Styblo M, Hughes M F, Thomas D J
Curriculum in Toxicology, University of North Carolina at Chapel Hill, 27514, USA.
J Chromatogr B Biomed Appl. 1996 Feb 23;677(1):161-6. doi: 10.1016/0378-4347(95)00490-4.
Protein-bound arsenicals were liberated from binding sites on liver cytosolic proteins by exposure to 0.1 M CuCl at pH 1. This method released greater than 90% of the arsenicals associated with biological matrices. Ultrafiltrates of CuCl-treated cytosols were subjected to thin-layer chromatography to speciate and quantify inorganic and methylated arsenicals. For rat liver cytosol in an in vitro methylation assay and for liver and kidney cytosols from arsenite-treated mice, most inorganic arsenic was protein bound. Appreciable fractions of the organoarsenical metabolites present in these cytosols were also protein bound. Therefore, CuCl treatment of cytosols releases protein-bound arsenicals, permitting more accurate estimates of the pattern and extent of arsenic methylation in vitro and in vivo.
通过在pH 1条件下暴露于0.1 M CuCl，从肝脏胞质蛋白的结合位点释放与蛋白质结合的砷化合物。该方法释放了与生物基质相关的90%以上的砷化合物。对经CuCl处理的胞质溶胶的超滤物进行薄层色谱分析，以鉴定和定量无机砷和甲基化砷。在体外甲基化试验中，对于大鼠肝脏胞质溶胶以及亚砷酸盐处理小鼠的肝脏和肾脏胞质溶胶，大多数无机砷与蛋白质结合。这些胞质溶胶中存在的有机砷代谢物的相当一部分也与蛋白质结合。因此，用CuCl处理胞质溶胶可释放与蛋白质结合的砷化合物，从而更准确地估计体外和体内砷甲基化的模式和程度。
