Preparative free-flow electrophoretic offline ESI-Fourier transform ion cyclotron resonance/MS analysis of Suwannee River fulvic acid

Free-flow electrophoresis (FFE), a preparative free zone electrophoretic method, was used offline in conjunction with ultrahigh-resolution FT/ion cyclotron resonance -MS to resolve the complexity of Suwannee River fulvic acid (SRFA). Before MS, the FFE separation conditions and the compatibility with ESI were optimized. The constituents in SRFA were effectively separated based on their charge states and sizes. The obtained mass spectra were compared by means of van Krevelen diagrams and the calculated aromaticity indices of the individual constituents were used to describe the distribution of aromatic/unsaturated structures across the FFE-fractionated samples. The consolidated number of ions observed within the individual SRFA fractions were much higher than those of the bulk samples alone, demonstrating extensive ion suppression effects in bulk SRFA likely also operating in the analysis of complex biogeochemical mixtures in flow injection mode. The FFE approach allows for producing sizable amounts of sample from dilute solutions, which can be easily fractionated into dozens of individual samples with the possibility of further in-depth characterization.