A SIMPLE (HPLCâ€“UV) METHOD FOR THE QUANTIFICATION OF COLCHICINE IN BULK AND ETHOSOMAL GEL NANO-FORMULATION AND ITS VALIDATION
Objective: To develop and validate a stability-indicating reversed phase high-performance liquid chromatography (RP-HPLC) method for the determination of colchicine in bulk and ethosomal gel nano-formulation.
Methods: The chromatographic conditions were optimized using stainless steel Hypersil Gold C-18 analytical column with the dimensions of 250 mm x 4.6 mm ID x 5 Âµm. The mobile phase consisted of acetonitrile and ammonium acetate buffer (20 mmol/l, pH=4.85) in the ratio of 32:68 v/v. The ï¬‚ow rate was set at 1 ml/min and the detection wavelength was 353 nm. The column was maintained at 30 Â°C and the injection volume was 10 Âµl. The stability of colchicine in different conditions was investigated by exposing the drug to stress degradation using acid, base, oxidation, heat and light.
Results: There was no interference from excipients, impurities, dissolution media or degradation products at the retention time of colchicine 5.9 min indicating the specificity of the method. The limit of detection (LOD) and the limit of quantification (LOQ) were 8.64 ng/ml and 26.17 ng/ml respectively. The drug showed good stability under heat, acid, oxidation and light, but substantial degradation was observed under alkali condition. The procedure was validated for specificity, linearity, accuracy and precision.
Conclusion: A simple, rapid, specific and stability-indicating HPLCâ€“UV method for the determination of colchicine in the pure and ethosomal gel was successfully developed. The developed method was statistically confirmed to be accurate, precise, and reproducible.
2. Echteld IV, Wechalekar MD, Schlesinger N, Buchbinder R, Aletaha D. Colchicine for acute gout. Cochrane Database Syst Rev 2014;8:1-41.
3. Terkeltaub RA. Colchicine update: 2008. Semin Arthritis Rheum 2009;38:411-9.
4. Naseri A, Karami M, Nadoushan MJ. Failing of information transmission by dorsal hippocampus due to microinjection of colchicine in ratâ€™s cortical area 1. Asian J Pharm Clin Res 2015;4:160-3.
5. Kesselheim AS, Solomon DH. Incentives for the drug development the curious case of colchicine. N Engl J Med 2010;362:2045-7.
6. Abdulbaqi IM, Darwis Y, Khan NAK, Assi RA, Khan AA. Ethosomal nanocarriers: the impact of constituents and formulation techniques on ethosomal properties, in vivo studies, and clinical trials. Int J Nanomed 2016;11:2279-304.
7. Shaji J, Garude S. Transethosomes and ethosomes for enhanced transdermal delivery of ketorolac tromethamine: a comparative assessment. Int J Curr Pharm Res 2014;6:88-93.
8. Toppo FA, Pawar RS. Novel drug delivery strategies and approaches for wound healing managements. J Crit Rev 2015;2:12-20.
9. Fleischmann W, Price HG, Fleischmann SK. Fate of intraperitoneally administered colchicine in the Mongolian gerbil. Pharmacology 1967;17:323-6.
10. Arai T, Okuyama T. Fluorometric assay of the tubulin-colchicine complex. Anal Biochem 1975;69:443-50.
11. Boudene C, Duprey F, Bohuon C. Radioimmunoassay of colchicine. Biochem J 1975;151:413-5.
12. Sarg TM, El-Domiaty MM, Bishr MM, Salama OM, El-Gindy AR. Thin-layer chromatographic scanner, spectrophotometric and high-performance liquid chromatographic methods for the determination of colchicine. Analyst 1989;114:575-8.
13. Bodoki E, Oprean R, Vlase L, Tamas M, Sandulescu R. Fast determination of colchicine by TLC-densitometry from pharmaceuticals and vegetal extracts. J Pharm Biomed Anal 2005;37:971-7.
14. Samanidou VF, Sarantis GA, Papadoyannis IN. Development and validation of a rapid HPLC method for the direct determination of colchicine in pharmaceuticals and biological fluids. J Liq Chromatogr Relat Technol 2006;29:1-13.
15. Nirmala K, Raju RR. A novel method development for validation and detection of colchicine drug by RP-HPLC. Rasayan J Chem 2012;5:106-11.
16. Hadad GM, Badr JM, El-Nahriry K, Hassanean HA. Validated HPLC and HPTLC methods for simultaneous determination of colchicine and khellin in pharmaceutical formulations. J Chromatogr Sci 2013;51:258-65.
17. Chandrethiya GD, Shelat PK, Zaveri MN. RP-HPLC method development and validation for estimation of colchicine in gelatin nanoparticles formulation. Int J Pharm Pharm Sci 2013;5 Suppl 1:228-31
18. Klein AE, Davis PJ. Determination of colchicine and colchiceine in microbial cultures by high-performance liquid chromatography. Anal Chem 1980;52:2432-5.
19. Lhermitte M, Bernier JL, Mathieu D, Mathieu-Nolf M, Erb F, Roussel P. Colchicine quantitation by high-performance liquid chromatography in human plasma and urine. J Chromatogr B Biomed Sci Appl 1985;342:416-23.
20. Fernandez P, Bermejo AM, Tabernero MJ, LÃ³pez-Rivadulla M, Cruz A. Determination of colchicine in biological fluids by reverse-phase HPLC. Variation of colchicine levels in rats. Forensic Sci Int 1993;59:15-8.
21. Chen QH, Hou S, Gan LC, Li YB, Song X, Cai Z. Determination of colchicine in mouse plasma by a high-performance liquid chromatographic method with UV detection and its application to pharmacokinetic studies. Yakugaku Zasshi 2007;127:1485-90.
22. Tracqui A, Kintz P, Ludes B, RougÃ© C, Douibi H, Mangin P. High-performance liquid chromatography coupled to ion spray mass spectrometry for the determination of colchicine at ppb levels in human biofluids. J Chromatogr B Biomed Sci 1996;675:235-42.
23. Jones GR, Singer PP, Bannach B. Application of LC-MS analysis to a colchicine fatality. J Anal Toxicol 2002;26:365-9.
24. Abe E, Lemaire-Hurtel A, Duverneuil C, Etting I, Guillot E, Mazancourt P, et al. A novel LC-ESI-MS-MS method for sensitive quantification of colchicine in human plasma: application to two case reports. J Anal Toxicol 2006;30:210-5.
25. Hamscher G, Priess B, Nau H, Panariti E. Determination of colchicine residues in sheep serum and milk using high-performance liquid chromatography combined with electrospray ionization ion trap tandem mass spectrometry. Anal Chem 2005;77:2421-5.
26. ChÃ¨ze M, Deveaux M, PÃ©pin G. Liquid chromatography-tandem mass spectrometry for the determination of colchicine in postmortem body fluids. Case report of two fatalities and review of the literature. J Anal Toxicol 2006;30:593-8.
27. Jiang Y, Wang J, Wang Y, Li H, Fawcett JP, Gu J. Rapid and sensitive liquid chromatographyâ€“tandem mass spectrometry method for the quantitation of colchicine in human plasma. J Chromatogr B 2007;850:564-8.
28. Clevenger CV, August TF, Shaw LM. Colchicine poisoning: report of a fatal case with body fluid analysis by GC/MS and histopathologic examination of postmortem tissues. J Anal Toxicol 1991;15:151-4.
29. 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.
30. Almeida AM, Castel-Branco MM, FalcÃ£o AC. Linear regression for calibration lines revisited: weighting schemes for bioanalytical methods. J Chromatogr B 2002;774:215-22.
31. Tijare LK, Rangari NT, Mahajan UN. A review on bioanalytical method development and validation. Asian J Pharm Clin Res 2016;9 Suppl 3:6-10.