OBJECTIVE To derive reference ranges for serum methylmalonic acid (MMA) for the diagnosis of cobalt/vitamin B12-responsiveness in lambs and critique existing serum vitamin B12 reference ranges. METHODS Individual animal data from earlier supplementation trials, involving 225 ewes, 106 suckling lambs, 301 lambs during the suckling and post-weaning periods and 414 weaned lambs, for which weight gain to supplementation was observed, were used to derive relationships between serum vitamin B12 and MMA, and liveweight gain. RESULTS Serum MMA concentrations were rarely elevated above the norm of <2 micromol/L when serum vitamin B12 concentrations were >375 pmol/L, and not elevated into the range where a liveweight response to supplementation occurred (>10 micromol/L) unless serum vitamin B12 concentrations were below 200 pmol/L. Suckling lambs were able to maintain high growth rates despite elevated serum MMA concentrations (>20 micromol/L). CONCLUSIONS The current reference ranges used in New Zealand for serum vitamin B12 are set conservatively high. Serum MMA concentrations appear to allow better differentiation of a responsive condition than vitamin B12 concentrations. Serum MMA concentrations >13 micromol/L indicate responsiveness to supplementation whilst concentrations <7 micromol/L indicate unresponsiveness. In the range 7-13 micromol/L, variation in response was observed and predictability of response is less certain, but supplementation is advisable. CONCLUSIONS The current reference ranges for vitamin B12 responsiveness are conservatively high and lead to over-diagnosis of vitamin B12 deficiency in ill-thriftiness of sheep.