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Current Pharmaceutical Analysis


ISSN (Print): 1573-4129
ISSN (Online): 1875-676X

Research Article

Quantitation of Pregabalin by HPLC-UV Method using Ninhydrin Derivatization: Development and Validation

Author(s): Fathiy Mutalabisin, Abul Bashar Mohammed Helaluddin, Pinaki Sengupta, Farahidah Mohamed and Bappaditya Chatterjee*

Volume 17, Issue 1, 2021

Published on: 14 November, 2019

Page: [165 - 171] Pages: 7

DOI: 10.2174/1573412916666191114120213

Price: $65


Introduction: A simple and reliable high performance liquid chromatographic method has been developed for the quantitative determination of pregabalin in bulk and dosage form. Pregabalin, a γ amino butyric acid analogue, has negligible sensitivity to UV or fluorescence detection. Hence, it has been derivatized by ninhydrin to form a chromophoric complex that could be quantified by UV detection.

Materials and Methods: The concentration of ninhydrin was set to 5 mg/ml and a phosphate buffer solution (pH 7.4) was used as a solvent for the reaction. The resultant complex was separated by HPLC and detected by a UV detector at 569nm wavelength.

Results: The developed method showed a linear response within 50 to 600 μg/mL of pregabalin. The method was accurate with mean recovery values within 100 ± 2%. The repeatability of the method was established by intra-day and inter-day precision study. Finally, a commercial pregabalin capsule was assayed by the developed HPLC method including ninhydrin derivatization. The result of the mean assay was found to be 100.37 ±2.94 %.

Conclusion: This is the first time we are reporting pregabalin analysis using ninhydrin derivatization for HPLC analysis. Therefore, the developed method can be considered as a significant improvement in pregabalin quantitation and it can be easily applied for routine quality control tests of pregabalin.

Keywords: Pregabalin, ninhydrin, precolumn derivatization, HPLC, UV detection, analytical method development, analytical method validation.

Graphical Abstract
Bali, A.; Gaur, P. A novel method for spectrophotometric determination of pregabalin in pure form and in capsules. Chem. Cent. J., 2011, 5(59), 59.
[] [PMID: 21982305]
Yoshikawa, N.; Naito, T.; Yagi, T.; Kawakami, J. A validated fluorometric method for the rapid determination of pregabalin in human plasma applied to patients with pain. Ther. Drug Monit., 2016, 38(5), 628-633.
[] [PMID: 27465975]
National Centre for Biotechnology Information. Compound Summary for CID 5486971
Kostić, N.; Dotsikas, Y.; Jović, N.; Stevanović, G.; Malenović, A.; Medenica, M. Quantitation of pregabalin in dried blood spots and dried plasma spots by validated LC-MS/MS methods. J. Pharm. Biomed. Anal., 2015, 109, 79-84.
[] [PMID: 25767905]
Mandal, U.; Sarkar, A.K.; Gowda, K.V.; Agarwal, S.; Bose, A.; Bhaumik, U.; Ghosh, D.; Pal, T.K. determination of pregabalin in human plasma using LC-MS-MS. Chromatographia, 2008, 67(3–4), 237-243.
Vermeij, T.A.C.; Edelbroek, P.M. Simultaneous high-performance liquid chromatographic analysis of pregabalin, gabapentin and vigabatrin in human serum by precolumn derivatization with o-phtaldialdehyde and fluorescence detection. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci., 2004, 810(2), 297-303.
[] [PMID: 15380728]
Jadhav, A. S.; Pathare, D. B.; Shingare, M. S. Validated Enantioselective LC Method , with Precolumn Derivatization with Marfey’s Reagent , for Analysis of the Antiepileptic Drug Pregabalin in Bulk Drug Samples. 2007, (3), 253-256.
Patil, D.D.; Patil, M.S.; Wani, Y.B. spectrophotometric method for pregabalin determination: an experimental design approach for method development. Journal of the Association of Arab Universities for Basic and Applied Sciences, 2016, 21, 31-37.
Derayea, S.M.; Attia, T.Z.; Elnady, M. Development of spectrofluorimetric method for determination of certain antiepileptic drugs through condensation with ninhydrin and phenyl acetaldehyde. Spectrochim. Acta A Mol. Biomol. Spectrosc., 2018, 204, 48-54.
[] [PMID: 29906646]
Guideline, I.C.H.H.T. Validation of Analytical Procedures: Text and Methodology Q2 (R1). International Conference on Harmonization, Geneva, Switzerland2005, pp. 11-12.
Adib, N.A.M.; Mandal, U.K.; Mohamed, F.; Chatterjee, B. fast and simple gas chromatographic method for simultaneous estimation of camphor, menthol and methyl salicylate in analgesic ointment: application in stability study. J. Pharm. Investig., 2017, 47(3), 275-285.
Shabir, G.A. Step-by-Step Analytical Methods Validation and Protocol in the Quality System; Institute of Validation Technology, 2005.
Siddiqui, F.A.; Sher, N.; Shafi, N.; Shamshad, H.; Zubair, A. Kinetic and Thermodynamic Spectrophotometric Technique to Estimate Gabapentin in Pharmaceutical Formulations Using Ninhydrin. J. Anal. Sci. Technol., 2013, 4(1), 17.
Friedman, M. Applications of the ninhydrin reaction for analysis of amino acids, peptides, and proteins to agricultural and biomedical sciences. J. Agric. Food Chem., 2004, 52(3), 385-406.
[] [PMID: 14759124]
Siddiqui, F.A.; Arayne, M.S.; Sultana, N.; Qureshi, F.; Mirza, A.Z.; Zuberi, M.H.; Bahadur, S.S.; Afridi, N.S.; Shamshad, H.; Rehman, N. Spectrophotometric determination of gabapentin in pharmaceutical formulations using ninhydrin and π-acceptors. Eur. J. Med. Chem., 2010, 45(7), 2761-2767.
[] [PMID: 20381213]
ICH Harmonised Tripartite Guideline.. 2005.
Geetha, G.; Raju, K.N.G.; Kumar, B.V.; Raja, M.G. analytical method validation : an updated review. international journal of advances in pharmacy. Biol. Chem., 2012, 1(1), 64-71.

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