Taxol purification with Centrifugal Partition Chromatography

Taxols are anti-cancer chemotherapy drugs. 10-Deacetyl Baccatin III (10-DAB III) is an important intermediate for both Paclitaxel and Docetaxel, two of the best anti-cancer drugs ever invented. 10-DAB III is one of the several taxols, which can be easily extracted from Yew Tree leaves (Taxus baccata), so it became an important starting raw material in the semi-synthetic manufacturing of these drugs, because of its particularly high concentration in Yew tree leaves.

The centrifugal partition chromatography (CPC) is a typical preparative chromatography technique, where both phases (mobile and stationary) are liquids, so both phases can be optimized for different separation methods. As no silica gel needed, and the used amount of solvent is lower the operational costs significantly decrease. The liquid static phase is immobilized by a strong centrifugal field, so high flowrate of mobile phase can be used, which means fast separation. Several references can be found in literature about successful taxol purification by CPC technique [1].

In the course of our work we measured 10-DAB III samples by centrifugal partition chromatography. By the separation of main component and impurities we wanted to create highly purified product. First, we tested several solvent systems, than we determined partition coefficient of taxol components by high performance liquid chromatography (HPLC). Next step was the optimization of CPC separation method with suitable partition coefficient solvent systems. Selectivity of all compounds are over 1,25. Finally, with repeated CPC purifications we collected purified fractions, which were measured by HPLC. CPC separations showed good result, fraction analysis has shown over 99% purity with 95% recovery possible. Further we were able to develop a CPC method, which is scalable to industrial levels, and is able to provide 10-DAB III with specified purity at high yields. On the whole, we determined a quick and low-cost method for 10-DAB III purification.

It is very important to find a green chemsitry technique which cause less ecological damage, since it is necessary to take 3000 Yew trees to produce only 1 kilogram of mentioned drugs [2]. CPC technique has several economic and environmental advantages such as low production cost from the reuse of solvents, consistent purity and yield, high sample loading (2,0 gramm of injection to the 250 mL column), and quick separation (one cycle takes around 1.5 hours) against other techniques, like MPLC and HPLC purification techniques.

References:

[1] Foucault AO. Centrifugal Partition Chromatography. New York, CRC press 1994. 335 – 350

[2] Malik S, Cusidó RM, Mirjalili MH, Moyano E, Palazón J, Bonfill M. Production of the anticancer drug Taxol in Taxus baccata suspension cultures: A review. Process Biochem 2011; 46: 23 – 34