The well-documented presence of fumonisin myco-toxins B₁ and B₂ (FB₁ and FB₂) in corn raises the possibility that these toxins are carried over into the milk of animals fed with contaminated feed. The presence of FB₁ and FB₂ in milk has not been assessed because of the lack of sensitive analytical techniques for this matrix. Two methods, liquid chromatography (LC) and enzyme-linked immunosorbent assay (ELISA), were adapted for the analysis of milk. The ELISA, produced commercially for screening corn, required no sample preparation and was reproducible but was of low sensitivity [concentration that inhibits color development by 50% (IC₅₀), 1200-1600 ng FB₁/mL]. The more sensitive LC method involves serial extraction of milk with methanol-acetone and strong anion exchange followed by derivatization with naphthalene-2,3-dicarboxaldehyde. Recoveries of 50 ng FB₁ and FB₂/mL from unpasteurized and un-homogenized milk were 84 and 83%, respectively (limit of detection, 5 ng/mL). Recoveries of FB₁ from whole homogenized milk (76%) were slightly lower. Heating milk for 30 min at 62°C, to mimic pasteurization, did not significantly reduce FB₁ or FB₂ recovery, nor did storing milk for 11 days at 4°C. The LC method was applied to 165 samples of milk, only 1 of which was positive. This finding suggests that exposure of humans to FB₁ and FB₂ from milk is low.