HPLC法测定远志中总皂苷的含量

HPLC determination of total saponins in Radix Polygalae

目的:建立远志药材中总皂苷的含量测定方法。方法:采用远志提取物碱水解方法制备远志总皂苷的次级苷—细叶远志皂苷,采用高效液相色谱法测定细叶远志皂苷的含量。色谱条件为 Alltima C_(18)色谱柱(4.6 mm×250 mm,5 μm),流动相为甲醇0.05%磷酸溶液(65:35),流速1.0 mL·min^(-1),检测波长202 nm。结果:细叶远志皂苷的理论塔板数为2500。回归方程 Y=4.242×10~6X-1.068×10~4(r=0.9999),线性范围100～1000μg·mL^(-1)。平均回收率(n=6)为101.2%(RSD=3.6%)。不同来源14批远志药材含量测定结果表明,远志中含总皂苷以细叶远志皂苷计为2.42%～3.71%;不同采收期样品分析表明,以3～6月份采收含量较高;1～3年生样品分析表明,以2年生和3年生药材含量为高。结论:该方法简便、准确,重复性好,可作为远志药材及含远志复方制剂质量控制方法。建议远志含总皂苷以细叶远志皂苷计应不低于2.5%,采收时间以春季为佳,生长期以2年以上为佳。

Objective：To develop a quantitative method for the determination of total saponins in Radix Polygalae by HPLC. Methods：An HPLC method was applied to separate and quantify tenuifolin in the products after alkaline hydrolysis. The samples were separated by an Alhima C18 column （4. 6 mm × 250 mm, 5 μm）using methanol - 0. 05% phosphoric acid（65：35 ）as mobile phase. The flow rate was 1.0 mL · min^-7 ,and the detection wavelength was set at 202 nm. Results： The number of theoretical plates calculated by tenuifolin peak was 2500. The regression equation of tenuifolin was Y=4. 242 × 10^6X - 1. 068 × 10^4（r =0. 9999） and the linear range was 100 - 1000 μg · mL^-1. The average recovery（n =6）of tenuifolin was 101.2% （RSD =3.6% ）. As the results shown,the content of total saponins calculated with the reference to tenuifolin in Radix Polygalae were 2. 42% - 3.71% from 14 different habitats, and they were relatively higher in the samples collected in March ,April, May and June than those in other months. The tenuifolin contents in 2 - 3 years cultivated samples were more than those in 1 year cultivated samples. Conclusion：The developed method is sensitive, rapid and accurate ,which can be used as quality control for Radix Polygalae and its preparations. It is suggested that the content of total saponins calculated with the reference to tenuifolin in Radix Polygalae is not less than 2. 5 %, and the sample should be harvested in spring after being euhivated more than two years.