| Effects of Xiaoqu on the saccharification of purple corn were investigated in this paper. Wine color, total polyphenol index and composition and content of anthocyanins were detected in the alcohol fermentation with Saccharomyces cerevisiae. After fermentation, purple corn wine (PCW) was co-pigmented with ferulic acid and caffeic acid during aging. Effects of these co-pigments on the color and anthocyanins of purple corn wine were studied. And then the simulations of co-pigmentation system were established with ferulic acid and caffeic acid. The color changes of thse simulation systems were determined during storage. The variations of anthocyanins and their derivatives were discussed and their formation mechanism was revealed in this paper. PCW anthocyanins were separated and purificated, their antioxidant and antitumor activeity were compared. The main results and findings were summarized as follows:1. Wine starch A, which was from Shanxi, Hu County, was suitable for the saccharification of purple corn. After saccharification, the reducing sugar and total anthocyanins reached 232.29 mg/g and 433.7μg/g. They were significantly higher than which was saccharified by Suzhou wine starch (209.45 mg/g and 329.5μg/g).Purple corn wine was fermented by traditional xiaoqu technology. Change of color and total anthocyanin content during brewing of purple corn wine were investigated. The results showed that during the brewing, the content of total anthocyanins decreased and the anthocyanins degraded. Total anthocyanins content reached 150.3μg/ml and 142.3μg/ml at the end of alcohol fermentation. Lightness and Chroma increased during the 7 days' fermentation. Tatal anthocyanins positively correlated with Chroma, and negatively correlated with hue, but had no correlation with lightness. HPLC-MS was used to detect the anthocyanins content and compositions during brewing. Five anthocyanins were detected and they were characterized as cyanidin-3-glucoside, pelargonidin-3-glucoside, peonidin-3-glucoside, cyanidin-3-(6"-malonylglucoside) and peonidin-3-(6"-malonylglucoside). The relative content of these five anthocyanins were 29.46%,4.59%,22.36%,29.96% and 13.63% in the purple corn wine with wine starch A and 32.97%,4.01%,23.46%,2.38% and 17.18% in the wine with wine starch B. The content of each anthocyanin decreased and the non-acylated anthocyanins degraded significantly.2. Purple corn wine was aging at 15℃in duckness. The change of color and anthcoyanins were studied in the aging of purple corn wine. The results showed that total anthocyanins continuously degraded during aging. The color of purple corn wine turned from red to orange. After 140 day's aging, total anthocyanin in the samples with 0.1 mg/ml of ferulic acid and caffeic acid was 180.32 and 157.51μg/ml, which was increased by 38.48% and 20.97%, respectively more than those of contrast. Moreover, the more co-pigments were added, the higher total anthocyanins content were detected and the better co-pigmentation was obtained.HPLC-DAD results showed that the maximum absorption peak of these two kinds of wine shifted hypsochromically. New anthocyanins derivatives in purple corn wine were detected. After co-pigmentation with ferulic acid, four kinds of new anthocyanins derivatives were detected. Their retention time were 31.26,33.37,34.17 and 38.23 min, respectively, and their maximum absorption peak in visible spectrum were 506,506,502 and 506 nm, respectively. Five anthocyanins derivatives were found in the wine co-pigmented with caffeic acid. Their retention time were 28.53,30.62,31.41,34.41 and 38.43 min, respectively, and their maximum absorption peak in visible spectrum were 506,506,502,506 and 503nm.3. simulating the pH and alcohol degree of purple corn wine, the co-pigmentation of purple corn anthocyanins were established. The change of color and anthocyanins were researched during the storaged. The results showed that in the simulations of co-pigmentation system, the lightness decreased significantly after 140 day's storaged. The co-pigmented anthocyanins solution still showed red, while the control turned to colorless and its maximum absorption peak disappeared after 80 d.The HPLC-DAD results showed that the maximum absorption peak in both of the two simulations changed from 515 nm to 505 nm. Three anthocyanins derivatives were detected in these simulations after adding ferulic acid and caffeic acid. This also proved the co-pigmentation in purple corn wine. The results of HPLC-MS suggested that both ferulic acid and caffeic acid can directly react with purple corn anthocyanins to form anthocyanins derivatives. The anthocyanins derivatives after co-pigmentation with ferulic acid were characterized as Cyanidin-3-glucoside-vinylguaiacol, Pelargonidin-3-glucosede-vinylguaiacol and Peonidin-3-glucosede-vinylguaiacol. The anthocyanins derivatives after co-pigmentation with caffeic acid were characterized as Cyanidin-3-glucosede-vinylcatechol, Pelargonidin-3-glucosede-vinylcatechol and Peonidin-3-glucosede-vinylcatechol。The formation of anthocyanins derivatives was associated with the type of co-pigments and the reaction time (aging time of wine). Through the variation of anthocyanins and their derivatives, the anthocyanins were degraded significantly in the simulations with ferulic acid, and their derivatives were detected after 50 day'storage. These anthocyanins derivatives in the caffeic acid simulation were detected after 80 day's storage. It was suggested that ferulic acid was easier to reaction with anthocyanins to produce anthocyanins derivatives than caffeic acid.4. The isolation and purification of purple corn wine anthocyanins were isolated and purificated by macroporous resin HP-20 and Scphadex LH-20. The results showed that relative retention areas of Cy-3-G and Pn-3-G separated from purple corn anthocyanins arrived at 86.45% and 96.23%, respectively. The main groups in these two co-pigmented simulations were Cyanidin-3-glucoside-vinylguaiacol and Cyanidin-3-glucosede-vinylcatechol, their relative retention areas reached 69.70% and 70.43%, respectively.5. In the five antioxidant system, Cy-3-G showed higher antioxidant ability among these five anthocyanins groups. The total antioxidant capacity of anthocyanins derivatives Fa and Ca reached 15.01 and 13.90 U/g, respectively. And their superoxide radical-scavenging activity arrived at 80.65 and 87.56 U/g, respectively. DPPH radical-scavenging activity of anthocyanins derivatives Fa and Ca were 66.95% and 60.33% which was significantly higher than that of Pn-3-G. Hydroxyl radical scavenging activities of anthocyanins derivatives Fa and Ca reached 319.53 and 582.84 U/mg, respectively, which was significantly lower than that of Pn-3-G. The reducing power, DPPH radical-scavenging activity, lipid Peroxidation Inhibitor and hydroxyl radical scavenging activities of anthocyanins derivatives Ca were lower than anthocyanins derivatives Fa, but its total antioxidant capacity was higher than that of anthocyanins derivatives Fa.6. MTT assay was used to measure the suppressive effects of of anthocyanins and their derivatives on human gastric cancer cell MGC-823 and human intestinal cancer cell HCT-116. The IC50 of Pn-3-G on MGC-823 and HCT-116 reached 107.9μg/ml and 72.3μg/ml, respectively, which was lower than that of Cy-3-G. It was also suggested that these two anthocyanins have a better suppressive effect on HCT-116 than on MGC-823, and Pn-3-G may have higher antitumor activity than Cy-3-G. The IC50 of Anthocyanins derivatives Fa and Ca were 74.5 and 75.0 g/ml, which was significantly lower than other anthocyanins groups. Anthocyanins derivatives Fa had a significant suppressive effect on HCT-116. And its IC50 was only 55.6 g/ml, which was lower than that of anthocyanins derivatives Ca. the suppressive effect of these two anthocyanins derivatives on HCT-116 was significant higher than that of Cy-3-G,Pn-3-G and PCW. Anthocyanins showed a dose-dependent manner on antitumor activity. After administration of anthocyanins derivatives, MGC-823 and HCT-116 cell line turn rounded and floated, proliferated slowly, and its cytomembrane collapse. Fluorescence staining results showed that HCT-116 performed karyopyknosis, inregular changes of caryotype and nuclear fragmentation after addition of anthocyanins derivatives.
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