反相高效液相色谱法分析穿山龙中薯蓣皂苷元的含量

RP-HPLC analysis of diosgenin in Rhizoma Dioscoreae Nipponicae

目的:建立测定穿山龙中薯蓣皂苷元含量的反相高效液相色谱法,从而为控制穿山龙的质量提供参考。方法:采用双相酸水解法提取穿山龙中的薯蓣皂苷元;使用 Agilent Zorbax 色谱柱(4.6 mm×250 mm,5 μm),以乙腈-水(94:6)为流动相,检测波长为203 nm,流动相流速为1.0 mL·min^(-1),柱温为40℃。结果:样品中的薯蓣皂苷元可以与各干扰组分达到基线分离;用本法测定薯蓣皂苷元的线性范围为1.055～21.10 μg;重复性试验(n=6)RSD=1.7%;平均回收率(n=6)达到96.8%,RSD=1.5%;试验中测得穿山龙中薯蓣皂苷元的平均含量(n=6)为1.61%,RSD=1.7%。结论:本方法具有供试品溶液制备简便,检测灵敏,线性范围宽,重复性好等优点,适合于穿山龙中薯蓣皂苷元的含量测定。

Objective：To establish an RP - HPLC method for determination of diosgenin in Rhizoma Dioscoreae Nipponicae for quality control. Methods ： Diosgenin was extracted by two - phase acid hydrolysis from the dried material ; Agilent Zorbax Extend C18 column （4.6 mm × 250 mm, 5μm） was adopted at 40℃ with acetonitrile - pure water（94： 6）as mobile phase. The detection wavelength was 203 nm,and the flow rate was 1.0 mL· min^-1. Results：Diosgenin could be separated obviously from the intefferential components in the sample. The linearity of the calibration curve （r = 0. 9999）was good in the range of 1. 055 -21.10μg for diosgenin, RSD of repeated experiments （ n = 6） was 1.7 %, with the average recovery （ n = 6 ） reached 96.8 % （ RSD = 1.5 % ）. The average content （ n = 6） of diosgenin in Rhizoma Dioscoreae Nipponicae was 1.61% （ RSD = 1.7 % ）. Conclusions： This method can sensitively detect the diosgenin with a simple pretreatment, and has a broad linear range with good repeatability, which makes it being fit for determination of diosgenin in Rhizoma Dioscoreae Nipponicae.