Project Number: 92700A2 Project Title: Immunochemical Biological Monitoring for Occup Exp & Dis Division: DART Project Officer (PO): Cynthia Striley PO Degree: Ph.D. PO Area of Expertise (scientific discipline): Analytical Chemistry Project Start Date: 10/04 Project End Date: 09/09 Total Budget over the Project Period: $73,772 Goal/Sub-Goal: Chronic Effects of Pesticide Exposure/Exposure Assessment Research Challenge/Issue: NIOSH is committed to developing new methods to assess workplace exposures to toxicants. This project is designed to further the NORA Exposure Assessment Team goal to develop and validate biomonitoring methods to assess the internal and biologically effective dose of an occupational exposure. The Institute's field investigations of occupational hazards require assessment of exposures which might lead to injury or illness. Environmental monitoring can be a good control measure for making a safer workplace, but only biological monitoring can accurately assess an individual's exposure. Exposure of workers to occupational hazards often results in the appearance of the chemical or metabolites in the workers' blood, breath, urine, or other biological medium. This exposure can be demonstrated and quantified by analytical methods appropriate for the chemical or metabolite. Termed biological monitoring, this technique is complementary to workplace environmental monitoring and is especially useful for documenting occupational exposure situations where absorption through the skin and ingestion are significant uptake routes. Classical analytical techniques that employ chromatographic analysis can be time consuming, labor intensive and expensive. Immunochemically-based methods are significantly less expensive than classical methods, much less time consuming, and usually less labor intensive. Biological monitoring results are used (1) for hazard surveillance, to demonstrate the existence of an exposure problem so that it can be corrected, and (2) to demonstrate the adequacy of control technologies and intervention strategies (3) identify those individuals most at risk. Activities: This project will provide biological monitoring required by Institute health-hazard evaluations and other NIOSH field research investigations – only a portion of the research activities contained herein are Ag related. To support this work, this project will modify, develop and validate new immunochemical methods for field studies that require biological monitoring and apply previously developed methods to field studies. Prior work, conducted under the project Biomonitoring Methods for Agricultural Exposures demonstrated the applicability of immunochemical detection as a rapid, simple method for biological monitoring. This project will initially address a collaborative effort with NCEH to develop analytical methodology and universally used standards for a large-scale study to investigate acrylamide exposure for DART field studies and also NHANES with the National Center for Environmental Health (NCEH) and National Center for Health Statistics (NCHS). Using synthetic peptides and prepared antibodies, an immunochemical method for assessing long term exposure to acrylamide will be developed. This method will be used to test workers involved in the production of acrylamide polymers and
workers that use acrylamide in farming, mining and manufacturing, but it also has far more reaching implications, since acrylamide has also been found in various common foods. Current methodology is very time consuming and expensive. Complimentary to development of immunochemical methods for occupational exposure, this project will develop assays for biomarkers of effect to occupational hazards. By combining a diverse array of markers of biological response in blood, urine and other biological materials an exposure-response matrix will be developed. Specific markers of organ toxicity (renal, hepatic, etc), immunotoxicity, inflammation, cellular stress response, and xenobiotic metabolism and transcription factors will be measured in worker samples and a biological response matrix will be developed for specific occupational hazards such as fuels and solvents, metals, asphalt fume, and pesticides. Methods will be developed in a tiered approach. First, in vitro studies will be performed by exposing appropriate cells (lung, liver, blood) to the occupational hazard and multiple cellular responses measured. Second, in association with researchers at Wright Patterson AFB, NTP and others, biological fluids and tissues from rodents exposed to occupational hazards in vivo will be examined for those markers identified by cellular studies as most promising. Last, once a pattern of diverse biological markers of response has been identified by cellular and animal studies this suite will be applied to worker samples from an identical exposure. By identifying early, sensitive indicators of toxicity prior to manifestation of clinical disease timely intervention strategies can be put in place. Additionally, this project will develop rapid and reliable testing kits, devices and dipstick type assays (e.g., lateral flow immunoassays) to assess biomarkers of exposure or effect of exposure. These assays would use saliva or blood components to rapidly (10-20 minutes) give a yes/no type answer to the existence of an analyte determined to be associated to a disease or exposure outcome from the above-defined response profiles. This type of kit would be useful not only in an occupational setting, but has applicability during public health crises to assess exposure or an effect of exposure. Current completed activities of this project include the evaluation/validation/analysis of on-site sampling technologies for cotinine, acetylcholinesterase, and specific gravity for a second hand smoke HHE. In addition, evaluation/validation of method for styrene in exhaled breath for an upcoming IWSB/EPHB was completed. Also, development of a high performance liquid chromatography (HPLC) method for the evaluation the coumaphos metabolite: chlorferon in urine was completed and all collected HHE urine samples were analyzed. Outputs: The following peer reviewed journal article was published. Hines, CJ, Deddens, JA, Lu, C, Fenske, RA, and Striley CAF. "Mixed-Effect Models for Evaluating Multiple Measures of Atrazine Exposure among Custom Applicators." Journal of Occupational and Environmental Hygiene, 3: 274–283 (2006) NN: 20030196 Data analysis was provided for the tick eradication HHE (HETA-20020206) and the casino second-hand smoke HHE (HETA-20050201, HETA-20050076) Intermediate Outcomes: Rapid, inexpensive assays to evaluate occupational chemical exposures in workers and early biomarkers of injury were developed and validated. Because environmental sampling techniques
can give an incomplete exposure assessment, these biological monitoring methods will provide a measure of actual dose received. Development of simple, sensitive assays to evaluate exposure and sub-clinical markers of toxicity will improve worker health and safety by providing an impetus for early intervention. Analyses provided for NIOSH field studies and HHEs directly impacts workers and interventions where significant exposures are found. End Outcomes: None External Factors: N/A Future Directions: At the time this review was written, this was an ongoing project. Future directions are still in the scope of the ongoing and planned activities.