Peak broadening caused by using different micro-liquid chromatography detectors.

Advancements in column technology resulted in smaller particles and more efficient phases. In parallel, the use of columns with reduced dimensions is becoming more common. This means the effective column volume is also decreased, thereby making the systems more susceptible to effects of band broadening due to extra-column volume. Despite these trends and the fact that a growing number of miniaturized liquid chromatography systems are being offered commercially, manufacturers often stick to the modular concept with dedicated units for pumps, column oven, and detectors. This modular design results in long connection capillaries, which leads to extra-column band broadening and consequently prevents the exploitation of the intrinsic efficiency of state-of-the-art columns. In particular, band broadening post column has a considerable negative effect on efficiency. In this study, mass flow and concentration-dependent detectors were examined for their influence on band broadening using a micro-LC system. A mass spectrometric detector, an evaporative light scattering detector, two UV detectors, and a previously undescribed fluorescence detector were compared. The influence on efficiency is compared using plate height vs linear velocity data and peak variance. It is shown that an increase in the inner diameter after the post-column transfer capillary leads to significant loss in plate height. Comparing the UV detectors, it could be shown that the dispersion was reduced by 38% by the reduction of the post-column volume. The largest variance was found for the evaporative light scattering detector, which was 368% higher compared to the variance of the detector with the least effect on band broadening.