Extraction, Separation And Determination Of Taxanes In Taxus Chinensis Var. Mairei From Hunan

ObjectiveTo determine the contents of taxanes in Taxus chinensis var. mairei in different parts and different growth years with HPLC. The results will provide a basis for the further exploitation and utilization of Taxus chinensis var. mairei from Hunan; To explore the optimum condition for extraction, separation and purification of 10-DABⅢ.MethodsHPLC was used to determine the contents of taxanes in Taxus chinensis var. mairei with different parts and different growth years, the Hypersil BDS C18(200 mm×4.6 mm, particle size 5μm) column was used to determine the mobile phase methanol-water (60:40) of paclitaxel, determine 10-DABⅢand baccatinⅢof mobile phase was methanol-water (40:60). The flow rate was 1.0ml/min, the UV detection wavelength was 227nm, and the column temperature was 25℃. Studies on the extraction technology of taxanes in Taxus chinensis var. mairei have been done, ethanol was extractant, ultrasonic treatment was extract method, and the factors which may affect the technology were investigated with orthogonal design. Taxanes were separated by silica gel column chromatography, eluant was petroleum ether (60-90℃) and ethyl acetate system and with the method of gradient elution. Methanol was used for recrystallizing. Results(1) The linear range of paclitaxel was 168-840 ng(r=0.9994), the average recovery was 96.86% and the RSD was 2.01%; The linear range of 10-DABⅢwas 200-1000 ng(r=0.9999), the average recovery was 97.27% and the RSD was 1.73%; The linear range of baccatinⅢwas 232-1160 ng(r=0.9999), the average recovery was 96.07% and the RSD was 1.42%.(2) The optimum extraction technology have been explored with the results of orthogonal design.(3) At last, we obtained 10-DABⅢfrom Taxus chinensis var. mairei. Besides, two other compounds have been also obtained. From data of UV, IR,'H-NMR, 13C-NMR (DEPT) and MS, they were identified as (+)-catechin and 1-hydroxyl-7,9-dideacetyl baccatin I, respectively.Conclusion(1) HPLC was easy, accurate and had good precision, which was reliable and provided helpful evidence for collecting Taxus chinensis var. mairei.(2) Technology of extraction, separation and purification was advisable.(3) Silica gel column chromatography had advantages of simplicity of operator, high separation efficiency and lower energy consumption which could be used to separate 10-DABⅢ.