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建立了一种简便、快速、准确的测定喜树(CamptothecaacuminataDecne.)种子中喜树碱和10-羟基喜树碱含量的高效液相色谱分析方法;色谱条件为:采用日本KYAHIQsilC18柱(250mm×4.6mm,5μm),流速为1mL/min,梯度洗脱程序为:在前15min流动相乙腈-水的体积比由10%线性增加至40%,在随后的3min乙腈-水的体积比线性降至10%并保持恒定3min,在21min时停止该程序。检测器为二极管阵列检测器,喜树碱定量分析波长254nm,10-羟基喜树碱定量分析波长266nm,进样量10μL。样品制备以60%的乙醇作溶剂,在60℃下超声波提取喜树种子50min。利用以上方法分别测定了喜树种子胚乳、胚轴、子叶和种皮中的喜树碱和10-羟基喜树碱含量,喜树碱的含量是胚乳>胚轴,子叶>种皮,10-羟基喜树碱的含量是胚乳>种皮>胚轴>子叶。
A simple, rapid and accurate method for the determination of camptothecin and 10-hydroxycamptothecin in Camptotheca acuminata Decne seeds was established. The chromatographic conditions were as follows: a Japanese KYAHI Qsil C18 column (250 mm × 4.6 mm, 5μm) at a flow rate of 1mL / min. The gradient elution program was as follows: the volume ratio of acetonitrile to water was increased linearly from 10% to 40% in the first 15 min, 10% and kept constant for 3 min, stopping the procedure at 21 min. Detector for the diode array detector, camptothecin quantitative analysis wavelength 254nm, 10-hydroxycamptothecin quantitative analysis wavelength 266nm, injection volume 10μL. Sample Preparation Camptotheca acuminata seeds were extracted with 60% ethanol for 60 min at 60 ℃. The contents of camptothecin and 10-hydroxycamptothecin in the endosperm, hypocotyl, cotyledon and seed coat of Camptotheca acuminatum were determined by the above methods. The content of camptothecin was the content of endosperm> hypocotyl> cotyledon> The content of hydroxycamptothecin is endosperm> seed coat> hypocotyl> cotyledons.