Process utilized oligo-β-cyclodextrin substituted agarose gel medium for efficient purification of paclitaxel from Taxus cuspidata

An efficient and economical process utilized oligo-β-cyclodextrin substituted agarose gel medium has been developed for paclitaxel recovery and purification directly from the plant materials of Taxus cuspidata. The process is a combination of extraction, alkaline Al2O3 chromatography step, oligo-β-cyclodextrin substituted agarose gel column chromatography which employed a novel oligo-β-cyclodextrin-Sepharose HP as the packing material, and crystallization. The Al2O3 normal-phase chromatography was used to obtain a preliminary isolation of paclitaxel from largely unwanted compounds in crude extract. The highest selectivity for paclitaxel was obtained on the oligo-β-cyclodextrin-Sepharose HP column compared with other β-cyclodextrin coupled gel media columns. The initial separation extract on Al2O3 column was purified by the subsequent oligo-β-cyclodextrin-Sepharose HP column chromatography with a purity of about 95.3% and a recovery of 87.1% under the optimum mobile phase composed of methanol/acetonitrile/water = 32/20/48 (v/v/v). One-step crystallization can increase the product purity up to 97.3%. HPLC analysis and ESI-MS/MS spectrum were used for the detection and characterization of paclitaxel in isolated fraction.