低共熔溶液提取铁观音茶多酚工艺的响应面法优化

阮怿航; 吴亮宇; 鲁静; 黄秀红; 刘丽辰; 林金科

doi:10.19303/j.issn.1008-0384.2020.02.013

低共熔溶液提取铁观音茶多酚工艺的响应面法优化

doi: 10.19303/j.issn.1008-0384.2020.02.013

阮怿航^1,,
吴亮宇¹,
鲁静²,
黄秀红¹,
刘丽辰¹,
林金科^{1, 2, ,}

1.
福建农林大学园艺学院，福建　福州　350002
2.
福建农林大学安溪茶学院，福建　泉州　362000

基金项目: 福建省高校产学合作项目（2016N5001）；安溪县科技项目（KH1500790）；福建省安溪县现代农业产业园协同创新中心项目（KMD18003A）

详细信息

作者简介:
阮怿航（1994−），男，硕士研究生，研究方向：茶资源利用与保健功效（E-mail：ruanyihang2017@163.com）

通讯作者:
林金科（1967−），男，博士，教授，研究方向：茶叶深加工与资源利用（E-mail：ljk213@163.com）

中图分类号: TS 201.2
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出版历程
- 收稿日期: 2019-08-20
- 修回日期: 2019-10-21
- 刊出日期: 2020-02-01

Deep Eutectic Solvents Extraction of Polyphenols from Tieguanyin Tea Optimized by Response Surface Method

1.
College of Horticulture, Fujian Agriculture and Forest University, Fuzhou, Fujian　350002, China
2.
College of Anxi Tea Science, Fujian Agriculture and Forest University, Quanzhou, Fujian　362000, China

摘要

摘要: 目的探索一种绿色、环保、安全、高效的铁观音茶多酚提取新方法，为茶多酚绿色提取提供新的技术参考。方法以铁观音成品茶为原料，采用新型绿色溶剂——低共熔溶液提取茶多酚；首先筛选出最优的低共熔溶液提取体系，然后在单因素试验的基础上，通过响应面法优化，研究时间、温度、溶液含水率对提取率的影响，得出最佳提取条件并分析其主要成分；最后通过测定DPPH自由基清除率分析茶多酚的抗氧化能力。结果筛选出最适的茶多酚低共熔溶液提取体系为：乳酸-甜菜碱，其次得到最佳单因素提取条件为提取时间40 min、提取温度60℃、含水率30%、摩尔比2 ∶ 1、固液比1 ∶ 40（g : mL）；响应面优化分析得到最佳提取条件为：时间46.79 min、温度62.48℃、溶液含水率32.15%，在此条件下茶多酚提取率为15.42%。通过高效液相色谱分析茶多酚组分，其中没食子酸含量占1.40%，儿茶素类物质占84.99%，其他组分占13.61%。低共熔溶液法提取的茶多酚DPPH自由基半清除浓度（IC₅₀）值73.89 μg·mL⁻¹，比抗坏血酸提高了37.80%。结论采用响应面法优化得出的低共熔溶液提取茶多酚最佳工艺条件，可有效提高铁观音茶多酚提取率。
- 低共熔溶液 /
- 铁观音 /
- 茶多酚 /
- 抗氧化活性 /
- 成分分析
Abstract: Objective A green, environmentally friendly, safe and efficient process to extract polyphenols from tea was explored and optimized using the response surface method. Methods Tieguanyin tea was used for the extraction with deep eutectic solvents (DESs). After the DES was selected and a single factor experiment conducted, the time, temperature and water content in solvent were optimized for the process with respect of polyphenol extraction rate by a response surface experimentation. Antioxidant capacity of the extract was determined according to the DPPH radical scavenging rate. Results Lactic acid and betaine of a molar ratio of 2:1 with a water content of 30% and a substrate-to-solvent ratio of 1:40 (g:mL) were chosen for the extraction at 60^oC for 40 min. The response surface method optimized the process to apply a moisture content of 32.15% of DESs for a 46.79 min-extraction at 62.48℃ with a yield of polyphenols at 15.42%. The polyphenol extract was analyzed by HPLC to show a gallic acid content of 1.40%, catechins of 84.99%, and the remainders of 13.61%. The semi-scavenging concentration (IC₅₀) of the extract was 73.89 μg·mL^-1, which was 37.80% higher than that of ascorbic acid. Conclusion The newly developed DESs extraction significantly improved the polyphenol extraction rate from Tieguanyin tea over the conventional process.
- deep eutectic solvents /
- Tieguanyin /
- tea polyphenol /
- antioxidant activity /
- chemical analysis

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图 1 不同提取时间对茶多酚提取率的影响

注：不同大写字母表示差异极显著（P＜0.01），不同小写字母表示差异显著（P＜0.05），图2~5同。

Figure 1. Effect of processing time on polyphenol extraction rate

Note: Different uppercase and lowercase letters indicate extremely significant differences（P＜0.01） and significant differences（P＜0.05）. The same as Fig.2–5.

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图 2 不同提取温度对茶多酚提取率的影响

Figure 2. Effect of processing temperature on polyphenol extraction rate

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图 3 不同低共熔组分摩尔比对茶多酚提取率的影响

Figure 3. Effect of molar ratio of DESs on polyphenol extraction rate

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图 4 不同低共熔溶液含水率对茶多酚提取率的影响

Figure 4. Effect of moisture content of DESs on polyphenol extraction rate

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图 5 不同固液比对茶多酚提取率的影响

Figure 5. Effect of substrate-solvent ratio on polyphenol extraction rate

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图 6 各因素影响多酚提取率的响应面分析

Figure 6. Response surface diagram of factors affecting polyphenol extraction rate in DESs process

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图 7 茶多酚组分HPLC色谱分析

Figure 7. HPLC chromatogram of tea polyphenols

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图 8 儿茶素各组分含量

Figure 8. Contents of catechin components

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图 9 茶多酚提取物对DPPH自由基的清除作用

Figure 9. DPPH free radicals scavenging of tea polyphenol extract

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表 1 Box-Behnken试验因素水平及编码

Table 1. Factors and coding levels in Box-Behnken experiment

水平 Levels	因素 Factors
水平 Levels	A时间 Time/min	B温度 Temperature/℃	C溶液含水率 Water raito of solvent/%
−1	20	50	20
0	40	60	30
1	60	70	40

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表 2 不同低共熔溶液体系提取茶多酚

Table 2. DESs for polyphenol extraction

名称 Name	提取率 Extraction ratio/%	相对水提取提高 Aqueous extraction ratio exceeded by DES/%
水 Water	8.61±0.27Aa
果糖/氯化胆碱 Fructose/Choline chloride	12.47±0.74Bb	44.90
乳酸/氯化胆碱 Lacticacid/Choline chloride	12.69±0.81Bb	47.36
苹果酸/甜菜碱 Malicacid/Choline chloride	12.68±0.68 Bb	47.24
柠檬酸/甜菜碱 Citricacid/Betaine	12.79±0.54Bb	48.59
乳酸/甜菜碱 Lacticacid/Betaine	14.40±0.53 Cc	67.29
注：同列数据后不同大、小写字母表示差异极显著（P＜0.01）或差异显著（P＜0.05）。 Note: Different uppercase and lowercase letters in the same column indicate extremely significant differences (P＜0.01) and significant differences (P＜0.05).

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表 3 响应面优化结果

Table 3. Response surface experiment and results

序号 Number	A时间 Time	B温度 Temperature	C含水率 Water ratio	Y提取率 Extraction ratio/%
1	0	0	0	15.46
2	−1	0	1	14.68
3	0	0	0	15.33
4	0	0	0	15.40
5	−1	0	−1	13.93
6	0	−1	−1	13.62
7	−1	−1	0	14.37
8	1	0	−1	14.63
9	0	0	0	15.27
10	1	1	0	15.07
11	1	−1	0	14.74
12	0	1	1	14.88
13	0	1	−1	14.57
14	−1	1	0	14.86
15	0	−1	1	14.76
16	0	0	0	15.39
17	1	0	1	15.01

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表 4 方差分析

Table 4. Regression statistical analysis

来源 Source	平方和 Sum of square	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value	显著性 Significance
模型 Model	4.13	9	0.46	48.03	＜0.000 1	**
A时间 Time	0.32	1	0.32	33.89	0.000 6	**
B温度 Temperature	0.45	1	0.45	46.7	0.000 2	**
C含水率 Water raito	0.83	1	0.83	87.03	＜0.000 1	**
AB	6.40×10⁻³	1	6.40×10⁻³	0.67	0.440 2
AC	0.034	1	0.034	3.58	0.100 4
BC	0.17	1	0.17	18.01	0.003 8	**
A²	0.27	1	0.27	28.08	0.001 1	**
B²	0.54	1	0.54	56.29	0.000 1	**
C²	1.3	1	1.3	135.65	＜0.000 1	**
残差 Resdual	0.067	7	9.561×10⁻³
失拟项 Lack of fit	0.046	3	0.015	2.92	0.164
纯误差 Pure error	0.021	4	5.250×10⁻³
总和 Cor total	4.2	16
注：，P＜0.05，表示显著差异；，P＜0.01，表示极显著差异。 Note: , P＜0.05, indicating significant difference; **, P＜0.01, indicating extremely significant difference.

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