蜂胶油提物的制备及抗氧化活性研究

doi:10.3969/j.issn.1004-1524.20231418

浙江农业学报 ›› 2024, Vol. 36 ›› Issue (10): 2338-2346.DOI: 10.3969/j.issn.1004-1524.20231418

蜂胶油提物的制备及抗氧化活性研究

彭正菊¹^,²(), 谌迪²(), 张岑², 卢文静², 喻宏应¹^,², 郭慧媛³, 蒋晗¹^,^*(), 肖朝耿²^,^*()

1.中国计量大学生命科学学院,浙江杭州 310018
2.浙江省农业科学院食品科学研究所,浙江杭州 310021
3.中国农业大学食品科学与营养工程学院,北京 100083

收稿日期:2023-12-20 出版日期:2024-10-25 发布日期:2024-10-30
作者简介:彭正菊(1999—),女,重庆人,硕士研究生,研究方向为蜂胶的活性研究与应用。E-mail: pengzj1127@163.com;
湛迪(1987—),男,湖北武汉人,博士,助理研究员,研究方向为功能性食品配料。E-mail:cd0sindy@163.com第一联系人：
^#为共同第一作者
通讯作者: ^*蒋晗,E-mail: jianghan825@126.com;肖朝耿,E-mail: xiaochaogeng@163.com
基金资助:
国家自然科学基金(32201930);浙江省农业科学院国际合作/港澳台专项经费资助项目(蜂胶油凝胶的制备及功效机理研究)

Preparation and antioxidant activity of propolis oil extracts

PENG Zhengju¹^,²(), CHEN Di²(), ZHANG Cen², LU Wenjing², YU Hongying¹^,², GUO Huiyuan³, JIANG Han¹^,^*(), XIAO Chaogeng²^,^*()

1. College of Life Sciences, China Jiliang University, Hangzhou 310018, China
2. Institute of Food Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
3. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083,China

Received:2023-12-20 Online:2024-10-25 Published:2024-10-30

摘要/Abstract

摘要：

蜂胶是一种良好的药食两用天然产品,具有广泛的生物活性。为探索山茶油提取对蜂胶中活性成分及抗氧化活性的影响,该研究以总黄酮为评价指标,通过单因素及响应面实验获得蜂胶提取最佳工艺条件,并对油提物的DPPH清除能力和铁离子还原能力等抗氧化活性进行了评价,最后以蜂胶中典型的脂溶性高良姜素为例,利用分子对接对其抗氧化活性机制进行了探究。结果表明,提取的最佳工艺为: 温度41 ℃,液固比16∶1 mL·g^-1, 提取时间31 h,此时总黄酮的含量为35.59 mg·g^-1。在该工艺下获得的油提物对DPPH清除率的IC₅₀值为12.48 μg·mL^-1,总抗氧化能力(FRAP)为3.70 mmol·g^-1。此外,分子对接结果表明,蜂胶中典型脂溶性成分高良姜素与蜂胶的抗氧化作用密切相关。

关键词: 蜂胶, 总黄酮, 响应面, 抗氧化, 分子对接

Abstract:

Propolis is a good medicinal and edible natural product, which have a wide range of biological activities. The purpose of this study was to explore the effect of camellia oil extraction on the active components and antioxidant activity in propolis. Firstly, total flavonoids were used as the evaluation index, this study obtained the optimal process conditions for propolis extraction by one-way and response surface experiments. Then, we evaluated the antioxidant activity of the oil extracts including DPPH scavenging ability and ferric iron reduction ability. Finally, the mechanism of the antioxidant activity of galangin, a typical fat-soluble compound in propolis, was investigated using molecular docking was explored. The results showed that the optimal process was as follows: extraction temperature 41 ℃, liquid-solid ratio 16∶1 mL·g^-1, extraction time 31 h, at which the content of total flavonoids was 35.59 mg·g^-1. The IC₅₀ value of the oil extracts obtained under this process was 12.48 μg·mL^-1 for DPPH scavenging, and the total antioxidant capacity(FRAP) was 3.70 mmol·g^-1. In addition, the molecular docking results indicated that galangin, a typical fat-soluble component of propolis, was closely related to the antioxidant effect of propolis.

Key words: propolis, total flavonoids, Box-Behnken response surface, antioxidant, molecular docking

中图分类号:

TS201

彭正菊, 谌迪, 张岑, 卢文静, 喻宏应, 郭慧媛, 蒋晗, 肖朝耿. 蜂胶油提物的制备及抗氧化活性研究[J]. 浙江农业学报, 2024, 36(10): 2338-2346.

PENG Zhengju, CHEN Di, ZHANG Cen, LU Wenjing, YU Hongying, GUO Huiyuan, JIANG Han, XIAO Chaogeng. Preparation and antioxidant activity of propolis oil extracts[J]. Acta Agriculturae Zhejiangensis, 2024, 36(10): 2338-2346.

图/表 13

表1 单因素试验设计

Table 1 Single factor experimental design

水平 Level	因素Factor
水平 Level	提取温度 Extraction temperature/℃	液固比 Liquid-solid ratio/(mL·g^-1)	提取时间 Extraction time/h
1	25	5∶1	18
2	30	10∶1	24
3	35	15∶1	30
4	40	20∶1	36
5	45	25∶1	42

表2 响应面试验设计

Table 2 Response surface experimental design

水平 Level	因素Factor
水平 Level	A:提取温度 Extraction temperature/℃	B:液固比 Liquid-solid ratio/(mL·g^-1)	C:提取时间 Extraction time/h
-1	35	10∶1	24
0	40	15∶1	30
1	45	20∶1	36

图1 槲皮素标准曲线

Fig.1 Quercetin standard curve

图2 温度对总黄酮含量的影响

Fig.2 Effect of temperature on total flavonoids content

图3 液固比对总黄酮含量的影响

Fig.3 Effect of liquid-solid ratio on total flavonoids content

图4 时间对总黄酮含量的影响

Fig.4 Effect of time on total flavonoids content

表3 响应面试验的设计及结果

Table 3 Design and results of the response surface test

序号 ID	A:温度 θ/℃	B:液固比 Liquid-solid ratio/(mL·g^-1)	C:时间 t/h	R总黄酮含量 Total flavonoids content/(mg·g^-1)
1	0	-1	1	26.07
2	0	1	-1	35.78
3	-1	-1	0	35.47
4	-1	1	0	30.50
5	0	0	0	29.63
6	0	0	0	33.08
7	0	0	0	30.07
8	0	-1	-1	35.15
9	1	-1	0	29.25
10	1	1	0	27.18
11	-1	0	1	35.36
12	1	0	-1	30.23
13	1	0	1	32.16
14	0	0	0	34.48
15	0	0	0	30.13
16	0	1	1	32.34
17	-1	0	-1	25.52

表4 方差分析及显著性结果

Table 4 ANOVA and significance results

变异源 Source	平方和 Sum of squares	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	171.19	9	19.0	64.65	<0.000 1	**
A-温度θ/℃	5.38	1	5.38	18.30	0.003 7	**
B-液固比Liquid-solid ratio/(mL·g^-1)	27.68	1	27.68	94.06	<0.000 1	**
C-时间t/h	5.02	1	5.02	17.08	0.004 4	**
AB	11.36	1	11.36	38.60	0.000 4	**
AC	2.28	1	2.28	7.75	0.027 2	*
BC	0.98	1	0.98	3.33	0.110 7	不显著Not significant
A²	13.86	1	13.86	47.09	0.000 2	**
B²	64.83	1	64.83	220.33	<0.000 1	**
C²	28.77	1	28.77	97.78	<0.000 1	**
残差 Residual	2.06	7	0.29	-	-	-
失拟项 Lack of Fit	1.11	3	0.37	1.58	0.327 0	不显著Not significant

图5 各因素交互作用的响应面图

Fig.5 Response surface plots for the interaction of factors

图6 不同质量浓度蜂胶油提取物对DPPH自由基清除率的影响

Fig.6 Effect of different concentrations of propolis oil extracts on the scavenging rate of DPPH radicals

图7 Fe2+标准曲线

Fig.7 Fe2+ standard curve

图8 蜂胶提取物的总抗氧化能力吸光度

Fig.8 Absorbance variation of total antioxidant capacity of propolis extracts

图9 高良姜素与NOS3和NOS2对接情况 A是NOS3的蛋白结构图,B、C分别表示高良姜素与NOS3结合的2D和3D图;D是NOS2的蛋白结构图,E、F分别表示高良姜素与NOS2结合的2D和3D图。

Fig.9 Docking of galangin with NOS3and NOS2 A is the protein structure of NOS3,B and C represent 2D and 3D plots of galangin binding to NOS3; D is the protein structure of NOS2, E and F represent 2D and 3D plots of galangin binding to NOS2.

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蜂胶油提物的制备及抗氧化活性研究

Preparation and antioxidant activity of propolis oil extracts

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