Fourier transformation liquid chromatography: increasing sensitivity by a factor of 50

Abstract

Methods that increase sensitivity are a constant topic in research. To increase the sensitivity for high-performance liquid chromatography (HPLC) a continuous injection method with a sinusoidal analyte concentration profile was developed. The sinusoidal analyte concentration profile is obtained by a sinusoidal variation of the relative volume contents between two solvent reservoirs, one containing a pure solvent and the other an analyte stock solution prepared with the same solvent. Discrete Fourier transformation enables the analyte-specific phase angle shift to be calculated from the chromatograms. The proposed method is established and evaluated for size exclusion chromatography (SEC). Accordingly, the phase angle shift is used for molecular weight determination by establishing a molecular weight calibration using different narrowly distributed polystyrene (PS) calibration standards. In a comparison with conventional SEC, the Signal-to-Noise ratio (S/N), normalized to the square root of the time of the evaluated data set, increases by a factor of approximately 50.