Hyphenation of Supercritical Fluid Chromatography and NMR with In-Line Sample Concentration

[Image: see text] By coupling supercritical fluid chromatography (SFC) and nuclear magnetic resonance (NMR) in-line, a powerful analytical method arises that enables chemically specific analysis of a broad range of complex mixtures. However, during chromatography, the compounds are diluted in the mobile phase, in this case supercritical CO(2) (scCO(2)), often resulting in concentrations that are too low to be detected by NMR spectroscopy or at least requiring excessive signal averaging. We present a hyphenated SFC-NMR setup with an integrated approach for concentrating samples in-line, which are diluted in scCO(2) during chromatography. This in-line concentration is achieved by controlled in-line expansion of the scCO(2). As a proof of concept four isomers of vitamin E (tocopherol) were isolated by SFC, concentrated in-line by expanding CO(2) from 120 to 50 bar, and finally shuttled to the NMR spectrometer fitted with a dedicated probehead for spectroscopic characterization of microfluidic samples. The abundant isomers were readily detected, supporting the viability of SFC-NMR as a powerful analytical tool.