Abstract
Taxol and 10-Deacetyl baccatin III are major taxanes in the bark, needles, and endophytes of Taxus baccata. The current study aimed to develop a process for their separation from different matrices. Crude taxoid was prepared by extraction of samples with methanol, followed by partitioning with dichloromethane and precipitation with hexane. Analytical high-performance liquid chromatography involved isocratic elution on C18 column (4.6 × 250mm, 5μm) with methanol-water (70:30 v/v) at a flow rate of 1ml/min. Injection volume was 20μl and detection was carried out at 227nm. The content of Taxol and 10-Deacetyl baccatin III in bark, needles and endophytic culture broth was 11.19 and 1.75μg/mg; 11.19 and 1.75μg/mg; and 2.80 and 0.22μg/L, respectively. Preparative high-performance liquid chromatography was done on C18 column (10 × 250mm, 5μm) at a flow rate of 10ml/min. About 20g crude taxoid was processed in<3h with a recovery of about 90% for both the analytes. The purity of recovered Taxol and 10-Deacetyl baccatin III determined by ultra-high-performance liquid chromatography-mass spectrometry was found to be 95.78±3.63% and 99.72±0.18%, respectively. The structure of recovered Taxol was confirmed by nuclear magnetic resonance. The method can find use in biotransformation studies.
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