Bahri Meriem, Hance Philippe, Grec Sébastien, Quillet Marie-Christine, Trotin Francis, Hilbert Jean-Louis, Hendriks Theo
Université Lille 1, Sciences et Technologies, UMR INRA 1281 "Stress Abiotiques et Différenciation des Végétaux Cultivés", GIS GENOCHIC, Bâtiment SN2, 3ème Étage, 59655 Villeneuve d'Ascq Cedex, France.
ScientificWorldJournal. 2012;2012:142983. doi: 10.1100/2012/142983. Epub 2012 Dec 5.
A "novel" protocol is presented for easy and reliable estimation of soluble hydroxycinnamate levels in Cichorium intybus L. leaf tissue in large-scale experiments. Samples were standardized by punching 6 discs per leaf, and hydroxycinnamates were extracted by submerging the discs in 80% ethanol with 5% acetic acid for at least 48 h in the darkness at 4°C. Residual dry mass of the discs was used for a posteriori correction of compound levels. Chlorophyll was eliminated by chloroform, and the aqueous phases were transferred to microplates, dried, and dissolved in 50% methanol for HPLC analysis and storage. An HPLC program of 8 min was developed for the analysis of the extracts. Comparisons with extractions of liquid nitrogen powders indicated that the novel extraction method was reliable. No degradation of the major hydroxycinnamates-caftaric, chlorogenic, and chicoric acids-was observed, during maceration at ambient temperatures, or after storage for 1 year.
本文介绍了一种“新颖”的方案，用于在大规模实验中轻松、可靠地估算菊苣叶组织中可溶性羟基肉桂酸的含量。通过在每片叶子上冲压6个圆片对样品进行标准化处理，将圆片置于含有5%乙酸的80%乙醇中，在4°C黑暗条件下浸泡至少48小时来提取羟基肉桂酸。圆片的残余干重用于对化合物含量进行事后校正。用氯仿去除叶绿素，将水相转移至微孔板中，干燥后溶于50%甲醇中用于高效液相色谱（HPLC）分析和储存。开发了一个8分钟的HPLC程序用于分析提取物。与液氮粉末提取物的比较表明，这种新颖的提取方法是可靠的。在常温浸渍过程中或储存1年后，未观察到主要羟基肉桂酸——咖啡酰酒石酸、绿原酸和菊苣酸——发生降解。
