Using thin-layer chromatography, celite column chromatography and recrystallization methods, articular (AR) and growth plate (GP) cartilage tissues and cells from prepubertal rabbits were shown to convert testosterone (T) into at least three main metabolites: dihydrotestosterone (DHT), delta 4-androstenedione and androstanediols. In tissue incubation experiments the amount of each newly formed metabolite per mg of tissue was always greater in AR than in GP cartilage. After a 24 h incubation with AR or GP cartilage tissues, T was mainly converted to DHT and delta 4-androstenedione in approximately equal amounts. The amount of androstanediol metabolites formed was much lower. In a time-course experiment, the conversion of T to DHT and delta 4-androstenedione was shown to increase in a linear fashion, while the conversion to androstanediols was more variable. Using cultured AR cartilage cell incubations, similar results were obtained. In addition, DHT was shown to be the sole metabolite which accumulated in the cellular pool during the first 3 h incubation, as well as during the 24 h incubation when maximum cellular uptake of radioactivity was observed. At this time, the intracellular amount of unmetabolized [3H]T (88 pmoles/100 micrograms DNA) was similar to the amount of [3H]DHT (70 pmoles/100 micrograms DNA) accumulated in the chondrocytes. For both delta 4-androstenedione and androstanediols, 99% of radioactivity was extracted from the incubation medium.