反相HPLC法同时测定牛黄消炎丸中华蟾酥毒基、酯蟾酥配基的含量

Simultaneous RP-HPLC Determination of Cinobufagin and Resibufogenin in Niuhang Xiaoyan Pills

目的:建立用反相HPLC法同时测定牛黄清炎丸中华蟾酥毒基、酯蟾毒配基含量的方法。方法:采用反相HPLC法测定华蟾酥毒基、酯蟾毒配基的含量,色谱柱:以十八烷基硅烷键合硅胶为填充剂(25mm×5μm,麦科菲);乙腈-0.5%磷酸二氢钾溶液(40:60)(用磷酸调pH值至3.2)为流动相。检测波长为296nm,柱温为400℃。结果:华蟾酥毒基进样量在0.2578μg～1.5468μg范围内,与峰面积呈良好的线性关系,r=0.9999,回归方程Y=768131x-16329,平均回收率为97.67%(n=6);酯蟾毒配基进样量在0.2482μg～1.4892μg范围内,与峰面积呈良好的线性关系,r=0.9999,回归方程y=727051x-18342,平均回收率为100.07%(n=6)。结论:定量方法操作简便、结果可靠,可用于牛黄消炎丸中蟾酥的质量控制。

Objective：To establish an RP -HPLC method for determination of cinobufagin and recibufogenin in in Niuhuang Xiaoyan Pills. Methods ： Using Kromasil C 18 column （ 250mm × 4.6mm, 5 μm）, mobile phase ： acetonitrile - 0.5 % potassium dihydrogen phosphate （ 50 ： 50,adjusted to pit 3.2 with phosphoric acid）, flow rate： 1. 0 mL · min^-1, detection wavelength： 296nm, column temperature： 40℃ . Results： The linear ranges of the determination of cinobufagin was （0. 2578 - 1. 5468 ） μg and recibufogenin was 0. 2482 - 1. 4892μg. The average recovery of cinobufagin was 97.67% and RSD was 1.78% ,the average recovery of recibufogenin was 100.07% and RSD was 1.89% . Conclusions： The method is simple and accurate for the determination of cinobufagin and recibufogenin in Niuhang Xiaoyan Pills. It can he used as quality control in medicinal preparation.