Richards M P
US Department of Agriculture, Livestock and Poultry Sciences Institute (LPSI), Nonruminant Animal Nutrition Laboratory, Beltsville, MD 20705-2350.
J Chromatogr B Biomed Appl. 1994 Jul 15;657(2):345-55. doi: 10.1016/0378-4347(94)00033-6.
In this study a fused-silica capillary treated internally with a polyamine coating which reverses electroosmotic flow in the direction of the anode was evaluated for its ability to resolve metallothionein (MT) isoforms. Analysis of different MTs purified from liver and kidney tissue revealed the following numbers of putative isoform peaks resolved: rabbit (3-6); horse (3-5); rat (2-3), chicken (1); human MT-1 (5-6); sheep (4-5) and pig (4-5). The greater degree of MT isoform heterogeneity detected in this study using the polyamine-coated capillary suggested a higher resolving capacity for capillary zone electrophoresis conducted with this capillary compared to an uncoated one. Using the single isoform of chicken MT (cMT) as a reference standard, relative standard deviations of 2.53, 1.85 and 2.21% for peak migration time, area and height, respectively, were observed for eight consecutive runs. A standard curve for cMT established linearity (r2 = 0.99) for integrated peak area over three log units of cMT concentration with a lower limit of detection estimated to be < or = 5 micrograms/ml. Acetonitrile extracts of chick liver tissue homogenates were successfully analyzed for the presence of MT isoforms from both control and zinc-injected animals. Based on our initial evaluation, capillary zone electrophoresis using the polyamine-coated capillary appears to be a very useful analytical method for the separation and quantification of individual MT isoforms.
在本研究中，对一根内部经多胺涂层处理、能使电渗流反向流向阳极的熔融石英毛细管，评估了其分离金属硫蛋白（MT）异构体的能力。对从肝脏和肾脏组织中纯化的不同MT进行分析，结果显示分辨出的推定异构体峰数量如下：兔（3 - 6个）；马（3 - 5个）；大鼠（2 - 3个），鸡（1个）；人MT - 1（5 - 6个）；绵羊（4 - 5个）和猪（4 - 5个）。本研究中使用多胺涂层毛细管检测到的MT异构体异质性程度更高，这表明与未涂层毛细管相比，使用该毛细管进行毛细管区带电泳时具有更高的分离能力。以鸡MT（cMT）的单一异构体作为参考标准，连续八次进样观察到峰迁移时间、峰面积和峰高的相对标准偏差分别为2.53%、1.85%和2.21%。cMT的标准曲线在三个对数单位的cMT浓度范围内，积分峰面积呈现线性关系（r2 = 0.99），估计检测下限为≤5微克/毫升。成功分析了来自对照和注射锌的动物的鸡肝脏组织匀浆的乙腈提取物中MT异构体的存在情况。基于我们的初步评估，使用多胺涂层毛细管进行毛细管区带电泳似乎是一种非常有用的分析方法，可用于分离和定量单个MT异构体。
