目的 建立蟾酥原浆HPLC指纹图谱及原浆中8种成分含量测定方法，对不同产地蟾酥原浆进行成分含量差异比较，同时进行质量评价。方法 以乙腈-水为流动相，梯度洗脱，检测波长296 nm，体积流量1.2 mL/min，柱温30℃，进样量10 μL；建立蟾酥原浆指纹图谱，测定8个成分含量，对结果进行聚类分析和主成分分析。结果 18批样品指纹图谱中有12个共有峰，各样品相似度有所差异，8个成分在各自范围内线性关系良好（r²≥0.9995），精密度、重复性RSD<0.5%，在30 h内稳定性较好（RSD<0.7%），日蟾毒它灵、沙蟾毒精、远华蟾蜍精、蟾毒它灵、华蟾毒它灵、蟾毒灵、华蟾酥毒基和脂蟾毒配基回收率平均值分别为103.7%、103.0%、102.9%、103.0%、103.9%、100.3%、103.4%、103.2%，且RSD均小于1.2%，8个成分含量测定结果及聚类分析和主成分分析结果可知不同产地原浆质量有所差异。结论 该方法简便可靠，为蟾酥原浆质量把控提供一定依据及参考。

Objective To establish the HPLC fingerprint of Bufonis Venenum pulp and the determination methods of eight components in Bufonis Venenum pulp, and compare the differences of components of Bufonis Venenum pulp in different origins for quality evaluation. Methods The mobile phase was acetonitrile and water with a gradient elution program. The detection wavelength was set at 296 nm. The flow rate was 1.2 mL/min. The column temperature was set at 30℃, and 10 μL of the test solution was injected. HPLC fingerprint of Bufonis Venenum pulp was established and eight components were determined. The results were analyzed by cluster analysis and principal component analysis. Results There were 12 common peaks in the HPLC fingerprints of 18 batches of samples, and the similarity of each sample was different. The linear relationship of eight components was good (r² > 0.999 5), RSD of precision and repeatability was less than 0.5%, and the stability was also good within 30 h (RSD < 0.7%). The average recoveries of gamabufotalin, arenobufagin, telocinobufagin, bufotaline, cinobufotalin, bufalin, cinobufagin and resibufogenin were 103.7%, 103.0%, 102.9%, 103.0%, 103.9%, 100.3%, 103.4%, and 103.2% respectively, and RSD was all less than 1.2%. The results of the content determination, cluster analysis and principal component analysis of eight components showed that Bufonis Venenum pulp in different habitats were different from each other. Conclusion The method is simple and reliable, which can provide some basis and reference for quality control of Bufonis Venenum pulp.

R286.2

浙江省大学生科技创新活动计划暨新苗人才计划项目（2017R410043）