METHODOLOGY FOR THE IDENTIFICATION AND PURIFICATION OF TAXOL AND CEPHALOMANNINE FROM TAXUS-CALLUS CULTURES

In this study we have developed a new protocol for the extraction and purification of taxol and cephalomannine from plant tissue culture sam ples. This protocol allows large numbers of samples to be processed wi th a minimum use of solvents and can be completed in a relatively-shor t period of time, compared to existing published protocols. The proced ure involves a methanol extraction followed by the use of C18 Sep-pak cartridges, before analysis by isocratic high performance liquid chrom atography (HPLC) using a reverse-phase analytical phenyl column. Our p urification protocol consisted of a 65% methanol : chloroform partitio ning step followed by C18 bonded silica column chromatography and fina lly, C18 preparatory HPLC.