嘉宝果叶片总多酚提取工艺优化及其体外降糖活性

林宝妹; 邱珊莲; 郑开斌; 张帅; 洪佳敏; 张少平

doi:10.19303/j.issn.1008-0384.2019.05.012

嘉宝果叶片总多酚提取工艺优化及其体外降糖活性

doi: 10.19303/j.issn.1008-0384.2019.05.012

福建省农业科学院亚热带农业研究所, 福建漳州 363005

基金项目:

福建省科技计划项目——省属公益类科研院所基本科研专项 2018R1024-6

福建省农业科学院科技创新项目 PC2018-9-6

福建省农业科学院青年英才计划项目 YC2015-19

福建省农业科学院青年创新团队项目 STIT2017-3-4

福建省农业科学院生产性工程化实验室中试项目 AG2017-5

详细信息

作者简介:
林宝妹(1988-), 女, 硕士, 研究实习员, 研究方向:天然产物化学(E-mail:yogobm@qq.com)

通讯作者:
邱珊莲(1979-), 女, 博士, 副研究员, 研究方向:天然产物化学(E-mail:slqiu79@163.com)

中图分类号: Q946.8
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- 被引次数: 0
出版历程
- 收稿日期: 2018-12-25
- 修回日期: 2019-03-12
- 刊出日期: 2019-05-28

Process Optimization and in vitro Hypoglycemic Effect of Polyphenols Extracted from Jaboticaba Leaves

Institute of Subtropical Agriculture, Fujian Academy of Agricultural Sciences, Zhangzhou, Fujian 363005, China

摘要

摘要: 目的优化嘉宝果叶片总多酚的提取工艺，检测优化提取后的嘉宝果叶片提取物对α-葡萄糖苷酶的抑制作用。方法利用超声波辅助与乙醇提取相结合的方法，以总多酚提取量为考察指标，采用单因素及正交试验，研究功率、时间、乙醇体积分数、料液比和温度对嘉宝果叶片总多酚提取量的影响，并采用酶动力学及紫外可见吸收光谱法探讨叶片提取物体外抑制α-葡萄糖苷酶的活性。结果嘉宝果叶片总多酚的最佳提取工艺为：功率120 W，时间10 min，乙醇体积分数40%，料液比1:160（g:mL），温度40℃，在此条件下，嘉宝果叶片总多酚质量分数可达33.84%。优化提取后的嘉宝果叶片提取物对α-葡萄糖苷酶具有较强的抑制作用，半数抑制浓度IC₅₀为0.003 1 mg·mL^-1，阿卡波糖对α-葡萄糖苷酶的半数抑制浓度IC₅₀为8.110 8 mg·mL^-1；Lineweaver-Burk双倒数作图显示嘉宝果叶片多酚对α-葡萄糖苷酶的抑制类型为反竞争性抑制。结论正交实验法优化嘉宝果叶片总多酚提取具有可行性，嘉宝果叶片总多酚对α-葡萄糖苷酶具有较强的抑制作用，可作为降血糖功能因子进行开发利用。
- 嘉宝果 /
- 多酚 /
- α-葡萄糖苷酶 /
- 正交试验
Abstract: Objective To optimize the polyphenol extraction from leaves of jaboticaba and determine the α-glucosidase inhibition by the extract. Method The ethanol extraction with ultrasound application was optimized by an orthogonal experiment. Effects of the power and time of ultrasonic application as well as the ethanol volume fraction, material-solvent ratio, and processing temperature on the total polyphenols extracted from the leaves were analyzed. The inhibition on α-glucosidase activity by the extract was determined in vitro by an enzyme kinetic study and uv-vis absorption spectrometry. Result The optimal extraction conditions included the use of 120 W ultrasonic wave for 10 min, 40% ethanol volume fraction, 1:160(g:mL) material-solvent ratio, and 40℃ processing temperature to achieve a polyphenol extraction rate of 33.84%. The α-glucosidase activity was significantly inhibited by the extract with a low IC₅₀ of 0.0031 mg·mL^-1, as compared to IC₅₀ of 8.110 8 mg·mL^-1 on acarbose. The kinetic study by the Lineweaver-Burk plot showed that the polyphenol extract was an anti-competitive inhibitor of α-glucosidase. Conclusion The polyphenol extract from jaboticaba leaves significantly inhibited α-glucosidase and could be explored for use as a hypoglycemic agent.
- Jaboticaba /
- polyphenol /
- α-glucosidase /
- orthogonal test

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图 1 功率对总多酚提取量的影响

Figure 1. Effect of ultrasonic wave power on polyphenol extraction

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图 2 时间对总多酚提取量的影响

Figure 2. Effect of ultrasonic wave application duration on polyphenol extraction

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图 3 乙醇体积分数对总多酚提取量的影响

Figure 3. Effect of ethanol volume fraction on polyphenol extraction

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图 4 料液比对总多酚提取量的影响

Figure 4. Effect of material-solvent ratio on polyphenol extraction

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图 5 温度对总多酚提取量的影响

Figure 5. Effect of processing temperature on polyphenol extraction

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图 6 嘉宝果叶片总多酚提取物(A)及阿卡波糖(B)对α-葡萄糖苷酶的抑制作用

Figure 6. Effects of polyphenol extract (A) and acarbose (B) on α-glucosidase

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图 7 嘉宝果叶片总多酚提取物对α-葡萄糖苷酶抑制作用类型

Figure 7. Inhibition type of polyphenol extract on α-glucosidase

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图 8 嘉宝果叶片总多酚提取物对α-葡萄糖苷酶抑制作用的Lineweaver-Burk曲线

Figure 8. Lineweaver-Burk plot on inhibition of polyphenol extract on α-glucosidase

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表 1 正交试验因素水平

Table 1. Factors and levels of orthogonal experiment

水平 Level	乙醇体积分数 Ethanol volume fraction/%	料液比 Material-solvent ratio/(g:mL)	温度 Temperature/℃
1	20	1:120	20
2	40	1:160	30
3	60	1:200	40

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表 2 正交试验设计与结果

Table 2. Orthogonal design, results and range analysis

试验号 Numbers	因素				总多酚提取量 Total polyphenol/%
试验号 Numbers	A/%	B/(g:mL)	C/℃	D	总多酚提取量 Total polyphenol/%
1	1(20)	1(1:120)	1(20)	1	30.13
2	1	2(1:160)	2(30)	2	31.91
3	1	3(1:200)	3(40)	3	31.59
4	2(40)	1	2	3	32.67
5	2	2	3	1	34.52
6	2	3	1	2	34.06
7	3(60)	1	3	2	32.22
8	3	2	1	3	33.14
9	3	3	2	1	33.20
k1	93.63	95.02	97.33	97.85
k2	101.25	99.57	97.78	98.19
k3	98.56	98.85	98.33	97.40
R	7.62	4.55	1.00	0.79
注：表中因素A为乙醇体积分数，B为液料比，C为温度，D为空列。表 3同。 Note：A.Ethanol volume fraction，B.Material-solvent ratio，C.Temperature，D.Vacant column.The same as table 3.

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表 3 正交试验结果方差分析

Table 3. Analysis of variance on results of orthogonal experiment

变异来源 Variations	偏差平方和 Sum of square	自由度 Freedom	F值 F value	F临界值 F critical value	P值 P value
A	9.956	2	94.819	19.000	＜0.05
B	3.988	2	37.981	19.000	＜0.05
C	0.167	2	1.590	19.000	>0.05
D	0.105	2	1.000	19.000	>0.05
误差Errors	0.105	2

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表 4 嘉宝果总多酚提取物对α-葡萄糖苷酶抑制作用的Lineweaver-Burk曲线方程

Table 4. Lineweaver-Burk plot equation for inhibition of polyphenol extract on α-glucosidase

项目 Items	质量浓度 Mass concentration/(mg·mL^-1)	线性回归方程Linear regression equation		R²	V_max/(OD·min^-1)	K_m/(mmol·L^-1)
项目 Items	质量浓度 Mass concentration/(mg·mL^-1)	斜率Slope	截距Intercept	R²	V_max/(OD·min^-1)	K_m/(mmol·L^-1)
对照组Contrast	0	5.7203	8.0232	0.9976	0.1246	0.7130
嘉宝果总多酚 Total polyphenol	0.003	5.8844	22.6842	0.9974	0.0441	0.2594
嘉宝果总多酚 Total polyphenol	0.006	5.8105	61.6144	0.9976	0.0162	0.0943

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