Qualitative and Quantitative Analysis of Three-Dimensional Fluorescence Spectra by Improved Parallel Factor Analysis...

Area of Science:

• Analytical Chemistry
• Spectroscopy
• Chemometrics

Background:

• Parallel factor analysis (PARAFAC) with internal standards is used for analyzing complex fluorescence spectra.
• Discrepancies in fluorophore numbers between internal standard and test samples can affect PARAFAC accuracy.
• A need exists for a robust PARAFAC strategy that accommodates varying fluorophore compositions.

Purpose of the Study:

• To establish and validate the internal standard sample embedding parallel factor analysis (ISSE-PARAFAC) framework.
• To investigate the impact of fluorophore number disparity on PARAFAC qualitative and quantitative outcomes.
• To enhance PARAFAC by aggregating spectrally similar fluorophores to reduce correlation.

Main Methods:

• Systematic establishment of the ISSE-PARAFAC strategy.
• Application of ISSE-PARAFAC to six datasets (two scenarios, one real-world) with varying fluorophore compositions.
• Incorporation of fluorophore aggregation based on emission wavelength peaks to mitigate spectral overlap.

Main Results:

• The ISSE-PARAFAC framework effectively handles disparities in fluorophore numbers between internal standard and test samples.
• Irrelevant fluorophores in the internal standard set do not significantly impact the analysis of target fluorophores.
• The enhanced ISSE-PARAFAC enables accurate qualitative decomposition and quantitative analysis of mixed fluorophores down to ppm levels.

Conclusions:

• ISSE-PARAFAC provides a robust and reliable method for the qualitative and quantitative analysis of three-dimensional fluorescence spectra.
• The method demonstrates high accuracy and precision, even when dealing with complex mixtures and varying sample compositions.
• This approach offers significant improvements for analyzing unknown mixed fluorophores in various scientific applications.