Cold plasma ashing improves the trace element detection of single Daphnia specimens by total reflection X-ray fluorescence spectrometry

The recently developed dry method for the element determination of single freshwater microcrustacean specimens (Daphnia) using total reflection X-ray fluorescence (TXRF) spectrometry showed that inhomogeneities of the biological material on the glass carriers resulted in some cases in high background and hampered the detection of certain trace elements (e.g. Cr, Ni). The aim of this study was to test how inhomogeneities of the biological material can be reduced using cold plasma ashing (CPA) techniques. For that, single specimens of the microcrustacean Daphnia pulex prepared according to the dry method were measured by TXRF before and after CPA. To determine the efficiency of the removal of organic matrix, the background and signal-to-background relationship of 28 samples were analyzed. The results showed (1) a highly significant reduction of the background by CPA fluctuating between 26 and 46% (all elements) and (2) a significant increase of the signal-to-background relationship by the factor 1.5–2.5 (all elements) and a much better detection of Cr, Pb, As and Se. The element concentrations (with exception of Cr, Ni and Pb) after ashing were in the same range or slightly higher than that before ashing. No significant differences between the two treatments were observed for Mn, As, Pb, Se (November), Sr (November), Cr (March) and Pb (March). The element concentration of P, K, Ca, Cu, Zn, Cr (November), Fe and Rb were significantly higher after ashing. In general, they increased by 1.5–13.6% and were highest for Rb (March) and P (November). In contrast, the element concentration of Ni and Cr (only March) decreased significantly after ashing (Ni: 91.6–92.1%, Cr: 91.3%). We recommend the use of CPA for biological material in the microgram-range as a routine method for TXRF analysis, especially when trace elements in minute concentrations are of interest.