Reverse-phase HPLC method for measuring polarity distributions of natural organic matter

A reverse-phase high-pressure liquid chromatography (RP-HPLC) method was developed to measure the polarity distribution of natural organic matter (NOM) samples. The polarity distribution is obtained by calibrating an octadecyl bonded silica phase column and polar eluent with compounds of known octanol-water partition coefficient (Kow) and using this calibration curve to transform NOM retention times into an equivalent Kow. Polarity distributions treat the NOM samples as a complex mixture rather than summarizing the polarity in a single number. The method is sensitive, with UV detection allowing quantitation of samples with <5 mg of C/L. Individual chromatograms are acquired in <20 min, allowing much faster analysis on smaller samples than XAD resin separation or 13C NMR. Polarity distributions of 10 representative NOM isolates and 2 whole water samples indicate that NOM is generally hydrophilic in nature (log Kow < 2), although XAD-8 isolates are more hydrophobic than RO isolates from the same source. Hydrophilicity, as indicated by recovery from the HPLC column, is correlated to the elemental oxygen/carbon ratio but does not correlate strongly with molecular weight or 13C NMR aromaticity.