Profiling glucosinolate metabolites in human urine and plasma after broccoli consumption using non-targeted and targeted metabolomic analyses

Broccoli is a popular brassica vegetable and its consumption may decrease the occurrence of cancer in certain populations. To gain insight into the metabolites that may induce physiological responses to broccoli intake, a non-targeted metabolomic approach and a targeted approach for analysis of glucosinolate metabolites were developed using high resolution accurate mass spectrometry. A human study was conducted in which 6 subjects consumed a single meal of 200 g of uncooked broccoli florets. The metabolomic analysis revealed changes in endogenous metabolites and a decrease in hippuric acid after broccoli consumption. Targeted analysis using high-resolution, accurate mass-mass spectrometry (HRAM-MS) enabled detection of low concentrations (nM) of glucosinolate metabolites in human urine and plasma. Glucosinolate metabolites were found in human urine (13) and plasma (8), respectively. Metabolites from methoxyl-indole glucosinolates, arising from broccoli consumption, are reported for the first time. Most glucosinolate metabolites reached their peak concentration in urine 2-4 h after consumption while, in plasma, peak maxima were achieved 2 h after intake. The results suggest that glucoraphanin metabolites (sulforaphane, sulforaphane cysteine, sulforaphane N-acetyl cysteine) and indole metabolites (ascorbigen and methoxyl ascorbigen from indole glucosinolates) may serve as marker compounds for the intake of broccoli.