title: Optimization of the fractional precipitation of paclitaxel from a Taxus chinensis cell culture using response surface methodology and its isolation by consecutive high-speed countercurrent chromatography.
abstract: A consecutive preparation method for the isolation and purification of paclitaxel from the Taxus Chinensis cell culture was developed in this study. The process involved alkaline Al2O3 chromatography, fractional precipitation, and high-speed countercurrent chromatography. The original cell culture materials were first extracted with methanol using ultrasound-assisted extraction, and then the extract (the content of paclitaxel is 1.5%) was separated by alkaline Al2O3 column chromatography. Subsequently, fractional precipitation was used to obtain paclitaxel. In particular, response surface methodology was used to optimize the factors of fractional precipitation (methanol concentration, material-to-solvent ratio, and precipitating time were optimized as 48.14%, 8.85 mg/mL, and 48.71 h, respectively) and the yield of fractional precipitation product was 30.64 ± 0.60 mg (the content of paclitaxel is 89.3%, 27.37 ± 0.54 mg) from a 100 mg fraction by Al2O3 column separation (the content of paclitaxel is 32.4%). Then, the product was used for further isolation by high-speed countercurrent chromatography. About 1.00 g paclitaxel (200 ± 2 mg in each loading) with a purity up to 99.61% was isolated from 1.25 g of fractional precipitation product with a solvent system of n-hexane/ethyl acetate/methanol/water (1.2:1.8:1.5:1.5, v/v/v/v) in one run of five consecutive sample loadings without exchanging a new solvent system.
authors.affiliation: School of Pharmaceutical Sciences, Sun Yat-Sen University, Guangzhou, China; Guangdong Technology Research Centre for Advanced Chinese Medicine, Guangzhou, China.
authors.forename: Zhikun
authors.initials: Z
authors.lastname: Liang
authors.forename: Zhisheng
authors.initials: Z
authors.lastname: Xie
authors.forename: Shingchung
authors.initials: S
authors.lastname: Lam
authors.forename: Xinjun
authors.initials: X
authors.lastname: Xu
mesh_terms: D018929:Cell Culture Techniques; D003377:Countercurrent Distribution / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000737:chemistry / Q000302:isolation & purification*; D005590:Fractional Precipitation / Q000379:methods*; D017239:Paclitaxel / Q000737:chemistry / Q000302:isolation & purification*; D020946:Taxus / Q000737:chemistry* / Q000254:growth & development
chemical_list: D004365:Drugs, Chinese Herbal; D017239:Paclitaxel
keywords: Cell culture; Fractional precipitation; Isolation; Paclitaxel; Response surface methodology