DEVELOPMENT AND VALIDATION OF HPTLC METHOD FOR ESTIMATION OF LEFLUNOMIDE IN ITS PHARMACEUTICAL DOSAGE FORM
A specific, accurate, precise and robust HPTLC method has been developed for estimation of Leflunomide in its
pharmaceutical dosage form. The chromatographic separation was performed using Aluminum backed precoated with
silica gel 60F254 as stationary phase and toluene: ethyl acetate: glacial acetic acid (8:2:0.5 %, v/v/v) as mobile phase.
The quantification was carried out at 270 nm wavelength. The method was validated as per ICH Q2 (R1) guidelines.
The Rf value was found to be 0.53 ± 0.02. The linearity of method was satisfactory over the range 25-125 ng/spot
with correlation coefficient of 0.9960.The limit of detection was found to be 1.71 ng. The limit of quantitation was
found to be 5.19 ng. The recovery was found in the range 99.67-100.89%.The method was successfully applied to
marketed formulations of Leflunomide.
2. Maryadele J, Neil O. The Merck Index - Encyclopedia of Chemicals, Drugs anBiologicals. 14th ed. USA: Merck Research Laboratories. 2006;5432.
3. Indian Pharmacopoeia, volume II, The Indian Pharmacopoeia Commission, Ghaziabad, Govt.of India,Ministry of Health and Family Welfare. 2014; 2073-75.
4. British Pharmacopoeia, volume III, The Department of Health, Medicine and Health Care Product Regulatory Agency.2010; 1250-51.
5. United States Pharmacopoeia 34 National Formulary 29 Volume II, United States Pharmacopeial Convention Inc, Rockville MD. 2011; 2686-88.
6. Soneji P, Shah J, Moheswari D. Development and validation of analytical method for Simultaneous estimation of Leflunomide and Methotrexate in synthetic mixture by Q Absorbance ratio method. International Journal of Pharmacy & Technology. 2015; 6:7500-12.
7. Shanmugarathinam A. Development of difference spectrophotometric method for the estimation of Leflunomide in tablet dosage form. Chemical Industry & Chemical Engineering Quarterly. 2012;3:407-410.
8. Abbas S, Begawy L, Fattah L, Refaat H. Spectrophotometric stability-indicating method for the determination of Leflunomide in the presence of its degradates. Journal of AOAC International. 2006;89.
9. Van E, Yaska J, Raemaekers J, Th T, Jansen A, Wanrooy M, Brouwers J. A rapid and Simple determination of A771726 in human serum by high performance liquid chromatography and its application for optimization of Leflunomide therapy. Journal of Pharmaceutical and Biomedical Analysis. 2004;36:17-22.
10. Chan V, Chartless B, Tett S. Rapid determination of active Leflunomide metabolite A77172 in Human plasma by high-performance liquid chromatography. Journal of Chromatography B. 2004; 803:331-35.
11. Schmidt A, Schwind B, Grilles M, Kay B and B. Simultaneous determination of Leflunomide and its active metabolite A1771726 in human plasma by high Performance liquid chromatography.Biomedical Chromatography. 2003; 17:276-81.
12. Miron D, SoldattelliC, Schapaval E. HPLC with diode-array detection for determination of Leflunomide in tablet. Chromatographia. 2006; 63:5-6.
13. Muhammad N, Khan S, Mehboob M, Muhammad N. Development and validation of liquid chromatographic method for the determine of Leflunomide: application to in vitro drug metal interaction. Chinese Journal of Chemistry. 2011; 29:1933-38.
14. Yeniceli D, Dogrukol D, Tuncel M. Determination of Leflunomide in pharmaceutical tablet by flow-injection analysis. Journal of Liquid Chromatography & Related Technologies. 2005; 28:1693-1701.
15. YiLMAZ H, Cubak E. Investigation of chromatographic behavior of Leflunomide and its Identification in pharmaceutical dosage form by reversed phase liquid chromatography. Journal of Liquid Chromatography & Related Technologies. 2015; 38:97-103.
16. Sobhani K, Garrett D, Liu D, Rainey P. A rapid and simple high-performance liquid Chromatography assay for the Leflunomide method, Teriflunomide(A771726),in renal transplant recipients. Clinical Chemistry. 2010; 133:454-57.
17. Rao S, Sunanda K, Rao N, Rao A, Maheswari IL, Srinababu G. Development and validation of LC method For the determination Of Leflunomide in pharmaceutical formulations using an experimental design. African Journal of Pure and Applied Chemistry. 2008;2:10-17.
18. Sultana N, Mohammed S, Khan M, Nadir S. Development of liquid chromatography-UV method for simultaneous determination of Leflunomide and NSAIDS in API and pharmaceutical formulation : its application to In vitro interation studies. Medicinal Chemistry. 2013;3:262-70.
19. Kher G, Ram V, Dubal K, Bapodara A, Joshi H. Validation of stability- indicating LC method for assay of Leflunomide in tablet and for determination of content uniformity. International Journal of Chem Tech Research. 2011;3:523-30.
20. Saini B, Bansal G.Isolation and characterization of a degradation product in Leflunomide and a validated selective stability-indicating HPLC-UV method for the quantification.Journal of Pharmaceutical Analysis. 2014;10.
21. Kher G, Ram V, Pandya G, Joshi H. Development and validation of stability indicating UPLC assay method for determination of Leflunomide in tablet formulation. Der ChemicaSinica. 2011; 5:65-74.
22. Parekh J, Vaghela R, Sutariya D, Sanyal M, Yadav M, ShrivastavP. Chromatographic Separation and sensitive detection of teriflunomide an active metabolite of Leflunomide in human plasma by liquid chromatography- tandem mass spectrometry. Journal of Chromatography B. 2010; 878:2217-25.
23. Eliangiringakaale, Peter Risha, Thomas Layloff. TLC for pharmaceutical analysis in resorce limited countries Review Article. J. of Chroma. A. 2011;1218:2732-36.
24. Shewiyo DH, Kaale E, Risha PG, Dejaegher B, Smeyers-Verbeke J, Vander Heyden J. HPTLC methods to assay active ingreadients in pharmaceutical formulations: A review of the method development and validation: A review of the method development and validation steps. J. Pharma.Bio.Analysis. 2012;66:11-23.
25. Sherma J, Review of HPTLC in Drug Analysis: 1996-2009. J. Pharma. Bio.Analysis. 2012;66:11-23.
26. ICH. Validation of Analytical Procedures: Text and Methodology. International Conference on Harmonization, IFPMA, Geneva, Switzerland; 2005
The International Journal of Drug Regulatory affairs require a formal written transfer of copyright from the author(s) for each article published. We therefore ask you to complete and return this form, retaining a copy for your records. Your cooperation is essential and appreciated. Any delay will result in a delay in publication.
I/we have read and agree with the terms and conditions stated Page 2 of this agreement and I/we hereby confirm the transfer of all copyrights in and relating to the above-named manuscript, in all forms and media, now or hereafter known, to the International Journal of Drug Regulatory affairs, effective from the date stated below. I/we acknowledge that the IJDRA is relying on this agreement in publishing the above-named manuscript. However, this agreement will be null and void if the manuscript is not published in the IJDRA.
Download link for COPYRIGHT FORM