High-Performance Liquid Chromatographic Analysis of Etoposide in Plasma Using Fluorescence Detection

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Abstract
A rapid and sensitive high-performance liquid chromatographic assay was developed for the quantitation of the antineoplastic drug etoposide in plasma. Etoposide and the internal standard teniposide were extracted from plasma (1ml) with methylene chloride. The extracts were analyzed on an Alltech C18 Econosil column with a mobile phase of 40% methanol, 15% acetonitrile, 44% water, and 1 % acetic acid. The assay utilized fluorescence detection (EX: 230 nm and EM: 328 nm) which provided sensitivity and specificity sufficient to quantify ≥ 50 ng/ml etoposide in plasma. The correlation coefficient of the multilevel standard curve was = 0.999 over a concentration range of 50–1000 ng/ml. The interday and intraday coefficients of variation were < 7%. This method has been used by our laboratory to provide the unattended overnight analysis of etoposide in plasma obtained from bone marrow transplant patients.

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