Efficient purification of paclitaxel from yews using high-performance displacement chromatography technique
| Autor: | Jin-Ming Gao, Yasuo Konishi, Faustinus K. Yeboah, Jean Watchueng, Pierre Kamnaing, Taira Kiyota |
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| Rok vydání: | 2010 |
| Předmět: |
Paclitaxel
Ion chromatography Docetaxel Biochemistry Analytical Chemistry Acetic acid chemistry.chemical_compound Liquid–liquid extraction Sample preparation Chromatography High Pressure Liquid Acetic Acid Chromatography Aqueous solution Methanol Organic Chemistry Water General Medicine Displacement chromatography Antineoplastic Agents Phytogenic Solvent Hexane Plant Leaves chemistry Taxoids Taxus Hydrophobic and Hydrophilic Interactions |
| Zdroj: | Journal of chromatography. A. 1218(20) |
| ISSN: | 1873-3778 |
| Popis: | Paclitaxel was purified using high-performance displacement chromatography (HPDC) technique, but not by the mechanism of HPDC. On small scale, paclitaxel was extracted with methanol from dry needles of Taxus canadensis and was enriched by extracting with chloroform after removing water-soluble hydrophilic components and hexane-soluble hydrophobic components. Then, 93-99% purity of paclitaxel was obtained using the HPDC technique. On large scale, taxanes were enriched by solvent partitioning between acetic acid/MeOH/H(2)O and hexane and extracted with CH(2)Cl(2). Taxanes except paclitaxel were further removed by extracting with methanol-water-trifluoroacetic acid (1.0:98.9:0.1, v/v/v). Applying HPDC technique to water-insoluble substances is problematic as this method requires a highly aqueous solvent system. In order to overcome this incompatibility, a system was set up where paclitaxel, although in low concentration, was extracted by methanol-water-trifluoroacetic acid (10.0:89.9:0.1, v/v/v). Recycling the extracting solvent to ensure minimal volume, the extracted paclitaxel was adsorbed on a C(18) trap column. A C(18) column of 4.6mm internal diameter was then connected to the trap column. The HPDC technique was thus carried out using an isocratic acetonitrile-water-trifluoroacetic acid (30.0:69.9:0.1, v/v/v) mobile phase consisting of a displacer cetylpyridinium trifluoroacetate (3mg/mL). Paclitaxel was co-eluted with the displacer and spontaneously crystallized. The crystal (114mg) showed 99.4% purity and only 10% of paclitaxel in the starting crude extract was lost during the enrichment/purification processes. This large scale purification method was successfully applied to purify paclitaxel from Chinese yew in small scale, suggesting general applicability of the method. This is the first report of purifying a water-insoluble natural product using HPDC technique. |
| Databáze: | OpenAIRE |
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