Separation Of Salvianolic Acid Compounds And Study On Antioxidant Activity

Radix salvia mitihorriza was one of the traditional Chinese medicine, and have effectson promoting blood flow, regulating menstruation, and relieving pain, clearing hot andcalming the nerves. A great deal of pharmacology researches discovered that thewater-soluble composition played an important role in effects of Radix Salviae Miltiorrhirae,they had strong scavenging activities of radical、antioxidation activities and related variousbioactivities, and they have been widely used in curing the disease of cardiovascular systemclinically. Under the guidence of HPLC fingerprinting, we selected a method for separatingsalvianolic acid compounds by color spectrum modern separation techniques andinvestigated the antioxidative activity of single compound and the relation between theantioxidant activities and concentration by five vitro-antio xidative methods.Through the method which combined extraction and silica、CG161、Sephadex LH-20preparative chromatography, we established a method for separating salvianolic acidcompounds, and got three salvianolic acid compounds, they are Rosmarinic acid、salvianolicacid B、salvianolic acid A by identifing their structrues according to IR、MS、NMR spectrumand so on.We chosed the systems of Fenton、pyrogallol antioxidation、DPPH、reducing power andyolk lipid peroxidation to evaluate the antioxidation activity of three salvianolic acidcompounds and the relation between the antioxidant activities and concentration of salvianolicacid compounds by spectroscopy. The results showed that The scavenging activities of threesalvianolic acid compounds to hydroxyl radical、superoxide anion and DPPH, reducingpower and the inhibitory effect on lipid peroxidation were very strong, and the antioxidantactivities depended on salvianolic acid compounds content, in a remarkable concentration-dependent manner. Apart from pyrogallol antioxidation, their were much stronger than thesame concentration Vc and rutin in other four antioxidative assays, the order of antioxidativeactivity is successively: salvianolic acid A>Rosmarinic acid>salvianolic acid B. The relationbetween the scavenging activities and concentration and concentration scavenging50%of·OH of salvianolic acid B、Rosmarinic acid、salvianolic acid A was y_{Sal B}=225.33x-24.721, r²=0.9919, IC₅₀=0.3371g/L; y_{Ros A}=496.37x-36.545, r²=0.9813, IC₅₀=0.1742g/L; y_{Sal A}=583.5x-29.411, r²=0.9944, IC₅₀=0.1363g/L respectively; the relation between the scavengingactivities and concentration and the concentration scavenging50%of O₂^-· was y_{Sal B}=12.034x+0.3919, r²=0.9708, IC₅₀=4.1223g/L; y_{Ros A}=12.403x+1.6514, r²=0.9869, IC₅₀=3.8984g/L; y_{Sal A}=11.865x+10.432, r²=0.9878, IC₅₀=3.3342g/L respectively; the relation between thescavenging activities and concentration and the concentration scavenging50%of DPPH· wasy_{Sal B}=3513.8x+1.567, r²=0.9990, IC₅₀=0.0138g/L; y_{Ros A}=3699.6x+4.086, r²=0.9906, IC₅₀=0.0124g/L; y_{Sal A}=4255x+3.18, r²=0.9995, IC₅₀=0.0110g/L respectively; the relation betweenthe inhibitory effect and concentration and the concentration inhibiting50%of lipidperoxidation was y_{Sal B}=218.95x-6.672, r²=0.9833, IC₅₀=0.2588g/L; y_{Ros A}=279.04x-7.358,r²=0.9873, IC₅₀=0.2056g/L; y_{Sal A}=210.3x+27.128, r²=0.9856, IC₅₀=0.1088g/L respectively;the relation between the reducing power and concentration was y_{Sal B}=7.41x+0.0699，r²=0.992；y_{Ros A}=10.355x+0.0746，r²=0.9997；y_{Sal A}=11.65x+0.0821，r²=0.9918respectively.The inhibitory effect on yolk lipid peroxidation and reducing power of three salvianolicacid compounds are related to the scavenging radical activities, the better scavenging radicalswas and the better reducing power and the inhibitory effect on lipid peroxidation was.