Improved quantification for non-transferrin-bound iron measurement using high-performance liquid chromatography by reducing iron contamination
Improved quantification for non-transferrin-bound iron measurement using high-performance liquid chromatography by reducing iron contamination
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DOI：
10.3892/mmr.2011.518
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发表时间：
2011-09-01
影响因子：
3.4
通讯作者：
Kohgo, Yutaka
中科院分区：
文献类型：
--
作者：
Sasaki, Katsunori;Ikuta, Katsuya;Kohgo, Yutaka
非转铁蛋白结合铁（NTBI）是指血浆中与转铁蛋白以外的配体结合的所有形式的铁，被认为是铁毒性的标志物。已经报道了多种测量 NTBI 的分析方法；然而，尚未达到临床相关的敏感性水平。此外，一些患者和健康受试者的 NTBI 值不足，导致试剂可能受到背景铁污染的假设。本研究针对每个步骤中铁污染的潜在点重新评估了测定的分析程序。制备 NTA 和三碳钴酸盐 (III) 溶液，去除铁污染，然后进行 NTBI 定量。结果，通过成功减少背景铁污染，提高了基于高效液相色谱 (HPLC) 的 NTBI 方法的灵敏度。应用我们改进的方法证明，即使在健康志愿者中也能检测到 NTBI，尽管浓度极低； NTBI 平均水平为 0.206 +/- 0.091 muM（男性，n=20）和 0.212 +/- 0.095 muM（女性，n=16）。因此，我们对 NTBI 测定的修改可能具有临床意义，并且可能有助于理解相对较低但升高的 NTBI 浓度在除典型铁超载以外的疾病中的临床意义。
Non-transferrin-bound iron (NTBI) refers to all forms of iron in the plasma that bind to ligands other than transferrin, and is considered to be a marker of iron toxicity. A variety of analytical approaches for measuring NTBI have been reported; however, a clinically relevant level of sensitivity has yet to be achieved. In addition, insufficient values of NTBI in some patients and healthy subjects have led to the assumption that there may be contamination of reagents with background iron. The present study re-evaluated the analytical procedures of the assay with regard to the potential points of iron contamination in each step. NTA and tris carbonatocobaltate (III) solutions were prepared with removal of iron contamination, and then quantification of NTBI was performed. As a result, the sensitivity of the high-performance liquid chromatography (HPLC)-based NTBI method was improved by the successful reduction of background iron contamination. Application of our modified method proved that NTBI was detected even in healthy volunteers, although the concentrations were extremely low; the average NTBI levels were 0.206 +/- 0.091 mu M (males, n=20) and 0.212 +/- 0.095 mu M (females, n=16). Thus, our modification of the NTBI assay may be clinically meaningful, and may contribute to the understanding of the clinical significance of relatively low, but elevated concentrations of NTBI in diseases other than typical iron overload.