超声波辅助单宁酶法提取五倍子没食子酸及其抗氧化作用研究

李振; 李萍

doi:10.3969/j.issn.1006-267x.2019.08.045

动物营养学报 >

2019 , Vol. 31 >Issue 8: 3830 - 3842

DOI: https://doi.org/10.3969/j.issn.1006-267x.2019.08.045

饲料科学与技术 Feed science and technology

超声波辅助单宁酶法提取五倍子没食子酸及其抗氧化作用研究

李振 ,
李萍

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1. 临沂大学药学院, 临沂 276005;
2. 山东省鲁南中药材资源开发工程技术研究中心, 临沂 276005;
3. 临沂市人民医院, 临沂 276003

李振(1971-),男,山东临沂人,副教授,博士,主要从事天然产物研究与开发工作。E-mail:Lizhen8217@126.com

收稿日期: 2019-01-16

网络出版日期: 2019-08-19

基金资助

山东省自然科学基金项目（ZR2015PH029）；临沂大学博士科研启动基金项目（LYDX2016BS082）

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Study on Gallic Acid from Galla chinensis by Ultrasonic-Assisted Tannin Enzymatic Method and Its Antioxygenation

LI Zhen ,
LI Ping

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1. College of Pharmacy, Linyi University, Linyi 276005, China;
2. Shandong Lunan Chinese Medicinal Materials Resources Development Engineering Technology Research Center, Linyi 276005, China;
3. Linyi People's Hospital, Linyi 276003, China

Received date: 2019-01-16

Online published: 2019-08-19

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摘要

本研究旨在探索超声波辅助单宁酶法提取五倍子没食子酸的最佳提取工艺参数及其抗氧化作用。采用单因素试验设计，以没食子酸得率为指标，研究单宁酶添加量、酶解温度、酶解时间、pH、超声功率5个因素对五倍子提取没食子酸得率的影响，并采用响应面法优化五倍子没食子酸的最佳提取工艺参数。选择健康、体重（BW）（20±2）g的昆明小鼠40只（雌雄各1/2），随机分为4组，每组10只。空白对照组按每10 g BW 0.1 mL生理盐水灌胃，试验组分别按50（低剂量组）、75（中剂量组）、100 mg/kg BW（高剂量组）剂量灌服同体积没食子酸溶液，每天1次，试验期21 d。结果表明，没食子酸最佳提取工艺参数为：单宁酶添加量11 U/g，酶解温度45℃，酶解时间4.8 h，pH 6，超声功率380 W，此条件下没食子酸得率为（64.73±1.61）%；所得没食子酸采用结晶和离子交换树脂方法进行分离纯化，纯度为（99.01±2.47）%。在没食子酸质量浓度为60 μg/mL时，还原力为0.85±0.04，对1，1-二苯基-2-苦肼基自由基（DPPH·）、超氧阴离子自由基（O₂^-·）清除率分别为（92.11±2.38）%、（98.62±2.75）%。与空白对照组比较，中、高剂量组肝脏中超氧化物歧化酶（SOD）、过氧化氢酶（CAT）及谷胱甘肽过氧化物酶（GSH-Px）活性显著或极显著升高（P<0.05或P<0.01），低、中、高剂量组肝脏中丙二醛（MDA）含量显著或极显著降低（P<0.05或P<0.01）。由此可见，超声波辅助单宁酶法能够提高没食子酸得率，没食子酸可以有效地清除自由基，具有良好的抗氧化作用。

关键词： 五倍子; 没食子酸; 超声波; 单宁酶; 响应面法; 抗氧化

本文引用格式

李振 , 李萍 . 超声波辅助单宁酶法提取五倍子没食子酸及其抗氧化作用研究[J]. 动物营养学报, 2019 , 31(8) : 3830 -3842 . DOI: 10.3969/j.issn.1006-267x.2019.08.045

Abstract

This study was conducted to explore the optimum extraction process conditions of gallic acid from Galla chinensist by ultrasonic-assisted tannin enzymatic method and its antioxygenation. A single factor design was adopted in the experiment, to study the effects of tannase enzyme addition amount, enzymolysis temperature, enzymolysis time, pH, ultrasonic power on yield of gallic acid, and the response surface method was used to optimize the extraction process condition of gallic acid from Galla chinensis. Forty healthy Kunming mice (half male and half female) with the body weight (BW) of (20±2) g were selected and divided into 4 groups with 10 mice per group. Mice in the blank control group were gavage 10 g BW 0.1 mL normal saline, and mice in the experimental groups were gavage 50 (low dose group), 75 (medium dose group) and 100 mg/kg BW (high dose group) gallic acid solution of the same volume, respectively, one times a day, and the experiment last for 21 days. The results showed that the optimum extraction process conditions of gallic acid were as follows:the tannase enzyme addition amount was 11 U/g, the enzymolysis temperature was 45℃,the enzymolysis time was 4.8 h, the pH was 6 and the ultrasonic power was 380 W. Under the above conditions, the yield of gallic acid was (64.73±1.61)%. The purity of gallic acid was (99.01±2.47)% which puritied by crystallization method and ion exchange resin method. On the mass concentration of 60 μg/mL, the reducing force was 0.85±0.04, and the scavenging rates of 1,1-diphenyl-2-picrylhydrazine radical (DPPH·) and superoxide anion radical (O₂^-·) were (92.11±2.38)% and (98.62±2.75)%, respectively. Compared with the blank control group, the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in liver of medium and high dose groups were significantly increased (P<0.05 and P<0.01), and the liver malondialdehyde (MDA) content of low, medium and high dose groups was significantly decreased (P<0.05 and P<0.01). Therefore, the ultrasonic-assisted tannin enzymatic method can improve the yield of gallic acid, the gallic acid can scavenge free radical effectively and has good antioxygenation.

Key words： Galla chinensis; gallic acid; ultrasonic; tannase enzyme; response surface method; antioxidant

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