Ground beef was fried at 200 degrees C (392 degrees F) to a well-done, non-charred state, and the extracted organic base fraction was found to be highly mutagenic in Salmonella strain TA1538 (6300 revertants/100 g equivalent, gE, fresh weight). The neutral and acidic extracts showed no mutagenic activity in any of the 5 standard strains of Salmonella. A new procedure based upon extraction and protein precipitation with acetone is described, which is simpler and more efficient than previously described methods. The organic base fraction was mutagenic only in Salmonella strains TA1537, TA1538, and TA98, all sensitive to frameshift mutations. Strains sensitive to base-substitution mutations showed no activity. Metabolic activation was an absolute requirement for mutagenesis; however cell toxicity was decreased by the presence of S9 activation mixture. After normal cooking, more than 20 times as much mutagenic material remained in the meat as was recovered in the pan grease and vapors. The results confirm that mutagens are formed under conventional frying conditions, and show that mutagen can be isolated by an improved extraction method.