Enhanced quantification of chlorophyll a and its degradation products in olive oil using time-resolved laser-induced fluorescence fingerprint analysis.

ABSTRACT

Potential errors in the fluorescence analysis of chlorophylls and their degradation products, primarily due to spectral overlap and inner filter, are widely acknowledged. This study aimed to devise a sensitivity-enhanced technique for the concurrent quantification of chlorophyll a and its degradation products while minimizing effects from type-B chlorophylls. Initially, a time-resolved laser-induced fluorescence spectroscopic system was designed and tested on stardard chlorophyll samples. The origins, implications, and mitigation strategies of spectral overlap and the inner filter effect on the measured fluorescence intensity were thoroughly examined. Then, this methodology was proved to be efficacious within complex liquid matrices derived from olive oil. The experimental outcomes not only shed additional light on the mechanisms of chlorophyll fluorescence overlap and the inner filter effect, but also establish a general framework for developing spectrally and timely resolved fluorescence fingerprint analysis for the simultaneous quantification of chlorophylls and their degradation products at high concentrations.