Analysis of glucosinolates, isothiocyanates, and amine degradation products in vegetable extracts and blood plasma by LC-MS/MS.
Dietary glucosinolates are under intensive investigation as precursors of cancer-preventive isothiocyanates. Quantitation of the dose and bioavailability of glucosinolates and isothiocyanates requires a comprehensive analysis of the major dietary glucosinolates, isothiocyanates, and related metabolites. We report a liquid chromatography with tandem mass spectrometric detection (LC-MS/MS) analytical method for the comprehensive analysis of the seven major dietary glucosinolates, related isothiocyanates, and putative amine degradation products. The parent glucosinolates were sinigrin, gluconapin, progoitrin, glucoiberin, glucoraphanin, glucoalyssin, and gluconasturtiin. The LC-MS/MS analysis method for these compounds was developed and validated; a standard addition analysis protocol was used generally to avoid the requirement for stable isotopic standards. Where stable isotopic standards were available, internal standardization with these gave estimates in agreement with those obtained by the standard addition analysis protocol. For glucosinolates, negative ion electrospray LC-MS/MS analysis was performed. Isothiocyanates and amines were prederivatized to the corresponding thiourea and N-acetamides, respectively, and were quantified by positive ion electrospray LC-MS/MS. The limits of detection were 0.5-2 pmol; the recoveries for glucosinolates, isothiocyanates, and amines were 85-90%, 50-85%, and 60-70%, respectively; and the intra- and interbatch coefficients of variation were 1-4% and 3-10%, respectively. These methods provide facile access to comprehensive analytical data on the major dietary glucosinolates and related metabolites to quantify inputs and metabolic formation of these compounds in cancer prevention and related studies.
Song
Lijiang
L
Disease Mechanisms and Therapeutic Research Group, Department of Biological Sciences, University of Essex, Wivenhoe Park, Colchester, Essex CO4 3SQ, UK.
Morrison
John J
JJ
Botting
Nigel P
NP
Thornalley
Paul J
PJ
D000588:Amines / Q000032:analysis* / Q000097:blood; D001774:Blood Chemical Analysis / Q000379:methods / Q000592:standards; D001937:Brassica / Q000737:chemistry; D002623:Chemistry Techniques, Analytical; D002853:Chromatography, Liquid / Q000379:methods; D005961:Glucosinolates / Q000032:analysis* / Q000097:blood; D006801:Humans; D017879:Isothiocyanates / Q000032:analysis* / Q000097:blood; D013058:Mass Spectrometry / Q000379:methods* / Q000592:standards; D010936:Plant Extracts / Q000737:chemistry
D000588:Amines; D005961:Glucosinolates; D017879:Isothiocyanates; D010936:Plant Extracts