An improved monoclonal antibody immunoaffinity chromatography/high-pressure liquid chromatography/ fluorescence detection method was developed to measure aflatoxin (AF) exposure by quantifying AFM1 in human and rat urine samples. Analysis of different amounts of various AF metabolites showed that the immunoaffinity resin was highly selective for aflatoxin B1 (AFB1), AFB2, and AFM1. Recovery of added AFs increased with the amount of immunoaffinity resin and was virtually complete within the range of 0.01-10 ng of AFM1 by using 7 ml of resin. The detection limit of this method is 0.5 pg/ml urine. Rats dosed with tritiated AFB1 excreted in their urine tritiated AFM1, among other AF metabolites, as indicated by chemical derivative confirmation and cochromatography with authentic AFM1 and agreement of radioactivity and fluorescence quantitation. A linear dose-response relationship was found over the range of 0.05-50 micrograms/kg of body weight/day. Two humans dosed with 1.0 microgram of pure AFB1 excreted 6-7% of the dose as urinary AFM1 over 5-7 days. Pooled urine samples from 30 men from each of 69 rural counties in mainland China and 16 survey areas in Taiwan, with two villages per county or area, were analyzed with this improved method (170 villages total). The correlation coefficient of urinary excretion of AFM1 compared between villages within all 85 survey areas was 0.50 (P < 0.001). Sixty-five % of the samples contained detectable concentrations of AFM1 with an average excretion of 3.1 ng/12 h. Assuming an excretion rate of 2-6%, this AFM1 excretion corresponds to a very low average daily AF consumption of 0.1-0.3 microgram/day (possible range, 0-11 micrograms/day). Patterns of urinary excretion of AFM1 were similar in mainland China and Taiwan.