Móricz Agnes M, Ott Péter G, Alberti Agnes, Böszörményi Andrea, Lemberkovics Eva, Szoke Eva, Kéry Agnes, Mincsovics Emil
J AOAC Int. 2013 Nov-Dec;96(6):1214-21. doi: 10.5740/jaoacint.sgemoricz.
In situ sample preparation and preparative overpressured layer chromatography (OPLC) fractionation on a 0.5 mm thick adsorbent layer of chamomile flower methanol extract prepurified by conventional gravitation accelerated column chromatography were applied in searching for bioactive components. Sample cleanup in situ on the adsorbent layer subsequent to sample application was performed using mobile phase flow in the opposite direction (the input and output of the eluent was exchanged). The antibacterial effect of the fractions obtained from the stepwise gradient OPLC separation with the flow in the normal direction was evaluated by direct bioautography against two Gram-negative bacteria: the luminescence gene tagged plant pathogenic Pseudomonas syringae pv. maculicola, and the naturally luminescent marine bacterium Vibrio fischeri. The fractions having strong activity were analyzed by SPME-GC/MS and HPLC/MS/MS. Mainly essential oil components, coumarins, flavonoids, phenolic acids, and fatty acids were tentatively identified in the fractions.
采用原位样品制备和制备型超压薄层色谱（OPLC）分级分离法，在通过常规重力加速柱色谱预纯化的洋甘菊花甲醇提取物的0.5毫米厚吸附剂层上进行，以寻找生物活性成分。在样品施加后，利用流动相沿相反方向流动（洗脱液的输入和输出进行交换）在吸附剂层上原位进行样品净化。通过直接生物自显影法，针对两种革兰氏阴性菌评估了在正常方向流动下通过逐步梯度OPLC分离获得的馏分的抗菌效果：发光基因标记的植物病原性丁香假单胞菌番茄斑点变种，以及天然发光的海洋细菌费氏弧菌。对具有强活性的馏分进行了固相微萃取-气相色谱/质谱联用（SPME-GC/MS）和高效液相色谱/串联质谱联用（HPLC/MS/MS）分析。在这些馏分中初步鉴定出主要为精油成分、香豆素、黄酮类化合物、酚酸和脂肪酸。
