Elucidation of production mechanism and functionality of the decolorized compounds from acylated anthocyanins under interstine condition

采用XAD-2000、PVP、制备型高效液相色谱等柱层析分离纯化相结合的方法，在较短时间内较好地实现了紫色甘薯贮藏根和红色牵牛花花瓣的大规模纯色素制备。这些被证实是酰基化花青素与3-槐花苷-5-葡萄糖苷作为一个共同框架。将分离得到的酰基化花青素放置于肠道环境（pH 6.8, 37℃，黑暗）24h后，其褪色（不稳定）程度依次为YGM-5b > SOA-4 > YGM-3和6 > SOA-6。这一结果不同于AGH抑制活性的顺序（SOA-4 > SOA-6 > YGM-6和3），并且不相关使得不稳定性和AGH抑制活性清晰。在该反应溶液中，发现了三种被认为是无氢碱(A)、假碱(B)和查尔酮(C)的成分。反应的弧长为A→B→C，其弧长为A:B:C弧长为1:1:2,24小时后YGM-5b和SOA-6的弧长为…所有色素都含有咖啡酸。接下来，在肠道环境下对SOA-4进行大量褪色，并尝试用制备型高效液相色谱法分离纯化C。因此，当使用中性磷酸盐缓冲溶液体系作为洗脱液时，可以将C与其他组分分离，并且C的分离具有足够的重现性。此外，质谱分析结果强烈提示组分C是由SOA-4水合异构化生成的查尔酮。目前正计划进行脱盐和核磁共振结构分析。此外，对剩余组分A和b的分离也进行了研究。认为SOA-4、-6和YGM-3、-6所共有的3-酰基化糖侧链结构（6-咖啡因-树马糖部分）对AGH抑制活性的表达是必不可少的，因为SOA-4的活性不随褪色而改变（糖基部分的结构转变）。少

Large-scale preparation of pure pigments was able to be comparatively performed from the purple sweet potato storage roots and the red morning glory petals in a short time by combining isolation and purification by column chromatographies such as XAD-2000, PVP and preparative HPLC. These were confirmed to be acylated anthocyanins with 3-sophoroside-5-glucoside as a common frame. As a result of putting the isolated acylated anthocyanins for 24 hours on the intestinal tract environment (pH 6.8, 37℃, darkness), the degree of fading (instability) was the order of YGM-5b > SOA-4 > YGM-3 and 6 > SOA-6. This result differs from the order (SOA-4 > SOA-6 > YGM-6 and 3) of AGH inhibitory activity, and not correlating made instability and AGH inhibitory activity clear.In this reaction solution, three kinds of components considered to be an anhydrobase (A), a pseudobase (B), and a chalcone (C) were found. The reaction progressed like A→B→C with time and the compositions led to A:B:C≒1:1:2 except f … More or YGM-5b and SOA-6 after 24 hours. All pigments were holding caffeic acid.Next, SOA-4 was faded under the intestinal tract environment in large quantities, and isolation and purification of C were tried using preparative HPLC. Consequently, when a neutral phosphate buffer solution system was used as eluent, C and other components could be separated, and C has been isolated with sufficient reproducibility. Moreover, it was strongly suggested as a result of mass analysis that component C is a chalcone which was generated from SOA-4 through hydrationand isomerization. Now, the desalting and the structural analysis by NMR are planned. Moreover, it is under examination also about isolation of the remaining components A and B.It was thought that its 3-acylateded sugar side-chain structure (6-cafffeoylshophorose moiety) which SOA-4, -6 and YGM-3, and -6 have in common was indispensable to the expression of AGH inhibitory activity since the activity of SOA-4 doesnot change with fading (structural transformation of the aglycon part). Less