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通过电感耦合等离子体质谱法和尺寸排阻色谱-电感耦合等离子体质谱法探究给予小鼠六价铬后的金属动态平衡效应。

Probing the metal-homeostatis effects of the administration of chromium(vi) to mice by ICP MS and size-exclusion chromatography-ICP MS.

机构信息

Hacettepe University, Chemistry Department, Analytical Chemistry Division, Beytepe, Ankara, Turkey.

出版信息

Metallomics. 2010 Aug;2(8):549-55. doi: 10.1039/c004508j. Epub 2010 Jul 29.

Abstract

Concentrations of chromium, copper, iron, manganese and zinc were determined in liver, kidney, brain, lung, heart and testis of mouse following intraperitoneal injection of hexavalent chromium [Cr(vi)] at a single dose of 8.0 mg Cr/kg. As result, chromium concentrations increased ca. 40-fold in liver and kidney and by a factor of 3-5 in all the other tissues. The homeostasis of Cu, Fe, Mn and Zn was also affected. The element molecular weight distribution was evaluated in the cytosols of the different mouse organs by size-exclusion chromatography (Superdex-75) with UV-VIS and ICP-MS detection. The administration of Cr(vi) resulted in differences in the elution profiles of Fe, Mn, Cu and Zn-protein complexes. Bioinduced Mn, Fe and Zn-binding proteins could be detected in some tissues, especially in liver and kidney. Different molecular weight fractions containing chromium were heartcut and submitted to tryptic digestion prior to MALDI MS analysis. Cr-peptide complexes could be obtained both in non-denaturing and in denaturing (in the presence of urea and DTT) conditions. They were isolated by size-exclusion chromatography with a smaller separation range (Superdex Peptide) but could not be identified by MALDI MS.

摘要

采用腹腔注射方式一次性给予小鼠 8.0mg/kg 六价铬[Cr(VI)]后,测定其肝脏、肾脏、大脑、肺脏、心脏和睾丸中的铬、铜、铁、锰和锌浓度。结果显示,铬在肝脏和肾脏中的浓度分别增加了约 40 倍和 3-5 倍,其他组织中的浓度也有所增加。铜、铁、锰和锌的体内平衡也受到了影响。采用尺寸排阻色谱(Superdex-75)结合紫外-可见和 ICP-MS 检测方法,评估了不同小鼠器官细胞溶质中元素分子量分布。Cr(VI)的给药导致 Fe、Mn、Cu 和 Zn-蛋白复合物的洗脱曲线出现差异。在一些组织中,特别是在肝脏和肾脏中,可以检测到生物诱导的 Mn、Fe 和 Zn 结合蛋白。对含铬的不同分子量级分进行心切割,然后进行胰蛋白酶消化,再进行 MALDI MS 分析。可以在非变性和变性(存在尿素和 DTT 的情况下)条件下获得 Cr-肽复合物。它们通过尺寸排阻色谱(Superdex Peptide)进行分离,分离范围较小,但无法通过 MALDI MS 进行鉴定。

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