DEVELOPMENT AND VALIDATION OF STABILITY INDICATING HPTLC METHOD FOR SIMULTANEOUS DETERMINATION OF LINAGLIPTIN AND METFORMIN
Keywords:Linagliptin, Metformin, HPTLC, Stability
Objective: A simple, precise, and accurate stability indicating high-performance thin-layer chromatography (HPTLC) method was developed and validated for simultaneous estimation of linagliptin and metformin active pharmaceutical ingredients and fixed dose combination.
Methods: Linagliptin and metformin densitograms were developed on silica gel 60 F254 HPTLC plates with acetone: methanol: chloroform: formic acid (3:1:5:1v/v) as the mobile phase. Densitometric quantification was performed at 230 nm.
Results: For linagliptin and metformin RF values were found as 0.72 and 0.19, respectively. The method was validated for precision, accuracy, specificity, and robustness. The linearity curves were obtained in the concentration range of 100â€“600 ng per spot by area with correlation coefficients of 0.999 and 0.99 for linagliptin and metformin, respectively. Limit of detection was found to be 5.19 and 8.72 ng per spot for linagliptin and metformin, respectively; lowest possible quantity to be quantified by the proposed method was found to be 15.74 and 26.44 ng per spot for linagliptin and metformin, respectively. From stability studies, the noninterference of the linagliptin and metformin degradants with drugs demonstrated the suitability of the developed method.
Conclusion: The developed method was validated and found to be selective, specific and suitable for application in pharmaceutical analysis of these drugs in bulk and fixed dose combination.
Vaishali D, Srikanta S, Shalini M, Shiva P, Russel J, Shweta U, and Kamran S. Safety and efficacy of the initial combination of linagliptin and metformin in patients with type 2 diabetes: A subgroup analysis of Indian patients from a randomized, double-blind, placebo-controlled study. Indian J Endocrinol Metab 2015;19:256â€“61.
Haak T. Combination of linagliptin and metformin for the treatment of patients with type 2 diabetes. Clin Med Insights Endocrinol Diabetes 2015;8:1-6.
Zhong GP, Bi HC, Zhou S, Chen X, Huang M. Simultaneous determination of Metformin and gliclazide in human plasma by liquid chromatography tandem mass spectrometry: application to a bioequivalence study of two formulations in healthy volunteers. J Mass Spectrom 2005;40:1462â€“71.
Marlice ASM, Alcenir SS, Olivia WP, Pedro TWB, Douglas PP, Milton FF. Simple and rapid method determination for Metformin in human plasma using high-performance liquid chromatography-tandem mass spectrometry: Application to pharmacokinetic studies. J Chromatogr B 2007;852:308â€“16.
Heinig K, Bucheli F. Fast liquid chromatographic-tandem mass spectrometric (LCâ€“MSâ€“MS) determination of Metformin in plasma samples. J Pharm Biomed Anal 2004;34:1005â€“11.
Porta V, Schramm SG, Kano EK, Koono EE, Armando YP, Fukuda K. HPLC-UV determination of Metformin in human plasma for application in pharmacokinetics and bioequivalence studies. J Pharm Biomed Anal 2008;46:143â€“7.
Raniah Q, Gabr RS, Padwal D, Brocks R. Determination of Metformin in Human Plasma and Urine by High-Performance Liquid Chromatography Using Small Sample Volume and Conventional Octadecyl Silane Column. J Pharm Pharm Sci 2010;13:486â€“94.
Zarghi A, Foroutan SM, Shafaati A, Khoddam A. Rapid determination of Metformin in human plasma using ion-pair HPLC. J Pharm Biomed Anal 2003;31:197â€“200.
AbuRuz S, Millership J, McElnay J. Determination of metformin in plasma using a new ion pair solid phase extraction technique and ion-pair liquid chromatography. J Chromatogr B: Anal Technol Biomed Life Sci 2003;798:203â€“9.
El-Bagary RI, Elkady EF, Ayoub BM. Spectrophotometric methods for the determination of linagliptin in binary mixture with metformin hydrochloride and simultaneous determination of linagliptin and metformin hydrochloride using high-performance liquid chromatography. Int J Biomed Sci 2013;9:41â€“7.
Kavitha KY, Geetha G, Hariprasad R, Kaviarasu M, Venkatnarayanan R. Development and validation of stability indicating RP-HPLC method for the simultaneous estimation of Linagliptin and metformin in pure and pharmaceutical dosage form. J Chem Pharm Res 2013;5:230â€“5.
International Conference on Harmonization, Validation of analytical procedure: Methodology (Q2B), ICH Harmonized Tripartite Guidelines, Geneva; 1996.
International Conference on Harmonization, Validation of Analytical Procedures: Methodology, Federal Register, Geneva; 1996.
Rajasekaran R Kavitha, R Arivukkarasu. Development and validation of HPTLC method for simultaneous estimation and stability indicating study of metformin HCl and linagliptin in a pharmaceutical formulation. World J Pharm Sci 2014;2:317-27.