title: Liquid chromatographic/electrospray ionization tandem mass spectrometric study of the phenolic composition of cocoa (Theobroma cacao).
abstract: Liquid chromatography coupled with ionspray mass spectrometry in the tandem mode (LC/MS/MS) with negative ion detection was used for the identification of a variety of phenolic compounds in a cocoa sample. Gradient elution with water and acetonitrile, both containing 0.1% HCOOH, was used. Standard solutions of 31 phenolic compounds, including benzoic and cinnamic acids and flavonoid compounds, were studied in the negative ion mode using MS/MS product ion scans. At low collisional activation, the deprotonated molecule [M - H](-) was observed for all the compounds studied. For cinnamic and benzoic acids, losses of CO(2) or formation of [M - CH(3)](-*) in the case of methoxylated compounds were observed. However, for flavonol and flavone glycosides, the spectra present both the deprotonated molecule [M - H](-) of the glycoside and the ion corresponding to the deprotonated aglycone [A - H](-). The latter ion is formed by loss of the rhamnose, glucose, galactose or arabinose residue from the glycosides. Different fragmentation patterns were observed in MS/MS experiments for flavone-C-glycosides which showed fragmentation in the sugar part. Fragmentation of aglycones provided characteristic ions for each family of flavonoids. The optimum LC/MS/MS conditions were applied to the characterization of a cocoa sample that had been subjected to an extraction/clean-up procedure which involved chromatography on Sephadex LH20 and thin-layer chromatographic monitoring. In addition to compounds described in the literature, such as epicatechin and catechin, quercetin, isoquercitrin (quercetin-3-O-glucoside) and quercetin-3-O-arabinose, other compounds were identified for the first time in cocoa samples, such as hyperoside (quercetin-3-O-galactoside), naringenin, luteolin, apigenin and some O-glucosides and C-glucosides of these compounds.
authors.affiliation: Department of Natural Products, University of Barcelona, Spain.
authors.forename: Ferran
authors.initials: F
authors.lastname: Sánchez-Rabaneda
authors.forename: Olga
authors.initials: O
authors.lastname: Jáuregui
authors.forename: Isidre
authors.initials: I
authors.lastname: Casals
authors.forename: Cristina
authors.initials: C
authors.lastname: Andrés-Lacueva
authors.forename: Maria
authors.initials: M
authors.lastname: Izquierdo-Pulido
authors.forename: Rosa M
authors.initials: RM
authors.lastname: Lamuela-Raventós
mesh_terms: D002099:Cacao / Q000737:chemistry*; D002853:Chromatography, Liquid; D005419:Flavonoids / Q000032:analysis* / Q000737:chemistry; D010636:Phenols / Q000032:analysis* / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D005419:Flavonoids; D010636:Phenols