Advanced glycation end products (AGEs) are an abnormal modification of the collagenous matrix in bone, and their accumulation contributes to alteration of mechanical properties. Using a mouse model of focal external radiotherapy, we quantified the time-dependent changes in the glycation of bone collagen after 4 daily fractions of 5 Gy exposure to unilateral hindlimb. Fluorometric analysis of decalcified femurs demonstrated a significant and transient increase in the quantity of pentosidine, pyridinolines and nonspecific AGEs per unit of collagen at one week postirradiation. These differences did not persist at 4, 8, 12 or 26 weeks postirradiation. Radiation had no effect on bone collagen content. We hypothesize that following the transient increase in glycation products, these crosslinks are then removed as a result of increased postirradiation osteoclast activity and continued mineralization of the bone.