Optimization of the Extraction Process of Peristrophe roxburghiana Polysaccharide and Analysis of Its Antioxidant Activity
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摘要: 为探究红蓝草多糖的最佳提取工艺及其体外抗氧化活性。试验以红蓝草为原料，探究料液比、浸提温度、浸提时间、浸提次数对红蓝草多糖得率的影响，并结合响应面法优化红蓝草多糖提取工艺，通过测定红蓝草多糖对DPPH·、·OH、ABTS+·等自由基的清除能力探究其抗氧化活性。结果表明：在料液比1:31 g/mL、浸提温度85 ℃、浸提时间118 min、提取3次的条件下，红蓝草多糖得率最高，可达11.05%。在一定范围内，红蓝草多糖清除自由基能力与多糖的浓度呈量效关系，红蓝草多糖溶液清除DPPH · 、 · OH和ABTS + ·等自由基的IC50值分别为0.18、0.73、0.64 mg/mL，说明红蓝草粗多糖具有一定的抗氧化活性。通过探究红蓝草多糖提取的最佳工艺及抗氧化活性，为今后红蓝草多糖的进一步开发与应用提供理论基础和参考。Abstract: In order to explore the optimum extraction process of polysaccharide and antioxidant activity in vitro in Peristrophe roxburghiana. In this study, Peristrophe roxburghiana was used as raw material to explore the effects of solid-liquid ratio, extraction temperature, time and frequency on the yield of polysaccharide. The extraction process of polysaccharide in Peristrophe roxburghiana was optimized by response surface methodology and the antioxidant activity of the polysaccharide was investigated by measuring its scavenging ability on DPPH·, ·OH, ABTS+· and other free radicals. The results showed that we could get the most quantity of polysaccharide extracted from Peristrophe roxburghiana, which reaches up to 11.05%, in these conditions: The solid-liquid ratio was 1:31 g/mL, the extraction temperature was 85 ℃, the extraction time was 118 min and the extraction frequency was 3 times. The scavenging ability of the polysaccharide against free radical was quantitatively related to the concentration of polysaccharide within a certain range. The IC50 values of DPPH·, ·OH and ABTS+· radicals scavenged by the polysaccharide solution were 0.18, 0.73 and 0.64 mg/mL, respectively, indicating that the polysaccharide extracted from Peristrophe roxburghiana was with certain antioxidant activity. By exploring the optimal process and antioxidant activity of the polysaccharide extracted from Peristrophe roxburghiana, we provide a theoretical basis and reference for the further development and application of the polysaccharide in the future.
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图 2 浸提温度对红蓝草多糖得率的影响
Figure 2. Effect of extraction temperature on Peristrophe roxburghiana polysaccharide yield rate
图 3 浸提时间对红蓝草多糖得率的影响
Figure 3. Effect of extraction time on Peristrophe roxburghiana polysaccharide yield rate
图 4 提取次数对红蓝草多糖得率的影响
Figure 4. Effect of extraction number on Peristrophe roxburghiana polysaccharide yield rate
图 5 不同因素交互作用对红蓝草多糖得率影响的响应面图和等高线图
Figure 5. Contour plot and response surface plot of the effect of different factors on the yield of Peristrophe roxburghiana polysaccharide
图 6 红蓝草多糖和VC对DPPH·的清除作用比较
Figure 6. The comparison of scavenging effect on DPPH free radical of Peristrophe roxburghiana polysaccharide and VC
图 7 红蓝草多糖和VC对·OH的清除作用比较
Figure 7. The comparison of scavenging effect on OH free radical of Peristrophe roxburghiana polysaccharide and VC
图 8 红蓝草多糖和VC对ABTS+·的清除作用比较
Figure 8. The comparison of scavenging effect on ABTS free radical of Peristrophe roxburghiana polysaccharide and VC
表 1 响应面试验设计因素与水平
Table 1. Response surface test design factors and levels
因素 水平 −1 0 1 A 料液比（g/mL） 1:25 1:30 1:35 B 浸提温度（℃） 80 85 90 C 浸提时间（min） 90 120 150 D 浸提次数 2 3 4 
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表 2 响应面试验设计及结果
Table 2. Response surface test design and results
试验号 A 料液比 B 浸提温度 C 浸提时间 D 浸提次数 得率（%） 1 −1 −1 0 0 8.95 2 1 −1 0 0 9.01 3 −1 1 0 0 7.93 4 1 1 0 0 9.46 5 0 0 −1 −1 9.01 6 0 0 1 −1 7.61 7 0 0 −1 1 8.38 8 0 0 1 1 8.33 9 −1 0 0 −1 7.56 10 1 0 0 −1 9.01 11 −1 0 0 1 8.25 12 1 0 0 1 8.08 13 0 −1 −1 0 9.48 14 0 1 −1 0 8.52 15 0 −1 1 0 8.88 16 0 1 1 0 8.97 17 −1 0 −1 0 8.70 18 1 0 −1 0 8.64 19 −1 0 1 0 7.37 20 1 0 1 0 9.16 21 0 −1 0 −1 9.09 22 0 1 0 −1 8.52 23 0 −1 0 1 9.57 24 0 1 0 1 9.78 25 0 0 0 0 11.02 26 0 0 0 0 11.03 27 0 0 0 0 11.03 28 0 0 0 0 10.54 29 0 0 0 0 10.98
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表 3 二次回归模型的方差分析数据显著性检验
Table 3. The ANOVA data significance test for the secondary regression model
方差来源 平方和 自由度 均方和 F值 P值 显著性 模型 29.26 14 2.09 21.07 <0.0001 ** A 1.76 1 1.76 17.78 0.0009 ** B 0.2700 1 0.2700 2.72 0.1212 C 0.4840 1 0.4840 4.88 0.0443 * D 0.2107 1 0.2107 2.12 0.1671 AB 0.5402 1 0.5402 5.45 0.0350 * AC 0.8556 1 0.8556 8.63 0.0108 * AD 0.6561 1 0.6561 6.62 0.0222 * BC 0.2756 1 0.2756 2.78 0.1177 BD 0.1521 1 0.1521 1.53 0.2359 CD 0.4556 1 0.4556 4.59 0.0501 A2 12.22 1 12.22 123.20 <0.0001 ** B2 2.47 1 2.47 24.94 0.0002 ** C2 10.24 1 10.24 103.21 <0.0001 ** D2 9.95 1 9.95 100.36 <0.0001 ** 残差 1.39 14 0.0992 失拟项 1.21 10 0.1206 2.65 0.1803 不显著 纯误差 0.1822 4 0.0455 总和 30.65 28 R2=0.9547 R2adj=0.9094 注：*表示差异显著（P<0.05）；**表示差异极显著（P<0.01）。
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