Abedini Amin, Chollet Sébastien, Angelis Apostolis, Borie Nicolas, Nuzillard Jean-Marc, Skaltsounis Alexios-Leandros, Reynaud Romain, Gangloff Sophie C, Renault Jean-Hugues, Hubert Jane
Institut de Chimie Moléculaire de Reims, UMR CNRS 7312, SFR CAP'santé, UFR de Pharmacie, Université de Reims Champagne-Ardenne, Reims, France; Biomatériaux et Inflammation en Site Osseux, EA 4691, SFR CAP'santé, UFR de Pharmacie, Université de Reims Champagne Ardenne, Reims, France.
Institut de Chimie Moléculaire de Reims, UMR CNRS 7312, SFR CAP'santé, UFR de Pharmacie, Université de Reims Champagne-Ardenne, Reims, France; CPC Engineering, Capacité SAS, France.
J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Sep 1;1029-1030:121-127. doi: 10.1016/j.jchromb.2016.07.021. Epub 2016 Jul 11.
Barks from conifers and broadleaved trees constitute abundant wastes generated from wood harvesting and logging activities. Extracts of such residues obtained from Alnus trees have been reported as interesting resources with potent antibacterial activities. The present study aims to determine the antimicrobial activity of a crude methanol extract prepared from the bark of Alnus glutinosa against a panel of 22 bacteria and yeasts and to optimize a purification method enabling the high production of the most active substances. Fractionation of the crude extract was performed by Centrifugal Partition Chromatography (CPC) using a three-phase solvent system composed of n-heptane, methyl-ter-butyl ether, acetonitrile and water. The major known compounds contained in the fractions produced by CPC were chemically profiled by (13)C NMR dereplication, resulting in the unambiguous identification of oregonin, hirsutanonol, betulinic acid, and alusenone 1a. The antibacterial evaluation of the fractions by bioautography on Staphylococcus aureus revealed that oregonin, in addition to being the major metabolite of the crude extract (∼32% w/w), was the most active with an antibacterial inhibitory effect comparable to antibiotics. The purification of oregonin was optimized at the laboratory-scale by CPC. A single injection of 3.7g of crude extract resulted in a recovery of 72% (850mg) of the available oregonin at purity higher than 94%.
针叶树和阔叶树的树皮是木材采伐和伐木活动产生的大量废弃物。据报道，从桤木中获得的此类残渣提取物是具有强大抗菌活性的有趣资源。本研究旨在测定从欧洲桤木树皮制备的粗甲醇提取物对一组22种细菌和酵母的抗菌活性，并优化一种纯化方法，以高产率生产最具活性的物质。使用由正庚烷、甲基叔丁基醚、乙腈和水组成的三相溶剂系统，通过离心分配色谱法（CPC）对粗提取物进行分馏。通过（13）C NMR去重复对CPC产生的馏分中所含的主要已知化合物进行化学分析，从而明确鉴定出俄勒冈宁、毛柳杉酚、桦木酸和桤木酮1a。通过对金黄色葡萄球菌的生物自显影法对馏分进行抗菌评估，结果表明，俄勒冈宁除了是粗提取物的主要代谢产物（约32% w/w）外，还是活性最强的物质，其抗菌抑制作用与抗生素相当。通过CPC在实验室规模上对俄勒冈宁的纯化进行了优化。单次注入3.7g粗提取物可回收72%（850mg）的可用俄勒冈宁，纯度高于94%。
