Lc-ms Technical Analysis Of The Phosphorus Chemistry Of Life, Applied Research,

Life science has been recognized as one of the most frontiers of the 21^st century. With the completion of the mapping of the human genome, life science is progressing into the proteomics age. Similar to the first gene mapping which was completed ahead of schedule using a capillary electrophoresis sequence technique, proteomics research will also develop quickly because of the introduction of the modern bio-mass spectrometry (bio-MS) technique. Together with modern separation techniques, the application scopes of the bio-MS technique will be extended, and high performance liquid chromatography/mass spectrometry (HPLC/MS) coupling technique is becoming a necessary support technique for life science research.In chapter two, in the solution of water, ethanol, carbon tetrachloride and triethylamine, the phosphoryl group was introduced into the peptide segments consisted of basic amino acids using the Atherton-Todd reaction with diethyloxyphosphite (DEPH) as a phosporylation reagent. Phosphorylation modification for amino acids, peptides and proteins were successfully achieved using DEPH. Using electrospray ionization mass spectrometry (ESI/MS), HPLC/MS and other analytical techniques, the phosphorylation products have been characterized. Of the 20 common amino acids, only lysine (Lys) and histidine (His) could be phosphorylated to produce diphosphorylation products. For the 10 peptides and 3 proteins of known sequence, it was found that the modification sites were on residues of Lys and His on the side-chains. According to the incremental value of the molecular weight, the numbers of the residues of Lys and His in peptide and protein could be determined. These findings were useful to peptide sequence determination and protein structure characterization. It also has potential value for research and development of peptide medicine.In chapter three, a study was conducted to establish a HPLC/ESI/MS methodology for simultaneously determining trace monoadenosine and diadenosine monophosphate in a model reaction between N- (O, O-diisopropyl) phosphoryl alanine(Dipp-Ala) and adenosine(A). The possible cleavage pathways were depicted and the specific ions were obtained. An optimal and practical HPLC/ESI/MS method was established for separating and determining the positional isomers of 2'-AMP, 3'-AMP, 5'-AMP and 3'-5' ApA from the complex reaction solution. We first investigated the aqueous reaction between Dipp-aa and the mixed...