Analysis of 4'-demethylepipodophyllotoxin-9-(4,6-O-ethylidene-beta-D-glucopyranoside) by high-pressure liquid chromatography

A rapid and specific high-pressure liquid chromatographic assay is described for the quantitative analysis of 4'-demethylepipodophyllotoxin-9-(4,6-O-ethylidene-beta-D-glucopyranoside) (I) in plasma. After chloroform extraction, I was analyzed by reversed-phase chromatography and UV detection (252nm). The maximum sensitivity was 0.1 microgram/ml. Quantitation was by relative response factor calibration using an integrating microcomputer. Over the concentration range of 0.5-90 microgram/ml, the average recovery of I from plasma was 95.4 +/- 3.8% (SD).