The amounts of colchicine present in two Jordanian species of Colchicum, namely, C. steveni Kunth and C. hierosolymitanum Feibrun (Colchicaceae), have been determined. An HPLC-UV (photodiode array) method employing gradient elution was developed and the results compared with those obtained using a simple TLC-spectrophotometric method. The levels of colchicine as measured by these methods were not significantly different (p < 0.05) indicating that the spectrophotometric method is an acceptable alternative to HPLC. With respect to C. steveni, the leaves contained the largest amount of colchicine (0.204/100 g), whilst in C. hierosolymitanum corms showed the highest colchicine content (0.126/100 g). As a source of colchicine, the two investigated species showed levels comparable with those found in C. autumnale, the traditional source of colchicine.