Minami Takeshi, Ichida Seiji, Kubo Kanenobu
Laboratory of Environmental Biology, Department of Life Science, School of Science and Engineering, Kinki University, 3-4-1 Kowakae, Higashiosaka, Osaka 577-8502, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Dec 5;781(1-2):303-11. doi: 10.1016/s1570-0232(02)00496-8.
Metallothioneins (MTs) have many different functions in tissues, but the roles of individual isoforms are still not entirely clear. Capillary zone electrophoresis (CZE) is a powerful method for the separation of substances because of its small sample requirement, rapid analysis, high sensitivity and high resolution. The separation and identification of mammalian MT-1, MT-2, and MT-3 and class III MTs by CZE has been reported. Uncoated and polyacrylamide-coated capillary tubes were recently used for the separation of MTs, and a UV detector is usually employed for observations of peaks of MTs. Small changes to the structure and metal components of MTs are reflected in the migration times of the peaks. N-acetylated and non-acetylated MTs can be separated and identified by CZE-mass spectrometry (MS). In addition, metal complexes with MTs can be characterized by CZE-proton-induced X-ray emission (PIXE) detector and CZE-inductively coupled plasma (ICP)-MS. For the quantification of an MT isoform, the peak area of UV absorption is used, but the technique has problems. One is lack of a purified isoform standard. The other is the need for a suitable internal standard substance. CZE-ICP-isotope dilution (ID)-MS is also reported to be able to quantify MT isoforms. CZE combined with other techniques is very effective for separation and quantitative and qualitative analyses of MT isoforms in biological materials.
金属硫蛋白（MTs）在组织中有许多不同功能，但各个异构体的作用仍不完全清楚。毛细管区带电泳（CZE）由于其样品需求量小、分析快速、灵敏度高和分辨率高，是一种强大的物质分离方法。已有报道通过CZE分离和鉴定哺乳动物的MT-1、MT-2、MT-3和III类MTs。未涂层和聚丙烯酰胺涂层的毛细管最近被用于MTs的分离，并且通常使用紫外检测器来观察MTs的峰。MTs结构和金属成分的微小变化反映在峰的迁移时间上。N-乙酰化和非乙酰化的MTs可以通过CZE-质谱（MS）进行分离和鉴定。此外，MTs的金属配合物可以通过CZE-质子诱导X射线发射（PIXE）检测器和CZE-电感耦合等离子体（ICP）-MS进行表征。对于MT异构体的定量，使用紫外吸收的峰面积，但该技术存在问题。一个问题是缺乏纯化的异构体标准品。另一个问题是需要合适的内标物质。也有报道称CZE-ICP-同位素稀释（ID）-MS能够定量MT异构体。CZE与其他技术相结合对于生物材料中MT异构体的分离以及定量和定性分析非常有效。
