Because of recent improvements in the serum vitamin B12 assay, literature criteria based on prior assay methods used in measuring B12 levels were evaluated. Of 1,708 B12 levels measured at Bellevue Hospital in a six-month period, 137 in 124 patients were below 200 pg/ml. Contrary to expectations, 81.6 percent of patients with low B12 levels had a mean corpuscular volume (MCV) below 95 fl. Literature-derived criteria missed 30 percent of patients with low B12 levels. Only three of 12 patients with megaloblastic bone marrow or an abnormal Schilling result had B12 levels that were low (below 100 pg/ml), and nine had values in an intermediate range (100 to 200 pg/ml). This suggests that the use of an MCV below 95 fl and a B12 level below 100 pg/ml as abnormal values may not detect clinically important B12 deficiency.