现有的荧光猝灭理论的应用混淆了猝灭剂的平衡浓度[Q]与初始浓度[Q]0之间的关系.基于这一问题,本文探讨了生物大分子与有机物小分子探针之间相互作用的静态荧光猝灭理论,推导了相关的表观结合常数K和结合位点数n的数学表达式,及其相关的定量测定关系式.推导结果表明,荧光体的荧光强度与荧光体和猝灭剂之间的相对浓度有直接的关系,在猝灭剂的初始浓度[Q]0远小于荧光体的初始浓度[P]0时,?F与猝灭剂初始浓度[Q]0在一定浓度范围内呈正比;而在猝灭剂的初始浓度[Q]0远大于荧光体的初始浓度[P]0时,lg1F与lg[Q]0呈正比.本文对上述推导结果进行了实验验证,结果表明,理论推导与实验结果相符,克服了现有的荧光猝灭理论存在的不足,不仅建立了静态荧光猝灭法测定生物大分子的更加科学的方法,而且对研究生物大分子与小分子荧光探针之间的相互作用具有一定的指导作用.The initial concentration and equilibrium concentration are usually confused in the application of existing theories of fluorescent static quenching.So in this article,the theory of fluorescent static quenching by the formation of compound of organic micromolecule with macrobiological molecule was studied.The relationship concerning the apparent formation constant K and the reacting locus number n are deduced and discussed,and the formulas of the quantitative determination are also deduced and proved by experiments.The deduced result as follows,⊿F is proportional to the initial concentration of quencher when the initial concentration of quencher is far less than the initial concentration of fluorescence probe,and lg1F is proportional to lg[Q]_0 when the initial concentration of quencher is far higher than the initial concentration of fluorescence probe.The experiment results proved that the theoretical results deduced in this paper are working.To some extent,these theoretical and experimental results overcome the disadvantages of the existing theoretical and experimental results.So these results not only establish a scientific method of quantitative determination,which can really describe the fluorescent static quenching,but also have instructions for the study on the action between micromolecule fluorescence probes and macrobiological molecules
