Determination of trace elements in placenta by total reflection X-ray fluorescence spectrometry: effects of sampling and sample preparation.

Placental elemental composition can serve as an indicator for neonatal health. Medical studies aiming at revealing such cause-and-effect relationships or studies monitoring potential environmental influences consist of large sample series to ensure statistically sufficient data. Several analytical techniques have been used to study trace metals in human placenta. However, most techniques require provision of clear liquid sample solutions and therefore time- and reagent-consuming total digestion of biological tissue is necessary. In total reflection X-ray fluorescence spectrometry (TXRF)-a straightforward multielement analytical technique-in contrast suspensions of minute sample amounts can be analyzed directly. Therefore, herein we report on a valid method to prepare homogenous sample suspensions for sustainable and fast TXRF analysis of large sample series. The optimized method requires only 10 mg of powdered placental tissue and 1 mL nitric acid. Suspensions are readily prepared within 30 min and the found mass fractions of major, minor, and trace elements are in good agreement in comparison to analysis of digests. In addition, possible effects on fixation time and the exact sampling location, i.e., maternal vs. fetal side of the placenta, were studied applying this method. Thereby, significant differences for fetal placenta tissue compared to maternal or intermediate tissue were observed revealing accumulation of trace elements in the fetal side of the placenta. Furthermore, considerable depletion of up to 60% mass fraction with longer fixation duration occurred in particular in fetal placenta tissue. These findings help to understand the large ranges of mass fraction of elements in placenta reported in the literature and at the same time indicate the necessity for more systematic investigation of non-homogenous elements distributed in placenta taking sampling and stabilization methods into account.