Amino Acids, Nucleotides And Related Substances Such As Liquid Chromatography Analysis

In nature, nucleotides have almost participated in every reaction of life chemistry processing. Amino acids are basic units of proteins, which are nearly correlated with life origin. They are materials of synthesizing hormone, enzyme and antibody. Meanwhile, amino acids take part in body's metabolisms, physiological functions. They also play a pivotal part in synthesizing proteins of organism and have been a significant investigation object. Recently, with the development of chromatographic techniques people have been paying more attention to amino acids and nucleotides. In this dissertation, three aspects on the studyof amino acids and nucleotides have been carried out.1. A practical method was set up to analyze N- phosphoryl aromatic amino acids and their corresponding amino acids. The influencing factors on the separation, such as the separation column, detection wavelength, mobile phase medium, elution mode and ionization mode, have been optimized and achieved. With this HPLC/ESI/MS method, the purity and stability of three N-phosphoryl aromatic amino acids were investigated.2. Based on the preceding investigation, a simple, fast, accurate and reproducible method has been developed for separating of twenty N-(O,O-Diisopropyl) phosphoryl amino acids using high-performance liquid chromatography(HPLC). The good relationship between the oil-water partition coefficient (LogP) and capacity factor (Logk') of Dipp-aa on NH₂ bonded phase was observed, which as expected could be applied to predict the LogP values of the amino compounds modified with N-(O,O-diisopropyl) phosphite in the future.3. This work reported the full scan electrospray ionization mass spectrometry (ESI-MS) and ESI-MS/MS for the four nucleotides(2'-AMP, 3'-AMP, 5'-AMP and 3'-5'-ApA) under both positive and negative polarity. Their specific ions were obtained, the optimal chromatographic condition for these relative nucleotides was achieved, and these compounds in the reaction mixture were successfully characterized and measured by HPLC/ESI-MS. This study provided an accurate and reliable method for the reference of the research on the related reaction system.