Studies On Evaluation Of Scavenging Superoxide Free Radial Activity For Salvianolic Acids

Water-soluble components such as salvianolic acids came from Radix Salviamiltiorrhiza had strong ability of scavenging free radials. However, more research about theability of scavenging superoxide free radial（O₂^-·） for monomer salvianolic acids componentshould been made. There were many methods for evaluation the ability of scavenging O₂^-· invitro. But different evaluation methods gived significant different results. Four kinds ofmethods for evaluation the ability of scavenging O₂^-· in vitro which referenced from literaturewere researched and improved In this paper. and they were respectively applied for theactivity assay for salvianolic acids. Meanwhile, the optimal evaluation system of salvianolicacid scavenging O₂^-· activity was confirmed.The ability of scavenging O₂^-· of SalB was assayed by means of four kinds ofevaluation methods in vitro respectively, such as NBT-illumination method, pyrogallolautoxidation, xanthine-xanthine oxidase-cytochrome C method （X-XOD-Cyt C） andNADH-PMS-NBT method. The determination conditions were optimized withspectrophotometric method in different system. Then, the reproducibility and sensitivity offour kinds of evaluation methods were compared. Finally, the ability of scavenging O₂^-· offive important monomer salvianolic acid such as SalvianolicA（SalA）, SalvianolicB（SalB）,Rosmarinic acid （RosA）, Protocatechuic aldehyde（Pca）, Lithospemic acid（Lith） wereevaluated systematically.The results indicated that：In each evaluation system, the results of the ability ofscavenging O₂^-· for SalB was effected greatly by determination conditions. Thereproducibility and sensitivity of four kinds of determination system was improved based onbuilding optimal evaluation system. The evaluation results indicated that t five kinds ofsalvianolic acid not only had strong ability on scavenging superoxide free radials, but alsoshowed significant dose-effect relationship. The diversity was confirmed in differentevaluation system by which activity comparison of five kinds of salvianolic acid, and thenecessity was proved that building optimal evaluation system to assay salvianolic acid.Comprehensive comparison with the reproducibility and sensitivity of four kinds of evaluation system, the optimal evaluation system were obtained as NBT-illumination method.It was suitable for evaluating the ability of scavenging O₂^-· for salvianolic acid, and it has thehighest reproducibility and sensitivity. The order of the ability of scavenging O₂^-· formonomer salvianolic acid were: RosA>SalA>SalB>Pca>Lith≈rutin.In addition, XOD activity was obviously inhibited by SalA，Lith and Ros A in theX-XOD-Cyt C system, which could provide evidence for drug development of hyperuricemiaand gout.