제조용 액체크로마토그래피를 이용한 taxol 분리공정의 개발

초록

Taxol, a powerful anticancer agent, was extracted by solvents from yew extracts and separated by RP-HPLC(Reverse-Phase High Performance Liquid Chromatography) and NP-HPLC (Normal-Phase High Performance Liquid Chromatography) in analytical and preparative scale. The solvent used in the extraction was methanol, and the extracts was further concentrated by the partition of water and methylene chloride. Such impurities as lipid, chlorophyl, pigment, et al. were removed in the pretreatment step with open column, where bare silica was packed. The mobile phase of MC(Methylene chloride)/acetone most clearly showed the developed band of the extract, and the elution time was fastest by the gradient mode (75/25 after 97/3 vol.% of MC/acetone). The artificial mixture of taxol, cephalomannine and 10-deacetyltaxol was resolved by the curosil-PFP column in water/acetonitrile/MeOH (56/39/5 vol.%) and the analytical Nova-Pak column in hexane/1-propanol/MeOH (96/2/2 vol.%) in an isocratic mode. In the preparative scale with larger particle size, the mobile phase composition was changed to hexane/IPA/MeOH (90/4/6 vol.%). The better resolution was achieved by the gradient mode : hexane/IPA/MeOH (86/4/10 vol.%), where the initial composition of mobile phase was maintained for 17min. and the final composition was step-changed to hexane/IPA/MeOH (92/2/6 vol.%). Under the experimental conditions, taxol was preparatively separated from yew extracts.