Hemodiafiltration on-line (on-line HDF) is a more efficient treatment than low-flux hemodialysis (HD). Unfortunately, it cannot be proposed to all patients. The aim of this study was to evaluate the safety, efficiency, and mechanisms of removal of toxins with high-flux HD vs. low-flux HD and on-line HDF. Randomized cross-over study designed to evaluate efficiency and tolerability of high-flux HD vs. low-flux HD in aged patients; to compare by means of biochemical and proteomic analyses the efficiency and mechanisms of removal of toxins with high-flux HD vs. on-line HDF. The removal of small toxins was similar with high-flux and low-flux HD. β2-microglobulin was removed only with high-flux HD, which had an excellent tolerability. The efficiency of high-flux HD was similar to on-line HDF. Proteomic analysis demonstrated that only high-flux membranes remove and adsorb small proteins. High-flux HD may be an efficient alternative to on-line HDF.