防风药材质量的多指标综合评价

Comprehensive quality evaluation of Saposhnikoviae radix based on multi-component contents determination

【目的】建立防风Saposhnikovia divaricata药材多指标成分同时定量分析的测定方法,并以多指标成分为评价指标,结合不同成分对防风药材抗炎活性的贡献,分析不同产地的防风药材质量,为防风药材质量的综合评价提供科学依据。【方法】采用高效液相色谱(HPLC)建立防风6种色原酮和4种香豆素成分同时定量的测定方法,采用聚类分析和主成分分析对防风药材质量进行综合评价;采用脂多糖诱导的RAW 264.7小鼠巨噬细胞炎症模型,分析防风10种化学成分与抗炎活性的相关关系。【结果】防风10种化学成分在各自范围内线性关系、准确度和分离度均良好。聚类分析和主成分分析把17批样品分为4类,其中以内蒙古S5和吉林S8~S11产地的2年生防风的综合质量相对较好;17批防风醇提物能不同程度地抑制脂多糖诱导的RAW264.7细胞分泌一氧化氮(NO)、白细胞介素6(IL-6)、白细胞介素1β(IL-1β);其中升麻素苷、5-O-甲基维斯阿米醇苷、升麻素、亥茅酚苷、5-O-甲基维斯阿米醇、欧前胡素与抗炎活性呈正相关;灰色关联度分析表明:防风抗炎活性是多种成分协同作用的结果,升麻素苷、5-O-甲基维斯阿米醇苷、升麻素、亥茅酚苷、5-O-甲基维斯阿米醇与3个细胞炎症指标(NO、IL-6和IL-1β)的关联度均大于0.8,是发挥抗炎作用的主要物质基础。【结论】本研究建立的防风多指标成分测定方法良好,可用于防风色原酮和香豆素成分的同时测定,结合抗炎活性与灰色关联度分析法建立了基于生物活性的防风药材质量综合评价模式,为建立防风药材质量综合评价的标准提供了新的方法和科学依据。

[Objective]To establish a method for quantitative analysis of multi-index components in Saposhnikoviae radix(Saposhnikovia divaricata),and for comprehensive quality evaluation of the medicinal materials from different areas,using the contents of multiple-index components as evaluation indexes,and the contribution of different components to the anti-inflammatory activity.[Method]The concentrations of 6 chromones and 4 coumarins from Saposhnikoviae radix were quantified by high-performance liquid chromatography(HPLC).The quality was comprehensively evaluated using cluster analysis and principal component analysis.The correlations between 10 chemical components and their anti-inflammatory activities were analyzed in a lipopolysaccharide-induced macrophage inflammation model in RAW 264.7 mice.[Result]The 10 kinds of chemical components had good linear relationship,accuracy,and separation in their respective ranges,respectively.17 batches of samples were grouped into 4 categories by cluster analysis and principal component analysis.The quality of 2-year-old S.divaricata grown in Inner Mongolia(S5)and Jilin(S8 to S11)were relatively better.17 batches of S.divaricata alcohol extracts differentially inhibited NO,IL-6,and IL-1βsecreted by lipopolysaccharide-induced RAW 264.7 cells.Prim-O-glucosylcimifugin,5-O-methylvisammioside,cimifugin,sec-O-glucosylhamaudol,5-O-methylvisamminol,and imperatorin were positively correlated with anti-inflammation.The anti-inflammatory effect of S.divaricata is the result of the combined action of several components,and the correlation degrees among prim-O-glucosylcimifugin,5-O-methylvisammioside,cimifugin,sec-O-glucosylhamaudol,and 5-O-methylvisamminol with three cellular indicators of inflammation(NO,IL-6,and IL-1β)were over 0.8,which were the main material basis for the anti-inflammatory activity,according to the gray correlation analysis.[Conclusion]A comprehensive evaluation model of the quality of S.divaricata based on biological activity and combining the anti-inflammatory activity and gray correlation analysis was established,which provided a new method and scientific basis for the establishment of quality standards for a comprehensive evaluation of the quality of S.divaricata.