HPLC法测定蟾皮中蟾毒配基类成分的含量

目的用HPLC法测定蟾皮中蟾蜍灵、脂蟾毒配基、华蟾蜍精等成分的含量。方法样品经乙醇室温浸渍提取,采用HPLC法测定。色谱柱为DiamonsilTMC18(250 mm×4.6 mm,5μm),流动相为甲醇-乙腈-水(体积比为4∶1∶5),流速为1.0 mL.min-1,检测波长为296 nm,柱温为40℃。结果蟾蜍灵、脂蟾毒配基和华蟾蜍精分别在0.0848~0.5088、0.0884~0.5304和0.2000~1.2000μg内与色谱峰面积线性关系良好,相关系数r分别为0.9999、0.9992和1.0000。蟾蜍灵、脂蟾毒配基和华蟾蜍精的平均回收率分别为99.3%、99.6%和99.0%,RSD分别为0.5%、0.2%和0.6%(n=9)。结论HPLC法可用于蟾皮中蟾蜍灵、脂蟾毒配基和华蟾蜍精的含量测定。

Determination of the content of bufogenin in toad skin by HPLC

Objective To determinate the contents of bufalin, resibufogenin and cinobufagin in toad skin by HPLC. Methods A DiamonsilTM C18 column was used; the mobile phase was methanol, acetonitrile and aqua which volume ratio was 4:1:5 and the UV detection wavelength was 296 nm; the flow rate was 1.0 mL/min; the column temperature was 40 ℃. Results The linear range of bufalin was 0.0848–0.5088 μg (r=0.999 9), the average recovery was 99.3 %, and RSD=0.5 % (n=9); the linear range of resibufogen- in was 0.088 4–0.5304 μg (r=0.999 2), the average recovery was 99.6%, and RSD=0.2 %(n=9); the linear range of cinobufagin was 0.200 0–1.200 0 μg(r=1), the average recovery was 99.0 %,and RSD=0.6 % (n=9). Conclusions The method is stable, precise, reproductive and can be applied to the determination of bufalin, resibufogenin and cinobufagin in toad skin.