Soldado Cabezuelo A B, Montes Bayón M, Blancon González E, Garcia Alonso J I, Sanz-Medel A
Department of Physical and Analytical Chemistry, University of Oviedo C/Julián Clavería 8, Spain.
Analyst. 1998 May;123(5):865-9. doi: 10.1039/a707669j.
The coupling of fast protein liquid chromatography (FPLC) with inductively couples plasma mass spectrometry (ICP-MS) was evaluated as a technique for studying aluminium bound to proteins present in human serum. Separation of human serum proteins was achieved on a MonQ (HR5/5) anion-exchange column using an ammonium acetate gradient (0-0.25 mol I-1) at the physiological pH of 7.4 (0.05 mol l-1 TRIS-HC1 buffer). Aluminium contamination was avoided with an on-line Al-chelating scavenger column. Proteins were detected spectrophotometrically at 295 nm and the Al detection was carried out on-line using both quadrupole ICP-MS and double-focusing ICP-MS systems. At metal basal levels in serum the latter detector proved to be adequate for this detection. Results obtained with the procedure developed confirmed clearly that transferrin is the only significant A1-binding proteins in unspiked uraemic serum. In addition, a high-resolution ICP-MS instrument was applied successfully as an A1-specific detector allowing for the first time A1 speciation studies in unspiked normal serum. The technique can also be used for studying the protein binding of elements other than A1.
快速蛋白质液相色谱（FPLC）与电感耦合等离子体质谱（ICP-MS）联用，作为一种研究与人类血清中蛋白质结合的铝的技术进行了评估。在生理pH值7.4（0.05 mol l-1 TRIS-HCl缓冲液）条件下，使用醋酸铵梯度（0-0.25 mol l-1），在MonQ（HR5/5）阴离子交换柱上实现了人血清蛋白的分离。通过在线铝螯合清除柱避免了铝污染。在295 nm处用分光光度法检测蛋白质，并使用四极杆ICP-MS和双聚焦ICP-MS系统在线进行铝的检测。在血清中金属基础水平下，后一种检测器被证明足以进行这种检测。用所开发的方法获得的结果清楚地证实，转铁蛋白是未加样的尿毒症血清中唯一重要的铝结合蛋白。此外，高分辨率ICP-MS仪器成功地用作铝特异性检测器，首次允许在未加样的正常血清中进行铝形态研究。该技术还可用于研究除铝以外的元素与蛋白质的结合。
