Prediction of polarity-dependent environmental behaviors of humic substances (HS) using a HS hydrophobicity index based on hydrophilic interaction chromatography

A variety of analytical methods have been applied to describe the properties of aquatic humic substances (HS). However, there are only a few methods available to probe HS hydrophobicity because of the heterogeneous character of HS. In this study, hydrophilic interaction chromatography (HILIC) equipped with a UV detector was employed to describe the heterogeneous distribution of HS with respect to its hydrophobicity and to suggest a representative HS hydrophobicity index. To this end, various mobile phases were explored to achieve the optimal separation capability of HILIC. The highest resolution was obtained with a mobile phase comprising acetonitrile and water at a ratio of 70:30 (v:v). A calibration curve was successfully constructed using eight different HS precursor compounds, which allowed for the successful conversion of the retention time (RT) into the octanol-water partition coefficient (K_ow) (log K_ow = −2.83 × (RT) + 17.44, R² = 0.950). Several possible HS hydrophobicity indices were derived from the HILIC chromatogram. Among those, the weight-average log K_OW value exhibited the strongest negative correlation with the well-known polarity index, (O + N)/C ratios, of seven reference HS samples. This new HILIC-based index (i.e., average log K_OW) also presented a good relationship with the HS binding coefficients with pyrene as well as the extent of HS adsorption onto kaolinite at a given solution chemistry (i.e., at a high ionic strength and a neutral pH), both of which are known to be largely governed by the hydrophobic nature of HS. This study demonstrated that the average K_OW value based on HILIC is an intuitive and robust HS hydrophobicity index to fully represent the heterogeneous distribution of hydrophobicity within a bulk HS and could be applied to predict many environmental behaviors related to HS hydrophobicity or HS polarity.