Formation And Bioactivity Evaluation Of Anthocyanin Derivatives In Mulberry Wine

As a kind of medicine and food,mulberry is attracting more and more attention.In particular,mulberry anthocyanin as the main active ingredient of mulberry,its anti-oxidation,anti-tumor and prevention of diabetes and other physiological activities were reported successively.Mulberry wine is an emerging fruit wine with high nutritional value,which can meet the modern people’s pursuit of nutritional and healthy diet.However,the anthocyanin is unstable,and it degrads in food processing.In this research,the effects of aging methods and copigmentation with phenolic aicd on the anthocyanins and derivatives in mulberry wine were studied,as well as the simulated gastri snd intestinal digestion and oxidative injury cell model were established.The bioactivity of anthocyanin derivatives were investigated.The main results were as follows.1.Mulberry wine was aged in stainless steel cans and oak barrels.The total phenol content,Scavenging DPPH·and ABTS·ability,color,anthocyanins and derivatives content were investigated in the aging.The results showed that there were two anthocyanin monomers in mulberry wine,namely cyanidin-3-glucoside and cyanidin-3-rutinose.In the stainless steel barrel,the cyanidin-3-glucoside and cyanidin-3-rutinoside decreased by 94.12%and 92.14%,respectively.In the oak barrel,the cyanidin-3-glucoside and cyanidin-3-rutinoside decreased respectively.96.82%,96.65%in aging 80 days.Barrel mulberry wine is beneficial to the retention of anthocyanin monomers in the early stage of aging.Two anthocyanin derivatives were detected in agedmulberrywine.Theywerevinylpyranocyanidin-3-glucoside-catecholand vinylpyranocyanidin-3-rutinoside-catechol.The content of vinylpyranocyanidin-3-glucoside-catechol was 1.36 mg cyanidin-3-glucoside/L on day 60.The color of mulberry wine depend on the content of polymeric anthocyanin.Stainless steel can make mulberry wine get better color.The antioxidant activity of mulberry wine was closely related to the content of total phenols and anthocyanin and derivatives.2.Caffeic acid,ferulic acid,sinapic acid and p-coumaric acid were used in the copigmentation of mulberry wine.The results indicated that the more stable derivatives were produce in the aging of mulberry wine with the selected four phenolic aicds,which can increased the color tonality of mulberry wine.After copigmentation,the polymeric anthocyanin content was significantlt increased than the control,the polymeric anthocyanin content reached 90%in the mulberry wine copigmented with sinapic acid.The vinylpyranocyanidin-3-glucoside-catechol and,vinylpyranocyanidin-3-rutinoside-catechol were the main anthocyanin derivatives in mulberry wine copigmented with caffeic acid,which were 9.2 and 13.7 times that of the control group;the main products were vinylpyranocyanidin-3-glucoside-guaiacol and vinylpyranocyanidin-3-rutinoside-guaiacol were detected in the mulberry wine copigmented with ferulic acid,and reached the highest value at 170 days,which was 13.29 mg cyanidin-3-glucoside/L and 6.14 mg cyanidin-3-rutinoside/L;The vinylpyranocyanidin-3-glucoside-（2,6-Dimethoxyphenol）was mainly produced by sinapic acid,and reached the highest value of 17.84 mg cyanidin-3-glucoside/L,vinylpyranocyanidin-3-rutinoside-（2,6-Dimethoxyphenol）,reaching the highest value of 8.84 mg mg cyanidin-3-rutinoside/L on 110 days;The main yields of vinylpyranocyanidin-3-glucoside-phenol and vinylpyranocyanidin-3-rutinoside-phenol after p-coumaric acid-assisted color reached the highest values at 170 d after aging,which were 4.74 mg cyanidin-3-glucoside/L and 1.82 mg cyanidin-3-rutinoside/L.3.The simulated stomach and intestinal digestion was established to investigate the digestibility characteristics of anthocyanin derivatives.The results showed that the anthocyanin and derivatives were relatively stable in the simulated gastric digestion.The content of total phenols,anthocyanins and derivatives was not siginificantly changed,but the antioxidant activity（scavenging ability of DPPH·and ABTS·and reducing power）both increased significantly from4.95 mg/100mL,8.99 mg/100mL,and 0.385 to 5.56 mg/100mL,9.27 mg/100mL,and 0.400,respectively,from 4.92 mg/kg.100mL,6.67 mg/100mL and 0.345 increased to 5.36 mg/100mL,7.40 mg/100mL and 0.374;In the simulated intestinal digestion,a large amount of anthocyanins were degraded in to caffeic acid,ferulic acid,sinapic acid and p-coumaric acid,while anthocyanin derivatives still exhibit high stability.In addition,both anthocyanins and derivatives showed good antioxidant ability during the simulated digestion.4.An oxidative stress injury model of vascular endothelial cells induced by H₂O₂ was established to investigate the protective effect of anthocyanin derivatives on oxidative stress damaged cells.The results showed that mulberry anthocyanins and derivatives reduced XO-1（xanthine oxidase-1,XO-1）and MDA（Malonic Dialdehyde,MDA）in oxidative stress damaged cells.Increase intracellular SOD activity,HO-1（Heme oxygenase-1,HO-1）content,increase oxidative stress injury cell mitochondrial membrane potential,and other pathways to reduce the level of ROS（Reactivexygen Specials,ROS）in Human Umbilical Vein Endothelial Cells（HUVEC）,inhibit apoptosis,Thus protecting HUVEC from oxidative stress caused by H₂O₂.The quantitative effects of anthocyanins and their derivatives in protecting cell oxidative stress were observed.