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ScientificWorldJournal. 2012;2012:142983. doi: 10.1100/2012/142983. Epub 2012 Dec 5.

A "novel" protocol for the analysis of hydroxycinnamic acids in leaf tissue of chicory (Cichorium intybus L., Asteraceae).

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  • 1Université 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.

Abstract

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.

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