Purification of Paclitaxel Isolated from Taxus baccata L. Cell Culture by Microwave‐Assisted Extraction and Two‐Dimensional Liquid Chromatography

Abstract

Microwave‐assisted extraction (MAE) has been applied for extraction of paclitaxel from a cell culture. Solid phase C₁₈ has been used for clean up of the sample obtained from the optimized condition of MAE. The factorial design shows that the temperature and ratio of methanol to water (as an extraction solvent) are effective and extraction time is not an essential parameter for MAE of paclitaxel from the cell culture. Recently, we introduced MAE for extraction of paclitaxel from the needle of Taxus baccata L. Paclitaxel was extracted from the cell culture and the needle of Taxus baccata L, have been purified by heart‐cutting two dimensional high performance liquid chromatography (2D‐HPLC) in analytical and semi‐preparative scales. The columns used in the analytical chromatography were a Eurospher‐100 C₈ and Nucleosil‐100 C₁₈, respectively, and paclitaxel was eluted at a flow rate of 1 mL min⁻¹ with 40∶60 v/v and 45∶55 v/v water/acetonitrile, respectively. These conditions used in analytical chromatography, have been extended to a semi‐preparative scale in which the first column was a Eurospher‐100 C₈ and the second was a Eurospher‐100 C₁₈, but the mobile phase used consisted of 40∶60 v/v water/acetonitrile eluted at a flow rate of 11 mL min⁻¹.

Detection has been carried out at a wavelength of 227 nm throughout the analysis in both methods. The yield and purity of paclitaxel obtained by this method were 89 and 89% for the cell culture and 85 and 83% for the needle of Taxus baccata L., respectively.

Keywords:

• Paclitaxel
• Cell culture
• Taxus baccata
• Microwave‐assisted extraction
• Two‐dimensional liquid chromatography
• Semi‐preparative HPLC