Spectrophotometric determination of serum iron at the submicrogram level with a new reagent (ferrozine).

Penicillamine therapy was discontinued for a period of 200 days in a patient with Wilson's disease. Copper reaccumulated in the body at the rate of about 0.43 mg/day. Urinary copper remain...

Hemolytic Anemia in Wilson's Disease

RAGAN, H. A., S. NACHT, G. R. LEE, C. R. BISHOP, AND G. E. CARTWRIGHT. Effect of ceruloplasmin on plasma iron in copper-deficient swine. Am. J. Physiol. 217(5) : 1320-1323. 1969.-The administration of homologous ceruloplasmin to copper-deficient swine with adequate iron stores resulted in a rapid, marked, and sustained increase in plasma iron concentration. An amount of inorganic copper equivalent to that contained in the ceruloplasmin produced only a minimal, transient increase in plasma iron. Ferrokinetic studies following ceruloplasmin infusion indicated that the increase in serum iron resulted from an increase in the rate of flow of iron into plasma. These studies support the hypothesis that ceruloplasmin, possibly because of its ferroxidase activity, directly affects the transfer of iron from cells to plasma.

Effect of ceruloplasmin on plasma iron in copper-deficient swine.

Five patients with Wilson's disease were treated initially with potassium sulfide and penicillamine and maintained on penicillamine for 9 to 13 years. All patients are now clinically well....

Long-Term Therapy of Wilson's Disease

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.

https://jcp.bmj.com/content/jclinpath/22/2/136.full.pdf

Micro determination of plasma and erythrocyte copper by atomic absorption spectrophotometry

Methods are described for the determination of microgram quantities of biological copper in aqueous medium as the intensely coloured oxalyldihydrazide-acetaldehyde complex (molar extinction coefficient 23,000 to 23,500). The methods are applicable on a routine basis to any biological material, in particular to serum, urine, or tissues, such as liver, brain, or kidney. In the case of urine a simple semi-quantitative screening method is also described. For quantitative work copper is liberated from serum by acid extraction and protein precipitation, and from urine and tissues by a rapid wet-ashing procedure. Recoveries of added copper from urine are quantitative. Precision is high although day-to-day control with standards and blanks is desirable. The methods are applied in an investigation of the diurnal variations in a normal man and in two cases of Wilson's disease.

https://jcp.bmj.com/content/jclinpath/17/3/260.full.pdf

R. N. Beale

Papers

The microdetermination of biological copper with oxalyldihydrazide