Lisa Jo Melnyk

Review of Pesticide Urinary Biomarker Measurements from Selected US EPA Children’s Observational Exposure Studies

Children are exposed to a wide variety of pesticides originating from both outdoor and indoor sources. Several studies were conducted or funded by the EPA over the past decade to investigate children’s exposure to organophosphate and pyrethroid pesticides and the factors that impact their exposures. Urinary metabolite concentration measurements from these studies are consolidated here to identify trends, spatial and temporal patterns, and areas where further research is required. Namely, concentrations of the metabolites of chlorpyrifos (3,5,6-trichloro-2-pyridinol or TCPy), diazinon (2-isopropyl-6-methyl-4-pyrimidinol or IMP), and permethrin (3-phenoxybenzoic acid or 3-PBA) are presented. Information on the kinetic parameters describing absorption and elimination in humans is also presented to aid in interpretation. Metabolite concentrations varied more dramatically across studies for 3-PBA and IMP than for TCPy, with TCPy concentrations about an order of magnitude higher than the 3-PBA concentrations. Temporal variability was high for all metabolites with urinary 3-PBA concentrations slightly more consistent over time than the TCPy concentrations. Urinary biomarker levels provided only limited evidence of applications. The observed relationships between urinary metabolite levels and estimates of pesticide intake may be affected by differences in the contribution of each exposure route to total intake, which may vary with exposure intensity and across individuals.

Contribution of children's activities to lead contamination of food

This study evaluates the relationship of childrens hygiene habits and food-handling behaviors on lead levels on hands and handled foods for toddlers living in lead-contaminated homes. Forty-eight inner city toddlers previously identified as having elevated blood lead levels participated in three consecutive days of designated food-handling activities. During the visits, duplicate diets were obtained, the child handled a banana, a hot dog, and had his/her hands wiped with a moist towelette. In addition, wipe samples were collected from the kitchen floor, and food items were deposited on and subsequently collected from the kitchen floor. All samples were analyzed for lead. The childs caregiver completed a questionnaire, which addressed the childs hygiene and eating behaviors. It was demonstrated that childrens contact with residential dust containing lead can transfer lead to food. Both lead in the home and on the childrens hands contribute to the contamination of food, and hence potential dietary exposure. Mean lead in handled bananas was 26 μg/kg and on hot dogs 65 μg/kg, and mean lead values on cheese and apple slices that had been on the floor were 119 and 215 μg/kg. In addition, the childs hygiene habits as reported by the parent indicate that lack of basic hygiene patterns within a high lead environment can contribute to childrens dietary exposure to lead.

Dietary exposure of children in lead-laden environments.

Children are the most susceptible population to lead exposure because of three interacting factors; they have more opportunity for contact with lead sources due to their activities, lead absorption occurs more readily in a child compared to an adult, and the childs development is more vulnerable to lead than adults. Low levels of lead in the blood have been shown to cause adverse health effects; the level of concern for children is currently 10 µg/dl. The contribution of dietary exposure of lead to increased blood lead levels (PbB) is not well characterized. This study was conducted to measure potential dietary lead intakes of children 2 to 3 years of age who live in homes contaminated with environmental lead. Objectives were to estimate lead intakes for children consuming food in contaminated environments, recognizing unstructured eating patterns and to investigate if correlations exist between daily dietary exposure and measured PbB. Dietary exposure was evaluated by collecting samples that were typical of the foods the young children ate in their homes. A 24-h duplicate of all foods plus sentinel foods, i.e., individual items used to represent foods contaminated during handling, were collected from 48 children. Ten homes were revisited to obtain information on the variation in daily dietary intakes. Drinking water was evaluated both as part of the segregated beverage sample composite and by itself. Additional information collected included lead concentrations from hand wipes, floor wipes, and venous blood, and questionnaire responses from the caregiver on activities potentially related to exposure. Activities and hygiene practices of the children and contamination of foods in their environment influences total dietary intake. Estimated mean dietary intakes of lead (29.2 µg Pb/day) were more than three times the measured 24-h duplicate-diet levels (8.37 µg Pb/day), which were almost six times higher than current national estimates (1.40 µg Pb/day). Statistically significant correlations were observed between floor wipes and foods contacting contaminated surfaces, hand wipes and foods contacting contaminated hands and surfaces, and hand wipes and floor wipes. This study indicates that the dietary pathway of exposure to lead is impacted by eating activities of children living in lead-contaminated environments and that analysis of foods themselves is not enough to determine excess dietary exposures that are occurring.

Transfer efficiencies of pesticides from household flooring surfaces to foods

The transfer of pesticides from household surfaces to foods was measured to determine the degree of excess dietary exposure that occurs when childrens foods contact contaminated surfaces prior to being eaten. Three household flooring surfaces (ceramic tile, hardwood, and carpet) were contaminated with an aqueous emulsion of commercially available pesticides (diazinon, heptachlor, malathion, chlorpyrifos, isofenphos, and cis- and trans-permethrin) frequently found in residential environments. A surface wipe method, as typically used in residential exposure studies, was used to measure the pesticides available on the surfaces as a basis for calculating transfer efficiency to the foods. Three foods (apple, bologna, and cheese) routinely handled by children before eating were placed on the contaminated surfaces and transfers of pesticides were measured after 10 min contact. Other contact durations (1 and 60 min) and applying additional contact force (1500 g) to the foods were evaluated for their impact on transferred pesticides. More pesticides transferred to the foods from the hard surfaces, that is, ceramic tile and hardwood flooring, than from carpet. Mean transfer efficiencies for all pesticides to the three foods ranged from 24% to 40% from ceramic tile and 15% to 29% from hardwood, as compared to mostly non-detectable transfers from carpet. Contact duration and applied force notably increased pesticide transfer. The mean transfer efficiency for the seven pesticides increased from around 1% at 1 min to 55– 83% when contact duration was increased to 60 min for the three foods contacting hardwood flooring. Mean transfer efficiency for 10-min contact increased from 15% to 70% when a 1500 g force was applied to bologna placed on hardwood flooring. Contamination of food occurs from contact with pesticide-laden surfaces, thus increasing the potential for excess dietary exposure of children.

Development of an analytical scheme for the determination of pyrethroid pesticides in composite diet samples.

Analysis of an individuals total daily food intake may be used to determine aggregate dietary ingestion of given compounds. However, the resulting composite sample represents a complex mixture, and measurement of such can often prove to be difficult. In this work, an analytical scheme was developed for the determination of 12 select pyrethroid pesticides in dietary samples. In the first phase of the study, several cleanup steps were investigated for their effectiveness in removing interferences in samples with a range of fat content (1-10%). Food samples were homogenized in the laboratory, and preparatory techniques were evaluated through recoveries from fortified samples. The selected final procedure consisted of a lyophilization step prior to sample extraction. A sequential 2-fold cleanup procedure of the extract included diatomaceous earth for removal of lipid components followed with a combination of deactivated alumina and C(18) for the simultaneous removal of polar and nonpolar interferences. Recoveries from fortified composite diet samples (10 microg kg(-1)) ranged from 50.2 to 147%. In the second phase of this work, three instrumental techniques [gas chromatography-microelectron capture detection (GC-microECD), GC-quadrupole mass spectrometry (GC-quadrupole-MS), and GC-ion trap-MS/MS] were compared for greatest sensitivity. GC-quadrupole-MS operated in selective ion monitoring (SIM) mode proved to be most sensitive, yielding method detection limits of approximately 1 microg kg(-1). The developed extraction/instrumental scheme was applied to samples collected in an exposure measurement field study. The samples were fortified and analyte recoveries were acceptable (75.9-125%); however, compounds coextracted from the food matrix prevented quantitation of four of the pyrethroid analytes in two of the samples considered.

Collecting urine samples from young children using cotton gauze for pesticide studies.

To estimate pesticide exposure, urine samples are often needed to analyze pesticide metabolites. However, this is difficult for children wearing diapers because simple and feasible techniques suitable for field collection are not available. The objectives of this study were to test the validity of using cotton gauze pad as a medium for collecting urine samples from young children and to examine the stability of the recoveries for creatinine and pesticide metabolites over 24 h. Urine spiked with a pesticide and four metabolites, 2,4-dichlorophenoxyacetic acid (which is mainly eliminated from urine unchanged), 3-phenoxybenzoic acid (metabolite for synthetic pyrethroids), atrazine mercapturate (metabolite for atrazine), malathion dicarboxylic acid (metabolite for malathion), and 2-isopropyl-4-methyl-6-hydroxypyrimidine (metabolite for diazinon) was added to the gauze pads and kept in jars at 37°C in a water bath. Urine was expressed from the gauze pads immediately and after 1, 2, 4, 8, and 24 h, then analyzed. The recoveries, calculated as the percentage of concentration in expressed urine divided by that of the control urine sample, were within a range of 70–130%. The metabolite and creatinine concentrations did not change with time in either expressed urine samples or controls. The results suggest that cotton gauze pad is a promising candidate for collecting urine samples from young children wearing diapers for studies in which these five urinary pesticide metabolites are to be analyzed.

Use of Pharmacokinetic Modeling to Design Studies for Pathway-Specific Exposure Model Evaluation

Validating an exposure pathway model is difficult because the biomarker, which is often used to evaluate the model prediction, is an integrated measure for exposures from all the exposure routes and pathways. The purpose of this article is to demonstrate a method to use pharmacokinetic (PK) modeling and computer simulation to guide the design of field studies to validate pathway models. The children’s dietary intake model is discussed in detail as an example. Three important aspects are identified for a successful design to evaluate the children’s dietary intake model: a) longitudinally designed study with significant changes in the exposure for the route/pathway of interest, b) short biologic half-life of the selected chemical, and c) surface loading of the selected chemical at sufficient levels. Using PK modeling to guide a study design allowed a path-specific exposure model to be evaluated using urinary metabolite biomarkers.

Dietary intakes of pesticides based on community duplicate diet samples

The calculation of dietary intake of selected pesticides was accomplished using food samples collected from individual representatives of a defined demographic community using a community duplicate diet approach. A community of nine participants was identified in Apopka, FL from which intake assessments of organophosphate (OP) and pyrethroid pesticides were made. From these nine participants, sixty-seven individual samples were collected and subsequently analyzed by gas chromatography/mass spectrometry. Measured concentrations were used to estimate dietary intakes for individuals and for the community. Individual intakes of total OP and pyrethroid pesticides ranged from 6.7 to 996 ng and 1.2 to 16,000 ng, respectively. The community intake was 256 ng for OPs and 3430 ng for pyrethroid pesticides. The most commonly detected pesticide was permethrin, but the highest overall intake was of bifenthrin followed by esfenvalerate. These data indicate that the community in Apopka, FL, as represented by the nine individuals, was potentially exposed to both OP and pyrethroid pesticides at levels consistent with a dietary model and other field studies in which standard duplicate diet samples were collected. Higher levels of pyrethroid pesticides were measured than OPs, which is consistent with decreased usage of OPs. The diversity of pyrethroid pesticides detected in food samples was greater than expected. Continually changing pesticide usage patterns need to be considered when determining analytes of interest for large scale epidemiology studies. The Community Duplicate Diet Methodology is a tool for researchers to meet emerging exposure measurement needs that will lead to more accurate assessments of intake which may enhance decisions for chemical regulation. Successfully determining the intake of pesticides through the dietary route will allow for accurate assessments of pesticide exposures to a community of individuals, thereby significantly enhancing the research benefit realized from epidemiological exposure studies.

Pesticides on Household Surfaces May Influence Dietary Intake of Children

The physical and chemical environment influences childrens exposures to pesticides in and around the home. Childrens activities, which increase their potential for exposure especially during eating, have been captured in the Childrens Dietary Intake Model (CDIM). In addition to the chemical exposure associated with the food itself, this model incorporates excess dietary exposures due to handling of food during consumption. To stochastically evaluate CDIM, distributions of measured, and in some cases estimated, model factors were determined from measurements of permethrin, chlorpyrifos, and diazinon derived from assembled databases and laboratory experiments. Using the distributions of these factors, Monte Carlo simulations were performed to obtain distributions of total dietary intake of pesticides. To target the sources of pesticide contamination that were influencing total dietary intake, each factor was evaluated. We found pesticide surface concentration to be highly influential. By excluding surface concentration, we were also able to determine the influence of the other factors based on the F-statistic. Transfer efficiencies, followed by pesticide residue in consumed foods and amount of food consumed, were the next most influential factors within the model. With these distributions for model inputs, CDIM has the potential to more accurately predict total dietary intake of a contaminant by a child.

Surface-to-food pesticide transfer as a function of moisture and fat content.

Transfer of pesticides from household surfaces to foods may result in excess dietary exposure in children (i.e., beyond that inherent in foods due to agricultural application). In this study, transfer was evaluated as a function of the moisture and fat content of various foods. Surfaces chosen for investigation were those commonly found in homes and included Formica®, ceramic tile, plastic, carpet, and upholstery fabric. Each surface type was sprayed with an aqueous emulsion of organophosphates, fipronil, and synthetic pyrethroids. In the first phase of the study, multiple foods (apples, watermelon, wheat crackers, graham crackers, white bread, flour tortillas, bologna, fat-free bologna, sugar cookies, ham, Fruit Roll-ups®, pancakes, and processed American cheese) were categorized with respect to moisture and fat content. All were evaluated for potential removal of applied pesticides from a Formica surface. In the second phase of the study, representative foods from each classification were investigated for their potential for pesticide transfer with an additional four surfaces: ceramic tile, plastic, upholstery, and carpet. Moisture content, not fat, was found to be a determining factor in most transfers. For nearly all surfaces, more efficient transfer occurred with increased hardness (Formica and ceramic tile). Comparatively, the polymer composition of the plastic delivered overall lower transfer efficiencies, presumably due to an attraction between it and the organic pesticides of interest.