MICRO-HPLC AND LC/MS METHODS FOR NEURAL SYSTEMS

该研究项目的目标是开发和利用 高灵敏的鉴定和定量方法 磷脂、鞘糖脂和低聚糖的分析 加强研究工作，以了解 神经系统。质谱学可以提供特定的 结构信息和高度特异和灵敏的检测。 这项拟议的研究是为了利用强大的 高质量量程质谱仪的分析能力 结合了微反相色谱柱的高分辨率。这个 该提案的具体目标是： 1)建立液-质联用的条件 未衍生磷脂的LC/MS光谱分析 类别和分子物种。我们将利用内部源代码 移动皮带和仪器能力，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)建立两种物质的LC/MS分析方法 未衍生化和衍生化的鞘糖脂。我们将探索 LC/MS联用分析神经节苷脂的潜力 界面和负离子检测。全甲基化的 衍生品提供了极好的结果与直接的化学 电离和将这种方法扩展到LC/MS将是 承担了。 3)开发未衍生产品的LC/MS分析程序 和衍生的低聚糖。 4)测试电离和LC/MS的替代方法 具有高灵敏定量潜力的接口 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、等离子喷雾和动态 (连续流动)FAB，用于离线和在线LC/MS分析。 拟议工作的总体意义在于使 糖偶联物和磷脂的分析达到 特异度和敏感度将允许化学- 细胞水平上的形态相关性。这样的能力 应该会让我们更深入地理解 正常和病理组织中的糖偶联物和磷脂 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.