Integrating Biolayer Interferometry, Atomic Force Microscopy, and Density Functional Theory Calculation Studies on the Affinity between Humic Acid Fractions and Graphene Oxide.

The interactions between nanoparticles and humic acid (HA) are critical to understanding the environmental risks and applications of nanoparticles. However, the interactions between HA fractions and graphene oxide (GO, a popular carbon nanosheet) at the molecular level remain largely unclear. Four HA fractions with molecular weights ranging from 4.6 to 23.8 kDa were separated, and the large HA fractions presented low oxygen contents and many aromatic structures. The binding constants of the large HA fractions on GO were 2.6- to 3551-fold higher than those of the small HA fractions, while the maximum adsorption capacities of the larger HA fractions onto GO were much higher. Atomic force microscopy (AFM) found that the small and large HA fractions were spread over the center and the edge of the GO nanosheets, respectively. Density functional theory (DFT) simulation and nuclear magnetic resonance spectroscopy confirmed the above phenomena (three adsorption patterns, "vs", "ps", and "pea") and revealed that HA bonded to the GO nanosheets mainly through van der Waals force and π-π interactions. The integrating analysis of binding affinity, AFM, and DFT provides new insights into the environmental behavior of GO and the applications of GO in pollutant removal under exposure from HA.