Process Optimization and Quality Analysis of Lycopus lucidus Fermented Wine
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摘要: 目的：以新鲜的地参为原料，研究地参发酵酒的最佳工艺。方法：采用单因素实验和Box-Behnken响应面优化地参发酵酒的工艺条件，以酒精度及感官评分为响应值，考察发酵温度、酵母接种量和初始糖度对发酵酒的影响，采用UPLC法对地参发酵酒的酚酸类物质进行了分析。结果：地参发酵酒最佳工艺条件为：酵母接种量为5.1%，初始糖度为22.8%，发酵温度为26 ℃，发酵时间为9 d。在此条件地参发酵酒酒精度达到11.23%vol，感官评分为85.50分，多糖、总酚、总酸和总黄酮含量分别为6.54 g/L、384.31 mg/L、20.67 g/L和204.59 mg/L。地参发酵酒的ABTS+、DPPH和对羟基自由基的清除率分别为90.53%、81.51%和55.86%，说明其具有一定的抗氧化活性；地参发酵酒含有没食子酸、丹参素、原儿茶酸、对羟基苯甲酸和咖啡酸5种酚酸类物质，其中没食子酸含量最高（11.20 mg/L）。结论：优化工艺后的地参发酵酒色泽呈橘黄色，澄清透明，酒体醇厚丰满，不仅具有独特的地参酒风味，同时较好的保留了地参发酵酒多酚等活性成分。Abstract: Objective: In this research, to study the best technology of fermented wine with fresh Lycopus lucidus as raw material. Methods: According to the results of single factor test, Box-Behnken response surface was used to optimize the technological conditions of L. lucidus fermented wine, and the effects of fermentation temperature, yeast inoculation amount and initial sugar content on fermented wine were investigated with alcohol content and sensory score as response values. Meanwhile, the phenolic acids in L. lucidus fermented wine were analyzed by UPLC. Results: In this study, the optimum technological conditions of L. lucidus fermentation wine were found to be yeast inoculation of 5.1%, initial sugar content of 22.8%, fermentation temperature of 26 ℃ and fermentation time of 9 days. The alcohol content of the fermented wine reached 11.23%vol, the sensory score was 85.50, and the contents of polysaccharide, total phenol, total acid and total flavone were 6.54 g/L, 384.31 mg/L, 20.67 g/L and 204.59 mg/L. The scavenging rates of ABTS+, DPPH and hydroxyl free radicals of L. lucidus fermented wine were 90.53%, 81.51% and 55.86%, indicating that it had certain antioxidant activity. L. lucidus fermented wine contains five phenolic acids including gallic acid, tanshinsu, protocatechuic acid, p-hydroxybenzoic acid and caffeic acid, of which gallic acid content was the highest (11.20 mg/L). Conclusion: The optimized technology of L. lucidus fermented wine was orange, clear and transparent, and the wine was mellow and full. It not only had a unique flavor of L. lucidus fermented wine, but also retained the active components such as polyphenols.
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Key words:
- Lycopus lucidus /
- fermented wine /
- process optimization /
- phenolic acids /
- quality analysis /
- antioxidant activity
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图 2 酵母接种量对地参发酵酒酒精度和残糖量的影响
Figure 2. Effects of yeast inoculation on the alcohol content and residual sugar of L. lucidus fermented wine
图 3 发酵温度对地参发酵酒酒精度和残糖量的影响
Figure 3. Effects of fermented temperature on the alcohol content and residual sugar of L. lucidus fermented wine
图 4 初始糖度对地参发酵酒酒精度和残糖量的影响
Figure 4. Effects of initial sugar content on alcohol content and residual sugar of L. lucidus fermented wine
图 5 发酵时间对地参发酵酒酒精度和残糖量的影响
Figure 5. Effects of fermented time on the alcohol content and residual sugar of L. lucidus fermented wine
图 6 各因素交互作用对酒精度和感官评分影响的响应面图
Figure 6. Response surface view of the effect of factor interaction on alcohol content and sensory score
图 8 混合对照品（A）及样品（B）的UPLC图谱
Figure 8. UPLC spectra of mixed reference substance (A) and sample (B)
注：A-酚酸标准品；B-样品；1-没食子酸；2-丹参素；3-原儿茶酸；4-对羟基苯甲酸；5-咖啡酸。
表 1 响应面试验因素水平表
Table 1. Factors and levels for response surface experiment
水平 因素 A发酵温度（℃） B酵母接种量（%） C初始糖度（%） ?1 22 4 20 0 25 5 22 1 28 6 24 
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表 2 地参发酵酒感官评分标准
Table 2. Standard of sensory evaluation for Lycopus lucidus fermented wine
指标 评分标准 分值 外观（10分） 澄清透明，橘黄色有光泽，无絮状物
澄清透明，橘黄色光泽较柔，无絮状物
淡橘黄色，无光泽，浑浊9～10
5～8
0～4香气（30分） 有愉悦的地参香和酒香，无异味
地参香气味较浓，酒香一般，无异味
香气味混杂，有异味21～30
11～20
0～10滋味（40分） 酒体醇厚丰满或酒体爽口，风味协调
酸甜适当，酒体较柔和，口感较佳
酒体味道寡淡，口感较差31～40
21～30
0～20典型性（20分） 具独特的地参酒风味，酒香纯正优雅
地参香味不明显，酒香不优雅
无风味，缺乏典型性16～20
11～15
0～10
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表 3 地参发酵酒工艺优化响应面试验设计及结果
Table 3. Experimental design and results of optimized response surface of Lycopus lucidus fermentation wine process
实验号 A B C 酒精度（%vol） 感官评分（分） 1 0 0 0 10.93±0.06 84.70±1.89 2 0 1 ?1 9.30±0.10 73.80±2.53 3 0 ?1 ?1 9.73±0.12 75.50±1.65 4 0 0 0 11.17±0.15 85.10±1.73 5 ?1 1 0 9.97±0.12 76.80±2.62 6 ?1 ?1 0 10.07±0.12 80.10±2.77 7 0 0 0 10.93±0.15 84.00±2.11 8 ?1 0 ?1 9.57±0.06 75.00±2.05 9 1 ?1 0 9.93±0.15 80.20±1.93 10 1 0 ?1 9.77±0.06 75.10±2.77 11 0 0 0 11.13±0.12 84.10±1.73 12 1 1 0 10.70±0.10 83.00±2.26 13 0 1 1 10.23±0.06 81.40±1.72 14 1 0 1 10.87±0.15 82.50±2.27 15 ?1 0 1 10.37±0.15 81.30±2.18 16 0 ?1 1 10.33±0.12 80.90±1.28 17 0 0 0 11.20±0.10 84.80±2.10 
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表 4 响应面试验方差分析
Table 4. Variance analysis of response surface methodology
来源 自由度 酒精度 感官评分 平方和 均方 F值 P值 平方和 均方 F值 P值 模型 9 5.64 0.63 16.36 0.0007** 233.46 25.94 43.86 < 0.0001** A 1 0.21 0.21 5.43 0.0526 7.22 7.22 12.21 0.0101* B 1 0.00245 0.00245 0.064 0.8077 0.36 0.36 0.61 0.4601 C 1 1.47 1.47 38.37 0.0004** 89.11 89.11 150.69 < 0.0001** AB 1 0.19 0.19 4.94 0.0617 9.30 9.3 15.73 0.0054** AC 1 0.022 0.022 0.59 0.4686 0.30 0.3 0.51 0.4976 BC 1 0.027 0.027 0.71 0.4272 1.21 1.21 2.05 0.1957 A2 1 0.45 0.45 11.85 0.0108* 16.34 16.34 27.63 0.0012** B2 1 1.4 1.4 36.45 0.0005** 27.27 27.27 46.12 0.0003** C2 1 1.51 1.51 39.35 0.0004** 70.61 70.61 119.4 < 0.0001** 残差 7 0.27 0.038 4.14 0.59 失拟项 3 0.2 0.066 3.8 0.1149 3.25 1.08 4.85 0.0805 纯误差 4 0.07 0.017 0.89 0.22 总离差 16 5.91 237.60 注：*表示在P<0.05 水平上显著，**表示在P<0.01 水平上极显著。 
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表 5 地参发酵酒5种酚酸类物质含量
Table 5. Contents of five phenolic acids in Lycopus lucidus fermented wine
化合物 回归方程 决定系数R2 含量（mg·L?1） 没食子酸 y=25813x?17372 0.9999 11.20±0.15 丹参素 y=3479x?2008 0.9997 4.38±0.06 原儿茶酸 y=7758x?4332 0.9998 3.74±0.09 对羟基苯甲酸 y=6719x+8443 0.9999 3.40±0.04 咖啡酸 y=15408x?12628 0.9998 11.00±0.11 总计 33.72 
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