control of in-house developed tablets, and can be applied for the industrial use.
2. AhrÃ©n B. Emerging dipeptidyl peptidase-4 inhibitors for the treatment of diabetes. Expert Opin Emerging Drugs 2008; 13:593-607.
3. McCall AL. Clinical review of glimepiride. Exp Opin Pharmacoth 2001;2:699-713.
4. Hadi MA, Babu BL, Pal N. Formulation and evaluation of sustained release matrix tablets of glimepiride based on combination of hydrophilic and hydrophobic polymers. J Appl Pharma Sci 2012;2:101-7.
5. Rosenstock J, Aguilar Salinas C, Klein E. Effect of saxagliptin monotherapy in treatment-naive patients with type 2 diabetes. Curr Med Res Opin 2009;25:2401-11.
6. Jadzinsky M, PfÃ¼tzner A, Paz Pacheco E. Saxagliptin given in combination with metformin as initial therapy improves glycemic control in patients with type 2 diabetes compared with either monotherapy: a randomized controlled trial. Diabetes Obes Metab 2009;11:611-22.
7. Rosenstock J, Sankoh S, List JF. Glucose-lowering activity of the dipeptidyl peptidase-4 inhibitor saxagliptin in drug-naive patients with type 2 diabetes. Diabetes Obes Metab 2008;10:376-86.
8. DeFronzo RA, Hissa M, Garber AJ, Luiz Gross J, Yuyan Duan R, Ravichandran S, et al. The efficacy and safety of saxagliptin when added to metformin therapy in patients with inadequately controlled type 2 diabetes on metformin alone. Diabetes Care 2009;32:1649-55.
9. Chacra AR, Tan GH, Apanovitch A, Ravichandran S, List J, Chen R. Saxagliptin added to a submaximal dose of sulphonylurea improves glycaemic control compared with uptitration of sulphonylurea in patients with type 2 diabetes: a randomised controlled trial. Int J Clin Pract 2009;63:1395-406.
10. Hollander P, Li J, Allen E, Chen R. Saxagliptin added to a thiazolidinedione improves glycemic control in patients with type 2 diabetes and inadequate control on thiazolidinedione alone. J Clin Endocrinol Metab 2009;94:4810-9.
11. Kulasa K, Edelman S. Saxagliptin: the evidence for its place in the treatment of type 2 diabetes mellitus. Core Evid 2010;5:23-37.
12. Dave DJ. Saxagliptin: a dipeptidyl peptidase-4 inhibitor in the treatment of type 2 diabetes mellitus. J Pharmacol Pharmacother 2011;2:230-5.
13. Chacra AR. Saxagliptin for type 2 diabetes. Diabetes Metab Syndr Obes: Targets Ther 2010;3:325-35.
14. http://www.fda.gov/downloads/ScienceResearch/FieldScience/UCM092147.pdf. [Last accessed on 10 Jan 2018]
15. Kalaichelvi R, Jayachandran E. Validated spectroscopic method for estimation of saxagliptin in pure and from tablet formulation. Int J Pharm Pharm Sci 2011;3:179-80.
16. Moneeb MS. Spectrophotometric and spectrofluorimetric methods for the determination of saxagliptin and vildagliptin in bulk and pharmaceutical preparations. Bull Faculty Pharm Cairo University 2013;51:139-50.
17. Jeyabalan G, Nyola N. Analytical method development and validation of saxagliptine a novel dipeptidyl peptidase IV inhibitors in pure and tablet dosage form by UV-VIS spectroscopy. Int J Instit Pharma Life Sci 2012;2:144-50.
18. Nyola N, Jeyabalan GS. Development and validation of uv-vis specctroscopy method for simultaneous estimation of saxagliptin hydrochloride and metformin hydrochloride in active pharmaceutical ingrident. J Pharm Educ Res 2012;3:19-23.
19. Bhargavi S, Suryasagar G, Sowmya DK, Ashok K, Nama S. UV spectrophotometric method for determination of glimepiride in pharmaceutical dosage forms. Int J Pharm Sci Rev Res 2013;21:131-3.
20. Kishore L, Kaur N. Estimation of pioglitazone and glimipride in its pharmaceutical dosage form by spectrophotometric methods. Pharm Lett 2011;3:276-84.
21. Pravin CR, Vasudevan M, Deecaraman. A validated RP-HPLC method for simultaneous estimation of metformin and saxagliptin in tablets. Ras J Chem 2012;5:137-41.
22. Mohammad MA, ElKady EF, Fouad MA. Development and validation of a reversed-phase column liquid chromatographic method for simultaneous determination of two novel gliptins in their binary mixtures with metformin. Eur J Chem 2012;3:152-5.
23. Prakash PP, Kalkotwar RS, Patil VV, Jadhav VB, Patil NP. A new RP-HPLC method for determination of metformin HCl and saxagliptin in tablet dosage form. Int J Pharma Biol Sci 2012;2:161-7.
24. Sarat M, Murali KP, Rambabu C. RP-HPLC method for simultaneous estimation of saxagliptin and pioglitazone in tablets. Int Reas J Pharm 2012;3:399-402.
25. Dhaneshwar SR, Salunkhe JV, Bhusari VK. Validated HPTLC method for simultaneous estimation of metformin hydrochloride, atorvastatin and glimepiride in bulk drug and formulation. J Anal Bioanal Tech 2010;1:1-5.
26. Devi R, Mohammad H. Stability indicating RP-HPLC method for the simultaneous determination of atorvastatin calcium, metformin hydrochloride, and glimepiride in bulk and combined tablet dosage form. Int Scho Res Notices 2014:1-8.
27. Raja P, Thejaswini JC, Gurupadayya BM, Sowjanya K. Determination and validation of metformin, glimepiride, pioglitazone using atorvastatin as an internal standardin bulk drug and pharmaceutical dosage form. J Appl Chem Res 2011;18:61-8.
28. Samala S, Tatipamula SR, Veeresham C. Determination of glimepiride in rat serum by RP-HPLC method. Am J Anal Chem 2011;2:152-7.
29. Lakshmi KS, Rajesh T, Sharma S, Lakshmi S. Development and validation of liquid chromatographic and UV derivative spectrophotometric methods for the determination of metformin, pioglitazone and glimepiride in pharmaceutical formulations. Pharm Chem 2009;1:238-46.
30. Pawar S, Meshram G, Jadhav R, Bansal Y. Simultaneous determination of glimepiride and metformin hydrochloride impurities in sustained release pharmaceutical drug product by HPLC. Scholars Res Library: Pharm Chem 2010;2:157-68.
31. Sakuntala MSV, Prasad SVUM, Devi SS, Yadav SK, Reddy KS. A RP-HPLC method development and validation for the simultaneous estimation of glimepiride and pioglitazone HCl in tablet dosage forms. J Chem Pharm Res 2012;4:154-9.
32. Xu SX, Demers R, Gu H, Christopher LJ, Su H, Cojocaru L, et al. Liquid chromatography and tandem mass spectrometry method for the quantitative determination of saxagliptin and its major pharmacologically active 5-monohydroxy metabolite in human plasma: method validation and overcoming specific and non-specific binding at low concentrations. J Chromatogr B 2012;889:77-86.
33. Polagania SR, Pillib NR, Gajulab R, Ganduc V. Simultaneous determination of atorvastatin, metformin and glimepiride in human plasma by LCâ€“MS/MS and its application to a human pharmacokinetic study. J Pharm Anal 2013;3:9-19.
34. Patel H, Gohel M. A review on development of multifunctional co-processed excipient. J Crit Rev 2016;3:48-54.
35. Kumar D, Keshavshetti GG, Mogale P, Bhalke N. Design and evaluation of fast dissolving tablets of ergotamine tartarate. Int J Curr Pharm Res 2015;7:101-4.
36. Medina JR, Aguilar E, Hurtado M. Dissolution behavior of carbamazepine suspensions using the usp dissolution apparatus 2 and the flow-through cell method with simulated gi fluids. Int J Pharm Pharm Sci 2017;l9:111-6.
37. Hiremath SP, Makanapur C. Formlation and evaluation of orodispersible tablets of a model antihypertensive drug. Int J Pharm Pharm Sci 2017;9:34-8.
38. CPMP/ICH/281/95, Note for guidance on validation of analytical procedures: methodology, ICH Topic Q2B validation of analytical procedures: methodology, step 4 (CPMP Adopted December 96); 1996.
39. Sharma M, Mahindroo N. Development and validation of stability indication RP-HPLC method for determination of Î²-acetyldigoxin. Int J Appl Pharm 2017;9:54-9.
40. Tijare LK, Rangari NT, Mahajan UN. A review on bioanalytical method development and validation. Asian J Pharm Clin Res 2016;9:6-10.