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

Document Type: Research article

Authors

1 Department of Chemistry, Amirkabir University of Technology, Tehran, Iran.

2 Department of Phytochemistry, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University,Tehran, Iran.

3 Department of Phytochemistry, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University,Tehran, Iran

Abstract

There is no other naturally occurring defense agent against cancer that has a stronger effect
than paclitaxel, commonly known under the brand name of Taxol®. The major drawback for the
more widespread use of paclitaxel and its precious precursor, 10-deacetylbaccatin III (10-DAB
III), 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 hydrophilic
interaction/ reversed-phase chromatography was used to introduce a semi-preparative
treatment 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 effective
method for large-scale production of taxanes. This process avoids the technical problem of
two-dimensional preparative liquid chromatography. The first stage of the process involves
discarding co-extractive polar compounds including chlorophylls and pigments using a nonpolar
synthetic hydrophobic absorbent, Diaion® HP-20. Extract was then loaded on to a silica
based hydrophilic interaction solid phase extraction (silica 40-60 micron). Taxanes was eluted
using a mixture of water and methanol at the optimized ratio of 70:30. Finally, the fraction
containing taxanes was applied to semi-preparative reversed phase HPLC. The results revealed
that 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.

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