A Combined Approach Employing Soxhlet Extraction and Linear Gradient Elution Reversed-Phase HPLC for the Fingerprinting of Soil Organic Matter According to Hydrophobicity

Abstract

This paper reports the results of a study carried out to evaluate the potentiality of combining selective solvent extraction and linear gradient elution reversed-phase high performance liquid chromatography (RP-HPLC) for characterizing soil organic matter on the basis of polarity of its constituting organic compounds. Such approach comprises the sequential extraction of soil organic matter with organic solvents of increasing polarity in a Soxhlet extractor and the subsequent separation of each extract by RP-HPLC. Accordingly, each soil sample has been subjected to cycles of sequential extraction with n-hexane, dichloromethane, ethyl acetate, and methanol. Each sample extracted by one of the four solvents have been dried in a rotary evaporator device at 40°C and then dissolved again in the proper volume of extraction solvent to obtaining sample solutions of concentration ranging from 5.0 to 15 mg ml⁻¹. These sample solutions have been subjected to RP-HPLC separation using a Supelcosil LC-ABZ column that has been eluted by a linear acetonitrile gradient in water, having the same profile for all samples. The study has evidenced the possibility of producing highly repeatable chromatographic profiles, which are correlated to the polarity of organic solvents employed for extracting the organic matter in the Soxhlet extractor.