Maximizing the peak production rate in off-line comprehensive two-dimensional liquid chromatography with mass spectrometry detection

This article describes an optimization strategy to obtain the best possible performance in the shortest analysis time—called the peak production rate—for comprehensive off-line two-dimensional liquid chromatography. The demands on column technology (particle size and column length) and LC conditions (including sampling time, gradient time and slope, and column-equilibration time) are discussed. The potential of off-line 2D-LC coupled to tandem mass spectrometry detection is demonstrated with the analysis of proteomics samples of various complexities and a trade-off is made between the 1D-LC and off-line 2D-LC performance.