FLUOROIMMUNOASSAY OF ABUSED DRUGS IN BIOLOGICAL FLUIDS

一种高选择性、高灵敏度的血清药物分析技术 尿液也会被开发出来。相分辨技术 荧光免疫分析(PRFIA)，是一种均相免疫分析技术 不需要从抗体结合的标记抗原中分离出游离的。 相反，自由标记的抗原与抗体结合的抗原是不同的 基于荧光寿命差异的标记抗原 在两个不同的微环境中的荧光标记。 相分辨荧光光谱(PRFS)，其中相敏 检测用于选择性地测量物种的荧光 用正弦调制光激发，将用于同时 测定样品中游离态和结合态抗原百分比 标准解决方案。血清中胆红素荧光的干扰 可通过将胆红素作为 解决方案和解决它的贡献，以及自由和 结合的标记抗原。这样就不需要减去 血清空白，即使是黄疸血清也是如此。高效毛细管电泳法在农药残留分析中的应用 尿液和全血也将被检查，治疗其他基质 以类似的方式进行干扰。 将优化PRFIA技术用于苯巴比妥的测定。 其他滥用药物的方法，包括四氢大麻酚， 苯环利定(PCP)、海洛因、可卡因、鸦片类药物和其他 完全发育成熟。 拟议项目的初步研究目前由NIH ADAMHA资助 小额助学金计划。这些研究的初步结果将是 描述，包括使用PRFS消除胆红素干扰 荧光素的测定，以及苯巴比妥的PRFIA初步结果。 建议的PRFIA与其他同类产品相比的优势 免疫分析技术包括程序简单、试剂最少 需求，以及多组件方法，从而消除 样品基质荧光团的干扰。

A technique for highly selective and sensitive analysis of drugs in serum and urine will be developed. The technique, Phase-Resolved Fluoroimmunoassay (PRFIA), is a homogeneous immunoassay technique which requires no separation of free from antibody-bound labelled antigen. Instead, the free labelled antigen is distinguished from the antibody-bound labelled antigen on the basis of the difference in fluorescence lifetimes of the fluorescent label in the two different microenvironments. Phase-resolved fluorescence spectroscopy (PRFS), in which phase-sensitive detection is used to selectively measure the fluorescence of species excited with sinusoidally-modulated light, will be used to simultaneously determine the percents of free and bound labelled antigen in sample and standard solutions. Interference due to bilirubin fluorescence in serum can be eliminated by treating the bilirubin as a separate component in the solution and resolving its contribution along with those of the free and bound labelled antigen. This eliminates the need for subtraction of a serum blank, even for icteric sera. Application of PRFIA to analysis of urine and of whole blood will also be investigated, treating other matrix interferents in a similar manner. The PRFIA technique will be optimized for determination of phenobarbital. Methods for other abused drugs, including tetrahydrocannabinol, phencyclidine (PCP), heroin, cocaine, opiates and others, will then be fully developed. Initial studies for the proposed project are currently funded by NIH ADAMHA Small Grants Program. Preliminary results of these studies will be described, including the use of PRFS to eliminate bilirubin interference in measurement of fluorescein, and initial results for PRFIA of phenobarbital. Advantages of the proposed PRFIA in comparison with other homogeneous immunoassay techniques include procedural simplicity, minimal reagent requirements, and a multicomponent approach which enables elimination of interference from sample matrix fluorophores.