Two‐dimensional Hydrophilic Interaction/Reversed‐phase Liquid Chromatography for the Preparative Separation of Polar and Non‐polar Taxanes

Abstract

Paclitaxel (Taxol®) is one of the best-known compounds of natural origin that has been used in the pharmaceutical industry. It is derived from taxanes extracted from different parts of yew trees (Taxaceae), cell cultures and taxane-producing endophytic fungi. Previous work reported a semi-preparative HPLC for the extraction of Paclitaxel (a non-polar taxane) from Taxus baccata L. and cell culture. In other work, 10-deacetylbaccatin III (a polar taxane) was separated from this medium at a preparative scale. To introduce a novel on-line two-dimensional heart-cut liquid chromatography using hydrophilic interaction/reversed-phase chromatography for preparative separation of polar and non-polar taxanes. To achieve this goal the system was developed from analytical to a larger preparative scale. In the first dimension of a semi-preparative column, taxanes were separated from polar compounds using a CN column, and in the second dimension the fraction containing taxanes was transferred to a C₁₈-column. Using this method polar and non-polar taxanes could be separated at analytical and semi-preparative scales. Combination of hydrophilic and reversed-phase chromatography in two-dimensional liquid chromatography (2D-LC) systems is a useful method for the separation of compounds with a large polarity range such as taxanes.