Analytical Method Development and Validation of Teriflunomide by RP- HPLC

INTRODUCTION: High-Performance Liquid Chromatography (HPLC)High-performance liquid chromatography, is a chromatographic technique used to separate the components in a mixture, to identify each component, and to quantify each component. The method involves a liquid sample being passed over a solid adsorbent material packed into a column using a flow of liquid solvent. Each analyte in the sample interacts slightly differently with the adsorbent material, thus retarding the flow of the analytes. If the interaction is weak, the analytes flow off the column in a short amount of time, and if the interaction is strong, then the elution time is long. AIM AND OBJECTIVE: The drug analysis plays an important role in the development, manufacture and therapeutic use of drugs. Most of the pharmaceutical industries do the quantitative chemical analysis to ensure that the raw material used and the final product thus obtained meet certain specification and to determine how much of each components are present in the final product. Standard analytical procedure for newer drugs or formulation may not be available in Pharmacopoeias; hence it is essential to develop newer analytical methods which are accurate, precise, specific, linear, simple and rapid. Aim: To develop and validate new RP HPLC method for the assay of Teriflunomide in tablet dosage. Objective: Literature survey has revealed that various method were reported for estimation of Teriflunomide. The existing methods are inadequate to meet the requirements; hence it is proposed to improve the existing methods and to develop new methods for the Estimation of Teriflunomide in pharmaceutical dosage forms. Hence, on the basis of literature survey it was thought to develop a precise, accurate, simple and reliable, less time consuming method for estimation. SUMMARY: A simple Reverse Phase High Performance Liquid Chromatographic method has been developed and subsequently validated for Teriflunomide tablets. The separation was carried out by using a Buffer : acetonitrile (65:35). The detection was carried out at 250nm. The column was Zorbax Eclipse XDB, C8,150 x 4.6mm, 5μl. The flow rate was selected as 1.5ml/min. The Retention time of Teriflunomide tablets was found to be 6.0. The asymmetry factor or tailing factor of Teriflunomide tablets was found to be 1.2, which indicates symmetrical nature of the peak. The number of theoretical plates of Teriflunomide tablets was found to be 7391, which indicates the efficient performance of the column. These parameters represent the specificity of the method. From the linearity studies, specified concentration levels were determined. It was observed that Teriflunomide tablets were linear in the range of 5% to 150% for the target concentration by RP-HPLC. The linearity range of Teriflunomide tablets 5% to 150% was found to obey linearity with a correlation coefficient of 0.999. The validation of the proposed method was verified by system precision and method precision by RP-HPLC. The %RSD of system suitability for Teriflunomide tablets was found to be 0.25. The validation of the proposed method was verified by recovery studies. The percentage recovery range was found to be satisfied which represent in results. The robustness studies were performed by changing the flow rate, filters and wavelength. The ruggedness study was also performed. CONCLUSION: A HPLC method for Teriflunomide tablets was developed and validated in tablet dosage form as per ICH guide lines. The results of this validation are as summarized in the report. The results are found to be complying with the acceptance criteria for each of the parameter. Waters Alliance HPLC (Empower software with PDA detector) with Zorbax Eclipse XDB, C8,150 x 4.6mm, 5μ column, Injection volume of 10 μl is injected and eluted with the Mobile phase (Buffer and ACN, in the ratio of 65:35) which was pumped at a flow rate of 1.5 ml at 250 nm. The peak of Teriflunomide was found well separated at 6.0 min. The developed method was validated for various parameters as per ICH guidelines like system suitability, accuracy, precision, linearity, specificity, ruggedness, robustness and solution stability. Hence it is concluded that the assay method is found to be valid in terms of reliability, precision, accuracy and specificity and hence it is suitable for routine analysis as well as for stability analysis