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Abstract

SIMULTANEOUS ESTIMATION OF BENZYL CHLORIDE AND BENZYL BROMIDE IN ENTECAVIR BY USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Vidyagauri V. Lele* and Uttam Prabhanand Dalvi

ABSTRACT

A highly sensitive analytical method for the simultaneous quantification of trace level impurities of Benzyl chloride and Benzyl bromide in the active pharmaceutical ingredient Entecavir has been developed. These genotoxic impurities have demonstrated to induce genetic mutation, chromosomal breaks or chromosomal rearrangement and have potential to cause cancer. The analysis was accomplished on an Inertsil poroshell C18 column (50mm x 4.6mm, 3μ) using phosphate buffer and acetonitrile as the organic solvent. The flow rate was set at 1.2 mL/ minute with a total analysis time of about 6 minutes. The method was validated for the analytical parameters such as system suitability, specificity, linearity and range, accuracy, limit of detection (LOD), limit of quantification (LOQ), precision, and solution stability. The limit of detection and limit of quantification were found to be 0.05μg/mL and 0.2μg/mL respectively with respect to Entecavir sample concentration (2μg/mL).

Keywords: Benzyl bromide, Benzyl chloride, Mutation, Genotoxic, Antiviral.


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