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

人类接触杀虫剂氯菊酯的生物标志物的识别和开发

• 批准号: 8362754
• 负责人: BRUCE D HAMMOCK
• 金额: $ 16.38万
• 依托单位: UNIVERSITY OF CALIF-LAWRNC LVRMR NAT LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362754
• 关键词: Acids; Agricultural Workers; Alcohols; Antibodies; Biological Assay; Biological Markers; Biological Markers; Blood; C14 isotope; Chemicals; Clinical; Collection; Data; Dermal; Detection; Development; Dose; Exposure to; Funding; Glucuronides; Glycine; Grant; Half-Life; Haptens; Health; High Pressure Liquid Chromatography; Hour; Human; Immunoassay; Insect Repellents; Insecticides; Knowledge; Lice; Liquid Chromatography; Liquid substance; Manuscripts; Mass Spectrum Analysis; Measures; Methods; Military Personnel; Monitor; National Center for Research Resources; Pattern; Permethrin; Pesticides; Preparation; Principal Investigator; Publications; Radiolabeled; Research; Research Infrastructure; Resource Development; Resources; Review Literature; Route; Saliva; Sampling; Scintillation Counting; Soaps; Source; System; United States National Institutes of Health; Urine; Validation; accelerator mass spectrometry; analytical method; base; cost; exposed human population; liquid chromatography mass spectrometry; metabolic abnormality assessment; prevent; pyrethroid; radiotracer; tool; ultraviolet; urinary

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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 STATUS/RESULTS TO DATE: Literature review has been completed and putative major metabolites identified. All of the major metabolites have been synthesized or acquired. Using the metabolite standards a high performance liquid chromatography method for their analysis has been developed. This method will later be used to identify metabolites found in human urine samples and the liquid chromatography-mass spectrophotometric method used for validation. Using the chemical knowledge from the preparation of metabolites, synthesis of haptens for immunoassay detection of these molecules is complete. Immunoassays for 3-phenoxybenzoic acid, the glycine conjugate of 3-phenoxybenzoic acid, the glycine conjugate of dichlorovinylchrysanthemic acid (DCCA) and the glucuronide conjugate of 3-phenoxybenzyl alcohol (see publication below) have been developed. An assay for free DCCA is in progress (manuscript in preparation). The assay for 3-phenoxybenzoic acid has been adapted to a sensitive, high throughput method (see publication below). Clinical exposures have been completed. All samples have been measured by AMS for total carbon-14. An estimate of the total dose absorbed (for 4 subjects) ranged from 0.06 to 0.27%. The permethrin is eliminated from the blood with a half life of about 12-24 hours hours. The urinary half life averaged 24 hours. Saliva was sampled, but permethrin does not appear to be excreted by that route. Liquid chromatography analysis of the metabolite pattern in urine is underway. 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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 农业工作者，园丁和房主通常会暴露于杀虫剂氯仿。 此外，使用DOD驱虫系统和非处方虱子肥皂使用氯菊酯作为活性成分时，军事人员会暴露于苄氯菊酯。苄氯菊酯的尿代谢产物很丰富并且相对稳定，可能是暴露于这种农药的理想生物标志物。 另外，根据给药途径，一种代谢物与另一种代谢物的比率可能有所不同。这项研究的结果将用于确定基于快速，敏感的免疫化学分析方法开发的候选者，该方法可用于常规监测人类对苄氯菊酯的接触。 目的：本研究的目的是确定皮肤暴露后尿液中氯菊酯的人类代谢产物，其丰富性最大，并且是最稳定的。加速器质谱法是一种测量比液体闪烁计数更敏感的14C水平的方法。使用这种高灵敏度，我们将在生物学相关剂量上进行人类代谢研究。 具体目的：I。开发出氯菊酯分离及其推定的人类代谢产物的LC/MS方法。 ii。 使用加速器质谱法（AMS）确定氯仿的人代谢物谱。 iii。 对目标II中鉴定为人类对苄氯菊酯的生物标志物的关键代谢产物开发免疫测定。 方法论：对于特定目的I，合成代谢物标准；开发一种HPLC方法，使用紫外线检测分离推定的拟除虫菊酯代谢产物；确定HPLC/质谱法用于分析拟除虫菊酯代谢物的可行性。 对于特定的目标II，人类临床暴露于放射性标记的苄氯菊酯真皮和尿液，血液和唾液的收集；通过在特定目的I中开发的方法和通过加速器质谱分离样品分析样品的分离；从结果数据中鉴定出最普遍的代谢产物。对于特定的目的III，合成触觉；抗体的开发；在为最普遍的代谢物的免疫测定开发中使用触觉和抗体；验证免疫测定。 预期产品（里程碑）：氯菊酯推定的人类代谢产物的文献综述；少量的苄氯菊酯合成代谢产物；一种用于分离人类尿液或唾液中苄氯菊酯代谢物的HPLC方法；通过加速器质谱法鉴定最丰富的人类代谢产物；一种免疫测定方法，可检测到氯菊酯的靶向人代谢产物 迄今为止的状态/结果：文献综述已经完成，并确定了主要的主要代谢物。 所有主要的代谢物均已合成或获得。 使用代谢物标准，已经开发了一种高性能液相色谱法进行分析。 该方法后来将用于鉴定人类尿液样品中发现的代谢产物以及用于验证的液相色谱 - 质量分光光度计方法。利用从代谢产物制备的化学知识，完成了对这些分子的免疫测定检测的合成。 三氧基苯甲酸的免疫测定，3-苯氧基苯甲酸的甘氨酸结合物，二氯丙烯基丙烯酸的甘氨酸偶联（DCCA）和3-磷酸苯甲酰醇的葡萄糖醛酸（DCCA）的出版物（见下文）。 正在进行免费DCCA的测定法（准备中的手稿）。 3-苯氧苯甲酸的测定已适用于一种敏感的高通量方法（请参见下面的出版物）。临床暴露已经完成。所有样品均已通过AMS测量总碳14。吸收的总剂量（4个受试者）的估计值范围为0.06至0.27％。苄氯菊酯以大约12-24小时的半衰期从血液中消除。尿半寿命平均24小时。 对唾液进行了采样，但苄氯菊酯似乎并未被该路线排出。尿液中代谢物模式的液相色谱分析正在进行中。 结论：这项研究的结果将用于确定候选者的发展，以开发快速，敏感的基于免疫化学的分析方法，该方法可用于常规监测人类对苄苄苄氯菊酯的暴露。仔细监测吸收剂量的苄氯菊酯的能力将是一种有用的工具，可以防止因苄氯菊酯暴露而引起的人类健康影响。