High Performance Liquid Chromatographic Studies of Teriflunomide: Method Development and Validation for Drug Development and Formulation

Abstract

Aim: There is an unmet analytical need to develop a rapid, robust, and precise method for teriflunomide in drug
development and research in pharmaceutical formulations. The main spotlight of this research work is to develop
a precise, simple, and accurate method for the application of analytical research development of teriflunomide
and its pharmaceutical formulations. Materials and Methods: The quantification and analytical validation
of teriflunomide was developed with a stationary phase XBridge column C18 (4.6 mm × 250 mm, 5 µm) and
LC1120 Agilent high performance liquid chromatographic instrument equipped with variable wavelength detector
at 294 nm and Acetonitrile: Buffer containing 20 mM of Ammonium acetate and 5 mL glacial acetic acid with pH
adjustment 4.48 (60:40, v/v) was used as mobile phase passed at a flow rate 1 mL/min. Elution takes place at a
retention time of 2.853 min. Results and Discussion: Validation of the method was performed as per International
Council for Harmonisation guidelines, which shows linearity concentration range from 10 to 60 µg/mL amid
correlation coefficient = 0.999 with Y = 78737 × +47703 regression equation was obtained. Accuracy of the
proposed method was established to 99.03–99.13% with relative standard deviation (% RSD) values <2%. Method
precision, system precision, and reproducibility % RSD value were 0.14%, 0.23%, and 0.14%, respectively. The
limit of detection and limit of quantitation of teriflunomide were 0.06 µg/mL and 0.2 µg/mL, correspondingly.
The intermediate precision was carried out and the results of Teriflunomide were achieved <2% RSD. Robustness
was performed by deliberate changes in wavelength and flow rate and the results of robustness were achieved
<2% RSD. Conclusion: The developed method was precise, simple, and accurate for the application of analytical
research development of teriflunomide and its pharmaceutical formulations.