Preparation of Herbal Tea Fermented by Lactobacillus from Lonicera japonica leaves and Analysis of Its Antioxidant Activity
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摘要: 为获得一款具强抗氧化活性的高保健性金银花叶发酵凉茶,以金银花叶为原材料,通过单因素实验和正交试验,研究了发酵时间、发酵温度、保加利亚乳酸杆菌(L. plantarum 758)接种量及糖的添加量(葡萄糖与蔗糖质量比为3:2)对金银花叶凉茶抗氧化活性的影响并对工艺条件进行了优化,还对比了金银花叶凉茶与市场上常见三种凉茶在抗氧化性及挥发性成分上的差异。结果表明,乳酸杆菌发酵金银花叶凉茶的最佳发酵条件为:发酵时间24 h、发酵温度38 ℃、接种量4%及糖添加量7%,在此发酵条件下所获得的金银花叶凉茶绿原酸含量达到7.11%,总黄酮含量为22.51%,DPPH自由基清除率为84.23%±3.32%,ABTS自由基清除率为85.30%±2.14%,均显著高于市场上的三种凉茶(P<0.05);金银花叶凉茶中共有挥发性物质32种,其种类和含量都较其它对比组高。总体表明,保加利亚乳酸杆菌(L. plantarum 758)发酵的金银花叶凉茶抗氧化能力突出,具有较高的营养价值和实用功能性。Abstract: In order to obtain a kind of Lonicera japonica leaves herbal tea with strong antioxidant activity and high health care, an orthogonal array design based on single factor experiments was employed to examine the influence of time, temperature, inoculum amount of L. plantarum 758 and the concentration of sugar (The mass ratio of glucose to sucrose was 3:2) on the fermentation process. The antioxidant and volatile components of Lonicera japonica leaves herbal tea were compared with those of three common herbal tea in the market. Results showed that the optimal fermentation conditions were determined as fermentation of Lonicera japonica leaves extract containing 7% sugar added at 38 ℃ for 24 h with an inoculum amount of 4%. Under these conditions, the chlorogenic acid content of Lonicera japonica leaves herbal tea reached 7.11%, the total flavonoids content was 22.51%, the DPPH free radical clearance rate was 84.23%±3.32%, and the ABTS free radical clearance rate was 85.30%±2.14%, which were significantly higher than those of the three kinds of herbal tea on the market (P<0.05). There were 32 kinds of volatile substances in Lonicera japonica leaves herbal tea, and their types and contents were higher than those in other comparison groups. Overall, the antioxidant capacity of Lonicera japonica leaves herbal tea fermented by L.plantarum 758 is outstanding, and it has higher nutritional value and practical function.
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表 1 驯化培养基的制备
Table 1. Preparation of domestication medium
培养编号 1 2 3 4 5 6 金银花叶浸提液添加量 0 10% 20% 30% 40% 50% 10%脱脂乳粉添加量 50% 40% 30% 20% 10% 0 注:培养基中按葡萄糖:蔗糖=3:2的比例添加6%的糖。 表 2 正交试验因素与水平
Table 2. Factors and levels of orthogonal design
水平 A发酵时间(h) B发酵温度(℃) C接种量(%) D糖添加量(%) 1 22 36 2 5 2 24 37 3 6 3 26 38 4 7 表 3 金银花叶凉茶发酵工艺条件正交试验设计及结果
Table 3. Orthogonal experimental design and results of Lonicera japonica leaves herbal tea fermentation
试验号 A发酵
时间B发酵
温度C接种量 D糖添
加量乳酸含
量(%)DPPH自由基
清除率(%)1 1 1 1 1 0.53 78.10 2 1 2 2 2 0.49 79.30 3 1 3 3 3 0.58 80.40 4 2 1 2 3 0.44 80.10 5 2 2 3 1 0.50 80.49 6 2 3 1 2 0.54 79.76 7 3 1 3 2 0.55 79.61 8 3 2 1 3 0.52 78.34 9 3 3 2 1 0.51 79.02 乳酸含量 k1 0.517 0.490 0.513 0.497 k2 0.493 0.503 0.480 0.527 k3 0.527 0.543 0.543 0.513 R 0.034 0.053 0.063 0.030 DPPH自由
基清除率k1 79.267 79.267 78.733 79.200 k2 80.133 79.367 79.467 79.567 k3 78.967 79.733 80.167 79.600 R 1.166 0.466 1.434 0.400 表 4 金银花叶凉茶与市场上三种凉茶活性成分与抗氧化活性对比
Table 4. Comparison of active ingredients and antioxidant activities between Lonicera japonica leaves herbal tea and three kinds of herbal tea on the market
种类 绿原酸
(%)总黄酮
(%)DPPH自由基
清除率(%)ABTS自由基
清除率(%)金银花叶凉茶 7.11±0.01a 22.51±1.52a 84.23±3.32a 85.30±2.14a 茶1 5.32±0.02c 17.42±1.34b 65.23±2.10c 78.41±2.35c 茶2 5.67±0.01b 17.28±1.60b 73.29±2.54b 80.12±2.03b 茶3 3.66±0.03d 12.66±1.22c 60.86±2.33d 75.38±1.98d 注:表中同一列不同字母代表差异显著(P<0.05)。 表 5 金银花叶凉茶与市场上三种凉茶挥发性成分比较
Table 5. Comparison of volatile components between Lonicera japonica leaves herbal tea and three kinds of herbal tea on the market
序号 挥发性化合物 金银花叶茶
(mg/L)茶1
(mg/L)茶2
(mg/L)茶3
(mg/L)A1 3-甲硫基丙醇 3.34 3.40 3.44 3.44 A2 苯乙醇 3.28 3.14 3.24 3.28 A3 1-辛醇 3.23 3.08 3.23 3.23 A4 2,3-丁二醇 − 3.23 3.08 6.45 A5 2-乙基己醇 2.94 − − 2.94 A6 苯甲醇 3.31 − − − A7 1-壬醇 3.19 − − − A8 丙醇 2.76 − − − A9 新薄荷醇 2.76 − − − A10 正己醇 1.78 − − − A11 (R)-1,2-丙二醇 − 3.05 − − A12 正丁醇 − 1.71 − − A13 3-甲基-1-戊醇 − − 3.05 − A14 庚酸乙酯 3.10 3.28 3.34 2.76 A15 辛酸乙酯 2.67 2.67 2.67 5.71 A16 正己酸乙酯 2.05 2.03 2.03 1.78 A17 乙酸异戊酯 1.93 1.93 1.93 1.78 A18 癸酸乙酯 3.31 3.38 3.38 − A19 月桂酸乙酯 2.01 2.10 2.08 − A20 棕榈酸乙酯 1.99 2.08 1.99 − A21 乙酸乙酯 3.89 − 3.89 3.77 A22 乙酸苯乙酯 1.93 1.93 − 1.93 A23 壬酸乙酯 2.49 2.92 − − A24 乙酸己酯 − 1.84 1.84 − A25 反式-2-己烯酸乙酯 2.79 − − − A26 乙酸苄酯 2.05 − − − A27 癸酸3-甲基丁酯 − 2.76 − − A28 丙酸-2-苯乙酯 − − 2.86 − A29 乙酸 1.93 1.95 1.95 1.95 A30 辛酸 2.80 2.80 − 2.46 A31 苯甲酸 6.00 − − − A32 癸酸 − 2.76 − − A33 2-甲基四氢噻吩-3-酮 − 3.37 − A34 十二甲基环六硅氧烷 5.75 2.86 2.67 2.55 A35 六甲基环三硅氧烷 2.80 8.29 5.97 5.97 A36 十甲基环五硅氧烷 2.67 5.59 2.80 5.31 A37 十四烷 5.68 2.49 2.80 − A38 十二烷 2.95 − 8.29 5.28 A39 八甲基环四硅氧烷 2.97 3.10 − − A40 十六烷 − 2.80 − − A41 二十七烷 3.10 − − − A42 2-亚甲基戊二腈 2.87 − − − 表 6 金银花叶凉茶主成分特征值及贡献率
Table 6. Characteristic values and contribution rates of principal components of Lonicera japonica leaves herbal tea
主成分 特征值 贡献率(%) 1 13.44 44.9 2 9.70 32.0 -
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