Sulforaphane is a cognate isothiocyanate of glucoraphanin, released by degradation of glucosinolate through myrosinase enzyme, when Brassica tissues are crushed or chewed. C 8 analytical column and mixture of water/acetonitrile 65/35 (v/v) as mobile phase were used to separate sulforaphane in broccoli tissues (e.g., 7-day-seedling, seed, floret and leave). Confirmation of sulforaphane in crude extract of broccoli tissues was monitored using gas chromatography-mass spectroscopy (GC-MS). The highest amount of sulforaphane (1216 µg/g dry weight) found in 7-day-seedling. This method is suitable for routine screening of plant materials and proposes a low cost and robust technique for the analysis of sulforaphane.