Estimation and Validation of Etoposide by RP-HPLC Method in Rat Plasma by Liquid- Liquid Extraction

The objective of this article is to develop a new validated RP-HPLC method for the estimation of Etoposide, an anti-cancer drug in rat plasma. Etoposide was estimated by a new RP-HPLC method developed on a phenomenex C18 (4.6 x 250 mm, 5μ i.d) column, by isocratic elution using mobile phase mixture of methanol: pH 6 phosphate buffer in a ratio of 80:20 % v/v, at a flow rate 0.9 ml/min and monitored at 273 nm with an average retention time of 4.7 ± 0.1 min. Extraction of the sample from rat plasma was performed by employing liquid-liquid extraction method using chloroform as a solvent. The evaporated chloroform extract was reconstituted by using methanol exhibits linearity in the concentration range of 2000- 5000 μg/ml. The LOD and LOQ were found to be 184 μg/ml and 561 μg/ml, respectively with a correlation coefficient of 0.9989 and slope of 1.3818. The method was proved to be more accurate, simple, precise and rapid by statistical validation, extraction and recovery studies.

Author(s): Magharla Dasaratha Dhanaraju Vankayalu Devendiran Sundar, Rajendran Vijayalakshmi and Vadapalli S Harsha Naveena

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