Factors Affecting Quantitative Applications of Fluorimetry

Eq. 6.5 may appear to be suggesting that the concentration determination using fluorescence is quite straight forward. Therefore, the following factors act as problem areas in the course of such quantitative analysis.

Self quenching

You would recall from the previous unit that the fluorinating molecules may lose their energy to other molecules through radiationless energy transfer during the collisions. This leads to the quenching of fluorescence. You will also recall in which the presence of paramagnetic species like oxygen also because quenching and thus proves to be of hindrance in quantitative determination. Therefore, the sample for fluorescence determination needs to be flushed with nitrogen.