A method for the detection of organochlorine pesticides and polychlorinated biphenyls (PCBs) in human milk is described. The method involves quantitative extraction of human milk with acetone and hexane. The pesticide and PCB residues are extracted from the milk fat with acetonitrile (ACN) and then partitioned back into hexane by aqueous dilution of the ACN extract. The ACN partitioning is followed by cleanup with Florisil column chromatography, eluting with 6 and 15% mixtures of ethyl ether-hexane. The 6% eluate is concentrated and transferred to a silicic acid column for separation of PCBs from pesticide residues. Compounds are quantitated by gas-liquid chromatography. Recovery studies demonstrated that the procedure provides 68–103% recovery for pesticides and PCBs.