Robust UHPLC-(ESI+)-MS/MS Method for Simultaneous Analysis of Glucoraphanin, Sulforaphane, and Sulforaphane Metabolites in Biological Samples

A UHPLC-(ESI+)-MS/MS method was developed and validatedfor thesimultaneous determination of glucoraphanin (GR), sulforaphane (SF),sulforaphane-glutathione (SF-GSH), sulforaphane-cysteine (SF-CYS),and sulforaphane-N-acetyl-cysteine (SF-NAC), in a range of biologicalmaterials (digesta, cell culture medium, cell lysates, urine) withno sample pretreatment. The limit of detection (LODs) ranged from0.03 to 1.95 & mu;g L-1. The intra- and interdayvariation was & LE;15% and recoveries were & GE;85%. Linearityranged from 3.90 to 1000 & mu;g L-1 (GR), 0.06-500 & mu;g L-1 (SF), 3.90-1000 & mu;g L-1 (SF-NAC), and 62.5-500 & mu;g L-1 for SF-CYS and SF-GSH. The method was validated in broccoli seedpowders, and by determining transport and metabolism of SF in Caco-2monolayers, and model digesta. Results indicated that 72.4% of SFwas absorbed, transported, and metabolized (SF-CYS, SF-GSH, SF-NAC)in the Caco-2 monolayers after 4 h of incubation. This is a robustand time- and cost-efficient method that is suitable for high throughputapplications.