| Safflower is the dried flowers of Carthamus tinctorius L. Safflower is a traditional medicine in activating blood and resolving hemostasis.Numerous studies show that the main pharmacological compunds in the safflower is the water-soluble safflor yellow. In recent years,it was found that the hydroxy safflower yellow A is the main pharmacological compound in SY. However, report on the other ingredients in SY is relatively small.Study of other compunds in SY has a certain role in exploring the efficacy of SY.This article includes the following aspects:First of all,the extraction process of SY was studied. The experiment is did First by single factor, then we determined the best extraction prccess through orthogonal test. Secondly,we selected the appropriate material for the separation of SY. One separation method is to divide SY into several components according to the different polarity,another separation method, after washed impurity by water, Eluted SY with a certain concentration of ethanol. The parameters of the two separation methods was examined totally, and purified group1and group3by preparative high performance liquid chromatography repeatedly,then we obtained HSYA and SYB. Finally,we compared the antioxidant capacity of different substances group and monomer. The aim is to seek antioxidant active ingredients in SY. Results are as follows1. The extraction process of SY:reference substance is HSYA, total SY content was determined by UV spectrophotometer, The contents of HSYA and SYB were determined by High performance liquid chromatography。 The effect of the different extraction conditions on the extraction efficiency of total SY was evaluated comprehensively。The extraction process:select distilled water as extraction for60min at70℃, extract two times,25times solvent each time, the extraction rate of total SY was89.55%, the extraction rate of HSYA was93.93%, the extraction rate of SYB was85.67%.2. Select LSA-21for material to separate SY:by single factor, orthogonal test, the best technology was:the concentration of sample is0.8g/mL,and the ratio of diameter to height is1:15,the quality ratio of crude drugs to medicinal resin is1:2.5. One of the separating method is:2BV water was used to elute impurity,then,4BV10%ethanol was used to elute HSYA component, after that2.5BV30%ethanol was used to elute intermediate component,last we used4BV40%ethanol to get SYB component. The content of HSYA in HSYA component could reach30%and the content of SYB in SYB component group could reach28%. The other separating method is:2BV water was used to elute impurity,then,we used70%ethanol3BV to elute SY component directly.the content of SY in SY component was30%,the recovery rate was85%.3.The SYB purification process parameters were as follows:the SYB crude product quality to the volume of materials was9.09mg/mL, the ratio of diameter to height of column was1:7, water was washed2BV,10%ethanol was washed2BV20%ethanol was washed2BV。After then, we used30%ethanol to wash6BV and get the active ingredients SYB. Concentrated under vacuum freeze-drying, the purity of SYB product was60%and the recovery rate was more than60%.4. The result of antioxidant activity of safflower yellow showed that:SY could protect the degradation of2-deoxyribose induced by hydroxyl radical. HSYA and SYB were two main active compounds in SY. SY could also eliminate DPPH-effectively, but they contained other ingredients which could remove DPPH radical better than HSYA and SYB The results of mechanism test showed that, apart from direct scavenging the hydroxyl radicals produced by the Fenton reaction, SYB could also chelated with iron ions and block Fenton reaction. |
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