Methodology for the Identification and Purification of Taxol and Cephalomannine from Taxus Callus Cultures

Abstract

In this study we have developed a new protocol for the extraction and purification of taxol and cephalomannine from plant tissue culture samples. This protocol allows large numbers of samples to be processed with a minimum use of solvents and can be completed in a relatively short period of time, compared to existing published protocols. The procedure involves a methanol extraction followed by the use of C₁₈ Sep-pak cartridges, before analysis by isocratic high performance liquid chromatography (HPLC) using a reverse-phase analytical phenyl column. Our purification protocol consisted of a 65% methanol: chloroform partitioning step followed by C₁₈ bonded silica column chromatography and finally, C₁₈ preparatory HPLC.