A rapid and convenient method for sample preparation and simultaneous densitometric quantification of hyoscyamine (1) and scopolamine (2) has been developed. The alkaloid profiles of plant samples, as well as the content of 1 and 2, determined by TLC-densitometry corresponded well with those obtained using a GC method. The proposed method is simple and sensitive and can be used for the routine assay of 1 and 2, and for screening for alkaloid-rich and biochemically-interesting transformed root cultures and intact solanaceous plants.