Total reflection X-ray fluorescence analysis (TXRF) was used to determine chromium, selenium and bromine concentrations in blood serum samples of 50 patients with parenteral nutrition treatment. The concentrations were measured two times, namely in the first day (I measurement) of the treatment and the seventh day (II measurement) after the chromium and selenium supplementation. For comparison purposes also serum samples of 50 patients without nutritional disorders, admitted to a planned surgical procedure to remove the gall bladder (cholecystectomy), were analyzed and treated as the control group. Descriptive statistics of measured concentrations of Cr, Se and Br both for the studied and control groups was determined. In order to check the effectiveness of Cr and Se supplementation, the results of the first and seventh day measurements for studied group were statistically compared with each other, with literature reference values and with the results of the control group (two-group comparison). These comparisons indicate the effectiveness of selenium supplementation in the applied treatment procedure. In the case of Cr and Br concentrations no statistically significant differences were observed. We conclude that monitoring of the concentration of the important trace elements in human serum should be standard procedure in parenteral nutrition treatment. In this monitoring the TXRF technique can be successfully used.