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 尿液会被显影 技术，相位分辨 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游离抗原。 相反，游离的标记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 在两种不同的微环境中的荧光标记。 相位分辨荧光光谱（PRFS），其中相位敏感 检测用于选择性地测量物质的荧光 用正弦调制光激发，将用于同时 测定样品中游离和结合标记抗原的效价， 标准溶液。 血清中胆红素荧光引起的干扰 可以通过将胆红素作为单独的组分处理来消除， 解决方案和解决其贡献沿着与那些自由和 结合标记抗原。 这消除了减去a的需要。 血清空白，甚至黄疸血清。 PRFIA技术在化学分析中的应用 还将研究尿液和全血，处理其他基质 以类似的方式干扰。 PRFIA技术将被优化用于测定苯巴比妥。 其他滥用药物，包括四氢大麻酚， 苯环己哌啶（PCP），海洛因，可卡因，阿片类药物和其他药物，然后将被 充分发展。 该项目的初步研究目前由NIH ADAMHA资助 小额赠款方案。 这些研究的初步结果将 描述，包括使用PRFS消除胆红素干扰， 荧光素的测量和苯巴比妥的PRFIA的初步结果。 与其他均质相比，建议的PRFIA的优点 免疫测定技术包括程序简单、最少试剂 要求，以及一个多成分的方法，使消除 来自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.