Fett Neto A G, DiCosmo F
Department of Botany, University of Toronto, Ontario, Canada.
Planta Med. 1992 Oct;58(5):464-6. doi: 10.1055/s-2006-961515.
Different fresh shoot parts of male and female plants of Taxus cuspidata were extracted and analysed for taxol concentration by high performance liquid chromatography (HPLC). Extracted parts included: young needles (first 10 top needle pairs of 30 cm long branches), old needles (last 10 needle pairs of 30 cm long branches), green bark, dark bark (with intense secondary growth), young wood (originally surrounded by green bark), wood (originally surrounded by dark bark), young stems (surrounded by green bark and devoid of needles), and mature male cones. Dichloromethane extracts were analysed by HPLC and diode array spectroscopy. Taxol identification was done by retention time, U.V. spectra, and spiking with an authentic taxol standard; 1H-NMR analysis was done for needle extracts. All parts except male cones had measurable amounts of taxol; no effect of plant sex on taxol levels of the plant parts analysed was observed. Results indicated that the bark accounted for almost all the taxol present in stems devoid of needles. Needles showed the highest levels of taxol (overall average of 0.035 +/- 0.006% of the extracted dry weight), significantly higher than those displayed by dark bark samples (0.012 +/- 0.001% of the extracted dry weight). Different needle post-harvesting procedures were evaluated in relation to taxol yields, 96 h dark incubation at -12 degrees C and 96 h dark incubation at 25 degrees C under vacuum gave taxol yields equivalent to those of freshly extracted samples. However, results obtained for 96 h dark incubation at 60 degrees C indicated some extent of taxol degradation.
对东北红豆杉雌雄植株不同的嫩梢部位进行提取,并通过高效液相色谱法(HPLC)分析紫杉醇浓度。提取的部位包括:嫩针叶(30厘米长枝条顶部的前10对针叶)、老针叶(30厘米长枝条底部的后10对针叶)、绿色树皮、深色树皮(次生生长旺盛)、嫩木质部(最初被绿色树皮包围)、木质部(最初被深色树皮包围)、嫩茎(被绿色树皮包围且无针叶)以及成熟雄球花。二氯甲烷提取物通过HPLC和二极管阵列光谱法进行分析。通过保留时间、紫外光谱以及与紫杉醇标准品加样来鉴定紫杉醇;对针叶提取物进行了¹H-NMR分析。除雄球花外的所有部位均含有可测量的紫杉醇;未观察到植株性别对所分析植株部位的紫杉醇水平有影响。结果表明,在无针叶的茎中,几乎所有的紫杉醇都存在于树皮中。针叶中的紫杉醇含量最高(占提取干重的总体平均值为0.035±0.006%),显著高于深色树皮样品(占提取干重的0.012±0.001%)。针对紫杉醇产量评估了不同的针叶采收后处理程序,在-12℃下黑暗孵育96小时以及在25℃真空条件下黑暗孵育96小时,所得紫杉醇产量与新鲜提取样品相当。然而,在60℃下黑暗孵育96小时的结果表明紫杉醇有一定程度的降解。