Non-aqueous size-exclusion chromatography coupled on-line to reversed-phase high-performance liquid chromatography

An interface has been developed which permits the on-line coupling of size-exclusion chromatography in tetrahydrofuran with aqueous reversed-phase high-performance liquid chromatography. The interface isolates the required size exclusion chromatography fraction and dilutes it with water to ensure reconcentration of analytes on the reversed-phase column prior to gradient elution. Operational parameters and the influence of analyte polarity have been examined in detail. A predictive system is presented for determining the applicability of the system to any analyte, based on solute retention times on an ODS phase eluted with a methanol—water gradient. The method is illustrated with examples of direct analyses of crude lipid extracts from a snack product for 2,6-di-tert.-4-methylphenol and from chocolate for dibutyl phthalate. Detection limits of ca.0.5 mg/kg have been achieved.