Analytical method development and validation for estimation of etoposide using reverse phase high performance liquid chromatography

Abstract

A simple and reliable reverse phase high-performance liquid chromatographic (RP-HPLC) method was described for the
determination of Etoposide dosage forms. Chromatographic separation was achieved on a Zodiac C18 column using mobile phase
consisting of a mixture of Triethylamine buffer (4.5): Acetonitrile (60:40 v/v), at 25 °C, with detection of 260 nm. Linearity was
observed in the range 20-100 µg /ml for Etoposide (r2 =0.9955) for the amount of drugs estimated by the proposed methods was in
good agreement with the label claim. For intermediate and precision percentage, the RSD values were found to be 1.49% and 0.88%
respectively, that falls within acceptance criteria, that is RSD NMT 2%. The accuracy limit is the percentage recovery should be in
the range of 98.0% - 102.0%. The total recovery was found to be 98.78%. The solutions were chromatographed at a constant flow
rate of 1.0 ml/min. The retention time was found to be 2.750 mins. The assay was performed and the % purity was found to be
99.56%. The validation of developed method shows that the accuracy is well within the limits and showed good accuracy and
reproducibility. The robustness limit for wavelength variation and flow rate variation are well within the limits. It was
concluded that, the method described for the determination of Etoposide dosage forms was found to be simple, precise, accurate
and high resolution and shorter retention time, makes this method more acceptable and cost effective.