โ Scribed by H. Irth; M. Lamoree; G.J. de Jong; U.A.Th. Brinkman; R.W. Frei; R.A. Kornfeldt; L. Persson
No coin nor oath required. For personal study only.
A detection system for D-myo-1,2,6-inositol trisphosphate (1 ,2,6-IP3) has been developed which is based on the strong complexing properties of the phosphate groups and which includes a post-column ligand-exchange reaction. The weakly fluorescent complex of Fe"' with methylcalcein blue (MCB) is used as a reagent. Analytes that form a stronger complex with Fe"' than MCB cause a ligand-exchange reaction; the amount of strongly fluorescent MCB released during this reaction is proportional to the analyte concentration and can be measured sensitively by direct fluorescence detection. The influence of the pH, the reagent concentration, the Fe"' to MCB ratio, the content of organic modifier and the dimensions of the post-column reaction system on the sensitivity of the detection system are discussed. In combination with ion-pair reversed-phase liquid chromatography, a detection limit of 3310 ng has been obtained for 1,2,6-IP3. The present system has been combined with a preconcentration technique, based on ion-pair formation, which allows the determination of lowppb concentrations of 1 ,2,6-IPj.
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