MICRO-HPLC & LC/MS METHODS FOR NEURAL SYSTEMS

该研究计划的目标是开发和利用 用于鉴定和定量的高灵敏度方法 磷脂、鞘糖脂和寡糖的分析 加强研究工作，以了解 神经系统 质谱法可以提供特定的 结构信息和高度特异性和灵敏的检测。 这项拟议中的研究是为了利用强大的 高质量范围质谱的分析能力， 结合微型HPLC的高分辨率。 的 该提案的具体目标是： 1)建立液相色谱/质谱条件 未衍生化磷脂的光谱（LC/MS）分析 类和分子种类。 我们会利用内部资源 移动带和仪器的能力，如快速原子 轰击（FAB），高分辨率和链接扫描，以开发 磷脂分子种类分析方法。 2)开发两种药物的LC/MS分析方法 未衍生化和衍生化的鞘糖脂。 我们将探讨 用LC/MS带进行FAB分析神经节苷脂的潜力 界面和负离子检测。 全甲基化 衍生物提供了优异的结果， 电离和这种方法的LC/MS的扩展将是 进行。 3)开发未衍生化的LC/MS分析程序 和衍生的寡糖。 4)测试电离和LC/MS的替代方法 具有高灵敏度定量潜力的接口 以及对感兴趣的化合物进行结构分析。 这些 包括热喷雾电离、等离子体喷雾和动态 （连续流）FAB用于离线和在线LC/MS分析。 拟议工作的总体意义在于， 分析糖缀合物和磷脂的水平， 特异性和敏感性，这将允许化学- 细胞水平上的形态学相关性。 这种能力 应导致更深入地了解的作用， 糖复合物和磷脂在正常和病理 流程.

The objectives of this research program are to develop and utilize highly sensitive methods for the identification and quantitative analysis of phospholipids, glycosphingolipids, and oligosaccharides to enhance research efforts directed toward understanding the nervous system. Mass spectrometry can provide specific structural information and highly specific and sensitive detection. The proposed research is an effort to exploit the powerful analytical capabilities of high mass range Mass Spectrometry in combination with the high resolving power of micro HPLC. The specific aims of the proposal are: 1) to establish conditions for the liquid chromatographic/mass spectrometric (LC/MS) analysis of underivatized phospholipid classes and molecular species. We will utilize the in-source moving belt and instrumentation capabilities, such as fast atom bombardment (FAB), high resolution and linked scans, to develop methods for phospholipid molecular species analysis. 2) To develop methods for the LC/MS analysis of both underivatized and derivatized glycosphingolipids. We will explore the potential of FAB analysis of gangliosides with the LC/MS belt interface and negative ion detection. The permethylated derivatives have provided excellent results with direct chemical ionization and the extension of this approach to LC/MS will be undertaken. 3) To develop procedures for the LC/MS analysis of underivatized and derivatized oligosaccharides. 4) To test alternative methods of ionization and LC/MS interfacing which have potential for highly sensitive quantitation and structural analysis for the compounds of interest. These include thermospray ionization, plasmaspray, and dynamic (continuous flow) FAB for off-line and on-line LC/MS analysis. The overall significance of the proposed work lies in bringing the analysis of glycoconjugates and phospholipids to a level of specificity and sensitivity that will permit chemical- morphological correlations at the cellular level. Such capability should lead to a deeper understanding of the roles of glycoconjugates and phospholipids in normal and pathological processes.