A stepwise stoichiometric representation to confirm the dependence of pesticide/humic acid interactions on salt concentration and to test the performance of a silica bonded humic acid column.

In a previous paper (André et al., in press), a novel chromatographic column was developed in our laboratory for studying the binding of pesticides with humic acid (HA), the main organic component in soil. It was demonstrated that this column supported a low fraction of organic modifier in the aqueous mobile phase (<0.25 (v/v)). To overcome this limitation for a practical use, a column in which the stationary phase was based on silica gel with chemically bonded humic acid was created. It was shown that this novel HA column supported a higher methanol fraction (<0.55 (v/v)). As well, the dependence of pesticide/humic acid interactions on salt (sodium chloride) concentration has been expressed in terms of a stepwise stoichiometric representation, which leads to a specific equation for the partition of the added salt between the pesticide molecule, the HA, and the pesticide/HA complex. Based on this novel equation, the dependence of the pesticide/humic acid association on the salt concentration can be formulated via a relation similar to the one of Tanford. In addition, for the first time, the calculation of the affinity energy distribution for different values of the salt concentration in the mobile phase confirmed the existence of several types of binding sites on the HA macromolecule.