油橄榄叶多糖的提取工艺优化及其理化性质和抗氧化活性

任一杰; 赵小亮; 王宝忠; 向紫骏; 杨超福; 王海利; 马君义; 张伟杰

doi:10.13386/j.issn1002-0306.2022030302

油橄榄叶多糖的提取工艺优化及其理化性质和抗氧化活性

doi: 10.13386/j.issn1002-0306.2022030302

任一杰^{1, 2,},
赵小亮^3, ,,
王宝忠³,
向紫骏³,
杨超福³,
王海利³,
马君义⁴,
张伟杰^3, ,

1.
甘肃陇神戎发药业股份有限公司，甘肃兰州 730102
2.
甘肃省中药固体分散制剂重点实验室，甘肃兰州 730102
3.
兰州理工大学生命科学与工程学院，甘肃兰州 730050
4.
西北师范大学生命科学学院，甘肃兰州 730070

基金项目: 国家自然科学基金（31860250，32260231）；甘肃省自然科学基金（18JR3RA148）；甘肃省基础研究创新群体计划项目（1506RJIA116）；甘肃省科技创新专项基金项目（2019ZX-05）。

详细信息

作者简介:
任一杰（1981−），男，本科，高级工程师，研究方向：天然产物化学，E-mail：471208356@qq.com

通讯作者:
赵小亮（1982−），男，博士，副教授，研究方向：糖化学与糖药物，E-mail：zhxl819@163.com

张伟杰（1975−），男，博士，教授，研究方向：糖基医用生物材料，E-mail：brossica@163.com

中图分类号: TS201.1
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出版历程
- 收稿日期: 2022-03-24
- 网络出版日期: 2022-10-21
- 刊出日期: 2022-11-23

Optimization of Extraction Process of Polysaccharides from Olea europaea L. Leaves and Its Physicochemical Properties and Antioxidant Activity

1.
Gansu Longshen Rongfa Pharmaceutical Industry Co., Ltd., Lanzhou 730102, China
2.
Gansu Provincial Key Laboratory of Solid Dispersible Preparations for Traditional Chinese Medicine, Lanzhou 730102, China
3.
School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China
4.
College of Life Sciences, Northwest Normal University, Lanzhou 730070, China

摘要

摘要: 为增加油橄榄叶多糖的开发利用价值，以油橄榄叶为实验材料，采用正交试验优化油橄榄叶多糖（OLP）的提取工艺，高效凝胶渗透色谱-多角度激光光散射仪联用（HPGPC-MALLS）测定分子量，PMP柱前衍生高效液相色谱法分析OLP的单糖组成，并评价其抗氧化活性。结果显示，OLP的最佳提取工艺条件为料液比1:27.5 g/mL，温度95 ℃，提取时间3.5 h，在此条件下OLP的得率为2.75%；OLP的重均分子量（Mw）为25.36 kDa，数均分子量（Mn）为19.32 kDa，多分散系数为1.313；OLP主要由葡萄糖（Glc）、半乳糖（Gal）和氨基半乳糖（GalN）组成，还含有鼠李糖（Rha）、阿拉伯糖（Ara）、木糖（Xyl）、甘露糖（Man）和氨基葡萄糖（GlcN）等单糖，各单糖的相对摩尔比为56.2:15.9:10.3:8.3:5.9:2.6:0.5:0.3；抗氧化活性实验结果发现，OLP具有较好的抗氧化活性，对羟自由基、超氧阴离子自由基和DPPH自由基的半数抑制浓度（IC₅₀）分别为0.422、0.302和0.268 mg/mL。本研究所得油橄榄叶多糖的提取工艺简单、得率高、抗氧化活性好，为油橄榄叶多糖的进一步研究和开发利用提供了重要参考。
- 油橄榄叶 /
- 多糖 /
- 正交试验 /
- 理化性质 /
- 抗氧化活性
Abstract: In order to increase the development and utilization value of polysaccharides from Olea europaea L. leaves, the extraction process of polysaccharides from Olea europaea L. leaves (OLP) was optimized by orthogonal experiment, the molecular weight was determined by high performance gel permeation chromatography combined with multi-angle laser light scattering (HPGPC-MALLS), and the monosaccharide composition of OLP was analyzed by PMP pre-column derivatization and its antioxidant activity was evaluated. The results showed that the optimal extraction conditions were material-liquid ratio of 1:27.5 g/mL, extraction temperature of 95 ℃ and extraction time of 3.5 h. Under these conditions, the yield of OLP was 2.75%. The average weight molecular weight (Mw) of OLP was 25.36 kDa, the average number molecular weight (Mn) was 19.32 kDa, and the polydispersity was 1.313. OLP was mainly composed of glucose (Glc), galactose (Gal) and galactose amino (GalN), but also contains rhamnose (Rha), arabinose (Ara), xylose (Xyl), mannose (Man) and glucosamine (GlcN), and the relative molar ratio of monosaccharide was 56.2:15.9:10.3:8.3:5.9:2.6:0.5:0.3. The results showed that OLP had good antioxidant activity, with IC₅₀ of 0.422, 0.302 and 0.268 mg/mL for hydroxyl free radical, superoxide anion free radical and DPPH free radical, respectively. The optimum technology of OLP is simple, the yield is high, and the antioxidant activity is good, which provides an essential reference for the further research, development and utilization of OLP.
- Olea europaea L. leaves /
- polysaccharide /
- orthogonal test /
- physicochemical properties /
- antioxidant activity

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图 1 料液比对多糖得率的影响

Figure 1. Effects of material-liquid ratio on the yield of polysaccharides

注：不同小写字母表示差异显著（P<0.05）；图2~图3同。

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图 2 温度对多糖得率的影响

Figure 2. Effects of temperature on the yield of polysaccharides

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图 3 时间对多糖得率的影响

Figure 3. Effects of time on the yield of polysaccharides

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图 4 OLP红外光谱图

Figure 4. Infrared spectrogram of OLP

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图 5 OLP单糖组成分析HPLC图

Figure 5. Monosaccharide composition analysis of OLP by HPLC

注：1. Man；2. GlcN；3. Rha；4. GlcA；5. GalN；6. GalA；7. Glc；8. Gal；9. Xyl；10. Ara。

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图 6 OLP对羟自由基的清除活性

Figure 6. Scavenging activity of OLP on hydroxyl free radical

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图 7 OLP对超氧阴离子自由基的清除活性

Figure 7. Scavenging activity of OLP on superoxide anion free radical

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图 8 OLP对DPPH自由基的清除活性

Figure 8. Scavenging activity of OLP on DPPH free radical

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表 1 正交试验因素水平

Table 1. Orthogonal assay factors and levels

水平	因素
水平	A 料液比（g/mL）	B 温度（℃）	C 时间（h）
1	1:27.5	85	2.5
2	1:30.0	90	3.0
3	1:32.5	95	3.5

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表 2 L₉（3⁴）正交实验及结果

Table 2. L₉(3⁴) orthogonal design and experiment results

序号	影响因素				多糖得率（%）
序号	A 料液比	B 温度	C 时间	D （空白列）	多糖得率（%）
1	1	1	1	1	2.10
2	1	2	2	2	2.51
3	1	3	3	3	2.74
4	2	1	2	3	2.07
5	2	2	3	1	2.42
6	2	3	1	2	2.24
7	3	1	3	2	2.19
8	3	2	1	3	2.44
9	3	3	2	1	2.46
K₁	7.35	6.36	6.78	6.98
K₂	6.73	7.37	7.04	6.94
K₃	7.09	7.44	7.35	7.25
k₁	2.45	2.12	2.26	2.33
k₂	2.24	2.46	2.35	2.31
k₃	2.36	2.48	2.45	2.42
R	0.21	0.36	0.19	0.11

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表 3 OLP分子量分析结果

Table 3. OLP molecular weight analysis results

OLP	Mw（kDa）	Mn（kDa）	多分散系数（Mw/Mn）
OLP	25.36	19.32	1.313

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表 4 OLP单糖组成及相对摩尔比分析结果

Table 4. The analysis results of monosaccharide composition and relative mole ratio

样品	单糖组成及其相对摩尔比
样品	Man	GlcN	Rha	GalN	Glc	Gal	Xyl	Ara
OLP	0.5	0.3	8.3	10.3	56.2	15.9	2.6	5.9

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