微量样品中喜树碱含量的高效液相色谱测定

为了测定微量喜树(Camptotheca acuminata Decne)样品中的喜树碱含量,建立了HPLC—荧光检测测定方法(采用Luna C18色谱柱(250 mm×4.6 mm,5μm),以乙腈∶水(体积比4∶6)为流动相,流速1.0 mL.min-1,荧光检测器激发波长380 nm,发射波长553 nm)。结果表明:喜树碱质量浓度在20～800μg.L-1范围内,与峰面积呈现良好的线性关系,r=0.999 6,平均加样回收率为98.41%,峰面积的相对标准偏差1.88%。这一方法仅需2 mg干质量植物样品即可实现对喜树碱含量的测定,且操作简单、重复性好、结果精确可靠,为微量植物材料中喜树碱含量的测定提供了有效方法。

Determination of Camptothecin in Milligram Samples by HPLC

To determine the content of camptothecin(CPT) in milligram samples of Camptotheca acuminata seedlings,a HPLC method with fluorescence detection was established.The analysis was carried out on a Luna C18 column(250 mm×4.6 mm,5 μm) with the mobile phase of acetonitrile/water(4∶6 by volume) at the flow rate of 1.0 mL·min-1.Fluorescence detector was used for the detection of CPT with excitation and emission wavelengths of 380 nm and 553 nm.The calibration curve was linear at the range of 20-800 ng·mL-1 for camptothecin(r=0.999 6).The average recovery was 98.41%,and the relative standard deviation was 1.88%.The method is simple,reproducible,reliable and sensitive for only 2 mg(dry weight) of the plant material.It is suitable for accurate determination of camptothecin content in milligram samples of C.acuminata seedlings.