The Comparison Of Antioxidative Ingredients In The Aqueous Extract Of Crude And Wine-processed P.cyrtonema Hua

Polygonatum cyrtonema Hua,a common medicinal and edible homologous plant,belongs to the genus Polygonatum Mill.(Liliaceae),which can be processed by nine steaming and nine processing processes with rice wine.At present,the comparison of pharmacological activity and composition difference of crude and wine-processed Polygonatum cyrtonema Hua is mainly focused on the composition of alcohol extract,but the comparative study of active composition related to aqueous extract is lacking.Therefore,in this paper,the changes of the antioxidant activity and related composition of crude and wine-processed P.cyrtonema Hua aqueous extract(hereinafter referred to as the aqueous extract before and after processing)were compared.In this paper,firstly,the aqueous extract of crude and wine-processed P.cyrtonema Hua was obtained by hot water extraction,and its antioxidant activity in vitro was compared.The total antioxidant capacity(T-AOC)and free radical scavenging ability of2,2-diphenyl-1-picrylhydrazyl(DPPH)of the aqueous extract were significantly increased after nine steam and nine processing.After that,the aqueous extract was precipitated by alcohol,and the supernatant was centrifuged,concentrated and dried to obtain the fat-soluble part.The alcohol precipitate was then deproteinized,dialysed and dried to obtain the polysaccharide part.By measuring the indexes of T-AOC and DPPH,it was found that the antioxidant activity of the fat-soluble and polysaccharide parts of wine-processed P.cyrtonema Hua aqueous extract were significantly higher than those of crude P.cyrtonema Hua aqueous extract.For fat-soluble part,the optimized HPLC analysis conditions were used.Chromatographic column was ZORBAX SB-C18(5μm,4.6×250 mm),injection was20μL,wavelength was 203 nm,column temperature was 30℃,flow rate was 1.0m L/min and gradient elution of mobile phase water and acetonitrile.Then,20 known compounds were found by HPLC-ESI-MS/MS.Among the known compounds,the content of Guanosine and L-tryptophan were significantly reduced while the content of Adenosine、Cyclo dileucyl(or isoleucyl)、Polygonatine B、4-(1-methyl-dodecyl)-benzene sulfonic acid and Linoleic acid were significantly improved after processing.For the polysaccharide part,HPGPC method was used to investigate the molecular weight of the polysaccharide before and after processing.It was found that the polysaccharide before processing(PCPC)had a uniform symmetrical peak with a molecular weight of 4842 Da.After processing,the polysaccharide(PCPW)had three overlapping peaks,and the molecular weights were 1.69×10~7Da,7.96×10~6Da and3.39×10~6.The analysis of monosaccharide composition showed that PCPC was mainly fructose(93.57%)and glucose(6.43%).PCPW was mainly fructose(95.35%),and it also contained glucose(3.00%),rhamnose(0.50%),galactose(0.05%),mannose(0.83%),galacturonic acid(0.15%)and glucuronic acid(0.14%).Further analysis on DEAE Cellulose-52 column showed that PCPC-1,a neutral polysaccharide eluted by distilled water,was mainly contained in PCPC.PCPW contained neutral polysaccharide PCPW-1 eluted by distilled water and acidic polysaccharide PCPW-2～PCPW-7 eluted by sodium chloride solution of different concentrations.In conclusion,the present study found that the antioxidant activity of P.cyrtonema Hua aqueous extract significantly increased after processing,which maight be related to the significant changes in the composition of fat-soluble part and polysaccharide part in P.cyrtonema Hua aqueous extract.