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Established methods to measure the concentration of trace elements in biological matrices often provide only single element information, are time-consuming, and require special sample preparation. Therefore, the development of straightforward and rapid analytical methods for enhanced, multi-trace element determination in biological samples is an important and raising field of trace element analysis. Herein, we report on the development and validation of a reliable method based on total reflection X-ray fluorescence (TXRF) analysis to precisely quantify iron and other trace metals in a variety of biological samples, such as the liver, parenchymal and non-parenchymal liver cells, and bone marrow\u2013derived macrophages. We show that TXRF allows fast and simple one-point calibration by addition of an internal standard and has the potential of multi-element analysis in minute sample amounts. The method was validated for iron by recovery experiments in homogenates in a wide concentration range from 1 to 1600\u00a0\u03bcg\/L applying well-established graphite furnace atomic absorption spectrometry (GFAAS) as a reference method. The recovery rate of 99.93\u2009\u00b1\u20090.14% reveals the absence of systematic errors. Furthermore, the standard reference material \u201cbovine liver\u201d (SRM 1577c, NIST) was investigated in order to validate the method for further biometals. Quantitative recoveries (92\u2013106%) of copper, iron, zinc, and manganese prove the suitability of the developed method. The limits of detection for the minute sample amounts are in the low picogram range.<\/jats:p>","DOI":"10.1007\/s00216-020-02614-8","type":"journal-article","created":{"date-parts":[[2020,4,26]],"date-time":"2020-04-26T20:02:22Z","timestamp":1587931342000},"page":"6419-6429","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":27,"title":["Total reflection X-ray fluorescence spectrometry for trace determination of iron and some additional elements in biological samples"],"prefix":"10.1007","volume":"412","author":[{"given":"Andreas","family":"Gruber","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Riccarda","family":"M\u00fcller","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessa","family":"Wagner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia","family":"Colucci","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maja Vuji\u0107","family":"Spasi\u0107","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kerstin","family":"Leopold","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,4,26]]},"reference":[{"key":"2614_CR1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3389\/fphar.2014.00042","volume":"5","author":"M Vujic Spasic","year":"2014","unstructured":"Vujic Spasic M. 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