ID AND DEV OF BIOLOGICAL MARKERS OF HUMAN EXPOSURE TO THE INSECTICIDE PERMETHRI

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 中心，但不一定是研究者所在的机构。 农业工人、园丁和房主经常接触杀虫剂氯菊酯。 此外，军事人员在使用国防部驱虫剂系统时会接触到氯菊酯，而非处方虱子肥皂则使用氯菊酯作为活性成分。氯菊酯的尿代谢产物丰度高，相对稳定，可能是暴露于该农药的理想生物标志物。 此外，根据给药途径，一种代谢物与另一种代谢物的比例可能不同。本研究的结果将用于确定候选人的快速，灵敏的免疫化学为基础的分析方法，可用于常规监测人类接触氯菊酯的发展。 目的：本研究的目的是确定皮肤接触后尿液中最丰富且最稳定的氯菊酯人体代谢物。加速器质谱是一种测量14 C水平的方法，比液体闪烁计数灵敏几个数量级。利用这种高灵敏度，我们将在生物学相关剂量下进行人体代谢研究。 具体目标： 开发用于分离氯菊酯及其推定的人体代谢物的LC/MS方法。 二. 使用加速器质谱法（AMS）测定氯菊酯的人体代谢物谱。 三. 开发目标II中确定的关键代谢物的免疫测定法，作为人类暴露于氯菊酯的生物标志物。 研究方法：对于特定目标I，合成代谢物标准品;开发HPLC方法，使用紫外检测分离推定的拟除虫菊酯代谢物;确定HPLC/质谱法分析拟除虫菊酯代谢物的可行性。 对于特定目标II，人体皮肤临床暴露于放射性标记的二氯苯醚菊酯，并采集尿液、血液和唾液;通过特定目标I中开发的方法分离样品，并通过加速器质谱法分析分离的样品;从所得数据中鉴定最常见的代谢物。对于特定目标III，半抗原的合成;抗体的开发;半抗原和抗体在最常见代谢物的免疫测定开发中的应用;免疫测定的验证。 预期产品（里程碑）：假定的二氯苯醚菊酯人体代谢物的文献综述;少量合成的二氯苯醚菊酯代谢物;用于分离人尿液或唾液中二氯苯醚菊酯代谢物的HPLC方法;通过加速器质谱法鉴定最丰富的人体代谢物;检测二氯苯醚菊酯靶向人体代谢物的免疫测定法 结论：本研究的结果将被用来确定候选人的快速，灵敏的免疫化学为基础的分析方法，可用于常规监测人类暴露于氯菊酯的发展。仔细监测氯菊酯吸收剂量的存在的能力将是一个有用的工具，以防止由于氯菊酯暴露对人类健康的影响的可能性。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Agricultural workers, gardeners and homeowners are routinely exposed to the insecticide permethrin. Also, military personnel are exposed to the permethrin when using the DOD Insect Repellent System and over-the-counter lice soaps use permethrin as the active ingredient. A urinary metabolite of permethrin, that is in high abundance and is relatively stable, may be an ideal biomarker of exposure to this pesticide. In addition, the ratio of one metabolite to another may vary, according to the route of administration. The results of this study would be used to identify candidates for the development of a rapid, sensitive immunochemical based analytical method that can be used to routinely monitor human exposure to permethrin. Objectives: The purpose of this study is to determine the human metabolite(s) of permethrin in urine following dermal exposure that are in greatest abundance and are the most stable. Accelerator mass spectrometry is a method for measuring levels of 14C several orders of magnitude more sensitive than liquid scintillation counting. With this high sensitivity we will conduct human metabolism studies at biologically relevant doses. SPECIFIC AIMS: I. Develop an LC/MS method for separation of permethrin and its putative human metabolites. II. Determine the human metabolite profile of permethrin using accelerator mass spectrometry (AMS). III. Develop an immunoassay to the key metabolite identified in Objective II as a biomarker of human exposure to permethrin. METHODOLOGY: For specific aim I, synthesize metabolite standards; develop an HPLC method to separate the putative pyrethroid metabolites using ultraviolet detection; determine the feasibility of an HPLC/mass spectrometry method for analysis of pyrethroid metabolites. For specific aim II, clinical exposure of humans to radiolabeled permethrin dermally and collection of urine, blood and saliva; separation of samples by methods developed in specific aim I and analysis of separated samples by accelerator mass spectrometry; identification of most prevalent metabolite from resultant data. For specific aim III, synthesis of haptens; development of antibodies; use of the haptens and antibodies in the development of an immunoassay for the most prevalent metabolite; validation of immunoassay. EXPECTED PRODUCTS (MILESTONES): Literature review of putative human metabolites of permethrin; small quantities of synthesized metabolites of permethrin; an HPLC method for separating permethrin metabolites in human urine or saliva; identification of the most abundant human metabolite(s) by accelerator mass spectrometry; an immunoassay to detect the targeted human metabolite of permethrin Conclusion: The results of this study will be used to identify candidates for the development of a rapid, sensitive immunochemical based analytical method that can be used to routinely monitor human exposure to permethrin. The ability to carefully monitor the presence of absorbed doses of permethrin will be a useful tool to prevent the possibility of human health effects due to permethrin exposure.