An animal study to evaluate dual-energy x-ray absorptiometry (DXA) and magnetic resonance (MR) imaging and spectroscopy for measurement of neonatal body composition was performed. Twenty-three piglets with body weights ranging from 848 to 7550 g were used. After measuring total body water, animals were killed and body composition was assessed using DXA and MR (1.5 T; MR imaging, T1-weighted sagittal spin-echo sequence; MR spectroscopy, three-dimensional chemical shift imaging) as well as chemical carcass analysis (standard methods) after homogenization. Body composition by chemical analysis (percent of body weight, mean +/- SD) was as follows: body water, 75.3 +/- 3.9%; total protein, 13.9 +/- 8.8%; and total fat, 6.5 +/- 3.7%. Absolute content of fat and total ash was 7-674 and 35-237 g, respectively. Mean hydration of fat-free mass was 0.804 +/- 0.011 g/kg and decreased with increasing body weight (r2 = 0.419) independent of age. Using DXA, bone mineral content was highly correlated with calcium content (r2 = 0.992), and calcium per bone mineral content was 44.1 +/- 4.2%. DXA fat mass correlated with total fat (r2 = 0.961). Using MR, spectroscopy and chemical analysis were highly correlated with fat-to-water ratio (r2 = 0.984) and absolute fat content (r2 = 0.988). Total fat by MR imaging volumetry showed a lower correlation (r2 = 0.913) and overestimated total fat by a factor of 2.46. Conversion equations for DXA were developed (total fat = 1.31 x fat mass measured by DXA--68.8; calcium = 0.402 x bone mineral content + 1.7), which improved precision and accuracy of DXA measurements. In conclusion, both DXA and MR spectroscopy give accurate and precise estimates of neonatal body composition and may become valuable tools for the noninvasive assessment of neonatal growth and nutritional status.