DEVELOPMENT AND VALIDATION OF NOVEL RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF GABAPENTIN AND AMITRIPTYLINE HYDROCHLORIDE IN BULK AND PHARMACEUTICAL DOSAGE FORMS
Objective: To develop a novel, accurate, precise and linear reverse phase high performance liquid chromatographic (RP-HPLC) method for simultaneous quantitative estimation of gabapentin and amitriptyline hydrochloride in gabantip-at tablet and validate as per international conference on harmonization (ICH) guidelines and to perform the force degradation studies using the developed method.
Methods: In the present work, good chromatographic separation was achieved isocratically using a shim-pack HPLC C18 column (4.6 x 250 mm, 5Î¼m) and mobile phase consisting of 0.05 M potassium dihydrogen orthophosphate pH 2.1 adjusted with orthophosphoric acid and acetonitrile in the ratio (55:45), at flow rate 1 ml/min and column temperature (25 Â°C). The effluents obtained were monitored at 221 nm with the UV-visible detector.
Results: The retention time of gabapentin and amitriptyline hydrochloride was found to be 1.959 min and 4.221 min respectively. The linearity of gabapentin was found in the range of 720-1680 ppm and that for amitriptyline hydrochloride was found to be 24-56 ppm. The correlation coefficient for gabapentin and amitriptyline hydrochloride were 0.999 and 0.9963 respectively. The high recovery values (98%-101%) indicate a satisfactory accuracy. The low percent relative standard deviation (% RSD) values in the precision study reveals that the method is precise.
Conclusion: The developed method is novel, simple, precise, rapid, accurate and reproducible for simultaneous estimation of gabapentin and amitriptyline hydrochloride tablet dosage form. Hence the proposed method may find practical applications as a quality-control tool in the simultaneous analysis of the two drugs in combined dosage forms in quality-control laboratories.
2. Rabie S, Ashraf MA. RP-HPLC determination of amitriptyline hydrochloride in tablet formulations and urine. Asian J Res Chem 2011;4:24-7.
3. Hyttel J, Christensen AV, Fjalland B. Neuropharmacological properties of amitriptyline, nortriptyline and their metabolites. Acta Pharmacol Toxicol (Copenh) 1980;47:53-7.
4. John TR, Leahy MR, Ian CM, Westhead TT. A comparison of nortriptyline and amitriptyline in depression. Br J Psychiatry 1965;111:1101-3.
5. Corrado B, Matthew H. Amitriptyline v. the rest: still the leading antidepressant after 40 y of randomised controlled trials. Br J Psychiatry 2001;178:129-44.
6. Das B. Estimation of gabapentin in human plasma using LC-MS/MS method. Asian J Pharm Clin Res 2013;6 Suppl 3:213-6.
7. Rao B, Maqdoom F, Nikalje AP. Determination of gabapentin in bulk drug and in pharmaceutical dosage form by HPLC method. J Chil Chem Soc 2009;54:424-7.
8. Faraat A, Singha GN, Sahua P, Nagarab R, Nagara M, Tyagi A. Application of an LC/HPLC method development and validation for the simultaneous estimation of amitriptyline hydrochloride and chlordiazepoxide in tablet dosage form by using a reverse phase technique. Pharm Lett 2015;7:172-7.
9. Srikantha D, Raju R. Method development and validation of chlordiazepoxide and amitriptyline hydrochloride in pharmaceutical formulations by RP-HPLC. Asian J Biomed Pharm Sci 2014;4:8-14.
10. Lakshmi B. RP-HPLC method development for the quantification of gabapentin in formulations. Exp Int J Sci Technol 2012;2:84-92.
11. Karchaliya CV, Patel PB. Development and validation of analytical methods for simultaneous estimation of amitriptyline hydrochloride and methylcobalamin in their tablet dosage form by UV spectrophotometric method. PharmaTutor Mag 2015;3:46-50.
12. Galande VR, Baheti KG, Dehghan MH. UV-visible spectrophotometric method for estimation of gabapentin and methylcobalamin in bulk and tablet. Int J ChemTech Res 2010;2:695-9.
13. Rabie S, Darwish M, Mohamed F, Hamdi A. New HPLC method to detect amitriptyline in the blood of rats on combination treatment. Int J Chem Anal Sci 2013;4:120-4.
14. Patel S, Patel NJ. Spectrophotometric and chromatographic simultaneous estimation of amitriptyline hydrochloride and chlordiazepoxide in tablet dosage forms. Indian J Pharm Sci 2009;71:472-6.
15. Saleh T. Assay of four psychotropic drugs chlorpromazine, clomipramine, amitriptyline and nortriptyline in tablets by a single HPLC method. Int J Pharm Pharm Sci 2016;8:182-8.
16. Sevak R, Sevak AD, Manan R, Vyas PJ. Analytical method development and validation for amitriptyline HCl (psychoactive drug) using HPLC instrument. IOSR J Appl Chem 2017;10:77-80.
17. Patel Y, Patel MB, Patel NK, Sakhreliya B. Development and validation of analytical method for simultaneous estimation of gabapentin and nortriptyline hydrochloride in pharmaceutical dosage form. J Pharm Sci Bio-Sci Res 2015;5:434-43.
18. Shah A, Kothari C, Patel N. Concurrent estimation of gabapentin and nortriptyline hydrochloride in their combined dosage form using OPA-Î²-mercaptoethanol derivatization by spectrophotometric and spectrofluorimetric methods. Curr Pharm Anal 2017;13:241-9.
19. Rawat T, Pandey IP. Forced degradation studies for drug substances and drug products-scientific and regulatory considerations. J Pharm Sci Res 2015;7:238-41.
20. ICH, Stability testing of new drug substances and products (Q1AR2), International Conference on Harmonization, IFPMA, Geneva; 2003.
21. Vemula VR, Sharma PK. RP-HPLC method development and validation for simultaneous estimation of diclofenac and tolperisone in tablet dosage form. Asian J Pharm Clin Res 2013;6 Suppl 3:186-9.
22. International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human use. Validation of Analytical Procedures: Text and Methodology ICH Q2(R1); 2005.
23. Sarat M, Murali PK, Rambabu C. Development and validation of RPHPLC method for simultaneous estimation of amlodipine besylate and benazepril Hcl in tablet dosage form. Int J Curr Pharm Res 2012;4:80-4.
24. FDA, Guidance R. Validation of chromatographic methods. centre for drug evaluation and research (CDER), Food and Drug Administration; 1994. p. 2.
25. FDA, ORA validation and verification guidance for human drug analytical methods. Food and Drug Administration; 2003. p. 1.