Separation and identification of flavonoids from complex samples using off-line two-dimensional liquid chromatography tandem mass spectrometry

In this study, a LC-based identification strategy was proposed and off-line two-dimensional liquid chromatography was developed for the separation and identification of flavonoids from complex samples. The highly orthogonal separation system was composed of a CD column, an OEG column and an XTerra C18 column. The CD column was employed for the first dimensional separation and the OEG column and XTerra C18 column were used for the second dimensional separation in parallel. A mixture of six traditional Chinese medicines was used as a complex sample in testing this method. Unknown peaks were identified by matching their retention times and accurate mass molecular weights with those of references. The identification efficacy of parallel unidimensional liquid chromatography and off-line two-dimensional liquid chromatography was compared, and the latter was demonstrated to be more efficient for the identification of target compounds—especially co-eluted compounds or minor compounds from complex samples.

► We developed a 2D-LC–MS based method for the identification of complex samples.
► Structurally similar or minor compounds can be well separated and identified.
► 2D-LC is more efficient than parallel 1D-LC for the identification of complex samples.