The Combination Process for Preparative Separation and Purification of Paclitaxel and 10-Deacetylbaccatin III Using Diaion® Hp-20 Followed by Hydrophilic Interaction Based Solid Phase Extraction

Abstract

There is no other naturally occurring defense agent against cancer that has a stronger effectthan paclitaxel, commonly known under the brand name of Taxol®. The major drawback for themore widespread use of paclitaxel and its precious precursor, 10-deacetylbaccatin III (10-DABIII), is that they require large-scale extraction from different parts of yew trees (Taxus species),cell cultures, taxane-producing endophytic fungi, and Corylus species. In our previous work,a novel online two-dimensional heart-cut liquid chromatography process using hydrophilicinteraction/ reversed-phase chromatography was used to introduce a semi-preparativetreatment for the separation of polar (10-deacetylbaccatin III) and non-polar (paclitaxel)taxanes from Taxus baccata L. In this work, a combination of the absorbent (Diaion® HP-20) and a silica based solid phase extraction is utilized as a new, efficient, and cost effectivemethod for large-scale production of taxanes. This process avoids the technical problem oftwo-dimensional preparative liquid chromatography. The first stage of the process involvesdiscarding co-extractive polar compounds including chlorophylls and pigments using a nonpolarsynthetic hydrophobic absorbent, Diaion® HP-20. Extract was then loaded on to a silicabased hydrophilic interaction solid phase extraction (silica 40-60 micron). Taxanes was elutedusing a mixture of water and methanol at the optimized ratio of 70:30. Finally, the fractioncontaining taxanes was applied to semi-preparative reversed phase HPLC. The results revealedthat using this procedure, paclitaxel and 10-DAB III could be obtained at 8 and 3 times more,respectively than by the traditional method of extraction.