AbstractMyrosinase activity in partially purified extracts of 12 cruciferous vegetables and an acetone powder preparation of Sinapis alba L. (white mustard) was determined by the initial rate of glucose formation from glucosinolate hydrolysis using a coupled assay. Of the species studied Raphanus sativus L. (radish, 12.8±0.7 μmol min−1g−1 powdered tissue) had the greatest myrosinase activity, and Brassica campestris L. ssp. rapifera (turnip) and Nasturtium officinalis R.Br. (watercress) (0.6±0.1 and 0.8±0.03 μmol min−1g−1 powdered tissue respectively) the least. The sub‐species of Brassica oleracea studied all had similar myrosinase activity (ca 2.5±0.2μmolmin−1g−1 powdered tissue) except B. oleracea L. var. gemmifera D.C. (Brussels sprouts) and B. oleracea L. var. capitata L. (white cabbage) which had higher activities (7.6±0.1 and 5.2±0.2μmol min−1g−1 powdered tissue respectively). The effect of ascorbate concentration upon the myrosinase activity of six of the crucifers studied and the white mustard preparation, revealed that the ascorbate concentration necessary to promote maximal activity varied with species. A concentration of 0.9mM ascorbate maximally activated radish and turnip myrosinase, while red cabbage, watercress, white mustard and Brussels sprouts were maximally activated at 2.0, 3.0, 5.0 and 0.7–1.0mM ascorbate respectively. Two peaks of maximal myrosinase activity, occurring between 0.9 and 1.0mM and at 3.0mM ascorbate, were found for B. oleracea L. var. botrytis L. subvar. cauliflora D.C. (cauliflower).