大孔吸附树脂纯化茶多酚的工艺优化及抗氧化活性研究

李晓洁; 刘金鑫; 李建华; 谈亚丽; 杜维力; 李啸

doi:10.13386/j.issn1002-0306.2022090164

大孔吸附树脂纯化茶多酚的工艺优化及抗氧化活性研究

doi: 10.13386/j.issn1002-0306.2022090164

李晓洁^1,,
刘金鑫¹,
李建华²,
谈亚丽³,
杜维力³,
李啸^{1, 3, ,}

1.
三峡大学生物与制药学院/中国轻工业功能酵母重点实验室，湖北宜昌 443002
2.
湖北安琪采花茶品科技有限公司，湖北宜昌 443413
3.
安琪生物科技有限公司，湖北宜昌 443000

基金项目: 国家重点研发计划（2021YFC2101100）

详细信息

作者简介:
李晓洁（1998−），女，硕士研究生，研究方向：茶叶深加工，E-mail：616451051@qq.com

通讯作者:
李啸（1969−），男，博士，教授，研究方向：微生物反应过程的优化与控制，E-mail：lx_6910@163.com

中图分类号: R284.2
计量
- 文章访问数: 36
- HTML全文浏览量: 18
- PDF下载量: 5
- 被引次数: 0
出版历程
- 收稿日期: 2022-09-15
- 网络出版日期: 2023-05-22
- 刊出日期: 2023-07-01

Optimization of Purification of Tea Polyphenols with Macroporous Adsorption Resin and Research of Their Antioxidant Activity

Xiaojie LI^1
,,
Jinxin LIU¹,
Jianhua LI²,
Yali TAN³,
Weili DU³,
Xiao LI^{1, 3
, ,}

1.
Key Laboratory of Functional Yeast of China National Light Industry, School of Biology & Pharmacy, China Three Gorges University, Yichang 443002, China
2.
Hubei Angel & Caihua Tea Processing Products Technology Co., Ltd., Yichang 443413, China
3.
Angel Biotechnology Co., Ltd., Yichang 443000, China

摘要

摘要: 为优化大孔吸附树脂纯化茶多酚的最佳工艺条件，通过对比14种不同类型大孔吸附树脂的静态吸附-解吸特性，在筛选出适宜的树脂型号后，利用单因素与响应面试验确定最佳提纯工艺要求，并进一步考察了树脂的重复使用和再生次数。同时，以V_C为对照采用体外实验考察纯化前、后茶多酚的抗氧化活性。结果表明，LX-8树脂对茶多酚的吸附-解吸效果最好，可重复使用5次、再生6次。其最佳纯化工艺条件：100 mL浓度为6.4 mg/mL，pH5.4的茶汤以1.0 mL/min流速上样至LX-8树脂后，经180 mL 76%乙醇溶液以1.0 mL/min流速解吸，在该条件下茶多酚的回收率为86.9%，纯度为74.6%。体外抗氧化活性试验结果表明，纯化后茶多酚的总抗氧化能力、清除DPPH·和·OH的能力均有显著性增加，且随着浓度的增大，其抗氧化能力增强，其总抗氧化能力（1 mg/mL）为80.59 U/mL，对DPPH·和·OH清除能力的IC₅₀值分别为0.0326和0.4167 mg/mL。虽然低于V_C的抗氧化活性，但均高于纯化前的茶多酚，说明通过该工艺，能够显著提高茶多酚的抗氧化活性，且当纯化后茶多酚浓度为1 mg/mL时，其DPPH·清除率已经接近V_C的DPPH·清除率，本研究为茶多酚的工业化生产和开发利用提供理论参考。
- 茶多酚 /
- 大孔吸附树脂 /
- 纯化 /
- 再生 /
- 抗氧化活性
Abstract: To optimize the technical condition for purification of tea polyphenols with macroporous adsorption resin, the static adsorption and desorption performance of macroporous adsorption resin of fourteen types were compared to select the best type of resin. The optimal purification process conditions were determined by single factor and response surface experiments, and the reuse and regeneration times of the resin were further investigated. Meanwhile, the antioxidant activities of tea polyphenols before and after purification were investigated by in vitro experiments with V_C as the control. The results showed that LX-8 resin had the best adsorption-desorption effect on tea polyphenols, which could be reused 5 times and regenerated 6 times. The optimum purification conditions were as below: The sample concentration of 6.4 mg/mL, the sample pH value of 5.4, the loading volume of 100 mL, the sample flow rate of 1.0 mL/min, ethanol concentration of 76% eluting agent volume of 180 mL, and desorption rate of 1.0 mL/min. Under these conditions, the recovery rate of tea polyphenols was 86.9% and the purity was 74.6%. In vitro antioxidant activity studies revealed that the total antioxidant capacity, DPPH· scavenging capacity and ·OH scavenging capacity of the purified tea polyphenols were significantly increased. And with the increase of polyphenol concentration, its antioxidant capacity enhanced. The total antioxidant capacity of the purified tea polyphenol (1 mg/mL) was 80.59 U/mL. The IC₅₀ of the purified tea polyphenol on scavenging ability of DPPH· and ·OH were 0.0326 and 0.4167 mg/mL respectively. Although the antioxidant activity of pure extract was lower than that of V_C, it was higher than that of crude extract, indicating that the antioxidant activity of tea polyphenols could be significantly improved by this process, and when the concentration of the purified tea polyphenol was 1 mg/mL, its DPPH· scavenging rate was close to that of V_C. This research provided a theoretical reference for the industrial production and utilization of tea polyphenols.
- tea polyphenols /
- macroporous adsorption resin /
- purification /
- regeneration /
- antioxidant activity

HTML全文

图 1 LX-8树脂对茶多酚的吸附等温曲线

Figure 1. Adsorption isotherm curves of tea polyphenols by LX-8 resin

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图 2 LX-8树脂对茶多酚的吸附动力学曲线

Figure 2. Adsorption kinetic curves of tea polyphenols by LX-8 resin

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图 3 pH对LX-8树脂吸附率的影响

Figure 3. Effect of pH on the adsorption rate of LX-8 resin

注：不同小写字母表示在不同水平之间存在显著性差异（P<0.05）；图4、图6同。

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图 4 茶汤浓度对LX-8树脂吸附率的影响

Figure 4. Effect of sample solution concentration on the adsorption rate of LX-8 resin

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图 5 不同吸附流速的泄漏曲线

Figure 5. The leakage curve of different flow velocity

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图 6 乙醇浓度对LX-8树脂解吸率的影响

Figure 6. Effect of ethanol concentration on the desorption rate of LX-8 resin

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图 7 不同解吸流速的解吸曲线

Figure 7. The desorption curves of different desorption flow rates

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图 8 pH、茶汤浓度和乙醇浓度对茶多酚回收率交互影响的响应面及等高线

Figure 8. Response surface plots and contour line of effects of interaction between pH, concentration of tea soup and ethanol concentration on the recovery rate of tea polyphenols

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图 9 LX-8树脂重复使用次数对吸附量和吸附率的影响

Figure 9. The effect of repeated use of LX-8 resin on adsorption capacity and adsorption rate

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图 10 LX-8树脂再生次数对吸附量和吸附率的影响

Figure 10. The effect of regeneration times of LX-8 resin on adsorption capacity and adsorption rate

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图 11 不同浓度样品的总抗氧化能力效果

Figure 11. Effects of total antioxidant capacity of samples with different concentrations

注：不同小写字母表示同一样品组内不同浓度间差异显著（P<0.05），图12、图13同。

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图 12 不同浓度样品对DPPH·的清除效果

Figure 12. Scavenging effects of DPPH· of samples with different concentrations

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图 13 不同浓度样品对·OH的清除效果

Figure 13. Scavenging effects of ·OH of samples with different concentrations

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表 1 Box-Behnken试验设计因素与水平

Table 1. Factors and levels of Box-Behnken experiments design

水平	A pH	B茶汤浓度（mg/mL）	C 乙醇浓度（%）
−1	4.37	5.16	60
0	5.37	6.24	70
1	6.37	7.32	80

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表 2 14种大孔树脂的静态吸附和解吸效果

Table 2. Static adsorption and desorption effects of 14 macroporous resins

树脂型号	极性	吸附量（mg/g）	吸附率（%）	解吸量（mg/g）	解吸率（%）
LX-B14	极性	39.20±0.55^b	90.85±1.27^a	28.52±0.65^b	72.77±0.86^ef
XDA-8G	极性	21.86±0.23^f	68.80±0.72^b	17.26±0.29^d	78.95±1.98^d
LX-8	极性	33.09±0.27^c	90.65±0.74^a	29.87±0.69^a	90.26±1.92^ab
SP207	极性	25.83±0.26^e	58.20±0.59^d	16.89±0.93^d	65.39±3.59^g
D301	极性	41.76±0.30^a	90.69±0.66^a	11.46±0.87^e	27.44±1.88^h
HZW635	极性	26.59±0.69^de	68.07±1.78^b	20.87±0.50^c	78.56±3.84^d
HZW636	极性	27.01±0.80^d	62.64±1.86^c	20.35±0.97^c	75.46±5.88^de
ADS-17	中极性	1.90±0.53^h	3.42±0.96^e	1.54±0.44^f	81.12±1.97^cd
HPD450	中极性	2.90±0.82^gh	4.69±1.32^e	1.96±0.52^f	67.84±1.72^fg
AB-8	弱极性	2.35±0.48^gh	4.87±0.98^e	2.11±0.38^f	90.26±4.24^ab
X-5	非极性	2.66±0.37^gh	4.39±0.62^e	2.04±0.32^f	76.72±2.23^de
D101	非极性	2.24±0.41^h	4.83±0.89^e	1.92±0.44^f	85.09±4.57^bc
H103	非极性	3.33±0.86^g	5.17±1.32^e	2.10±0.46^f	63.65±2.83^g
HPD100	非极性	2.89±0.56^gh	5.32±1.02^e	2.64±0.55^f	91.02±1.83^a
注：同列不同字母表示差异显著（P<0.05）。

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表 3 不同温度下的Langmuir模型拟合参数

Table 3. Fitting parameters of Langmuir models at different temperatures

温度（℃）	Langmuir方程	K_L	q_max	R²
25	C_e/q_e=0.0148C_e+0.0088	1.682	67.57	0.9984
35	C_e/q_e=0.0229C_e+0.0132	1.735	43.67	0.9931
45	C_e/q_e=0.0406C_e+0.0137	2.964	24.63	0.9823
注：q_e为饱和吸附量，mg/g；q_t为t时刻吸附量，mg/g；q_max为树脂最大吸附量，mg/g；C_e为吸附饱和的茶多酚浓度，mg/mL；K_L、K_F、n为特征常数，表4、表5同。

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表 4 不同温度下的Freundlich模型拟合参数

Table 4. Fitting parameters of Freundlich models at different temperatures

温度（℃）	Freundlich方程	K_F	1/n	R²
25	lnq_e=0.4806InC_e−3.4963	0.03	0.4806	0.9831
35	lnq_e=0.5678InC_e−3.2386	0.04	0.5678	0.9805
45	lnq_e=0.6557InC_e−2.814	0.06	0.6557	0.9235

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表 5 吸附动力学模型拟合参数

Table 5. Fitting parameters of adsorption kinetics models

拟合模型	拟合方程	q_e	k	R²
一级动力学	ln（q_e-q_t）=−0.3252t+1.2	3.32	0.325	0.846
二级动力学	t/q_t=0.1285t+2.2022	7.78	0.008	0.9996

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表 6 响应面试验设计及结果

Table 6. Design and results of response surface experiment

试验号	A pH	B 茶汤浓度	C 乙醇浓度	Y 回收率（%）
1	1	0	1	82.6±0.26
2	0	0	0	86.4±0.17
3	−1	0	−1	80.4±0.10
4	0	0	0	87.5±0.29
5	1	−1	0	72.8±0.26
6	1	0	−1	74.7±0.30
7	−1	0	1	81.2±0.44
8	−1	−1	0	77.9±0.40
9	0	1	1	83.8±0.46
10	0	0	0	87.2±0.17
11	0	−1	1	81.6±0.30
12	0	−1	−1	79.2±0.10
13	0	1	−1	82.7±0.20
14	1	1	0	77.4±0.36
15	−1	1	0	78.3±0.17
16	0	0	0	87.6±0.30
17	0	0	0	87.9±0.10

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表 7 回归模型方差分析

Table 7. Variance analysis of regression model

方差来源	平方和	自由度	均方	F值	P值	显著性
模型	341.23	9	37.91	51.93	< 0.0001	**
A	13.26	1	13.26	18.16	0.0037	**
B	14.31	1	14.31	19.60	0.0031	**
C	18.60	1	18.60	25.48	0.0015	**
AB	4.41	1	4.41	6.04	0.0436	*
AC	12.60	1	12.60	17.26	0.0043	**
BC	0.42	1	0.42	0.58	0.4717
A²	173.00	1	173.00	236.97	< 0.0001	**
B²	78.22	1	78.22	107.13	< 0.0001	**
C²	5.91	1	5.91	8.10	0.0248	*
残差	5.11	7	0.73
失拟项	3.80	3	1.27	3.88	0.1118	不显著
纯误差	1.31	4	0.33
总和	346.34	16
R²=0.9852				R²_adj=0.9663
注：“”表示对结果影响差异显著（P<0.05）；“*”表示对结果影响差异极显著（P<0.01）。

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