三尖杉悬浮培养细胞中hinokiol的定量分析

摘要

建立了高效液相色谱分析三尖杉悬浮培养细胞中次生代谢产物的方法,实现了次生代谢产物的分离以及hinokiol的定量分析.样品经甲醇提取后,再用氨水-氯仿萃取.采用Apollo C18色谱柱(250 mm×4.6 mm,5μm)进行分离,流动相为甲醇-水,梯度洗脱,柱温为30℃,检测波长为290 nm,流速为1 mL/min.在0.012 5～0.2 g/L范围内,hinokiol的色谱峰面积与质量浓度之间具有良好的线性关系.采用该方法测定了实际样品中hinokiol的含量,并进行了 3个水平的加标回收试验,其回收率为87.2％～94.7％,相对标准偏差(n=3)为0.9％～4.2％.该方法可靠、重现性好,适合对植物培养细胞中的hinokiol进行分析.%A high performance liquid chromatography (HPLC) method was developed for the separation of secondary metabolites and quantitative analysis of hinokiol from cell suspension cultures of Cephalotaxus fortunei. The samples were prepared by extraction using methanol followed by partitioning between ammonium hydroxide and chloroform. The HPLC separation was achieved on an Apollo C18 column (250 mm ×4. 6 mm, 5 μm) with gradient elution using methanol and water at 1 mL/min and 30 ℃. The detection was carried out at 290 nm. A good linear correlation between the hinokiol peak area and mass concentration was observed over the mass concentration range of 0. 012 5 -0. 2 g/L. The proposed method was applied to the determination of hinokiol in the actual samples with recoveries of 87. 2% - 94. 7% and with the relative standard deviations of 0. 9% - 4. 2%. This method is reliable and reproducible and is suitable for the analysis of hinokiol in plant cell cultures.