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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 农业工人、园丁和房主经常接触杀虫剂氯氰菊酯。还有，军事 使用国防部驱蚊系统和非处方药虱子肥皂时，人员暴露在氯氰菊酯中 使用氯氰菊酯作为活性成分。氯氰菊酯一种二氯氰菊酯的尿代谢物，含量高，且相对 稳定，可能是接触这种杀虫剂的理想生物标志物。此外，一种代谢物与另一种代谢物的比率可 根据给药途径的不同而有所不同。这项研究的结果将被用来确定候选人 建立一种快速、灵敏的免疫化学分析方法，可用于常规监测 人类接触二氯苯醚菊酯。 目的：研究氯氰菊酯在人体皮肤中的代谢产物S。 最丰富和最稳定的暴露。加速器质谱学是一种用于 测量14C的能级比液体闪烁计数灵敏几个数量级。有这么高的价格 我们将以生物相关剂量进行人体新陈代谢研究。 方法：我们将综合利用三种前沿分析技术来识别和开发 检测和定量人类接触氯氰菊酯的实验室和现场便携方法。 高压液相色谱与最先进的串联质谱仪(LC/MS)联用将用于 分离和鉴定氯菊酯的尿代谢物。加速器质谱学(AMS)是一种 测量14C的能级比液体闪烁计数灵敏几个数量级。有了这个 超灵敏，将使用反映人体实际暴露剂量的14C-氯菊酯来测定代谢物 具有最低辐射风险的配置文件。经LC/MS和AMS鉴定的代谢物将用于开发快速、灵敏的 二氯苯醚菊酯暴露人体常规监测的免疫分析。六名人类志愿者将暴露在低氧环境中 皮下注射14C-氯氰菊酯8小时。尿液和血液样本将在7天内收集，并将有更多 在最初的24小时内经常出现时间点。首先将进行一项利用一个对象的试验性研究，以测试这些方法 为分离预期代谢物而开发。使用其他5个受试者的暴露研究将提供信心。 观察到的代谢物模式可以作为一般人群的预测指标。 结果：将对尿代谢物进行鉴定和定量。基于这一信息最丰富、最稳定 将选择代谢物或代谢物结合物作为开发免疫分析的靶标。成功将会 包括适用于常规监测的免疫分析和暴露之间清晰的定量关系， 6个受试者的吸收剂量和尿中氯氰菊酯代谢物的测定。 结论：本研究的结果将用于快速、灵敏地确定候选人的发展 以免疫化学为基础的分析方法，可用于常规监测人类接触二氯氰菊酯。这个 仔细监测氯氰菊酯吸收剂量的存在的能力将是一个有用的工具，以防止 二氯氰菊酯暴露对人类健康的影响。

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. Methodology: We will utilize three leading edge analytical technologies in an integrated approach to identify and develop both laboratory-based and field portable methods for the detection and quantitation of human exposure to permethrin. High-pressure liquid chromatography coupled to a state-of-the-art-tandem mass spectrometer (LC/MS) will be used to fractionate and identify urinary metabolites of permethrin. Accelerator mass spectrometry (AMS) is a method for measuring levels of 14C several orders of magnitude more sensitive than liquid scintillation counting. With this ultrasensitivity, doses of 14C-permethrin reflective of actual human exposure will be used to determine the metabolite profile with minimal radiation risk. The metabolites identified by LC/MS and AMS will be used to develop a rapid, sensitive immunoassay for routine monitoring of human exposure to permethrin. Six human volunteers will be exposed to low dermal doses of 14C-permethrin for 8 hours. Urine and blood samples will be collected over a 7-day period with more frequent time points in the first 24 hours. A pilot study utilizing one subject will be conducted first to test the methods developed for separating anticipated metabolites. An exposure study utilizing 5 other subjects will provide confidence that the metabolite patterns observed can be a predictor for the general population. Outcome: The urinary metabolites will be identified and quantified. Based on this information the most abundant, stable metabolite or metabolite conjugate will be chosen for the target of the development of an immunoassay. Success will include an immunoassay suitable for routine monitoring and clear quantitative relation between the exposures, absorbed dose, and measured metabolites of permethrin in the urine among the six subjects. 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.