High-performance liquid chromatography (HPLC)-size exclusion chromatography (SEC) for qualitative detection of humic substances and dissolved organic matter in mineral soils and peats in Lithuania

Abstract

High-performance liquid chromatography (HPLC) with size exclusion (SEC) separation function was used to isolate and examine the molecular mass (MM) distributions and polydispersity of humic substances (HSs) and dissolved organic matter (DOM) from mineral soils and peats. The aim was to improve their detailed characterisation and to inform of their soil carbon (C) sequestration and environmental quality. This is the first study conducted in Lithuania in which HSs and DOM, separated from two soil types, have been used to characterise soil at the molecular level. The HPLC-SEC, as a separation method, was coupled with diode-array detection (DAD), thus enabling the separation of molecular fractions. Results showed that HPLC-SEC can be used to determine the MM of HSs in soil, provided that the relation between retention time and MM is known and a suitable method for fitting the HS peak is available. The UV-spectra analysis showed that DOM has a larger MM (Mw = 2439–3436 Da), which contains more aliphatic C. The HS fraction has a smaller MM (Mw = 2776 Da), with aromatic structures that reflect a higher aromaticity. Separated fractions had characteristic MMs of humic acid (HA) and fulvic acid (FA) and DOM. The HSs separated from peat samples were characterised by higher aromaticity, humification and stability. The HSs extracted from mineral soil samples showed a higher degradability level. The results also show the MM distribution and polydispersity of HS and DOM fractions (Mw/Mn = 1.009–1.252) are relatively homogenous in both soil types. Findings confirm that chromatographic and spectrometric parameters can be used for characterisation of both HSs and DOM, and for detecting changes in organic matter quality. Moreover, they can also be used for a further understanding the C-cycle and could be applied for enhancing soil C-sequestration and informing environmental quality management.