The compost’s humic acid (HA) content decreased when it was fertilized in coastal seawater with steelmaking slag, as confirmed. This study clarified the cause for this change by a detailed analysis of the structural changes in HAs based on the TMAH-py-GC/MS, HPSEC, and 3D-EEM spectra. An increase in the levels of pyrolysates of tannic acid with a low polymerization degree was attributed to the biodegradation of a high polymerized aromatic structure. Moreover, analyses of 3D-EEM, supported by HPSEC, indicated that approximately 20 kDa of the fluorescent matter was generated at the protein-like peaks (Ex/Em = 220/340 and 275/350 nm) in HAs derived from a mixture of compost with steelmaking slag. It would be caused due to the formation of HAs from the bacterial by-product by a catalytic reaction of the steelmaking slag. From these findings reported herein, we conclude that bio-degradation was a major reason for the decreased HA content, and the formation of HA from a part of the degradation products. This would be a reason for the structural modification of HA under the seawater condition.
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