Blomfield J, Macmahon R A
J Clin Pathol. 1969 Mar;22(2):136-43. doi: 10.1136/jcp.22.2.136.
The free and total plasma copper and total erythrocyte copper levels have been determined by simple, yet sensitive and highly specific methods, using atomic absorption spectrophotometry. For total copper determination, the copper was split from its protein combination in plasma or red cells by the action of hydrochloric acid at room temperature. The liberated copper was chelated by ammonium pyrrolidine dithiocarbamate and extracted into n-butyl acetate by shaking and the organic extract was aspirated into the atomic absorption spectrophotometer flame. The entire procedure was carried out in polypropylene centrifuge tubes, capped during shaking. For the free plasma copper measurement the hydrochloric acid step was omitted. Removal of the plasma or erythrocyte proteins was found to be unnecessary, and, in addition, the presence of trichloracetic acid caused an appreciable lowering of absorption. Using a double-beam atomic absorption spectrophotometer and scale expansion x 10, micro methods have been derived for determining the total copper of plasma or erythrocytes with 0.1 ml of sample, and the free copper of plasma with 0.5 ml. The macro plasma copper method requires 2 ml of plasma and is suitable for use with single-beam atomic absorption spectrophotometers. With blood from 50 blood donors, normal ranges of plasma and erythrocyte copper have been determined.
采用原子吸收分光光度法，通过简单、灵敏且高度特异的方法测定了游离和总血浆铜以及总红细胞铜水平。对于总铜的测定，在室温下通过盐酸的作用使血浆或红细胞中的铜从其与蛋白质的结合物中分离出来。游离出的铜用吡咯烷二硫代氨基甲酸铵螯合，通过振荡萃取到乙酸正丁酯中，然后将有机萃取物吸入原子吸收分光光度计的火焰中。整个操作过程在聚丙烯离心管中进行，振荡时加盖。对于游离血浆铜的测定，省略了盐酸步骤。发现去除血浆或红细胞蛋白质并无必要，此外，三氯乙酸的存在会导致吸光度明显降低。使用双光束原子吸收分光光度计并将量程扩展10倍，已推导出微量方法，用于测定0.1毫升样品中血浆或红细胞的总铜以及0.5毫升血浆中的游离铜。常量血浆铜方法需要2毫升血浆，适用于单光束原子吸收分光光度计。利用50名献血者的血液，测定了血浆和红细胞铜的正常范围。
