ANALYSIS OF ENDORPHINS BY LC-ELECTROCHEMISTRY

定位和量化内啡肽的努力需要技术 是高度敏感和具体的。目前的检测方法包括 生物测定、放射受体、放射免疫分析(RIA)和HIGH 缺少紫外检测的高效液相色谱(LC) 有足够的特异性或足够的敏感性，尤其是对 对较大的内啡肽的研究。LC已被用于分离 生物样本中的内啡肽，但要进行量化 通过放射免疫法对几个层析组份中的每一种内啡肽进行分析。 这个项目的目的是开发一种特定的和 小脑组织中内啡肽的定量测定方法 区域使用带电化学检测的LC(LC-EC)。至 这样就可以提高EC检测的灵敏度 使用包括替代电极在内的各种程序 配置(常规玻璃炭、石墨复合材料和 网状玻璃碳)，电极表面的变化 电极阳极氧化前处理的制备 最后是化学衍生化程序的改进 内啡肽的电活动。 然后将通过以下几种方法对内啡肽的LC-EC分析进行评估 分析技术，包括流体动力伏安法、峰 电流比、放射免疫分析和质谱学确证峰 纯度和巅峰身份。氨基酸序列对蛋白质合成的影响 N-末端酪氨酰的电活性将在#年进行评估 多种二肽和更长的多肽序列可供研究 位阻效应和诱导效应对氧化还原活性的影响。相遇--和 亮氨酸-脑啡肽、强啡肽和-内啡肽将被提取 从特定的大脑区域和获得的量化值 与放射免疫法测定的结果进行比较。 样本。这种LC-EC分析方法特别适用于 需要对内啡肽进行详细分析的研究 内容。

Efforts to localize and quantify endorphins require techniques that are highly sensitive and specific. Current assays including bioassays, radioreceptor, radioimmunoassay (RIA) and high performance liquid chromatography (LC) with UV detection lack either adequate specificity or adequate sensitivity, particularly for the study of the larger endorphins. LC has been used to separate endorphins in biological samples but quantification is performed by RIA for every endorphin in several chromatographic fractions. The purpose of this project is to develop a specific and quantitative method for the assay of endorphins from small brain regions using LC with electrochemical detection (LC-EC). To accomplish this, the sensitivity of EC detection will be improved using a variety of procedures including alternative electrode configurations (conventional glass carbon, graphite composite, and reticulated vitreous carbon), variation in electrode surface preparation, electrode pre-treatment with anodization procedures and lastly chemical derivatization procedures to improve endorphin electroactivity. The LC-EC assay for endorphins will then be evaluated by several analytical techniques including hydrodynamic voltammetry, peak current ratios, RIA and mass spectrometry to corroborate peak purity and peak identity. The effect of amino acid sequence upon the electroactivity of the N-terminal tyrosyl will be evaluated in a variety of dipeptides and longer peptide sequences to examine both steric and inductive effects upon redox activity. Met- and leu-enkephalin, the dynorphins and -endorphins will be extracted from specific brain regions and the quantitative values obtained with the LC-EC assay compared with those obtained by RIA of the samples. This LC-EC assays will be especially useful for those studies which require a detailed analysis of endorphin content.