Improved synthesis methods of standards used for quantitative determination of total isothiocyanates from broccoli in human urine

A well-known method for quantification of isothiocyanates (ITCs) and their metabolites is the condensation reaction with 1,2-benzenedithiole to produce 1,3-benzodithiole-2-thione, which can be quantified by high-performance liquid chromatography. Standards of an ITC metabolite and 1,3-benzodithiole-2-thione are required for this assay but are not commercially available. In the present study, we report on an improved synthesis of the ITC metabolite N-acetyl-S-(N-4-methylsulfinylbutylthiocarbamoyl)-L-cysteine and 1,3-benzodithiole-2-thione. The standards were used to quantify the urinary excretion of ITCs from 10 healthy subjects who consumed 350 g broccoli. The excretion was investigated throughout 48 h showing a cumulative urinary ITC excretion of 49.1+/-25.2% of the dose.