The amounts of collchicine present in two Jordanian species of Colchicum, namely, C. steveni Knuth and C. hierosolyvmitanum Feibrun (Coichicaceae), 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 collchicine (0.204/100 g), whilst in C. hierosolymitanum corms showed the highest collchicine 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. Copyright (C) 2004 John Wiley Sons, Ltd.