Separation of molecular constituents from a humic acid by solid-phase extraction following a transesterification reaction

Abstract

A selective removal of humic constituents involved in ester groups was conducted by a boron trifluoride–methanol transesterification reaction after removal of structurally unbound lipids. An analytical separation of subfractions containing specific classes of compound liberated from the humic matrix simplifies their identification by gas chromatography–mass spectrometry. We compared the traditional liquid–liquid separation into phenolic and aliphatic fractions with the modern and versatile fractionation technique using solid-phase extraction (SPE) on aminopropyl-bonded phases. Our results showed that both methods ensured separation of the same compounds, such as di- and tri-hydroxyalkanoic acids, α-, β- and ω-hydroxy fatty acids, alkanoic acids, α,ω-alkanedioic acids, n-alkanols, phenolic acids and sterols. Moreover, the SPE method not only provided a larger recovery of compounds, but involved smaller sample and solvent requirements, and larger ease and rapidity of sample handling than the traditional liquid–liquid separation. The SPE method should be thus recommended in structural studies of natural organic matter.