A panel of 18 rat x mouse somatic cell hybrid clones segregating individual rat chromosomes in different combinations was used to assign 23 biochemical loci to rat chromosomes. The chromosomal locations for these 23 loci were determined as follows: GOT1 on rat chromosome 1; HAGH on 2; ACP2, ADA, GANC, ITPA, and SORD on 3; LDHB on 4; PEPB on 7; GLB1 and HEXA on 8; IDH1 on 9; UMPH2 on 10; GUSB on 12; FH and PEPC on 13; PEPS on 14; ESD and NP on 15; DIA4 on 19; and PP on 20. In addition, ACP1 and GLO1 were reassigned to rat chromosomes 6 and 20, respectively. The chromosomal assignments of these loci extends the known syntenic homologies among rats, mice, and humans.