Quantitative studies of aluminium binding species in human uremic serum by fast protein liquid chromatography coupled with electrothermal atomic absorption spectrometry
Quantitative studies of aluminium binding species in human uremic serum by fast protein liquid chromatography coupled with electrothermal atomic absorption spectrometry
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DOI：
10.1039/a608258k
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发表时间：
1997-06-01
期刊：
影响因子：
4.2
通讯作者：
SanzMedel, A
中科院分区：
文献类型：
--
作者：
Cabezuelo, ABS;Gonzalez, EB;SanzMedel, A
快速蛋白液相色谱 (FPLC) 与电热原子吸收光谱 (ETAAS) 检测结合使用，对未加标的人尿毒症血清中的铝结合物质进行定量研究，在 Mono Q (HR 5/5) 阴离子交换柱上实现了人血清蛋白和其他铝结合物（柠檬酸盐和去铁胺）的快速、可重复分离 使用氯化钠梯度（0-0.25 mol l(-1)），生理人血清pH值为7.4（0.05 mol l(-1)缓冲液TRIS-HCl）。柱级分中的铝分布通过 ETAAS 测定。使用配备在线铝螯合清除剂柱（Kelex 100 浸渍二氧化硅 C-18）的惰性色谱系统避免了铝污染。所提出方法的灵敏度（血清中铝的检测限 = 5 mu g l(-1)）允许在临床相关浓度（透析患者的未加标血清）下进行铝形态研究。获得的结果证实，转铁蛋白是唯一结合铝的血清蛋白，其约占总血清铝的90%（洗脱后铝回收率= 105 +/- 5%），还证实在螯合药物去铁胺（DFO）存在的情况下，大部分血清铝（80%）与DFO结合。
Fast protein liquid chromatography (FPLC) was used with electrothermal atomic absorption spectrometric (ETAAS) detection for quantitative studies of aluminium binding species in unspiked human uremic serum, A rapid and reproducible separation of human serum proteins and other aluminium binders (citrate and desferroxiamine) was achieved on a Mono Q (HR 5/5) anion-exchange column using a sodium chloride gradient (0-0.25 mol l(-1)) at the physiological human serum pH of 7.4 (0.05 mol l(-1) buffer TRIS-HCl). The aluminium distribution in the column fractions was determined by ETAAS. Aluminium contamination was avoided by using an inert chromatographic system equipped with an on-line aluminium-chelating scavenger column (Kelex 100-impregnated silica C-18) The sensitivity of the proposed method (detection limit for Al in serum = 5 mu g l(-1)) allowed aluminium speciation studies at clinically relevant concentrations (unspiked serum from dialysis patients). The results obtained confirmed that transferrin is the only serum protein binding aluminium and it contains about 90% of total serum aluminium (post-elution aluminium recovery = 105 +/- 5%), It was also confirmed that in the presence of the chelating drug desferrioxamine (DFO) most of the serum aluminium (80%) is bound to DFO.