Objective To set up a simple and accurate method for the determination of cinobufagin in dry skin of toadby HPLC.Methods Cinobufagin was determined by HPLC.Chromatographic conditions concluded Waters Xbridge C18 (5 μm,250 mm×4.6 mm) column and the mobile phase was consisted with the mixture of acetonitril-0.5 mol/L-NaH2PO4 (52∶48) (adjusted to pH 3.2 by phosphoric acid).Cinobufagin was detected at 294 nm wavelength.Flow rate was 1 mL/min; Column temperature was setted to room temperature.Results The calibration curves of Cinobufagin were in good linearity over the range of 2.5-15.0 μg (r =0.9999).Linear regression equation was that,y =-4×107x-10 196 (r =0.9999); solution precision and repeatability were good,the stability of the best was within 12 h; solution of the recycling experiment investigation showed that,the average recovery was 97.4％,RSD =2.8％.the content of medicinal materials in 5 batch of dry toad skin were 0.012,0.009,0.013,0.011,and 0.007 mg/g,the average concentration was 0.010 mg/g.Conclusion Chromatographic conditions of this established method is scientific,rapid and stable,which can be used as quality control method for dry toad skin medicine internal.%目的 建立了干蟾皮中华蟾酥毒基的含量测定方法.方法 色谱法:采用Waters Xbridge C18反相色谱柱(4.6 mm×250 mm,5μm);流动相:0.5％磷酸二氢钠溶液:乙腈(52∶48)加磷酸调pH 3.2;检测波长为294 nm;流速:1 mL/min;柱温为室温.结果 华蟾酥毒基在进样量2.5～15.0 μg(r=0.9999)范围内呈良好的线性关系,线性回归方程y=-4×107x-10 196(r=0.9999);溶液精密度、重复性均良好;在12h内测定,其稳定性最佳;溶液加样回收实验考察得平均回收率为97.4％,RSD=2.8％.5批干蟾皮药材的含量分别为0.012、0.009、0.013、0.011、0.007 mg/g,平均含量为0.010 mg/g.结论 本方法建立的色谱条件科学、快速、稳定,可作为干蟾皮药材内在质量的控制方法.
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