Research On Synthesis Of Tb(Ⅲ) Chelate For New Solid-phase Time-resolved Fluoroimmunoassay

Time-resolved fluoroimmunoassay is a new immunoassay technology established in 1980s.It uses lanthanide chelates with a long fluorescence lifetime,big Stokes shift,and the characteristic sharp peaks as the fluorescence probes.It eliminates the interference of background fluorescence effectively through delaying the measurement time.Therefore TRFIA is more sensitive than other immunoassay methods.The synthesis of bifunctional chelate is the key of time-resolved fluorescence immunoassay.The synthesis of heterocycle molecule,groups of protein connection and rare earth chelate are the key to realize the bifunctionalization.In this dissertation,2,6-bis(3'-ethyl-1'-pyrazolyl)-4-aminopyridine and several important intermediates such as 3-methylpyrazole,3-pyrazole acid,3-ethyl pyrazole were prepared and characterized with the aim to meet the need of bifuncional chelatores for direct solid lanthanide time-resolved fluorescence immunoassay(DSLTRFIA). 2,6-bis(3'-ethyl-1'-pyrazolyl)-4-aminopyridine was prepared from 2,6-dibromopyridine by oxidation,nitration,reduction,nucleophilic substitution reactions,etc.During the experiment,the products were characterized by DSC,IR,~1H-NMR and elemental analysi??sides,the reaction conditions and mechanism of several steps were also studied.The nitrogen and oxygen atoms in the molecule of 2,6-bis (3'-ethyl-1'-pyrazolyl)-4-aminopyridine could form stable complex with lanthanide ions. The 4-amino group in this molecule could be substituted by other functional groups to attach to protein.