Shu-Chen Guo, Rui Guo, Yi-Teng Xia, Ting-Xia Dong, Huai-You Wang, Wah-Keung Karl Tsim
Shenzhen Key Laboratory of Edible and Medicinal Bioresources, Shenzhen Research Institute,Hong Kong University of Science and Technology Shenzhen 518057, China.
Division of Life Science and Center for Chinese Medicine, the Hong Kong University of Science and Technology Hong Kong, China.
Zhongguo Zhong Yao Za Zhi. 2020 Sep;45(17):4238-4245. doi: 10.19540/j.cnki.cjcmm.20200622.305.
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.
本研究旨在通过分析生物活性成分含量与其对病原菌抑制作用之间的相关性，预测植物乳杆菌发酵柴胡地上部分的抗菌成分。本研究建立了超高效液相色谱-串联质谱（UPLC-MS-MS）检测方法，用于测定8种黄酮类化合物（山柰酚-3-O-β-D-芸香糖苷、异槲皮苷、槲皮素、异鼠李素、芦丁、鸢尾苷、槲皮素-3-O-β-L-阿拉伯糖苷、山柰酚）和DL-3-苯乳酸，并监测其在发酵过程中的含量动态变化。同时，采用5种常见的非水生病原菌（金黄色葡萄球菌、大肠杆菌、铜绿假单胞菌、肺炎克雷伯菌和枯草芽孢杆菌）和4种常见的水生病原菌（嗜水气单胞菌、迟缓爱德华氏菌、溶藻弧菌和哈维氏弧菌），验证柴胡地上部分发酵物在不同发酵时间点的体外抗菌活性。最后，通过Pearson相关性分析预测柴胡地上部分发酵物的抗菌成分。所建立的UPLC-MS-MS方法线性良好，线性范围最宽为0.19 μg·mL⁻¹至50 μg·mL⁻¹。定量限和检测限分别为0.19 - 1.56 μg·mL⁻¹和0.1 - 0.78 μg·mL⁻¹。在48 h内的发酵过程中，柴胡地上部分发酵物中的3种黄酮类化合物（芦丁、槲皮素-3-O-β-L-阿拉伯糖苷、异槲皮苷）和DL-3-苯乳酸含量急剧增加。在发酵过程中，柴胡地上部分发酵物对水生病原菌的抗菌效果明显强于对非水生病原菌的抗菌效果。此外，Pearson相关性分析预测，异槲皮苷、芦丁、槲皮素-3-O-β-L-阿拉伯糖苷和DL-3-苯乳酸与4种水生病原菌呈显著相关性。本研究表明，柴胡地上部分发酵物对水生病原菌具有较高的敏感性，这可能是由于异槲皮苷、芦丁、槲皮素-3-O-β-L-阿拉伯糖苷和DL-3-苯乳酸含量增加所致。综上所述，本研究为大量废弃的柴胡地上部分的进一步开发提供了理论依据和新思路。
