MICRO-HPLC AND LC/MS METHODS FOR NEURAL SYSTEMS

神经学研究最终依赖于对 蜂窝组件。这项研究计划的目标是开发 将在线色谱-质谱学方法与 糖偶联物和脂类的最高灵敏度，以便 有助于了解神经系统。我们假定 细胞表面糖偶联物之间发育调节的相互作用 而互补的细胞表面或细胞外配体参与 特定形态发生事件的调节和膜 环境或细胞膜的脂质成分会影响这些 互动。生物系统，适用于神经研究 该机构正在调查的差异化包括： 发育中的小鼠新皮质的糖偶联物和细胞类别， 髓鞘磷脂合成的调节，发育研究 周围神经与组织培养细胞糖脂代谢的关系 神经起源。这些系统的性质决定了 追求分析技术，以获得定量的化学物质 信息的规模足够小，足以让有意义的 化学-形态关联。质谱学可以提供 特定的结构信息和/或高灵敏度测量。在……里面 追求这些化学极限，以下是糖脂的方法， 磷脂和糖蛋白高甘露糖低聚糖分析将 被调查：(1)建立液体的条件 色质联用(LC/MS)分析未烷化产品 磷脂的种类和分子种类，(2)发展方法 未烷化和/或全甲基化鞘糖脂的LC/MS分析 过苯甲酰基衍生物的分析条件，(3) 设计用于未烷化、全甲基化的 和N-三氟乙酰甲基化高甘露糖低聚糖和(4) 研究用于质谱学的电离的替代方法 可能对分析磷脂有用的分析， 糖脂和低聚糖。

Neurological research depends ultimately on the accurate analysis of cellular components. The objective of this research program is to develop combined on line chromatographic-mass spectrometric procedures with the highest achievable sensitivity for glycoconjugates and lipids in order to contribute to the understanding of the nervous system. We postulate that developmentally regulated interactions between cell-surface glycoconjugates and complementary cell-surface or extracellular ligands participate in the regulation of specific morphogenetic events and that the membrane environment or lipid composition of cellular membranes influences these interactions. Biological systems, suitable for studies of neural differentiation, under investigation in this institution include: glycoconjugates and cell classes of the developing mouse neocortex, regulation of myelin phospholipid synthesis, developmental studies of peripheral nerve, and glycolipid metabolism in tissue culture cells of neural origin. The nature of these systems dictate that the limits of analytical technology be pursued in order to obtain quantitative chemical information on a scale sufficiently small to allow meaningful chemical-morphological correlations. Mass spectrometry can provide specific structural information and/or high sensitivity measurements. In pursuit of these chemical limits, the following approaches for glycolipid, phospholipid and glycoprotein high mannose oligosaccharides analysis will be investigated: (1) Establish conditions for the liquid chromatographic-mass spectrometric (LC/MS) analysis of underivatized phospholipid classes and molecular species, (2) Develop methods for the LC/MS analysis of underivatized and/or permethylated glycosphingolipids and also conditions for the analysis of the perbenzoylated derivatives, (3) Devise procedures for the LC/MS analysis of underivatized, permethylated and N-trifluoroacetyl-methylated high mannose oligosaccharides and (4) Investigate alternative methods of ionization for mass spectrometric analysis which are potentially useful for analysis of phospholipids, glycolipids and oligosaccharides.