浙贝母化痰效应成分指数的建立与应用

Establishment and Application of Fritillariae Thunbergii Bulbus Resolving Phlegm Effect-constituent Index

目的:建立一种以化痰为主要效应的,既能关联浙贝母临床功效,又不增加质控难度的基于效应成分指数的浙贝母质量评价与控制新技术,使浙贝母品质评价与控制新技术更好地助力于浙贝母产业发展。方法:收集不同品种、不同产地的浙贝母药材,确定浙贝母药材中活性成分贝母素甲和贝母素乙含量的上下限,分析条件为Hypersil ODS-3色谱柱(4.6 mm×250 mm,5μm),流动相乙腈-水-二乙胺(70∶30∶0.03),流速0.8 m L·min^(-1);Alltech 2000-ELSD,漂移管温度85℃,载气流速2.2 L·min^(-1)。在确定的浓度范围内,配制不同浓度的贝母素甲、贝母素乙对照品溶液,采用小鼠气管酚红排痰实验法,以酚红浓度为化痰效应指标,将小鼠酚红排泌量与贝母素甲(X1),贝母素乙(X2)的质量浓度进行线性关系分析,拟合贝母素甲、贝母素乙与化痰效应指标之间的量-效关系方程f(Xi),构建浙贝母的化痰效应指数并验证。结果:浙贝母主要产区中贝母素甲的质量分数(以干燥品计)在0.046 5%~0.092 5%,贝母素乙的质量分数在0.017 5%~0.054 3%。给药溶液质量分数的范围为贝母素甲0.04%~0.10%,贝母素乙0.02%~0.06%。小鼠气管段酚红排痰实验结果表明,贝母素甲在0.09~0.23 g·L^(-1),贝母素乙在0.045~0.135 g·L^(-1)与小鼠气管酚红浓度有较好的线性关系(r甲=0.994 7,r乙=0.997 8)。根据拟合的量效方程构建的浙贝母化痰效应成分指数能很好地通过成分的量表征其祛痰的药效活性。结论:化痰效应成分指数的建立,不仅做到了量效关联,更实现了以量观效,对浙贝母品质评控、产业化发展及临床合理用药具有较好的指导意义和应用前景。

Objective： To establish a new technology for Fritillariae Thunbergii Bulbus quality control based on effect-constituent index,with resolving phlegm as the main effects and associated with Fritillariae Thunbergii Bulbus clinical efficacy,and make the new quality control method better promote the development of Fritillariae Thunbergii Bulbus industry. Method： Fritillariae Thunbergii Bulbus samples with different varieties and different origins were collected and the upper and lower limits of active ingredients contents（ peimine and peiminine） were determined. The analysis conditions were as follows： Hypersil ODS-3 column（ 4. 6 mm × 250 mm,5 μm）; with acetonitrile-water-diethylamine（ 70 ∶ 30 ∶ 0. 03） as the mobile phase; flow rate at 0. 8 m L·min^（-1）; the drift tube temperature of Alltech2000-ELSD detector was 85 ℃,carrier gas flow rate at 2. 2 L·min^（-1）. In the range of concentrations determined, different concentrations of peimine and peiminine standard solution were prepared. With the concentration of phenol red as the phlegm effect indicator in the mouse tracheal phenol red expectoration experiment, the linear relationship between the mouse phenol red excretion and the mass concentration of peimine and peiminine was analyzed to fit the dose-response equation f（ Xi） between peimine,peiminine and resolving phlegm effect indicators. Thus the resolving phlegm effect index was constructed and verified. Result： In the main producing areas of Fritillariae Thunbergii Bulbus,the content of peimine was between0. 046 5%-0. 092 5%,and the content of peiminine was between 0. 017 5%-0. 054 3%. Therefore,the range of dosing solution concentration was： peimine content in the range of 0. 04%-0. 10% and peiminine content in the range of 0. 02%-0. 06%. The results of mouse tracheal phenol red expectoration experiment showed that peimine in 0. 09-0. 23 g·L^（-1） and peiminine in 0. 045-0. 135 g · L^（-1） range had a good linear relationship with the concentration of mouse tracheal phenol red（ rpeimine= 0. 994 7,rpeiminine= 0. 997 8）. The established resolving phlegm effectconstituent index of Fritillariae Thunbergii Bulbus according to the fitted dose-response equation can characterize its expectorant efficacy through the contents of the compositions. Conclusion： The establishment of resolving phlegm effect-constituent index,has not only achieved dose-efficacy correlation,but also achieved the concept of observing effect by the composition contents. Therefore, it has better guiding significance and application prospects for Fritillariae Thunbergii Bulbus quality control assessment,industrial development and clinical rational drug use.