高效液相色谱-质谱联用法测定喉症丸中华蟾酥毒基和脂蟾毒配基

Determination of cinobufagin and resibufogenin in Houzheng pills by highperformance liquid chromatography-mass spectrometry

建立高效液相色谱-质谱联用法(HPLC-MS)测定喉症丸中华蟾酥毒基和脂蟾毒配基含量的方法。采用HPLC-MS正离子多反应监测模式进行测定,色谱柱为Thermo Hypersil BDS C18(250.0 mm×4.6 mm,5μm),流动相为乙腈-1%乙酸溶液(38∶62),流速为0.5 mL/min,检测波长为296 nm,柱温为20℃。质谱条件:鞘气流速40 arb,辅气流速5 arb,喷雾电压4.5 kV,毛细管温度300℃。结果显示,华蟾酥毒基和脂蟾毒配基的检测质量浓度线性范围均为50~300μg/mL(r=0.9992),重复性试验、精密度试验、稳定性(12 h)试验的RSD均小于1.00%(n=5或n=6),加样回收率分别为101.8%(RSD为2.61%,n=6)和100.9%(RSD为3.20%,n=6)。该方法灵敏度高、重复性好,能准确测定喉症丸中华蟾酥毒基和脂蟾毒配基的含量,可用于喉症丸的质量控制。

To establish a method for simultaneous determination of cinobufagin and resibufogenin in Houzheng pills.The sample was separated by Thermo-BDS Hypersil C18(250.0 mm×4.6 mm,5μm)and detected by liquid chromatography�mass spectrometry.The mobile phase was acetonitrile-1%acetic acid solution(38∶62),the flow rate was 0.5 mL/min,the column temperature was 20℃.Sheath gas flow rate was 40 arb and aux gas flow rate was 5 arb.Spray voltage was 4.5 kV,the capillary temperature was 300℃.The linear range of cinobufagin and resibufogenin was 50-300μg/mL(r=0.9992),the RSD of repeatability,precision and stability(12 h)experiments were all less than 1.00%(n=5 or n=6).The recovery rate of sample addition were 101.8%(RSD=2.61%)and 100.9%(RSD=3.20%),respectively.The method is sensitive and reproducible,and can be used for the simultaneous determination of cinobufagin and resibufogenin in Houzheng pills.