Salt-assisted liquid-liquid extraction and on-column concentration for chromatographic determination of phenolic compounds in beer

Abstract

Salt-assisted liquid-liquid extraction (SALLE) and on-column concentration were evaluated to simplify the chromatographic determination of phenolic compounds in beer. Resveratrol, flavonoids (quercetin and kaempferol), and phenolic acids (caffeic, p-coumaric, and trans-ferulic acids) were used as model compounds based on their concentrations reported in craft beers. SALLE with acetonitrile was effective for sample cleanup, with recoveries ranging from 84% to 112%. The on-column concentration was carried out in high performance liquid chromatography by starting the gradient elution with acidified water (sample carrier stream) in a reversed-phase mode. Consequently, the analytes were retained in a narrow band, and highly polar/ionic potentially interfering species were flushed from the column. The effectiveness of the approach was demonstrated by linear correlations between the peak areas and the injected volumes from 10 to 200 µL (r ≥ 0.999), without significant variations in peak width (< 10%) or retention times (< 0.7%), and peak symmetry (tailing factors within the range of 0.8–1.1). For all analytes, a linear response was achieved from 100 to 1000 µg L⁻¹ (r > 0.999), with limits of detection within the range of 9–82 µg L⁻¹. The approach was also effective for on-column concentrations of resveratrol, quercetin, and kaempferol in sequential injection chromatography (SIC), with injected volumes up to 750 µL and linear responses within 100–1000 µg L⁻¹ (r > 0.997). The proposal reduced consumption of organic solvents (with only 770 µL required per determination in SIC) and avoided solid-phase extraction cartridges for analyte preconcentration.