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。此外，军事人员在使用国防部驱蚊系统和非处方药虱子肥皂时会接触到氯菊酯，而非处方药中使用的是氯菊酯作为活性成分。氯菊酯的尿代谢物丰度高且相对稳定，可能是接触这种杀虫剂的理想生物标志物。此外，根据给药途径的不同，一种代谢物与另一种代谢物的比例可能会有所不同。这项研究的结果将被用来确定开发一种快速、灵敏的基于免疫化学的分析方法的候选对象，该方法可用于常规监测人类接触二氯氰菊酯。 目的：确定人体皮肤暴露后尿中含量最高、最稳定的二氯氰菊酯代谢物S。加速器质谱仪是一种测量14C水平的方法，比液体闪烁计数灵敏几个数量级。凭借这种高灵敏度，我们将在生物相关剂量下进行人体新陈代谢研究。 具体目的：1.建立氯氰菊酯及其可能的人体代谢物的LC/MS分离方法。使用加速器质谱仪(AMS)测定二氯氰菊酯的人体代谢物分布。建立目标II中确定的关键代谢物的免疫分析方法，作为人类接触氯氰菊酯的生物标记物。 方法：针对特定目标I，合成拟除虫菊酯类代谢物标准品；建立拟拟除虫菊酯类代谢物的紫外检测高效液相色谱分离方法；确定拟除虫菊酯类代谢物的高效液相色谱/质谱联用分析方法的可行性。针对特定目标二，人体经皮下暴露于放射性标记的氯菊酯，并采集尿液、血液和唾液；采用针对特定目标I开发的方法分离样品，并用加速器质谱仪分析分离样品；从所产生的数据中确定最普遍的代谢物。针对特定目的III，半抗原的合成；抗体的开发；半抗原和抗体在开发最普遍代谢物的免疫分析中的使用；免疫分析的验证。 预期产品(里程碑)：氯菊酯可能的人体代谢物的文献综述；少量合成的氯菊酯代谢物；高效液相色谱法分离人体尿液或唾液中的氯菊酯代谢物；加速器质谱仪鉴定最丰富的人体代谢物(S)；检测目标人体代谢物的免疫测定法 结论：这项研究的结果将被用来确定开发一种快速、灵敏的基于免疫化学的分析方法的候选对象，该方法可用于常规监测人类接触氯氰菊酯。能够仔细监测二氯氰菊酯吸收剂量的存在，将是防止因接触二氯氰菊酯而可能对人类健康造成影响的有用工具。

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.