Project Number: 9278351 Project Title: Biomonitoring Methods for Agricultural Exposures Division: DART Project Officer (PO): Cynthia Striley PO Degree: Ph.D. PO Area of Expertise (scientific discipline): Analytical Chemistry Project Start Date: 10/97 Project End Date: 09/04 Total Budget over the Project Period: $890,403 Goal/Sub-Goal: Chronic Effects of Pesticide Exposure/Exposure Assessment Research Challenge/Issue: Millions of pounds of pesticides are applied each year, putting a large number of workers at risk for exposure to potentially toxic chemicals. NIOSH currently has several studies investigating exposures to organophosphate insecticides and the herbicide metolachlor. These surveillance and intervention studies require sensitive and accurate methods to assess exposure of the workers, and since much of their exposure is through the skin, biological monitoring is needed to complement sampling of air, clothing, etc. Because conventional biomonitoring methods are relatively slow and expensive, new biomonitoring methods that employ ELISAs are being developed and evaluated. These new methods will be cheaper and easier to use than conventional assays, thereby allowing for quicker identification and correction of exposure risks. Activities: ELISA (Enzyme-linked Immunosorbent Assay) was coupled with microbead multiplexing to produce analyses for metolachor mercapturate, atrazine mercapturate, trichloropyridinol and glyphosate. This method was developed for a DSHEFS exposure assessment field study that evaluated pesticide exposure of small family farms. The study addressed the direct exposure of the farmers and secondary exposure of their families including children. An ELISA for trichloropyridinol was developed and validated and used to evaluate levels in urine from an orchardist sprayer study and a pesticide applicators study. This method included an additional step to liberate pesticide metabolite from an excreted conjugate. The method exhibited a high correlation to standard chromatographic methodology. Human hemoglobin adducts of acrylamide (a soil stabilization chemical) were prepared as standards for the evaluation of exposure to acrylamide. In addition a modified valine amino acid was synthesized. This modified amino acid was used to prepare alpha and beta hemoglobin chain peptides for use in the characterization and evaluation of acrylamide-adducted blood samples. Outputs: The following peer reviewed journal articles were published as a result of this research: Biagini, RE, Sammons, DL, Smith, JP, Snawder, JE, Striley, CAF, MacKenzie, BA, Robertson, SR. “Development of a Sensitivity Enhanced Multiplexed Fluorescence Covalent Microbead Immunosorbent Assay (FCMIA) for the Measurement of Glyphosate, Atrazine and Metolachlor Mercapturate in Water and Urine.” Analytical and Bioanalytical Chemistry, 379:368-374, 2004. NN 20026130
Hines, C. J., Deddens, J. A., Striley, C. A. F., Biagini, R. E., Shoemaker, D. A., Brown, K. K., MacKenzie, B. A., and Hull., R. D. "Biological Monitoring for Selected Herbicide Biomarkers in the Urine of Exposed Custom Applicators: Application of Mixed-Effect Models," Annals of Occupational Hygiene, 47, 503 (2003). NN: 20023365 Biagini, R.E., Murphy, D.M., Sammons, D.L., Smith, J.P., Striley, C.A.F. and MacKenzie B.A. "Development of Multiplexed Fluorescence Microbead Covalent Assays (FMCAs) for Pesticide Biomonitoring" Bull of Environ Contam and Toxicol 68, 470 (2002). NN: 20023004 MacKenzie, B.A., Striley, C.A.F., Biagini, R.E., Stettler L.E. and. Hines, C.J. "Improved Rapid Analytical Method for the Urinary Determination of 3,5,6-Trichloro-2-pyridinol, a metabolite of chlorpyrifos." Bull of Environ Contam and Toxicol 65:1, 1 (2000). [Cited 2 times] NN: 20020744 Striley, C.A.F., Biagini, R.E. Mastin, J.P., MacKenzie, B.A. and. Robertson, S.K. "Development and validation of an ELISA for metolachlor mercapturate in urine" Analytical Chimica Acta 399, 109 (1999). [Cited 4 times] Mastin, J.P., Striley, C.A.F., Biagini, R.E., Hines, C.J., Hull, R.D., MacKenzie B.A. and Robertson, S.K. "Use of Immunoassays for Biomonitoring of Herbicide Metabolites in Urine." Analytical Chimica Acta 376:1, 119 (1998). NN: 20027943 The following presentations and posters were presented as a result of this work Curwin, B.D., Hein, M.J., Sanderson, W.J., Striley, C., Heederik, D., Reynolds, S.J., Ward, E.M. and Alavanja, M.C., "Urinary pesticide levels in Iowa farm spouses and children." 15th Annual International Society of Exposure Analysis (ISEA) Conference, in Tucson AZ, Oct. 30 to Nov 3, 2005. Curwin, B.D., Hein, M.J., Sanderson, W.T., Striley, C.A., Heederik, D., Reynolds, S.J., Ward, E.M., Alavanja, M.C. “Urinary pesticide levels in Iowa farm spouses and children.” National Advisory Panel meeting for the Agricultural Health Study, Research Triangle Park, NC, March 3-4, 2005 J. Smith, D. Sammons, R. Biagini, J. Snawder, C. Striley, B. MacKenzie and L. Hollstegge. “Multiplexed (tetra-plex) urinalyses of pesticides using Fluorescence Microbead Covalent ImmunoAssays (FMCIAs).” Poster, 2005 Toxicology and Risk Assessment Conference, April 25-28, 2005, Fairborn, OH, April 25-28, 2005, Fairborn, OH. Hines, C.J., Deddens, J.A., Striley, C.A.F., Brown, K.K, Lu, C., MacKenzie, B.A., Biagini, R.E., Shoemaker, D.A., and Fenske, R.A., “Biological Monitoring for Atrazine and 2,4-D Among Custom Applicators I: Results of Exposure Determinant Modeling.” American Industrial Hygiene Conference & Exposition Atlanta (2004) Utrecht and X2004 -Exposure Assessment in a Changing Environment, Abstract No. 34-2, 2004, Utrecht, the Netherlands.
Hines, C.J., Deddens, J.A., Striley, C.A.F., Biagini, R.E., Shoemaker, D.A., Brown, K.K., MacKenzie, B.A. and Hull, R.D., “Biological Monitoring for Urinary Herbicide Biomarkers among Custom Applicators: Application of Mixed-Effect Models.” AICHE, (2003) Striley, C.A.F., Biagini R.E., Cheever, K. and Snawder J.E. "Development of an ELISA using polypeptide fragments of hemoglobin-acrylamide adducts for biological monitoring of acrylamide exposure.” NIOSH Salt Fork XII Interdivisional Meeting in Mineral Wells, WV. November (2003). Striley, C.A.F., Biagini R.E. and Snawder J.E., “Development of an ELISA using polypeptide fragments of hemoglobin-acrylamide adducts for biological monitoring of acrylamide exposure.” 226th ACS National Meeting Book of Abstracts, #AGRO 33, New York (2003). Smith, J.P., Sammons, D.L., Biagini, R.E., Snawder, J.E., Striley, C.A.F. and MacKenzie, B.A., “The Use of Fluorescence Microbead Immunosorbent Assays (FCMIAs) for Pesticide Biomonitoring.” 226th ACS National Meeting #AGRO 33, New York, (2003). Snawder J.E., MacKenzie, B.A., Biagini, R.E., Sammons, D.L., Smith, J.P. and Striley C.A.F., "Development and application of new and existing biological monitoring methods to assess workers' exposure to toxic chemicals and their role in hazard identification, evaluation and control. “ Abstract, Applying Biomarkers to Occupational Health Practice, Santa Fe, NM, March 24-25, (2003). Biagini, R.E., Sammons, D.L., Smith, J. P., Snawder, J.E., Striley, C. A. F., MacKenzie, B.A. and Hollstegge L. "Fluorescence Microbead Immunoasays: General Overview and an Example of Deep Multiplexing for Evaluating Vaccine Efficacy." NIOSH Salt Fork XI Interdivisional Meeting in Mineral Wells, WV. November, (2002). Smith, J.P., Sammons, D.L., Biagini, R.E., Snawder, J.E., Striley, C.A.F., MacKenzie, B.A. and Hollstegge L., "Multiplexed (tetra-plex) urinalyses of pesticides using Fluorescence Microbead Immunoasays." NIOSH Salt Fork XI Interdivisional Meeting in Mineral Wells, WV November, (2002). Biagini, R.E., Murphy, D.M., Striley C.A.F., Mackenzie, B.A., and Robertson, S.K., “The Use of Fluorescence Microbead Immunosorbent Assays (FMIAs) for Pesticide Biomonitoring." 2000 International Chemical Congress of Pacific Basin Societies, Honolulu Hawaii December 14-19, (2000), Striley, C.A.F. Biagini, R.E., Snawder, J.E., Hines, C.J. and Mackenzie, B.A., "Clinical Biological Monitoring Using Immunobiochemical Methods". Immunochemistry Summit VIII /219th American Chemical Society National Meeting, [Book of Abstracts, Part 1:106] San Francisco CA, March (2000). Striley, C.A.F. Biagini, R.E., Snawder, J.E., Hines, C.J. and Mackenzie, B.A., “Immunochemical Biological Monitoring at NIOSH." Invited seminar presentation at the Department of Chemistry: University at Albany-SUNY, Albany, NY, May (2000).
Striley, C.A.F. Biagini, R.E., Mastin, J. P., Hines, C. J., Mackenzie, B. A., and Zimmer H. "Pesticide Determination through Immunoassay". NIOSH Salt Fork Interdivisional Conference, Salt Fork State Park, Ohio. (1998). Striley C.A.F., Biagini, R.E., Mastin, J. P., Hines, C. J., Mackenzie, B. A., and Zimmer H. "Development and Validation of Immunochemical Methods for Biomonitoring." Immunochemistry Summit VII and Third Workshop on Biosensors and Biological Techniques in Environmental Analysis, Las Vegas, NV, December (1998). Intermediate Outcomes: The new methods developed as part of this project are novel assays which were used to monitor exposure to pesticides in several DSHEFS field studies so that levels can be decreased to prevent worker illness. Additionally, methods developed in this project were utilized for biomonitoring in the Farm Family field study that is addressing the scope and impact of agrochemical exposure on children. External Factors: N/A Future Directions: Agricultural chemical exposure and biological monitoring in worker blood and urine continues to be the subject of NIOSH field studies and health hazard evaluations. At the time this project ended, a new project that included agricultural exposure assessment was in the planning stages due to the need for such analyses.