Analytical Method Development and Validation of Etanercept by UV and RP-UFLC Methods

Abstract

The research work was carried out using Ultraviolet (UV)—visible spectroscopy and Reverse Phase-Ultra Fast Liquid Chromatography (RP-UFLC) for es-tablishing novel methods for the analysis and quantification of Biosimilar drug, Etanercept. The maximum absorbance of Etanercept was found to be 215 nm and it obeyed Beer-Lamberts law in the range of 5 to 200 µg/ml and 1 to 32 µg/ml for UV and RP-UFLC, respectively. The correlation coefficient (r 2 ) value was found to be between 0.999 and 0.9995. All the validation parameters like linearity, accuracy, and precision, Limit of Detection (LOD), Limit of Quan-titation (LOQ) and Robustness were found to be within acceptance criteria as per ICH guidelines. The results of accuracy studies (99.0% to 100.38%) indicated the methods to be accurate. The RSD % for interday and intraday precision studies was found to be less than 2%. Robustness and ruggedness were expressed in terms of RSD % which were also in the specified limits. LOD and LOQ of proposed method was calculated and found to be 1.257 and 3.809 μg/ml by UV, and 0.1073 μg/ml and 0.3251 μg/ml by RP-UFLC method, respectively. The developed methods were observed to be simple, rapid and cost-efficient. It can be easily applied for the estimation of Etanercept in the marketed formulations and for routine analysis of the Biosimilar drug.