Preparation And Sorghum Root Effective Part Enrichment Process Optimization

Based on the previous research,this paper was carried out in order to find the active component by analyzing the chemical elememts from the fractions,BES and WEAE,which are of anticoagulant activity of Sorghum root.The components in these two active fractions were separated and purified,and their structures were confirmed.Optimizing the effective fractions using 4-Hydroxybenzenecarboxylic acid and4-Hydroxy-3-methoxybenzoic acid as indexes.The anti-platelet aggregation activity of extract from sorghum root and compound evaluated respectively in vivo.Meanwhile,the antioxidant activity of extract were evaluated in vitro.The main results are as follows:1.TLC and HPLC methods found that there were two same chemical components in two active fractions(BES and WEAE).2.Two compounds coexisting in these two active fractions were obtained by column chromatography.They were identified as 4-Hydroxybenzenecarboxylic acid and 4-Hydroxy-3-methoxybenzoic acid respectively,also know as Vanillic acid,by MS,1HNMR and 13 CNMR spectra and comparing with related articles.These two compounds were isolated from Sorghum root for the first time.3.The result indicatied that the contents of 4-Hydroxybenzenecarboxylic acid and 4-Hydroxy-3-methoxybenzoic acid in two fractions,BES and WEAE,of fresh Sorghum root are lower than that of not fresh Sorghum root.4.Optimum enrichments of 4-Hydroxybenzenecarboxylic acid and4-Hydroxy-3-methoxybenzoic acid from BES and WEAE of Sorghum root respectively were obtained through the optimization tests using five macroporous adsorptive resins.The NKA-9 type of macroporous absorbent is suit able to separate these two compounds from the fractions of Sorghum root.The optimum parameters for the enrichment of 4-Hydroxybenzenecarboxylic acid and4-Hydroxy-3-methoxybenzoic acid from BES of Sorghum root are as follows:conditions of elution are 40% ethanol and 2 BV/h,weight ratio is 1:25,elution solvent volume is two times the volume of the column.The optimum parameters for another fraction are as follows: conditions of elution are distilled water and 3 BV/h,weight ratio is 1:50,elution solvent volume is two times the volume of the column.6.Studying on the methodology of these two fractions and establishing the HPLC specific chromatograms of BES and WEAE of ten varieties of Sorghum root.The experimental result indicated that the similarity of the chromatographic peak common to these two fractions was above 0.85.Moreover,the similarity of most peaks were above 0.9.5.Studying the inhibitory effect of the samples on platelet aggregation according to the maximum inhibition rates of platelet aggregation of blank group without any treatment and experimental group treated with samples using acetylsalicylic acid(aspirin)as positive control.Both of blank group and experimental group stimulated by ADP acting as aggregationantileptic.The inhibition rates of platelet aggregation was calculated by formula.It was showed that BES 10% ethanol fraction,WEAE pure water flow,WEAE 10% ethanol fraction,complexes containing mainly benzoic acid and vanillic acid can inhibit the platelet aggregation induced by ADP obviously.On the other hand,BES pure water flow,WEAE 30% ethanol fraction,40% ethanol fraction,75% ethanol fraction and WCE showed weak inhibitory effect on the platelet aggregation.7.Using Vc as positive control,study on the antioxidant activity of extract from Sorghum root was carried out via salicylic acid spectrophotometry,DPPH and colorimetric with potassium ferricyanide in vitro.The antioxidant activity of water extract was stronger than that of ethanol extract and correlated with the content of phenolic acids.