HPLC法测定红豆杉中紫杉醇及同系物的含量

A HPLC Method for Determination of Content of Taxol and Other Derivatives of Taxanes in the Branches and Leaves of Taxus spp.

建立了HPLC法测定红豆杉枝叶中10-去乙酰基巴卡亭Ⅲ（简称10-DABⅢ）、巴卡亭Ⅲ、三尖杉宁碱和紫杉醇的含量。采用二种流动相系统分别测定南方红豆杉枝叶中四种成分，以较大极性的甲醇-水（43：57）流动相系统测定10-去乙酰基巴卡亭Ⅲ和巴卡亭Ⅲ，以乙腈-水-甲醇（27：44：29）为流动相测定三尖杉宁碱和紫杉醇，采用Nova—Pak C18反相柱，流速为1mL／min，柱温35℃，紫外检测波长为227nm。10-去乙酰基巴卡亭Ⅲ、巴卡亭Ⅲ、三尖杉宁碱和紫杉醇的线性范围（n=5）分别为0．01～0．5μg（r=0．9998），0．01～1μg（r=0．9999），0．015～0．5μg（r：0．9990），0．01～0．5μg（r=0．9992）；样品用丙酮和水浸提，体积比为60：40，回收丙酮后，用二氯甲烷萃取得到上述四种成分。本实验方法操作简便，结果准确，重现性好，可作为不同地区红豆杉的质量控制方法之一。

In order to further determine the content of taxol and other derivatives of taxanes （ 10-deacetyl baccatin Ⅲ, baccatin Ⅲ, cephalomannine） in the branches and leaves of Taxus spp. , a HPLC method was established. The content of four components from the branches and leaves of Taxus chinensis var. mairei was analyzed respectively by two kinds of mobile phase system. The determination of 10-deaeetyl baceatin Ⅲ and baecatin Ⅲ was done by using Nova-Pak C18 column and the more polar mobile phase （43% methanol ： 57% water ）. The determination of cephalomannine and taxol was done by Nova-Pak C18 column and the proper proportion of mobile phase （ 27% acetonitrile ： 44% water ： 29% methanol）. The flow rate was 1.0 mL/min and the wavelength of UV detection was 227 nm. The assay displayed good linearity over the concentration ranges of 0.01-0.5 μg （ r = 0. 9998 ）, 0.01-1 μg （ r = 0. 9999 ）, 0. 015-0.5 μg （ r = 0.9990） and 0.01-0.5μg（ r = 0. 9992）, n = 5, respectively. The four components were extracted with 60% acetone and dichloromethane. This method is simple, accurate and with a good repeatability and providing a reference for quality control of Taxus spp. in different areas.