An invasive comparative study of some pharmacokinetic aspects of 99mTc-mercaptoacetyltriglycine (MAG3), 131I-orthoiodohippurate (OIH), and 125I-iothalamate (iothalamate) was performed in six pigs 0-150 min after a simultaneous single injection (SI) and during a subsequent 90 min of continuous infusion (CI). The total plasma clearance and the renal clearance of MAG3 were about 75% that of OIH. The renal clearance of MAG3 was about 2 1/2 times the glomerular filtration rate. The distribution volume of MAG3 was 71% that of iothalamate and only 47% that of OIH. There was a significant hepatic plasma clearance of MAG3 of 5.9 ml min-1 and 3.9% of the injected dose was excreted in the bile. HPLC analysis revealed that technetium was excreted in urine and bile mainly labelled to MAG3. The average red blood cell (RBC) binding after single injection/during continuous infusion was 1.0%/2.3% for MAG3, 13.5%/9.0% for OIH, and 3.1%/5.3% for iothalamate. The binding of OIH to RBC in arterial blood increased from 8% at 1 min post-injection to 21% at 150 min post-injection. The RBC binding was higher in the renal vein, indicating incomplete back diffusion from RBC to plasma. The protein binding was 90% for MAG3, 49% for OIH and 16% for iothalamate. The renal plasma extraction of MAG3 was constant but significantly smaller after SI (0.54) than during CI (0.62). Following SI, the renal plasma extraction of OIH decreased continuously from 0.85 to 0.52, 3-150 min post-injection. On the average there was no significant difference in renal plasma extraction after SI and during CI of either OIH (0.72 versus 0.77) or iothalamate (0.26 versus 0.27). It is concluded that MAG3 is preferential to OIH as a tracer for renal function studies using a single injection technique mainly due to the constant renal extraction of MAG3.