Limitations of Electrospray Ionization of Fulvic and Humic Acids as Visible from Size Exclusion Chromatography with Organic Carbon and Mass Spectrometric Detection

A method was developed to link size exclusion chromatography electrospray ionization mass spectrometry (SEC-ESI-MS) analyses of fulvic and humic acids with SEC and organic carbon detection (SEC-OCD), the latter providing an absolute measure of the amount of organic matter eluting from the SEC column. This approach allows us to determine which molecular weight fraction of the complex polydisperse mixtures is detectable by ESI-MS. It could be shown that the cone voltage setting for the ESI interface has strong impact on ESI-MS detection. Using conventional settings for low molecular weight compounds, the high molecular weight (HMW) compounds are hardly amenable to ESI-MS. With increasing cone voltage, an increasing signal intensity is obtained for the HMW fraction that elutes at shorter retention times. However, mostly fragment ions are obtained under these conditions. Thus, the range of compounds amenable to ESI-MS analysis is restricted by the limited stability of the fulvic and humic acid molecules of higher molecular weight in the electrospray process rather than by the mass spectrometer used. Compounds above 1000 amu are hardly visible as intact ions. However, insight into structural characteristics of these compounds can be gained by investigating their fragment ions by SEC-ESI-MS. The use of SEC-OCD parallel to SEC-MS helps to assess and optimize the detection potential of ESI-MS for polydisperse mixtures.