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藤茶()叶中抗菌化合物及其作用模式的液相色谱-电喷雾电离-四极杆飞行时间质谱表征

LC-ESI-QTOF/MS characterization of antimicrobial compounds with their action mode extracted from vine tea () leaves.

作者信息

Umair Muhammad, Sultana Tayyaba, Xiaoyu Zhu, Senan Ahmed M, Jabbar Saqib, Khan Labiba, Abid Muhammad, Murtaza Mian Anjum, Kuldeep Dhama, Al-Areqi Niyazi A S, Zhaoxin Lu

机构信息

College of Food Science and Technology Nanjing Agriculture University Nanjing China.

College of Public Administration Nanjing Agriculture University Nanjing China.

出版信息

Food Sci Nutr. 2022 Jan 19;10(2):422-435. doi: 10.1002/fsn3.2679. eCollection 2022 Feb.

Abstract

Vine tea () is a tea plant cultivated south of the Chinese Yangtze River. It has anti-inflammatory properties and is used to normalize blood circulation and detoxification. The leaves of vine tea are the most abundant source of flavonoids, such as dihydromyricetin and myricetin. However, as the main bioactive flavonoid in vine tea, dihydromyricetin was the main focus of previous research. This study aimed to explore the antibacterial activities of vine tea against selected foodborne pathogens. The antimicrobial activity of vine tea extract was evaluated by the agar well diffusion method. Cell membrane integrity and bactericidal kinetics, along with physical damage to the cell membrane, were also observed. The extract was analyzed using a high-performance liquid chromatography-diode array detector (HPLC-DAD), and the results were confirmed using a modified version of a previously published method that combined liquid chromatography and electrospray-ionized quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF/MS). Cell membrane integrity and bactericidal kinetics were determined by releasing intracellular material in suspension and monitoring it at 260 nm using an ultraviolet (UV) spectrophotometer. A scanning electron microscope (SEM) was used to detect morphological alterations and physical damage to the cell membrane. Six compounds were isolated successfully: (1) myricetin (CHO), (2) myricetin 3-O-rhamnoside (CHO), (3) 5,7,8,3,4-pentahydroxyisoflavone (CHO), (4) dihydroquercetin (CHO), (5) 6,8-dihydroxykaempferol (CHO), and (6) ellagic acid glucoside (CHO). Among these bioactive compounds, CHO was found to have vigorous antimicrobial activity against   (AS11846) and (CMCCB26003). A dose-dependent bactericidal kinetics with a higher degree of absorbance at optical density 260 (OD) was observed when the bacterial suspension was incubated with CHO for 8 h. Furthermore, a scanning electron microscope study revealed physical damage to the cell membrane. In addition, the action mode of CHO was on the cell wall of the target microorganism. Together, these results suggest that CHO has vigorous antimicrobial activity and can be used as a potent antimicrobial agent in the food processing industry.

摘要

藤茶是一种在中国长江以南地区种植的茶树。它具有抗炎特性,可用于促进血液循环和排毒。藤茶的叶子是黄酮类化合物最丰富的来源,如二氢杨梅素和杨梅素。然而,作为藤茶中的主要生物活性黄酮类化合物,二氢杨梅素是先前研究的主要焦点。本研究旨在探讨藤茶对选定食源性病原体的抗菌活性。采用琼脂扩散法评估藤茶提取物的抗菌活性。还观察了细胞膜完整性和杀菌动力学以及细胞膜的物理损伤。使用高效液相色谱 - 二极管阵列检测器(HPLC - DAD)对提取物进行分析,并使用先前发表的结合液相色谱和电喷雾电离四极杆飞行时间质谱(LC - ESI - QTOF/MS)方法的改进版本对结果进行确认。通过释放悬浮液中的细胞内物质并使用紫外(UV)分光光度计在260nm处进行监测来确定细胞膜完整性和杀菌动力学。使用扫描电子显微镜(SEM)检测细胞膜的形态变化和物理损伤。成功分离出六种化合物:(1)杨梅素(C₁₅H₁₀O₈),(2)杨梅素3 - O - 鼠李糖苷(C₂₁H₂₀O₁₂),(3)5,7,8,3',4' - 五羟基异黄酮(C₁₅H₁₂O₆),(4)二氢槲皮素(C₁₅H₁₂O₇),(5)6,8 - 二羟基山奈酚(C₁₅H₁₂O₆),以及(6)鞣花酸葡萄糖苷(C₂₀H₁₈O₁₂)。在这些生物活性化合物中,发现C₂₁H₂₀O₁₂对金黄色葡萄球菌(AS11846)和大肠杆菌(CMCCB26003)具有强烈的抗菌活性。当细菌悬浮液与C₂₁H₂₀O₁₂孵育8小时时,观察到在光密度260(OD)处具有更高吸光度的剂量依赖性杀菌动力学。此外,扫描电子显微镜研究揭示了细胞膜的物理损伤。此外,C₂₁H₂₀O₁₂的作用模式是作用于目标微生物的细胞壁。总之,这些结果表明C₂₁H₂₀O₁₂具有强烈的抗菌活性,可在食品加工行业中用作有效的抗菌剂。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d28/8825723/3ed25891e086/FSN3-10-422-g002.jpg

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