Separation And Determination Of ATP And Phosphate Based On Uranyl Complexes

In this paper, we mainly studied several new methods for the separation anddetermination of ATP and phosphate based on uranyl complexes. In the first part, theproperties and applications of ATP and phosphate are outlined. The researchdevelopments in the separation and determination of ATP and phosphate are reviewed.The structure characteristics and applications of uranium complexes are alsodescribed.In the second part, we report a double-receptor sandwich supramolecule sensingmethod for the determination of ATP. One receptor is a uranyl salophen complexwhich can bind the triphosphate group in ATP selectively, and another is ananti-adenosine aptamer which is a single-stranded oligonucleotide and can recognizethe adenosine group in ATP specifically. The uranyl salophen complex wasimmobilized on the surface of amino-silica gel particles and used as the solid phasereceptor of ATP. The anti-adenosine aptamer was labeled with a fluorescent group andused as the labeled receptor of ATP. In the procedure of ATP detection, ATP was firstcombined with the solid phase receptor and then conjugated with the labeled receptorto form a sandwich-type supramolecule. The conditions of fabricating solid phasereceptor and the influence of manifold variables on the determination were studied.The experimental results demonstrate that the method has a number of advantagessuch as high selectivity, high sensitivity, good stability and low cost. Under optimalconditions, the linear range for detection of ATP is0.2-5.0nmol/mL with a detectionlimit of0.037nmol/mL. The proposed method was successfully applied for thedetermination of ATP in real samples with the recoveries of96.8-103.3%.In the third part, we report a simple spectrophotometric method for the separationdetermination of phosphoate based on uranyl salophen complex ionophore polymer.The uranyl salophen complex was mixed with methacrylic acid (monomer), ethyleneglycol dimethacrylate (crosslinking monomer) and2,2′-azobisisobutyronitrile (initiator) by thermal polymerization. Phosphate anions were first combined with theuranyl salophen ionophore polymer particles and then were eluted with0.5mol/Lsulfuric acid. The phosphate anion content of eluent was determinedspectrophotometrically using molybdenum-blue method. We have investigated theinfluence of varying concentrations of sulfuric acid, ammonium molybdate andstannous chloride on the formation of phosphomolybdenum-blue and the colourstability. The experimental results demonstrate that this method has many advantagessuch as high selectivity, high sensitivity, good stability and low cost. Under optimalconditions, the linear range for detection of phosphate is1.0×10-6-3.6×10-5mol/L.The retention capacity of the uranyl salophen ionophore polymer particles was foundto be3.93×10-6mol of phosphate anion per gram of polymer. The proposed strategywas successfully applied to the determination of phosphate in real samples.