Department of Nutritional Physiology, Institute of Nutritional Sciences, Friedrich Schiller University Jena, 07743 Jena, Germany; TraceAge-DFG Research Unit on Interactions of Essential Trace Elements in Healthy and Diseased Elderly, Potsdam-Berlin-Jena, Germany.
Department of Nutritional Physiology, Institute of Nutritional Sciences, Friedrich Schiller University Jena, 07743 Jena, Germany.
J Trace Elem Med Biol. 2023 Jul;78:127167. doi: 10.1016/j.jtemb.2023.127167. Epub 2023 Mar 30.
Trace elements exhibit essential functions in many physiological processes. Thus, for research focusing on trace element homeostasis and metabolism analytical methods allowing for multi-element analyses are fundamental. Small sample amounts may be a big challenge in trace element analyses especially if also other end points want to be addressed in the same sample. Therefore, the aim of the present study was to examine trace elements (iron, copper, zinc, and selenium) in murine liver tissue prepared by a RIPA buffer-based lyses method.
After centrifugation, lysates and pellets were obtained and trace elements were analyzed with TXRF in liver lysates. The results were compared to that obtained by a standard microwave-assisted acidic digestion with subsequent ICP-MS/MS analysis of the same liver tissue, liver lysates, and remaining pellets. In addition, trace element concentrations, determined in murine serum with both methods, were compared. For serum samples, both TXRF and ICP-MS/MS provide similar and highly correlating results. Furthermore, in liver lysate samples prepared with RIPA buffer, comparable trace element concentrations were measured by TXRF as with the standard digestion technique and ICP-MS/MS. Only marginal amounts of trace elements were detected in the pellets.
Taken together, the results obtained by the present study indicate that the RIPA buffer-based method is suitable for sample preparation for trace element analyses via TXRF, at least for the here investigated murine liver samples.
微量元素在许多生理过程中发挥着重要作用。因此,对于专注于微量元素动态平衡和代谢的研究,能够进行多元素分析的分析方法是基础。小样本量可能是微量元素分析的一个重大挑战,特别是如果还希望在同一样品中解决其他终点问题。因此,本研究的目的是通过 RIPA 缓冲液基础裂解方法检测鼠肝组织中的微量元素(铁、铜、锌和硒)。
离心后,得到裂解物和沉淀物,并使用 TXRF 分析肝裂解物中的微量元素。将结果与用相同的肝组织、肝裂解物和剩余沉淀物进行的标准微波辅助酸性消化后的 ICP-MS/MS 分析结果进行比较。此外,还比较了两种方法在鼠血清中测定的微量元素浓度。对于血清样品,TXRF 和 ICP-MS/MS 均提供相似且高度相关的结果。此外,在使用 RIPA 缓冲液制备的肝裂解物样品中,通过 TXRF 测量的微量元素浓度与标准消化技术和 ICP-MS/MS 相当。在沉淀物中仅检测到微量的微量元素。
综上所述,本研究的结果表明,RIPA 缓冲液基础方法适用于通过 TXRF 进行微量元素分析的样品制备,至少对于本文研究的鼠肝样品是如此。