Increase in oxidizing agents in serum causing methaemoglobinaemia and a test for this condition

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Comme on le voit, les conditions de travail peuvent &tre ameliorees grandement d’une facon simple. C’est pourquoi nous avons cru utile de signaler le fait & ceux qui s’interessent au passionnant mais difficile probleme de l’electrophorbe sur papier.

Fig. 2. Au dessus: I’ essai a intensite variable; en dessous: 2’ essai a intensite constante m&me serum. La separation Clectrophoretique est de loin superieure dans le second cas.

avec le

BIBLIOGRAPHIE I R. AUDUBERT ET S. DE MENDE Les principes de l’ilectrophonke, Presses Universitaires de France, Paris, 1957. 2 J. M. FINE, J. GROWLADE, M. SAINT-PAUL ET R. J. TIZZANI,&O~. me'd.(Paris),45 (1956) 591. 3 C. CARAVEL, J. DUCLOZ ET J. GROULADE, Ann. Inst. Pasteur, 78 (1954) 227.

Regu le 4 avril1958

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INCREASE

IN OXIDIZING

METHAEMOGLOBINAEMIA

AGENTS

CAUSING

AND A TEST FOR THIS CONDITION

J. HOMOLKA Department

IN SERUM

*

of Pediatric Biochemistry, Charles University, Prague (Czechoslovakia)

When one part of serum is mixed with three parts of concentrated hydrochloric acid, a red colour may develop within one minute and this can be used photometrically for determination of bilirubin I. Occasionally, when the test is used for detection of bilirubin, a green colour is produced instead of the usual red; we have noticed this 15 times during the last three years. From the biochemical point of view, it is clear that in these particular cases bilirubin is oxidized to biliverdin along with other substances. The reaction may occur immediately or within one minute, but in the serum of other icteric patients it * Address: References $. 604

Zelinarska

‘4, Prague 15, Czechoslovakia.

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does not occur at all or only after 5 (or even more) minutes. Hence the sera which give the g,reen colour quickly have a greater oxidizing activity than the others. The intensity of the green colour naturally depends on the bilirubin concentration. The green colour is obtained most commonly from sera of newborn infants, much less often from older children ; presumably this is due to the raised serum bilirubin concentration during the period immediately after birth. In the last 8 cases in which te green colour was encountered we were able to demonstrate an increased concentration of methaemoglobin either spectroscopically in serum or, by a modified EVELYN-MALLOY method 2, in the red cells. Presumably the increased concentration of oxidizing agents causes methaemoglobinaemia , the methaemoglobin being found either in the red cells or, if there is haemolysis of the .. red cells as often occurs in the newborn, even in the serum. Such cases are often incorrectly diagnosed as icterus or fetal erythroblastosis. In many instances the power of forming the green colour is associated

with a

raised serum catalase activity 3. Thus suggesting, that the oxidising agent present in increased concentration may be of a peroxide character. In a test experiment I drop of 3”//0hydrogen peroxide was added to I ml of serum, after the addition of 3 ml concentrated hydrochlorid acid, a green colour developed within I minute. We suggest the following test in suspected methaemoglobinaemia. Mix one part of icteric serum with three parts of concentrated hydrochloric acid. If the serum to be tested is not icteric, add to it an icteric serum which does not, itself, give the green colour with hydrochloric acid. A positive result consists in the development of the green colour within one minute; most icteric sera give the colour after a prolonged time which should be ignored. Though seldom required since the condition is uncommon, we believe that this test may be valuable

in suitable

cases. REFERENCES

I L. HEILMEYER,

2 R. PODIVINSKY,

Biochem. Z., 296 (1938) 383.

Personal communication. 3 J. HOMOLKA AND V. MYDLIL, Lecture “Methaemoglobinaemia”

Congress of Pediatrics,

Received

Bmo, 1957.

March r8th,

1958