河北农业大学 硕士学位论文 蔷薇红景天中原花青素提取、精制及稳定性研究 姓名：孟玉彩 申请学位级别：硕士 专业：农产品加工及贮藏工程 指导教师：赵文 2009-06-13 摘 要 蔷薇红景天（Rhodiola rosea L.）为景天科红景天属植物，是一种具有优良保健功 效的珍稀药用植物资源，含有多种生理活性成分，如红景天苷、酪醇、黄酮类等。然而 国内外对蔷薇红景天中原花青素的研究报道却极少。为此，本文对蔷薇红景天中原花青 素的提取、精制工艺以及稳定性进行了较为系统的研究，为进一步工业化生产和精制原 花青素提供了理论基础。 本文建立了香草醛-盐酸法测定蔷薇红景天中原花青素的方法。分别采用两种不同 的方法从蔷薇红景天中提取原花青素， 通过单因素试验及正交试验设计确定了乙醇回流 提取法的最佳工艺条件为：提取溶剂为 50% (v/v)乙醇，提取温度 80，提取时间 120 min，料液比 1:25；超声波-微波协同萃取法的最佳工艺条件为：提取溶剂为 50% (v/v) 乙醇，料液比 1:25，在 200 W 的微波功率下提取 50 s，然后再在 50恒温条件下超声 波处理 20 min，两种方法在最佳提取条件下原花青素得率为分别为 5.43%、4.72%。 蔷薇红景天原花青素精制采用了大孔吸附树脂精制法，根据吸附解吸性能，从 5 种 树脂中筛选出 AB-8 树脂作为精制蔷薇红景天原花青素的树脂，并确定了其吸附和洗脱 条件。结果表明，经 AB-8 树脂固定床进行分离富集后，有效的去除了杂质，原花青素 纯度得到了提高，试验达到了预期目的。 通过对原花青素精制物进行 IR、 HPLC 分析， 明确了蔷薇红景天中的原花青素是以 表棓儿茶素-3-O-棓酸酯（EGCG）为基本结构单元的原翠雀定类原花青素。 蔷薇红景天原花青素的稳定性研究表明，光照可使原花青素含量逐渐下降，较低的 温度和较低的 pH 值条件下原花青素比较稳定；防腐剂苯甲酸钠、山梨酸钾对原花青素 的稳定性无太大影响；H2O2可导致原花青素的稳定性下降，而 Vc 对原花青素的稳定性 有保护作用；原花青素对 K+、Mg2+、Na+和 Cu2+表现出很好的稳定性，而 Fe3+、Sn2+、 Al3+可破坏原花青素的稳定性。 关键词：蔷薇红景天；原花青素；提取；大孔树脂；稳定性 Study on Extraction, Purification and Stability of proanthocyanidins from Rhodiola rosea Author: Meng Yu-cai Supervisor: Zhao Wen Major: Food processing and storage engineering Abstract Rhodiola rosea is a species which comes from Rhodiola genus of Crassulaceae. As a valuable resource of the medicinal plants, Rhodiola rosea has the superordinary efficiency in health. It contains many kinds of chemical composition with physiological activation, such as salidroside, tyrosol, flavonoids and so on. However, there are only few literatures about proanthocyanidins in Rhodiola rosea at home and abroad. In this paper, the optimum extracting condition and purification method of proanthocyanidins from Rhodiola rosea, and the stability of the purified proanthocyanidins were studied. The study laid theoretical foundation for large-scale industrialization manufacture and purification of proanthocyanidins. The method of analyzing proanthocyanidins of Rhodiola rosea by spectrophotometric determination of the vanillin-hydrochloric acid method was founded. Two different methods were adopted to extract the proanthocyanidins. Through the single factor experiment and the orthogonal experimental design, the optimum extraction conditions of proanthocyanidins from Rhodiola rosea by ethanol circumfluence were showed as follows: extraction solvent was 50% ethanol (v/v), extraction temperature was 80, the solid- solvent ratio was 1:25, extraction time was 120 min. The optimum extraction conditions of proanthocyanidins from Rhodiola rosea by ultrasonic-microwave synergistic extraction were obtained as follows: extraction solvent was 50% ethanol (v/v), the solid-solvent ratio was 1:25, the microwave power was 200 W, the microwave time was 50 s, then ultrasonic wave treatment 20 min under the 50 constant temperature. Under the optimum condition, the extracting ratio of the two methods were 5.43% and 4.72%, respectively. Proanthocyanidins of Rhodiola rosea was refined by macroreticular resins. According to the adsorption and desorption capacity, AB-8 was the best in five kinds of selected macroreticular resins. The adsorption and elution conditions were determined. The results showed that much impurity was removed and the purity of the proanthocyanidins was raised by separation and enriched with AB-8 resin fixed bed. The experiment was successful. The purified proanthocyanidins determined by IR and HPLC, the result showed that proanthocyanidins of Rhodiola rosea was mainly composed of (-)-epigallocatechin-3-O -gallate (EGCG), and it belonged to prodelphinidins. The stability of proanthocyanidins was studied. The results showed that the amount would decrease under the sunlight, the proanthocyanidins was likely stable at low temperature and low pH. The proanthocyanidins was stable against to antiseptic such as sodium benzoate and potassium sorbate. H2O2 had obvious effect on the stability of proanthocyanidins, on the contrary, Vc could protect its stability. Among different metal ions, K+, Mg2+, Na+ and Cu2+ had little influence on the stability of proanthocyanidins, but Fe3+, Sn2+ and Al3+ could destroy its stability. Key words: Rhodiola rosea; Proanthocyanidins; Extraction; Macroreticular Resin; Stability 缩略表 PC Proanthocyanidins 原花青素 OPC Oligomeric proanthocyanidins 低聚原花青素 PPC Polymeric proanthocyanidins 高聚原花青素 HPLC High performance liquid chromatography 高效液相色谱 IR Infrared Spectroscopy 红外光谱 EGCG Epigallocatechin-3-O-gallate 表棓儿茶素-3-O-棓酸酯 EGC Epigallocatechin 表棓儿茶素 GSPC Grape seed proanthocyanidins extract 葡萄籽原花青素提取物 TLC Thin-layer chromatography 薄层色谱 MAE Microwave assisted extraction 微波辅助提取 UAE Ultrasonic assisted extraction 超声波辅助提取 SFE Supercritical fluid extraction 超临界流体萃取 DAD Diode array detector 二极管阵列检测器 蔷薇红景天中原花青素提取、精制及稳定性研究 1 R1 = H or OH R2 = OH or 1 前言 1.1 原花青素概述 1.1.1 原花青素的化学结构及分类 原花青素（Proanthocyanidins 或 Procyanidins，简称 PC，又译原