Speciation of basal aluminium in human serum by fast protein liquid chromatography with inductively coupled plasma mass spectrometric detection.
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.
Department of Physical and Analytical Chemistry, University of Oviedo C/Julián Clavería 8, Spain.
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AB
Soldado Cabezuelo
M
M
Montes Bayón
E
E
Blancon González
J I
JI
Garcia Alonso
A
A
Sanz-Medel
D000535:Aluminum / Q000097:blood* / Q000378:metabolism; D001798:Blood Proteins / Q000378:metabolism; D002853:Chromatography, Liquid; D006801:Humans; D011485:Protein Binding; D013057:Spectrum Analysis
D001798:Blood Proteins; D000535:Aluminum