title: Effects of sample properties and mass spectroscopic parameters on electrospray ionization mass spectra of size-fractions from a soil humic acid.
abstract: Size-fractions from a soil humic acid were separated by preparative size-exclusion chromatography (SEC), desalted, and concentrated by ultrafiltration and vacuum centrifugation without being subjected to any freeze-drying process. After having assessed the lack of formation of any multiple-charged ions by high-resolution Fourier transform ion cyclotron resonance electrospray ionization (ESI) mass spectrometry (MS), the size-fractions were used by direct infusion to compare the molecular ion distribution by both atmospheric pressure chemical ionization (APCI)- and ESI-MS in negative mode. The weight- (Mw) and number-averaged (Mn) molecular weight obtained by ESI-MS were invariably larger than by APCI-MS for all size-fractions, thereby indicating that ESI is more efficient than APCI to evaluate the molecular mass distribution of humic samples. No substantial difference was observed when concentration and pH of unfreeze-dried humic size-fractions were varied. The negative mode was applied to assess the effect of cone voltage from -20 to -60 V on ESI of the humic size-fractions further separated through an on-line SEC column. The resulting mass spectra and Mw and Mn values suggested that the variation of cone voltage in ESI-MS affects the ionization potential of associated humic molecules more in solution rather than their fragmentation. These findings agree with previous observations which indicated a limitation of ESI in providing consistent mass detection for a complex mixture of heterogeneous humic molecules, especially when they are aggregated by a freeze-drying process.
authors.affiliation: Dipartimento di Scienze del Suolo, della Pianta, dell'Ambiente, delle Produzioni Animali, Università di Napoli Federico II, Via Università 100, 80055 Portici, Italy. alpiccol@unina.it
authors.forename: A
authors.initials: A
authors.lastname: Piccolo
authors.forename: M
authors.initials: M
authors.lastname: Spiteller
authors.forename: A
authors.initials: A
authors.lastname: Nebbioso
mesh_terms: D002850:Chromatography, Gel; D005583:Fourier Analysis; D006812:Humic Substances / Q000032:analysis*; D008970:Molecular Weight; D012987:Soil / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D006812:Humic Substances; D012987:Soil