A programme of metabolic and nutritional research is being undertaken in critically ill patients requiring intensive care. Central to this research is the measurement of the three nutritionally important compartments of body composition, protein, fat, and water by a combination of tritium dilution and prompt gamma in vivo neutron activation analysis (IVNAA). In this paper a calibration technique is presented that enables absolute estimates of total body nitrogen (TBN) to be made using prompt gamma IVNAA in critically ill patients with gross abnormalities in body composition, especially in their state of hydration. This technique, which is independent of skinfold anthropometry and does not make a priori assumptions about the ratios of major body compartments, has been applied to 41 normal volunteers and the derived values for nitrogen compared with values obtained by applying three currently used calibration methods to the same experimental data; the other methods are used at centers which also measure nitrogen by prompt gamma IVNAA. Close agreement is obtained between the four calibration methods, with correlation coefficients relative to the Auckland technique of 0.999, 0.998, and 0.994. In addition, the measured values of TBN are compared to values obtained for the same group of volunteers using sets of empirical equations from two centers, one of which uses prompt gamma IVNAA (Brookhaven National Laboratory, Long Island, USA) and the other delayed gamma IVNAA (Leeds, Britain). The empirical equations relate TBN in normal people to age, height, weight and sex. The mean ratios of experimental to predicted TBN (with SEMs) are 1.013 +/- 0.017 and 1.002 +/- 0.014, respectively. Mean values of the ratio of TBN to fat-free mass (0.0340 +/- 0.0004) and of total body water to fat-free mass (0.716 +/- 0.002) agree closely with values reported elsewhere for normals by a variety of techniques including chemical analysis. Finally, TBN results based on the four different calibration methods are presented for five surgical patients, demonstrating the importance of the calibration method on estimates of TBN in patients with abnormal body composition. It is concluded that this technique will provide accurate estimates of the total body content of protein, water, and fat in intensive care patients.