Syntheses Of Chelate Intermediates For Time-resolved Fluoroimmunoassay

Homogeneous time-resolved fluorescence immunoassay（HTRF）is an immunoassay technique that combines fluorescence resonance energy transfer with time-resolved fluoroimmunoassay.The most critical technique is the synthesis of chelators.Employing the Homogeneous time-resolved fluorescence immunoassay（HTRF）a new chelating agent was synthesized.For the synthesis of new chelating agent we used 2-chloro-6-methyl isonicotinic acid and p-toluenesulfonamide and other compounds as raw materials.The reaction steps involvedetherification,asymmetric coupling,oxidation,substitution,bromination,and reduction.Employing these reactions we obtained seven kinds of non-commercials chelate intermediates.The structure confirmation and purification was assessed by IR,MS,and GC-MS.A novel rare earth fluorescent chelate-Eu³⁺6,6’-{[4,13–diaza-18-crown-6-ether] bis（aminomethyl）}-2,2’-bipyridine-4-carboxylate was synthesized.The fluorometric studies revealed the fluorescence signal of the chelate in the range of 200-380 nm.Thephoto physicalproperties observed were363 nm maximum excitation wavelength,621 nm（5D0-7F2）,and 583 nm（5D0-7F0）,were main emissionwavelengths,258 nm stokes shift,and 1456 μs fluorescence lifetime as measured by time resolvedfluorescence spectra.The chelate was found to have strong fluorescence,soluble in water,methanol and DMF.The solubility and luminescence intensity was found better in DMF relative to methanol.Hence,our chelate is aluminescent material that provides important reference for the synthesis of HTRF chelating agent.