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

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• 作者：P. Franco^a ; S. Spinozzi^a ; E. Pagnotta^b ; L. Lazzeri^b ; L. Ugolini^b ; C. Camborata^a ; A. Roda^a ; ^{aldo.roda@unibo.it" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Glucosinolates and isothiocyanates ; Eruca sativa defatted seed meals ; HPLC&ndash ; ES-MS/MS ; Bakery products ; Functional food
• 刊名：Journal of Chromatography A
• 出版年：2016
• 出版时间：8 January 2016
• 年：2016
• 卷：1428
• 期：Complete
• 页码：154-161
• 全文大小：762 K

文摘

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 1–400 ng/mL for each analytes. Calibration curves were validated on concentration ranges from 0.05 to 50 μg/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.