中国中药杂志

2020, v.45(17) 4238-4245

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发酵柴胡地上部分定量分析及其抗菌活性成分预测
Quantitative analysis of fermented aerial part of Bupleurum chinense and prediction of their antimicrobial activity

郭舒臣;郭瑞;夏翊腾;董婷霞;王怀友;詹华强;
GUO Shu-chen;GUO Rui;XIA Yi-teng;DONG Ting-xia;WANG Huai-you;TSIM Wah-Keung Karl;Shenzhen Key Laboratory of Edible and Medicinal Bioresources, Shenzhen Research Institute,Hong Kong University of Science and Technology;Division of Life Science and Center for Chinese Medicine, the Hong Kong University of Science and Technology;

摘要(Abstract):

通过关联分析植物乳杆菌发酵的柴胡地上部分中活性成分的含量和抗菌活性结果,预测其抗菌活性成分。建立8种黄酮成分(山柰酚-3-O-芸香糖苷,异槲皮苷,槲皮素,异鼠李素,芦丁,野鹫尾苷,槲皮素-3-O-β-L-阿拉伯糖苷,山柰酚)和DL-3-苯基-2-羟丙酸的UPLC-MS-MS检测条件,并且对其发酵过程中含量变化进行动态监测。同时选用5种常见非水生病原菌(金黄色葡萄球菌,大肠杆菌,铜绿假单胞菌,肺炎克雷伯菌,枯草芽孢杆菌)和4种常见水生病原菌(嗜水气单胞菌,迟缓爱德华菌,解藻弧菌,哈维弧菌),考察柴胡地上部分不同发酵时间的体外抗菌活性,最终通过皮尔逊相关性分析预测柴胡地上部分发酵产物的抗菌活性成分。该研究所建立的UPLC-MS-MS分析方法线性良好,最宽的线性范围在0.19~50μg·mL~(-1),定量限在0.19~1.56μg·mL~(-1),检测限在0.10~0.78μg·mL~(-1)。在发酵柴胡地上48 h内,3种黄酮(芦丁,槲皮素-3-O-β-L-阿拉伯糖苷,异槲皮苷)和DL-3-苯基-2-羟丙酸的含量明显增加。发酵柴胡地上部分对水生病原菌的抑菌效果明显强于非水生病原菌;进一步通过皮尔逊相关性分析对发酵物活性成分的预测得出,对于这4种水生病原菌,异槲皮苷,芦丁,槲皮素-3-O-β-L-阿拉伯糖苷,DL-3-苯基-2-羟丙酸与抗菌活性呈现显著相关性。该研究表明发酵柴胡地上部分对水生病原菌有较高的敏感度,并且通过相关性分析得出,可能是由于异槲皮苷,芦丁,槲皮素-3-O-β-L-阿拉伯糖苷,DL-3-苯基-2-羟丙酸含量的增加所导致。该研究为进一步开发大量浪费的柴胡地上部分提供理论依据和新思路。
The aim of this study was to predict the anti-microbial components in the aerial part of Bupleurum chinense fermented by Lactobacillus plantarum through analyzing the correlation between contents of bioactive components and their inhibitory action for pathogenic bacteria. In this study, the UPLC-MS-MS detection method was established for eight flavonoids(kaempferol-3-O-β-D-rutinoside, isoquercitrin, quercetin, isorhamnetin, rutin, iridin, quercetin-3-O-β-L-arabinoside, kaempferol) and DL-3-phenyllactic acid, and the dynamic change of their contents at fermentation course were monitored. Meanwhile, the experiment employed five common no-naquatic pathogenic bacteria(Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Bacillus subtilis), and four common aquatic pathogenic bacteria(Aeruginosa hydrophila, Delayed Edwards, Vibrio alginolyticus, and Vibrio harveyi) to validate in vitro anti-microbial activity of the fermented aerial part of B. chinense at different fermentation time points. Finally, the Pearson correlation analysis was applied to predict the anti-microbial components of the fermented aerial part of B. chinense.The established UPLC-MS-MS method showed a good linearity and the widest linear range was from 0.19 μg·mL~(-1) to 50 μg·mL~(-1). The limit of quantitation and the limit of detection were 0.19-1.56 μg·mL~(-1) and 0.1-0.78 μg·mL~(-1) respectively. During the fermentation within 48 h, the contents of three flavonoids(rutin, quercetin-3-O-β-L-arabinoside, isoquercitrin) and DL-3-phenyllactic acid from the fermented aerial part of B. chinense increased sharply. In the process of fermentation, the anti-microbial effect of the fermented aerial part of B. chinense on aquatic pathogens was significantly stronger than that on non-aquatic pathogens. Furthermore, Pearson correlation analysis predicted that isoquercitrin, rutin, quercetin-3-O-β-L-arabinoside and DL-3-phenyllactic acid showed significant correlation with the four aquatic pathogens. This study revealed that the fermented aerial part of B. chinense had a high sensitivity to aquatic pathogens, which may be caused by the increased contents of isoquercitrin, rutin, quercetin-3-O-β-L-arabinoside and DL-3-phenyllactic acid. In conclusion, this study provides a theoretical basis and new idea for the further development of the large amount of wasteful aerial part of Bupleurum chinense.

关键词(KeyWords): 柴胡地上部分;植物乳杆菌发酵;UPLC-MS-MS分析;抗菌活性;皮尔逊相关性分析
aerial part of Bupleurum chinense;fermentation by Lactobacillus plantarum;UPLC-MS-MS analysis;anti-microbial activities;Pearson correlation analysis

Abstract:

Keywords:

基金项目(Foundation): 深圳市科技计划项目(CKFW2016082916015476,ZDSYS201707281432317);; 佛山科大专项(FSUST19-SRI10)

作者(Author): 郭舒臣;郭瑞;夏翊腾;董婷霞;王怀友;詹华强;
GUO Shu-chen;GUO Rui;XIA Yi-teng;DONG Ting-xia;WANG Huai-you;TSIM Wah-Keung Karl;Shenzhen Key Laboratory of Edible and Medicinal Bioresources, Shenzhen Research Institute,Hong Kong University of Science and Technology;Division of Life Science and Center for Chinese Medicine, the Hong Kong University of Science and Technology;

Email:

DOI: 10.19540/j.cnki.cjcmm.20200622.305

参考文献(References):

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