喜树种子中喜树碱和10-羟基喜树碱高效液相色谱法分析(英文)

DETERMINATION OF CAMPTOTHECIN AND 10-HYDROXYCAMPTOTHECIN IN CAMPTOTHECA ACUMINATA D. SEEDS BY HPLC

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 sensitive method for the determination of contents of camptothecin （CPT） and 10-hydroxycamptothecin （HCPT） in Camptotheca acuminata Decne. seeds by HPLC was established. The HPLC separation was performed on a KYA HIQ sil C18 column （250 mm×4.6 mm, 5 μm） at flow rate 1 mL/min, the solvent gradient program was as follow： acetonitrile-water increased linearly from 10 % to 40 % in the first 15 min, then decreased linearly to 10 % in 3 min; at time 21 min, stopped the program. The alkaloids were detected by a photodiode array detector and quantitatively analyzed at 254 nm for CPT and 266 nm for HCPT. CPT and HCPT were ultrasonically extracted by 60 % alcohol at 60 ℃ for 50 min. CPT and HCPT contents in different parts of C. acuminata D. seeds were determined by the above analyzing method. The results showed that CPT and HCPT levels were different in the four parts of C. acuminata D. seeds. CPT content was the highest in endosperm, lower in embryonic axis and cotyledon, and the lowest in seed coat, whereas HCPT content was the highest in endosperm, lower in seed coat, and the lowest in embryonic axis and cotyledon.