title: Development of a liquid chromatography-electrospray ionization-tandem mass spectrometry method for the simultaneous analysis of intact glucosinolates and isothiocyanates in Brassicaceae seeds and functional foods.
abstract: A new high pressure liquid chromatography-electrospray ionization-tandem mass spectrometry method for the simultaneous determination of glucosinolates, as glucoraphanin and glucoerucin, and the corresponding isothiocyanates, as sulforaphane and erucin, was developed and applied to quantify these compounds in Eruca sativa defatted seed meals and enriched functional foods. The method involved solvent extraction, separation was achieved in gradient mode using water with 0.5% formic acid and acetonitrile with 0.5% formic acid and using a reverse phase C18 column. The electrospray ion source operated in negative and positive mode for the detection of glucosinolates and isothiocyanates, respectively, and the multiple reaction monitoring (MRM) was selected as acquisition mode. The method was validated following the ICH guidelines. Replicate experiments demonstrated a good accuracy (bias%<10%) and precision (CV%<10%). Detection limits and quantification limits are in the range of 1-400ng/mL for each analytes. Calibration curves were validated on concentration ranges from 0.05 to 50μg/mL. The method proved to be suitable for glucosinolates and isothiocyanates determination both in biomasses and in complex matrices such as food products enriched with glucosinolates, or nutraceutical bakery products. In addition, the developed method was applied to the simultaneous determination of glucosinolates and isothiocyanates in bakery product enriched with glucosinolates, to evaluate their thermal stability after different industrial processes from cultivation phases to consumer processing.
authors.affiliation: Department of "Chemistry G. Ciamician", University of Bologna, Via Selmi 2, 40126 Bologna, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Franco
authors.affiliation: Department of "Chemistry G. Ciamician", University of Bologna, Via Selmi 2, 40126 Bologna, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Spinozzi
authors.affiliation: Consiglio per la ricerca in agricoltura e l'analisi dell'economia agraria, Centro di ricerca per le colture industriali (CRA-CIN), Via di Corticella 133, 40128 Bologna, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Pagnotta
authors.affiliation: Consiglio per la ricerca in agricoltura e l'analisi dell'economia agraria, Centro di ricerca per le colture industriali (CRA-CIN), Via di Corticella 133, 40128 Bologna, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Lazzeri
authors.affiliation: Consiglio per la ricerca in agricoltura e l'analisi dell'economia agraria, Centro di ricerca per le colture industriali (CRA-CIN), Via di Corticella 133, 40128 Bologna, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Ugolini
authors.affiliation: Department of "Chemistry G. Ciamician", University of Bologna, Via Selmi 2, 40126 Bologna, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Camborata
authors.affiliation: Department of "Chemistry G. Ciamician", University of Bologna, Via Selmi 2, 40126 Bologna, Italy. Electronic address: aldo.roda@unibo.it.
authors.forename: A
authors.initials: A
authors.lastname: Roda
mesh_terms: D019607:Brassicaceae / Q000737:chemistry*; D002853:Chromatography, Liquid; D005504:Food Analysis / Q000295:instrumentation / Q000379:methods*; D055951:Functional Food / Q000032:analysis*; D005961:Glucosinolates / Q000032:analysis*; D017879:Isothiocyanates / Q000032:analysis*; D057230:Limit of Detection; D012639:Seeds / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D005961:Glucosinolates; D017879:Isothiocyanates
keywords: Bakery products; Eruca sativa defatted seed meals; Functional food; Glucosinolates and isothiocyanates; HPLC–ES-MS/MS