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【目的】以基于茶多酚的思茅松树皮多聚原花青素降解反应中(-)-表儿茶素-(4β-8)-(-)-表没食子儿茶素3-O-没食子酸酯(化合物1)的含量作为评价指标,确定思茅松树皮多聚原花青素的最佳降解条件,为思茅松树皮多聚原花青素降解产物的进一步开发利用提供基础。【方法】采用HPLC定量分析方法测定各反应溶液中化合物1的含量,通过单因素试验考察反应温度(50~90℃)、反应时间(30~180 min)、盐酸浓度(0.1%~5%)和茶多酚/多聚原花青素比率(1∶3~3∶1,w/w)对化合物1生成的影响。采用4因素5水平中心组合旋转设计的响应面法优化其降解条件,以化合物1的含量为响应值,以上述4个因素为自变量,利用Design-Expert V8.0.6软件对28个试验点测定所得数据进行多元非线性分析,建立回归模型,并通过方差分析对模型进行显著性检测。【结果】反应温度、盐酸浓度和茶多酚/多聚原花青素比率明显影响降解反应中化合物1的生成,建立的回归模型极显著(P<0.000 1),且线性系数良好(R2=0.952 6),说明建立的数学模型能较好地描述试验结果,可用于分析和预测化合物1的生成。思茅松树皮多聚原花青素的最佳降解条件为反应温度70℃、反应时间60 min、盐酸浓度1%和茶多酚/多聚原花青素比率3∶2。经验证此条件下反应液中化合物1的浓度可达718.57 nmol·mL~(-1),与理论值(721.39 nmol·mL~(-1))较为接近。【结论】采用中心组合旋转设计的响应面法分析优化思茅松树皮多聚原花青素降解以获取主产物(-)-表儿茶素-(4β-8)-(-)-表没食子儿茶素3-O-没食子酸酯的方法可行。
【Objective】 The objective of this study was to investigate the effects of (-) - epicatechin- (4β-8) - (-) - epigallocatechin 3-O- Compound 1) content as the evaluation index to determine the optimal degradation conditions of polygonatum anthracnose bark, to provide the basis for the further development and utilization of the polyphenanthrene degradation products. 【Method】 The content of compound 1 in each reaction solution was determined by HPLC quantitative analysis. The reaction temperature (50-90 ℃), reaction time (30-180 min) and hydrochloric acid concentration (0.1-5%) were investigated by single factor test. And the effect of tea polyphenols / poly-procyanidins ratio (1: 3 ~ 3: 1, w / w) on the production of compound 1. The response surface methodology was used to optimize the degradation conditions using a 4-factor-5, 5-center combined rotation design. The compound 1 content was used as the response variable. Based on the 4 factors described above, 28 samples were determined by Design-Expert V8.0.6 software The data obtained were subjected to multivariate nonlinear analysis, regression model was established, and the model was tested by ANOVA. 【Result】 The results showed that the reaction temperature, the concentration of hydrochloric acid and the ratio of tea polyphenols to polyphenanthrene significantly affected the production of compound 1 in the degradation reaction. The established regression model was significantly (P <0.000 1) with a good linearity (R2 = 0.952 6) , Indicating that the established mathematical model can better describe the experimental results and can be used to analyze and predict the production of Compound 1. The optimal degradation conditions of polygonatum anthracnose bark were as follows: reaction temperature 70 ℃, reaction time 60 min, concentration of hydrochloric acid 1% and ratio of tea polyphenol to poly procyanidin 3: 2. The concentration of compound 1 in the reaction solution proved to be 718.57 nmol · mL -1, which is close to the theoretical value (721.39 nmol · mL -1). 【Conclusion】 Polymeric proanthocyanidins from bark of Pinus yunnanensis were analyzed by response surface methodology to optimize the yield of (-) - epicatechin - (4β-8) - (-) - epigallocatechin 3 -O-gallate method is feasible.