DEVELOPMENT AND VALIDATION OF STABILITY INDICATING REVERSED PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC METHOD FOR THE DETERMINATION OF RELATED SUBSTANCES IN FAMPRIDINE DRUG SUBSTANCE AND TABLET DOSAGE FORMS
Â Objective: The objective of this method is to develop a stability-indicating reversed phase high performance liquid chromatographic method for the quantification of related substances in the drug substance and tablet dosage form of Fampridine.
Methods: Inertsil ODS 3V, (150 mm Ã— 4.6 mm, 5 Î¼m particle size) column was used for the separation of analytes. Mobile phase A was prepared by dissolving 6.8 g of potassium dihydrogen orthophosphate (0.05 mol) and 1 g of 1-octane sulfonic acid into a 1000 ml of water, pH was adjusted to 4.0Â±0.05 with diluted orthophosphoric acid. Mobile phase B was prepared by mixing the above phosphate buffer (pH 4.0) and acetonitrile in 20:80 (% v/v). Gradient mode was used with the flow rate of 1.0 ml/minutes, and the peaks were monitored at 260 nm.
Results: Linearity results showed that the correlation coefficient (r2) is >0.995 for individual active drug substances as well as their related substances in the range of limit of quantification to 150% of the specification concentration (0.5% with respect to sample concentration of 0.4 mg/ml). Accuracy of the method was established with their recovery values in the range of 98.5-104.5% with the % RSD not more than 1.7%. The method was proved by highly precise (% RSD of intra-day and inter-day study was not more than 4.3%) and more robust.
Conclusion: Present method is able to separate two related compounds with each other and with the main drug substance with the resolution more than 2.0. The test standard solution and test solution were found to be stable in diluent up to 24 hrs. The mass balance of forced degradation of formulations is close to 99% made this method as a stability indicating method.
2. Korenke AR, Rivey MP, Allington DR. Sustained-release fampridine for symptomatic treatment of multiple sclerosis. Ann Pharmacother 2008;42(10):1458-65.
3. Madhumathi CH, Padmalatha M, Sneha S, Divya CB. Development and validation of UV spectrophotometry method for the estimation of dalfampridine in tablets. Int J Pharm Sci Res 2014;2:1019-26.
4. Fatatry HM, Hammad SF, Elagamy SH. Validated spectrofluorimetric determination of dalfampridine in its synthetic mixture and spiked human plasma through derivatization with fluorescamine. J Anal Tech 2013;3:23-6.
5. Dharani NR, Padmini K, Sumakala S. Stability indicating RP-HPLC method development and validation for estimation of dalfampridine in its bulk and formulation. Int J Adv Res 2016;4:184-91.
6. van der Horst A, de Goede PN, van Diemen HA, Polman CH, Martens HJ. Determination of 4-aminopyridine in serum by solid-phase extraction and high-performance liquid chromatography. J Chromatogr 1992;574(1):166-9.
7. Suneetha A, Rajeswari KR. A high throughput flow gradient LC-MS/MS method for simultaneous determination of fingolimod, fampridine and prednisone in rat plasma, application to in vivo perfusion study. J Pharm Biomed Anal 2016;120:10-8.
8. Thomas S, Bharti A, Sanjeev S, Ashutosh A. A stability indicating simultaneous dual wavelength UVâ€“HPLC method for the determination of potential impurities in fampridine active pharmaceutical ingredient. J Pharm Biomed Anal 2012;58(1):136-40.
9. Jain M, Srivastava V, Kumar R, Dangi V, Hiriyanna SG, Kumar A, et al. Determination of five potential genotoxic impurities in dalfampridine using liquid chromatography. J Pharm Biomed Anal 2016;133:27-31.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.