Tb^(3+)-钛铁试剂络合物荧光猝灭法测定含磷酸基辅酶

Determination of Coenzymes with Phosphates by Fluorescence Quenching of Tb^(3+)-Tiron Complex

含磷酸基辅酶磷酸吡哆醛(PALP)、烟酰胺腺嘌呤二核苷酸(NAD)、烟酰胺腺嘌呤磷酸二核苷酸(NADP)、黄素腺嘌呤二核苷酸(FAD)可显著猝灭铽-钛铁试剂(Tb3+-TR)络合物的荧光,其荧光猝灭值△F=F0-F分别与PALP、NAD、NADP和FAD的浓度呈线性关系。分别对测定条件进行了优化,并研究了干扰物质情况,建立了Tb3+-TR络合物荧光探针灵敏测定PALP、NAD、NADP和FAD的方法。PALP、NAD、NADP和FAD的检出限分别为:1.1×10-7mol/L、6.4×10-8mol/L、1.6×10-8mol/L和1.15×10-8mol/L。

The coenzymes,containing phosphate, usually are essential small biological molecules and play an important role in many biologial processes,such as pyidoxal phosphate （PALP）,nicotinamide adenine dinucleotide （NAD）,nicotinamide-adenine dinucleotide phosphate （NADP）, flavine-adenine dinucleotide （FAD）, and so on. They can prominently quench the fluorescence of the terbium-tiron （Tb^3＋-TR） complex. And the decrease value of the fluorescence,△F = F0- F, shows linear relationships with concentrations of PALP, NAD, NADP and FAD, respectively. By optimizing experiment conditions and researching interference influence, the sensitive determination method of PALP, NAD, NADP and FAD was developed with the Tb^3＋-TR complex as a fluorescent probe. The method detection limits were 1.1×10^-7 mol/L for PALP, 6.4×10^-8 mol/L for NAD, 1.6×10^-8 mol/L for NADP and 1.2×10^-8 mol/L for FAD.