We evaluated the precision and accuracy of peripheral dual-energy X-ray absorptiometry (DXA) for the measurement of bone mineral density (BMD) and bone mineral content (BMC) in intact and excised femurs and tibias from rats. Thirty-one Sprague-Dawley rats (18F/13M; 114-360 g) were used in the study. Precision and accuracy were determined in 23 rats and prediction equations were evaluated in an independent sample of 8 animals. Precision was determined by measuring the right hindquarter three times with repositioning between scans. The femur and tibia were then excised, cleaned, and scanned in triplicate, with repositioning. CVs ranged from 0.66 to 2.24%. Accuracy of BMC was determined by comparison to bone ash values. BMC values for the intact and excised femur significantly overestimated bone ash (p < 0.001) by 33% and 5.5%, respectively. BMC for the intact tibia overestimated ash by 37% (p < 0.001), whereas BMC for the excised tibia underestimated ash by 1% (p < 0.05). However, BMC and bone ash were highly related for both bones, whether BMC was measured in the intact animal or after excision (r2 > 0.99). Cross-validation of prediction equations in an independent sample showed that there were no significant differences between predicted ash (based on BMC from DXA) and measured bone ash. These results suggest the peripheral DXA is a useful tool for measuring intact and excised rat leg bones.