五味子木脂素超声提取工艺优化及其抗氧化和抗真菌的潜力

doi:10.3969/j.issn.1004-1524.2021.11.17

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (11): 2145-2154.DOI: 10.3969/j.issn.1004-1524.2021.11.17

五味子木脂素超声提取工艺优化及其抗氧化和抗真菌的潜力

李启思¹^,²(), 王雅玲¹^,²^,^*(), 邓玉华¹^,², 廖建萌¹^,², 叶林¹^,², 吴莉莉¹^,², 郑佳纯¹^,², 罗杏燕¹^,², 邓旗¹^,², 孙力军¹^,²

1.广东海洋大学食品科技学院,广东省水产品加工与安全重点实验室,广东普通高等学校水产品深加工重点实验室,广东湛江 524088
2.湛江市食品药品检验所,广东湛江 524037

收稿日期:2020-12-24 出版日期:2021-11-25 发布日期:2021-11-26
通讯作者: 王雅玲
作者简介:*王雅玲,E-mail: wangylchina@163.com
李启思(1995—),女,广东肇庆人,硕士研究生,研究方向为水产品质量与安全。E-mail: 190199230@qq.com
基金资助:
国家自然科学基金(31871898);广东省大学生创新创业训练计划(粤教高函〔2018〕42号);广东省现代农业产业技术体系创新团队(2019KJ149)

Optimization of ultrasonic extraction technology of Shisandra lignans and its anti-oxidant and anti-fungi potentials

LI Qisi¹^,²(), WANG Yaling¹^,²^,^*(), DENG Yuhua¹^,², LIAO Jianmeng¹^,², YE Lin¹^,², WU Lili¹^,², ZHENG Jiachun¹^,², LUO Xingyan¹^,², DENG Qi¹^,², SUN Lijun¹^,²

1. College of Food Science and Technology, Guangdong Ocean University, Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Key Laboratory of Advanced Processing of Aquatic Products of Guangdong Higher Education Institution, Zhanjiang 524088, China
2. Zhanjiang Institute for Food & Drug Control, Zhanjiang 524037, China

Received:2020-12-24 Online:2021-11-25 Published:2021-11-26
Contact: WANG Yaling

摘要/Abstract

摘要：

为探究五味子木脂素的超声提取工艺,及其抗氧化和抗真菌的能力,建立起一种基于液相色谱-质谱联用法(LC-MS/MS)同步检测五味子中5种木脂素含量的方法,在此基础上,基于4项单因素试验和中心复合设计-响应面法优化五味子木脂素的超声提取工艺,并评价其抗氧化和抗水产干制品中优势真菌的能力。结果表明:适宜超声提取五味子木脂素的工艺参数为乙醇体积分数70%,液料比12 mL·g^-1,提取2次,每次超声10 min。在此条件下,五味子木脂素提取量为(9.45±0.11) mg·g^-1。五味子木脂素提取液的总还原力,及对DPPH(1,1-二苯基-2-三硝基苯肼)、ABTS(2,2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸)自由基的清除能力分别是维生素C(V_C)的2/5、1/2和1/3,对黄曲霉(Aspergillus flavus)、黑曲霉(Aspergillus niger)、橘青霉(Penicillium citrinum)、尖孢镰孢菌(Fusarium oxysporium)和厚垣镰孢菌(Fusarium chlamydosporum)的最低抑菌浓度(MIC)约为1.17 mg·mL^-1,对哈茨木霉(Trichoderma harzianum)、扩展青霉(Penicillium expansum)和腐皮镰孢菌(Fusarium solani)的MIC约为2.34 mg·mL^-1,说明其具有较强的抗氧化和抗水产干制品中优势真菌的能力。

关键词: 五味子木脂素, 液质联用, 超声提取, 响应面, 抗氧化

Abstract:

To explore the suitable ultrasonic extraction technology of Schisandra lignans and its inhibitory effect against oxidant and dominant fungi in dried aquatic products, a synchronous detection methods for 5 kinds of lignans was established via LC-MS/MS (liquid chromatography-tandem mass spectrometry). On the basis, four single-factor tests, and the central composite response surface test were adopted to optimize the ultrasonic extraction parameters of Schisandra lignans, and the anti-oxidant and anti-fungi activities of Schisandra lignans were analyzed. It was shown that the suitable extraction parameters included ethanol volume fraction of 70%, liquid to material ratio of 12 mL·g ^-1, extraction times of twice, and ultrasound time of 10 min per time. Under this condition, the extraction amount of lignans was (9.45±0.11) mg·g^-1. The total reducing power, and DPPH (1,1-diphenyl-2-picrylhydrazyl), ABTS [2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate)] free radical scavenging ability of Schisandra lignans were 2/5, 1/2 and 1/3 of vitamin C (V_C), respectively. The minimum inhibitory concentration (MIC) of Schisandra lignans against Aspergillus flavus, Aspergillus niger, Penicillium citrinum, Fusarium oxysporium, Fusarium chlamydosporum was about 1.17 mg·mL ^-1, and the MIC of Schisandra lignans against Trichoderma harzianum, Penicillium expansum, Fusarium solani was about 2.34 mg·mL ^-1, which indicated that the extracted Schisandra lignans had strong inhibitory effect against oxidant and dominant fungi in dried aquatic products.

Key words: Schisandra lignans, liquid chromatography-tandem mass spectrometry, ultrasonic extraction, response surface, antioxidation

中图分类号:

TQ461
R917

李启思, 王雅玲, 邓玉华, 廖建萌, 叶林, 吴莉莉, 郑佳纯, 罗杏燕, 邓旗, 孙力军. 五味子木脂素超声提取工艺优化及其抗氧化和抗真菌的潜力[J]. 浙江农业学报, 2021, 33(11): 2145-2154.

LI Qisi, WANG Yaling, DENG Yuhua, LIAO Jianmeng, YE Lin, WU Lili, ZHENG Jiachun, LUO Xingyan, DENG Qi, SUN Lijun. Optimization of ultrasonic extraction technology of Shisandra lignans and its anti-oxidant and anti-fungi potentials[J]. Acta Agriculturae Zhejiangensis, 2021, 33(11): 2145-2154.

图/表 8

图1 五味子木脂素的色谱图 A,标准品;B,提取液。图A中5个峰从左到右依次为五味子醇甲、五味子醇乙、五味子酚、五味子甲素、五味子乙素。

Fig.1 Chromatograms of Schisandra lignans A, Reference; B, Extract. The five peaks from left to right in figure A are Schisandrol A, Schisandrol B, Schisanhenol, Deoxyschizandrin, γ-Schizandrin.

表1 五味子木脂素的质谱定性离子对及其保留时间

Table 1 Ion pairs and their retention time for mass spectrometry detection of Schisandra lignan

化合物 Compound	母离子 Parent ion	子离子 Daughter ion	保留时间 Retention time/min
五味子甲素	434.51	316.0/402.1	7.75
Deoxyschizandrin
五味子乙素γ-Schizandrin	418.46	300.0/331.0	8.50
五味子醇甲Schisandrol A	450.50	415.2/384.2	6.35
五味子醇乙Schisandrol B	434.77	399.2/368.1	6.80
五味子酚Schisanhenol	421.20	340.2/302.2	7.25

表2 五味子木脂素的回归方程

Table 2 Regression equation of Schisandra lignans

化合物 Compound	回归方程 Regression equation	R²	线性范围 Linear range/(μg·kg^-1)	LOD/ (μg·kg^-1)	LOQ/ (μg·kg^-1)
五味子甲素Deoxyschizandrin	y=4.595 5×10⁴x-5.121 2×10⁴	0.999	0~200	0.2	0.5
五味子乙素γ-Schizandrin	y=1.940 3×10⁵x-4.002 5×10⁵	0.998	0~200	0.1	0.4
五味子醇甲Schisandrol A	y=8.235 7×10⁴x-1.625 7×10⁵	0.998	0~200	0.3	0.6
五味子醇乙Schisandrol B	y=9.077 9×10⁴x-3.420 7×10⁵	0.998	0~200	0.2	0.6
五味子酚Schisanhenol	y=1.228 5×10⁵x-1.130 1×10⁵	0.999	0~200	0.1	0.5

图2 乙醇体积分数、超声时间、提取次数、液料比对五味子木脂素提取效果的影响柱上无相同字母的表示处理间差异显著(P<0.05)。

Fig.2 Effect of ethanol volume fraction, ultrasound time, extraction times, liquid to material ratio on extraction effect of Schisandra lignans Bars marked without the same letters indicated significant difference at P<0.05.

表3 响应面试验设计与结果

Table 3 Design and results of response surface experiment

试验号 Test number	A	B	C	Y/(mg·g^-1)
1	-1	-1	-1	6.12
2	1	-1	-1	6.71
3	0	0	-1	7.52
4	-1	1	-1	7.02
5	1	1	-1	7.84
6	0	-1	0	7.92
7	-1	0	0	8.59
8	0	0	0	9.35
9	0	0	0	9.58
10	1	0	0	8.81
11	0	1	0	9.31
12	-1	-1	1	7.85
13	1	-1	1	8.51
14	0	0	1	9.38
15	-1	1	1	8.26
16	1	1	1	9.00

图3 各因素交互作用的响应面和等高线图 A,乙醇体积分数;B,液料比;C,提取次数;Y,五味子木脂素提取量。

Fig.3 Response surface and contour map of interaction of various factors A, Ethanol volume fraction; B, Liquid to material ratio; C, Extraction times; Y, Schisandra lignans content.

图4 五味子木脂素的抗氧化能力同一物质不同浓度间无相同字母的表示差异显著(P<0.05)。

Fig.4 Antioxidant capacity of Schisandra lignans Points marked without the same letters indicate significant difference at P<0.05 within test concentrations of the same material.

表4 五味子木脂素对水产干制品中优势真菌的最低抑菌浓度(MIC)和最低杀菌浓度(MBC)

Table 4 Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Schisandra lignans against dominant fungi in dried aquatic products mg·mL-1

供试菌株Test strain	MIC	MBC
黄曲霉Aspergillus flavus	1.17	2.34
黑曲霉Aspergillus niger	1.17	2.34
橘青霉Penicillium citrinum	1.17	2.34
扩展青霉Penicillium expansum	2.34	4.69
哈茨木霉Trichoderma harzianum	2.34	4.69
腐皮镰孢菌Fusarium solani	2.34	4.69
尖孢镰孢菌Fusarium oxysporium	1.17	2.34
厚垣镰孢菌Fusarium chlamydosporum	1.17	2.34

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五味子木脂素超声提取工艺优化及其抗氧化和抗真菌的潜力

Optimization of ultrasonic extraction technology of Shisandra lignans and its anti-oxidant and anti-fungi potentials

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