Elemental Scientific, Inc., 7277 World Communications Dr., Omaha, NE, USA.
University of Münster, Institute of Inorganic and Analytical Chemistry, Corrensstrasse 30, 48149 Münster, Germany.
Metallomics. 2020 Sep 23;12(9):1348-1355. doi: 10.1039/d0mt00132e.
Copper is an essential element for biological functions within humans and animals. There are several known diseases associated with Cu deficiency or overload, such as Menkes disease and Wilson disease, respectively. A common clinical method for determining extractable Cu levels in serum, which is thought to be potentially dangerous if in excess, is to subtract the value of tightly incorporated Cu in ceruloplasmin from total serum Cu. In this work, an automated sample preparation and liquid chromatography (LC) system was combined with inductively coupled plasma-mass spectrometry (ICP-MS) to determine bound Cu and extractable Cu in serum. This LC-ICP-MS method took 250 s for sample preparation and analysis, followed by a column recondition/system reset, thus, a 6 minute sample-to-sample time including sample preparation. The method was validated using serum collected from either control (Atp7b+/-) or Wilson disease rats (Atp7b-/-). The extractable Cu was found to be 4.0 ± 2.3 μM Cu in healthy control rats, but 2.1 ± 0.6 μM Cu in healthy Wilson rats, and 27 ± 16 μM Cu in diseased Wilson rats, respectively. In addition, the extractable Cu/bound Cu ratio was found to be 6.4 ± 3.5%, 38 ± 29%, and 34 ± 22%, respectively. These results suggest that the developed method could be of diagnostic value for Wilson disease, and possibly other copper related diseases.
铜是人类和动物体内生物功能所必需的元素。有几种已知的疾病与 Cu 缺乏或过载有关，例如 Menkes 病和 Wilson 病。一种常见的临床方法是确定血清中可提取的 Cu 水平，如果过量，认为这是潜在危险的，方法是从总血清 Cu 中减去铜蓝蛋白中紧密结合的 Cu 值。在这项工作中，自动化样品制备和液相色谱（LC）系统与电感耦合等离子体质谱（ICP-MS）相结合，用于测定血清中的结合 Cu 和可提取 Cu。该 LC-ICP-MS 方法的样品制备和分析时间为 250 s，随后进行柱再生/系统重置，因此，包括样品制备在内，每个样品的分析时间为 6 分钟。使用来自健康对照（Atp7b+/-）或 Wilson 病大鼠（Atp7b-/-）的血清对该方法进行了验证。结果发现，健康对照组大鼠的可提取 Cu 为 4.0 ± 2.3 μM Cu，而健康 Wilson 大鼠的可提取 Cu 为 2.1 ± 0.6 μM Cu，患病 Wilson 大鼠的可提取 Cu 为 27 ± 16 μM Cu。此外，可提取 Cu/结合 Cu 比值分别为 6.4 ± 3.5%、38 ± 29%和 34 ± 22%。这些结果表明，所开发的方法可能对 Wilson 病，以及可能的其他铜相关疾病具有诊断价值。
