[关键词]
[摘要]
本文主要研究黑豆(Phaseolus vulgaris)种皮中原花青素的提取和纯化方法。通过使用超声波辅助提取法粗提原花青素(Procyanidins,PC),以AB-8型大孔树脂吸附法纯化粗提物,并对纯化产物进行了红外色谱定性分析。单因素提取实验结果显示:提取条件分别在乙醇的浓度为60%,提取时间在40 min,料液比为1:20以及提取温度为50 ℃时提取效率相对较高。随后正交试验结果显示了最优综合提取条件为:乙醇浓度为60%、提取时间为50 min、料液比为1:20 (m/V)、提取温度为60 ℃;确定了AB-8型大孔树脂纯化工艺的最佳动态吸附和解析条件为:以0.6 mg/mL的原花青素粗提液为上样液质量浓度,流速为1.0 mL/min,吸附平衡时间为4 h,60%的乙醇溶液作为洗脱液,洗脱流速1.5 mL/min,解析时间为3 h;且红外光谱定性分析显示以上条件纯化原花青素相对有效。
[Key word]
[Abstract]
In this study, extraction and purification methods of procyanidins from black bean seed coats was investigated. The procyanidins (PC) were extracted by ultrasonic assisted extraction and the crude extracts were purified by AB-8 macroporous resin adsorption method. The purified products were analyzed by infrared spectrum. The results of single factor experiments showed that the extraction method was more relatively effective when the concentration of ethanol was 60%, the extraction time was 50 mins, the solid-liquid ratio was 1:20 (m/V) and extraction temperature was 50℃. Whereafter, the results of orthogonal test showed that the optimal extraction conditions were as follows: the ethanol concentration of 60%, the extraction time of 50 mins, the solid-liquid ratio of 1:20 (m/V), and the extraction temperature of 60℃. The optimal dynamic adsorption and analytical conditions of the purification of AB-8 macroporous resin were as follows: 0.6 mg/mL procyanidins extract, the flow rate of 1 mL/min, the adsorption equilibrium time of 4 h, 60% ethanol solution as the eluent, elution flow rate of 1.5 mL/min, the elution balance time of 3 h. Qualitative analysis by infrared spectroscopy showed that the above conditions for the purification of proanthocyanidins were relatively effective.
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[基金项目]
江苏高校“青蓝工程”资助;国家自然科学基金青年科学基金项目(31301507);江苏省高校自然科学基金面上项目(13KJB550006)