基于主成分与止咳祛痰功效比较研究平贝母、伊贝母和川贝母

A Comparative Study on the Principal Component, Antitussive and Expectorant Effects of Chuan Fritillaria, Ping Fritillaria and Yi Fritillaria

目的:比较平贝母、伊贝母和川贝母的主成分和止咳祛痰功效的差异,并建立同时测定西贝母碱、贝母素乙、贝母素甲含量的方法。方法:采用HPLC-ELSD法,色谱柱为Hypersil Sino Chrom ODS-BPC18(4.6 mm×250 mm,5μm),流动相为甲醇(A)-0.03%二乙胺水溶液(B)梯度洗脱,柱温为25℃,流速为1.0 m L/min,漂移管温度为105℃,载气流速为2.0 L/min;采用小鼠氨水引咳及酚红祛痰实验,比较3种贝母止咳祛痰作用的差异。结果:西贝母碱、贝母素甲和贝母素乙分别在0.440~4.400μg(r=0.999,8),0.264~2.640μg(r=0.999,2),0.442~4.420μg(r=0.999,8)与色谱峰面积呈良好的线性关系,平均回收率分别为99.70%、98.26%和99.99%。川贝母与平贝母中可同时检出西贝母碱、贝母素甲及贝母素乙,含量分别为0.169,5、0.062,3、0.124,9 mg/g,0.049,3、0.031,6、0.025,9 mg/g,而伊贝母只能检出西贝母碱、贝母素甲,含量为0.401,8、0.083,4 mg/g;平贝母、伊贝母及川贝母的醇提物止咳祛痰作用比较,差异无统计学意义(P>0.05)。结论:所建含量测定方法精密度高,重复性好,为评价3种贝母内在质量提供科学依据。通过检测比较,3种贝母生物碱类主成分在其数和量上存在一定的差异,但其主要药效作用差异无统计学意义。

Objective： This paper reported the comparison of Ping Fritillaria, Yi Fritillaria and Chuan Fritillaria in the main component and antitussive and expectorant effects and the establishment of a method for simultaneous determination of sipeimine, peiminine and peimine contents in Chuan Fritillaria, Ping Fritillaria and Yi Fritillaria. Methods： The analyses were performed on an ultra-performance liquid chromatography with evaporative light scattering detection（UPLC-ELSD）. A Hypersil Sino Chrom ODS-BPC18（4.6×250 mm,5 μm） was used for all analysis. The investigated compounds were separated with gradient mobile phase consisting of methanol-0.03% diethylamine-water. The column temperature was set at 25 ℃; flow rate was 1.0 m L/min; the drift tube temperature of ELSD was set at 105 ℃ with the flow rate of 2.0 L/min. At the same time, three kinds of Fritillaria were compared through ammonium hydroxide causing cough and the amount of phlegm secreted in mouse.Results： The investigated compounds including sipeimine, peiminine and peimine had good linearity [0.440-4.400 μg（r=0.999,8）,0.264-2.640 μg（r=0.999,2）,0.442-4.420 μg（r=0.999,8）] over the tested ranges. The average recovery was 99.70%, 98.26% and 99.99%. Sipeimine, peimine and peiminine could be detected simultaneously in Chuan Fritillaria and Ping Fritillaria and the content was 0.169,5, 0.062,3, 0.124,9 mg/g; 0.049,3, 0.031,6, 0.025,9 mg/g.However, only the former two could be detected in Yi Fritillaria and the content was 0.401,8, 0.083,4 mg/g.Ethanol extracts of Ping Fritillaria, Yi Fritillaria and Chuan Fritillaria had similar eliminating cough and expec-torant activity, the difference was not significant（P〈0.05）. Conclusion： This method is accurate and have a good repeatability for the quality of fritillaria controlled. Three kinds of main fritillaria alkaloids components existed certain differences in the number and the quantity, but there was no statistical difference in the main pharmacological effects.