| 摘要: |
| 【目的】用大孔树脂纯化黑豆异黄酮粗提物，优化黑豆异黄酮最佳纯化工艺，并对纯化后的黑豆异黄酮组分进行鉴定。【方法】选用NKA-9、AB-8、D101、HPD100和DM301 5种大孔吸附树脂纯化黑豆异黄酮，从中筛选吸附效果最好的树脂，然后以上样液质量浓度、pH值和流速进行单因素试验；在此基础上进行响应面试验，对吸附工艺进行优化；并以洗脱剂乙醇体积分数和用量为试验因素，分析黑豆异黄酮的解吸优化条件；最后在优化的吸附、解吸条件下对黑豆异黄酮进行二次纯化。用超高效液相色谱（UPLC）对纯化后的黑豆异黄酮组分进行鉴定。【结果】AB-8型大孔吸附树脂对黑豆异黄酮具有最佳的吸附、解吸效果。最佳纯化工艺条件：黑豆异黄酮上样液质量浓度为7 mg/mL，pH值为2，上样液流速为2 mL/min，采用体积分数60%乙醇溶液进行洗脱，其用量为80 mL。响应面优化修正为上样液质量浓度7.47 mg/mL，pH值2.27，上样液流速2 mL/min，此时可以达到更优的纯化效果。在此优化工艺条件下，黑豆粗黄酮第1次通过AB 8型大孔吸附树脂，纯度可达72.42%，比未纯化前提高了近3倍，回收率为81.34%；将一次纯化后的黑豆异黄酮在同样条件下再次过柱，所得纯度为78.56%，较一次纯化提高了8.48%，回收率为81.66%。经UPLC对黑豆异黄酮组分进行分析，确定其中主要含有染料木苷、染料木素、大豆苷元和大豆苷4种成分。【结论】AB-8型大孔吸附树脂可有效地纯化黑豆异黄酮，并获得了最优的吸附和解吸条件。 |
| 关键词: 黑豆 异黄酮 大孔吸附树脂 响应面法 |
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| 基金项目:2013年吉林省教育厅资助课题“黑豆中活性成份的提取及药用研究”（吉教科合字［2013］第21号） |
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| Optimization of purification process of black beans isoflavones with macroporous adsorption resin |
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ZHANG Xing,DONG Xin,LIU Zhenchun,et al
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| Abstract: |
| 【Objective】This paper purified soybean isoflavone crude extracts by macroporous resin,and the purification process was optimized.【Method】Five macroporous resins including AB-8,D101,NKA-9,HPD100 and DM301 were selected to purify soybean isoflavones and the best adsorption resin was identified.Then,the sample solution concentration,pH value and flow rate were studied by single factor test and response surface experiments were used to optimize the purification processes.Using eluent volume fraction and ethanol amount as experiment factors,the resin adsorption and desorption conditions of black soybean isoflavone were analyzed.The soybean isoflavone was also purified for the second time under the optimal conditions.Ultra performance liquid chromatography (UPLC) was used to identify the components of purified black soybean isoflavone.【Result】AB-8 macroporous adsorption resin had the best effects for adsorption desorption of black bean isoflavones.The single factor experiments obtained the best purification process of the mass concentration of soybean isoflavone solution 7 mg/mL,the pH 2,and the sample flow rate 2 mL/min.The ethanol solution with volume fraction of 60% was used for elution with dosage of 80 mL.The response surface optimization showed that the optimal conditions were mass concentration of the sample liquid 7.47 mg/mL,pH 2.27,and sample velocity 2 mL/min.Under the optimized conditions,the purity of the soybean isoflavone reached 72.42% by AB-8 at the first time,increased by three times,and the recovery rate was 81.34%.A second round purification of the purified soybean isoflavone under same optimum conditions obtained the purity of 78.56% (increased by 8.48%) and the recovery rate was 81.66%.The components of black soybean isoflavone were analyzed by UPLC,and the four main components were genistein,genistein,daidzein and daidzein.【Conclusion】AB-8 macroporous resin can purify black soybean isoflavone effectively,and the optimum adsorption and desorption conditions were obtained. |
| Key words: black soybean isoflavone macroporous adsorption resin response surface method |
