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采用薄层色谱-直接生物自显影与 GC-MS、LC-MS 和 NMR 联用技术对抗临床重要抗生素耐药菌的抗菌精油成分进行表征和筛选。

Characterisation and screening of antimicrobial essential oil components against clinically important antibiotic-resistant bacteria using thin layer chromatography-direct bioautography hyphenated with GC-MS, LC-MS and NMR.

机构信息

Infectious Disease Research Group, School of Pharmacy, De Montfort University, Leicester, UK.

School of Pharmacy and Pharmaceutical Sciences, Cardiff University, Cardiff, UK.

出版信息

Phytochem Anal. 2019 Mar;30(2):121-131. doi: 10.1002/pca.2797. Epub 2018 Oct 2.

Abstract

INTRODUCTION

The antimicrobial activity of many essential oils (EOs) is well established, indicating that EOs may be a source of compounds for antimicrobial drug development. Thin layer chromatography-direct bioautography (TLC-DB) can quickly identify antimicrobial components in complex mixtures and can be applied to the screening of EOs for lead compounds.

OBJECTIVES

This study aimed to identify antimicrobial components of oregano, rosewood and cumin EOs against antibiotic-sensitive and -resistant bacteria using TLC-DB and a multi-faceted approach of GC-MS, LC-MS and NMR techniques to characterise bioactive compounds. The study also aimed to quantify the antimicrobial activity of bioactive compounds in order to evaluate their potential for the development of therapies against antibiotic-resistant bacteria.

MATERIALS AND METHODS

EOs were eluted on TLC plates and sprayed with a suspension of Staphylococcus aureus, Enterococcus faecium, Escherichia coli or Pseudomonas aeruginosa (antibiotic-sensitive and -resistant isolates). Zones of inhibition, visualised with iodonitrotetrazolium chloride, were subject to GC-MS, LC-MS and NMR to characterise the bioactive compounds.

RESULTS

Seven compounds were identified from the three EOs using GC-MS, while LC-MS and NMR failed to detect the presence of any further non-volatile or heat labile compounds. Carvacrol was most antimicrobial compound identified, with minimum inhibitory concentrations ranging 0.99-31.62 mM.

CONCLUSION

The identified antimicrobial compounds present in oregano, rosewood and cumin EOs including carvacrol may be candidates for the development of novel antimicrobial therapies against antibiotic-resistant bacteria.

摘要

简介

许多精油(EOs)的抗菌活性已得到充分证实,这表明 EOs 可能是抗菌药物开发的化合物来源。薄层层析-直接生物自显影法(TLC-DB)可快速鉴定复杂混合物中的抗菌成分,可用于筛选含抗菌成分的精油以寻找先导化合物。

目的

本研究旨在使用 TLC-DB 以及 GC-MS、LC-MS 和 NMR 技术的多方面方法,鉴定牛至、紫檀和孜然精油中的抗菌成分,以对抗生素敏感和耐药的细菌进行分析。本研究还旨在量化生物活性化合物的抗菌活性,以评估其用于开发针对抗生素耐药菌治疗方法的潜力。

材料与方法

将 EOs 洗脱在 TLC 板上,然后用金黄色葡萄球菌、屎肠球菌、大肠杆菌或铜绿假单胞菌(抗生素敏感和耐药分离株)的悬浮液喷洒。用碘硝基四唑氯化物可视化的抑制带,通过 GC-MS、LC-MS 和 NMR 对生物活性化合物进行鉴定。

结果

使用 GC-MS 从三种 EOs 中鉴定出 7 种化合物,而 LC-MS 和 NMR 未能检测到任何进一步的非挥发性或热不稳定化合物的存在。香芹酚是鉴定出的最具抗菌活性的化合物,最低抑菌浓度范围为 0.99-31.62mM。

结论

鉴定出的牛至、紫檀和孜然精油中的抗菌化合物,包括香芹酚,可能是针对抗生素耐药菌开发新型抗菌治疗方法的候选物。

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