Epidemiological studies have shown that consumption of Brassica vegetables decrease the risk of cancer. These associations are linked to dietary intake of glucosinolates and their metabolism to cancer preventive isothiocyanates. Bioavailability of glucosinolates and related isothiocyanates are influenced by storage and culinary processing of Brassica vegetables. In this work, the content of the 7 major glucosinolates in broccoli, Brussels sprouts, cauliflower and green cabbage and their stability under different storage and cooking conditions is examined. Glucosinolates and isothiocyanates were quantified by liquid chromatography with tandem mass spectrometric detection (LC-MS/MS). Isothiocyanates were detected with high sensitivity as the corresponding thiourea derivatives. Storage at ambient temperature and in a domestic refrigerator showed no significant difference and a minor loss (9-26%) of glucosinolate levels over 7 days. Vegetables shredded finely showed a marked decline of glucosinolate level with post-shredding dwell time - up to 75% over 6h. Glucosinolate losses were detected partly as isothiocyanates. Cooking by steaming, microwaving and stir-fry did not produce significant loss of glucosinolates whereas boiling showed significant losses by leaching into cooking water. Most of the loss of the glucosinolates (approximately 90%) was detected in the cooking water. Increased bioavailability of dietary isothiocyanates may be achieved by avoiding boiling of vegetables.