摘要
为充分开发蓝莓果实的潜在应用价值,主要采用柱色谱法及半制备高效液相色谱法系统研究蓝莓果实中花色苷(花青素的糖苷形式)单体的制备技术。蓝莓花色苷粗提液经超声辅助浸提、乙酸乙酯萃取2次,能够促进花色苷类物质的溶出,并有效去除溶液中的黄酮类杂质。经Amberlite XAD-7HP大孔树脂层析、Sep-Pak C18固相萃取,所得蓝莓花色苷粗品的纯度为62.49%。经Sephadex LH-20凝胶色谱柱分离,获得的3种花色苷纯化组分纯度在65%~75%之间。运用半制备型高效液相色谱技术从3种花色苷纯化组分中制备出两种蓝莓果实中含量较低的半乳糖苷化的花色苷单体,经分析型高效液相色谱鉴定为飞燕草素-3-O-半乳糖苷和锦葵色素-3-O-半乳糖苷,纯度分别为96.98%和95.63%。本研究为花色苷单体的规模化生产提供了技术参考,同时为实现蓝莓花青素高附加值产品的生产提供良好理论依据。
To fully develop the potential application of blueberry fruits, this paper is focused on the preparation of anthocyanin monomers from blueberry fruits using column chromatography and semi-preparative high performance liquid chromatography(HPLC). Crude blueberry anthocyanins were ultrasonically extracted with acidified 70% ethanol and further subjected to two cycles of ethyl acetate extraction for the purpose of facilitating the dissolution of anthocyanins and removing non-anthocyanin flavonoids. The purity of crude blueberry anthocyanins reached 62.49% after Amberlite XAD-7HP column chromatography separationand Sep-Pak C18 solid-phase extraction. Three anthocyanin fractions were obtained by Sephadex LH-20 column chromatography with purities ranging from 65% to 75%. Using semi-preparative HPLC, two pure anthocyanin monomers were successfully isolated from the three purified anthocyanin fractions, which were identified as delphinidin-3-O-galactoside and malvidin-3-O-galactoside by analytical HPLC, with purities of 96.98% and 95.63% respectively. The present study can provide technical references for large-scale production of anthocyanin glycoside monomers, and also provide a good theoretical basis for the production of high value-added products of blueberry anthocyanins.
引文
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