Relationship between trans-column eddy diffusion and retention in liquid chromatography: Theory and experimental evidence

Experimental results demonstrate that trans-column eddy diffusion depends on the retention of compounds. The combination of elution profiles recorded in different points of the exit column cross-section and of the height equivalent to a theoretical plate (HETP) of small molecules clearly show a strong link between retention and column performance in liquid chromatography. These results validate a new model of trans-column eddy diffusion in packed columns. The contribution to the column HETP of the trans-column eddy diffusion term decreases with increasing retention factor from k′ = 0 to k′ = 3 above which it becomes negligible. The best column performance in RPLC is observed for the most retained compounds. This is due to the combination of the lack of a residual trans-column eddy diffusion contribution and the vanishing contribution of the instrument to band broadening.