刺葡萄皮中花色苷的分离纯化与结构鉴定  

Isolation, purification and structure identification of anthocyanins from Vitis davidii Foex skin

作　　者：王维茜 邓洁红[1,2] 石星波[1,2] 刘永红[3] 

机构地区：[1]湖南农业大学食品科技学院,长沙410128 [2]食品科学与生物技术湖南省重点实验室,长沙410128 [3]湖南生物机电职业技术学院,长沙410127

出　　处：《农业工程学报》2016年第4期296-301,共6页Transactions of the Chinese Society of Agricultural Engineering

基　　金：国家自然科学基金(31271836);湖南省教育厅研究生科研创新课题(CX2015B264)

摘　　要：为研究刺葡萄花色苷的结构及其纯化分离的柱层析法,将刺葡萄色素粗提液依次经大孔树脂HP-20、聚酰胺树脂、葡聚糖凝胶Sephadex LH-20吸附纯化,利用超高效液相色谱三重四级杆飞行时间质谱联用技术对分离所得花色苷进行结构鉴定,并运用荧光光度法探索荧光图谱与花色苷结构的关系。研究发现,聚酰胺树脂对部分花色苷产生了吸附作用,而Sephadex LH-20凝胶能起到较好的分离作用,最终得到3种色素,经鉴定,确定色素I为锦葵素-3,5-O-双葡萄糖苷,通过质谱信息初步确定色素III可能为锦葵素-3,5-O-双葡萄糖苷-香豆酰,色素IV可能为飞燕草素-3-O-芸香糖苷和锦葵素-3-O-芸香糖苷的混合物,经高效液相色谱以归一法计算峰面积,色素I和色素III的纯度分别达到了98.64%、98.33%,得率分别为0.114%和0.076%。研究结果为花色苷的分离及鉴定提供参考。Anthocyanins are a widespread source of natural pigment of foods. Due to their bright attractive color, high solubility in water and health benefits, anthocyanins could be a replacement for synthetic colorant. In recent years, research on the preparation and purification of anthocyanins becomes the focus because of the requirements of bioactive and quantitative analyses. However, the effective methods for the purification of anthocyanins from natural products on a large scale are rarely reported. In this study, isolation and identification of anthocyanins from Vitis davidii Foex skin were investigated. Anthocyanins were extracted from Vitis davidii Foex skin with 70% ethanol containing 0.03% HCl, and then purified orderly by macroporous resin HP-20 column （eluted by 80% ethanol containing 0.05% HCl）, polyamide resin column （eluted by 50% methanol containing 0.1% TFA）, Sephadex LH-20 column chromatography （eluted by 50% methanol containing 0.1% TFA）. The anthocyanins were detected by the ultraviolet-visible （UV-vis） spectrophotometer and the high-performance liquid chromatography （HPLC） after dissolved in methanol containing 0.1% HCl, and then characterized by MSEmethod using UPLC-QTOF. Moreover, the relationship between the fluorescence spectrum and the structure of the anthocyanins was explored by fluorimetry. The results showed that some anthocyanin （elution time was 11.935 min） was absorbed by polyamide resin, causing the proportion of anthocyanin to change, while Sephadex LH-20 could be used to separate and purify the anthocyanin extracts fromVitis davidii Foex skin. In the process of separation, the column bed must be smooth and tight. The optimum conditions of Sephadex LH-20 column included that the volume of sample was 2 ml and the 50% methanol with 0.1% TFA was used as eluant at a flow rate of 1.5 mL/min. Four types of pigments were isolated fromVitis davidii Foex skin, which were Pigment I, II, III andⅣ, and the scanning result of UV-vis showed that Pigment II was not the anthoc

关 键 词：提取 纯化 柱层析 刺葡萄 花色苷 分离 鉴定 荧光分光光度法 

分 类 号：TS202.3[轻工技术与工程—食品科学]