Correction of inner filter effect in mirror coating cells for trace level fluorescence measurements.

ABSTRACT

The sensitivity of the spectrofluorometric technique can be improved by a factor of about 3.6 using a mirror coating cell. In the case of a large working range, the nonlinear relationship due to the absorbance of solutions between concentration of the analyte of interest and fluorescence intensity (called inner filter effect) must be corrected. This paper suggests a universal inner filter correction equation based on the physical absorbance phenomenon of a mirror coating cell that only depends on the solution absorbance spectra and the cell parameters. These parameters are determined with rhodamine b standard solutions and a simplex method-based mathematical fitting. The methodology has been successfully applied to the correction of classical and synchronous spectra in absorbent media. The partial least squares (PLS) quantification of a mixture of trace levels (approximately 1-10 microg L(-1)) of six polycyclic aromatic hydrocarbons (PAHs) by synchronous fluorescence is possible, even in an absorbent matrix. This simple method allows extension of the analytical field of fluorescence quantification to a large working range in absorbent solutions.