Analytical potential of total reflection X-ray fluorescence spectrometry for simultaneous determination of iron, copper and zinc in human blood serum and plasma.

Due to many roles of trace elements such as Fe, Cu and Zn in various physiological and pathophysiological processes, their determination in serum and plasma is of high clinical relevance. In the present study, for the first time, the effect of serum and plasma preparation parameters (dilution factor and sample deposition volume) on the quality of results obtained by TXRF analysis was evaluated by means of experimental design tools (response surface analysis). It was found that the best strategy was the direct analysis of both human fluids without a previous dilution step. The accuracy and precision of the proposed methods were evaluated by analysis of reference materials (ClinChek® Plasma Control Level II and Seronorm™ Trace Elements Serum L-1). TXRF results agreed with the reference values and no significant differences at 95% confidence level were found. Limits of detection for the elements of interest were also adequate, taking into account their typical concentration ranges in real serum and plasma samples. Finally, the developed TXRF methods were applied to a set of serum and plasma samples from patients with different genders, ages and diagnoses, previously analysed by ICP-OES and ICP-MS techniques. The results showed good agreement between both analytical approaches. These results suggest that the proposed TXRF method provides reliable results thus being suitable for plasma and serum analysis, but in a simpler and more sustainable way.