Aflatoxins are dietary contaminants that are hepatocarcinogenic and immunotoxic and cause growth retardation in animals, but there is little evidence concerning the latter two parameters in exposed human populations. Aflatoxin exposure of West African children is known to be high, so we conducted a longitudinal study over an 8-month period in Benin to assess the effects of exposure on growth. Two hundred children 16–37 months of age were recruited from four villages, two with high and two with low aflatoxin exposure (50 children per village). Serum aflatoxin–albumin (AF-alb) adducts, anthropometric parameters, information on food consumption, and various demographic data were measured at recruitment (February) and at two subsequent time points (June and October). Plasma levels of vitamin A and zinc were also measured. AF-alb adducts increased markedly between February and October in three of the four villages, with the largest increases in the villages with higher exposures. Children who were fully weaned at recruitment had higher AF-alb than did those still partially breast-fed (p < 0.0001); the major weaning food was a maize-based porridge. There was no association between AF-alb and micronutrient levels, suggesting that aflatoxin exposure was not accompanied by a general nutritional deficiency. There was, however, a strong negative correlation (p < 0.0001) between AF-alb and height increase over the 8-month follow-up after adjustment for age, sex, height at recruitment, socioeconomic status, village, and weaning status; the highest quartile of AF-alb was associated with a mean 1.7 cm reduction in growth over 8 months compared with the lowest quartile. This study emphasizes the association between aflatoxin and stunting, although the underlying mechanisms remain unclear. Aflatoxin exposure during the weaning period may be critical in terms of adverse health effects in West African children, and intervention measures to reduce exposure merit investigation.

Fumonisins are mycotoxins produced by Fusarium spp. and commonly contaminate maize and maize products worldwide. Fumonisins are rodent carcinogens and have been associated with human esophageal cancer. However, the lack of a valid exposure biomarker has hindered both the assessment of human exposure and the evaluation of disease risk. A sensitive liquid chromatography-mass spectrometry method to measure urinary fumonisin B1 (FB1) following extraction on Oasis MAX cartridges was established and applied to urine samples from women in a cohort recruited in Morelos County, Mexico. Urinary FB1 was compared with dietary information on tortilla consumption. FB1 recovery in spiked samples averaged 94% as judged by deuterium-labeled FB1 internal standard. Urinary FB1 was determined in 75 samples from women selected based on low, medium, or high consumption of maizebased tortillas. The geometric mean (95% confidence interval) of urinary FB1 was 35.0 (18.8-65.2), 63.1 (36.8-108.2), and 147.4 (87.6-248.0) pg/mL and the frequency of samples above the detection limit (set at 20 pg FB1/mL urine) was 45%, 80%, and 96% for the low, medium, and high groups, respectively. Women with high intake had a 3-fold higher average FB1 levels compared with the ''low intake'' group (F = 7.3; P = 0.0015). Urinary FB1 was correlated with maize intake (P trend = 0.001); the correlation remained significant after adjusting for age, education, and place of residence. This study suggests that measurement of urinary FB1 is sufficiently sensitive for fumonisin exposure assessment in human populations and could be a valuable tool in investigating the associated health effects of exposure. (Cancer Epidemiol Biomarkers Prev 2008;17(3):688 -94)

Essential to the conduct of epidemiologic studies examining aflatoxin exposure and the risk of heptocellular carcinoma, impaired growth, and acute toxicity has been the development of quantitative biomarkers of exposure to aflatoxins, particularly aflatoxin B 1 . In this study, identical serum sample sets were analyzed for aflatoxin-albumin adducts by ELISA, high-performance liquid chromatography (HPLC) with fluorescence detection (HPLC-f), and HPLC with isotope dilution mass spectrometry (IDMS). The human samples analyzed were from an acute aflatoxicosis outbreak in Kenya in 2004 (n = 102) and the measured values ranged from 0.018 to 67.0, nondetectable to 13.6, and 0.002 to 17.7 ng/mg albumin for the respective methods. The Deming regression slopes for the HPLC-f and ELISA concentrations as a function of the IDMS concentrations were 0.71 (r 2 = 0.95) and 3.3 (r 2 = 0.96), respectively. When the samples were classified as cases or controls, based on clinical diagnosis, all methods were predictive of outcome (P < 0.01). Further, to evaluate assay precision, duplicate samples were prepared at three levels by dilution of an exposed human sample and were analyzed on three separate days. Excluding one assay value by ELISA and one assay by HPLC-f, the overall relative SD were 8.7%, 10.5%, and 9.4% for IDMS, HPLC-f, and ELISA, respectively. IDMS was the most sensitive technique and HPLC-f was the least sensitive method. Overall, this study shows an excellent correlation between three independent methodologies conducted in different laboratories and supports the validation of these technologies for assessment of human exposure to this environmental toxin and carcinogen. (Cancer Epidemiol Biomarkers Prev 2008;17(7):1653 -7)

Metabolic activation of the hepatocarcinogenic mycotoxin aflatoxin B 1 (AFB 1 ) results in the covalent attachment of AFB 1 to serum albumin. Digestion of adducted albumin releases AFB 1 -lysine, a biomarker of exposure status. AFalbumin adducts have been most frequently measured in precipitated serum albumin using an immunoassay (ELISA); however, a sensitive and specific isotope dilution mass spectrometric (IDMS) assay for measurement of AFB 1 -lysine in serum has recently been developed. The ELISA and IDMS methods were compared using 20 human sera collected in Guinea, West Africa, where AF exposure is endemic. Measurement of AFB 1 -lysine adduct concentrations by IDMS in serum and albumin precipitated from the same sample revealed that precipitation has no effect on the measured adduct levels. The concentration of AF-albumin adducts measured by ELISA and AFB 1 -lysine measured by IDMS in 2 mg of albumin were well correlated (R = 0.88, P < 0.0001); however, AF-albumin adduct concentrations measured by ELISA were on average 2.6-fold greater than those of the AFB 1 -lysine adduct. Although these data suggest that the ELISA is measuring other AF adducts in addition to AFB 1 -lysine, these biomarkers are comparable in their ability to assess AF exposure at AF-albumin concentrations z3 pg AFB 1 -lysine equivalents/mg albumin. Identification of other adducts may clarify the mechanistic basis for using AFprotein biomarkers to assess exposure status in future epidemiologic studies of liver cancer. (Cancer Epidemiol Biomarkers Prev 2006;15(4):823 -6)

To assess the scale of the possible exposure by the breast-fed infant to potentially harmful substances in breast milk, methodologically robust studies are essential. Many studies in this field, however, do not report details of crucial issues such as recruitment and milk sampling. The aims of the study reported here were to develop robust methods for the study of contaminants in breast milk, and to develop a framework for future research and population monitoring. Three cohorts of women and babies were recruited by midwives from five sites in northern England. Cohort 1 (cross-sectional, n = 322) were asked to provide two milk samples, one at one week following birth and one at a subsequent time point. Cohort 2 (longitudinal, n = 54) were asked to provide five samples at specified time points over the first 12-16 weeks after birth. Cohort 3 (convenience, n = 18), mothers of babies in the Special Care Unit, were asked to donate surplus breast milk. A novel method of analysing fat concentration in small volumes was developed and tested. A randomly selected set of samples from different donors and stages of lactation was screened for organochlorine pesticide residues, polychlorinated biphenyls, dioxins/furans, phthalates and heavy metals. A total of 453 samples were donated. Cohort 3 was the least successful route of recruitment. Cohorts 1 and 2 combined were most representative of the population. Sample collection, transport and storage procedures, and the collection of data on life style and diet, were robust and acceptable to women. Midwifery involvement in recruitment was an essential component. This study offers a framework both for the conduct of future research studies, and for the establishment of regional and national monitoring mechanisms for contaminants in breast milk. Similar work on contaminants in formula as fed to babies is needed to inform risk assessment methods.