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Variability in potassium/hemoglobin ratios for hemolysis correction.

To the Editor:

Because the increase in plasma potassium in hemolyzed specimens is linearly related to hemoglobin concentration (1), knowledge of the potassium/ hemoglobin ratio can allow estimation of the magnitude of increase in plasma potassium. Previous estimates of the potassium/hemoglobin ratio have evaluated only a few apparently healthy volunteers (2-4). Interindividual differences, particularly in diseased populations, could produce variability in correction factor determination. This letter describes the interindividual variability in the potassium/hemoglobin ratio for in vitro hemolysis in 100 hospitalized patients.

Erythrocyte hemolysates were prepared by the procedure of Meites (5). Briefly, the erythrocytes from lithiumheparin-blood samples from 100 medical and surgical inpatients were separated by centrifugation at 25008 within 1 h of collection, washed three times with isotonic saline, and then lysed by the addition of an equal volume of distilled water. After freezing overnight and thawing, the hemolysates were centrifuged at 25008 for 30 min to remove the stroma.

For each of the 100 patient hemolysate samples, 100 [micro]L of hemolysate was added to 2 mL of a common pooled serum to produce a severely hemolyzed sample. This was diluted in the common pooled serum to form a series of nine equispaced, linearly related samples. Each series of the diluted samples was analyzed in duplicate (same batch) for potassium (reported to two decimal places with an indirect ion-selective electrode) and hemolysis index (multiwavelength spectrophotometry with saline as the reagent) on a Roche Diagnostics 917 clinical chemistry analyzer. Pearson correlation coefficient calculation and Deming model linear regression analysis by Analyse-It add-on software were performed for each dilution series, and the slope of the potassium/ hemoglobin relationship was noted.


The range of hemoglobin concentrations tested was 50-6000 mg/L. Within-batch CVs were as follows: potassium, 0.4%; hemoglobin, 2%. The range of correlation coefficients for the 100 dilution series was 0.99-1.00, confirming the linear relationship between potassium and hemoglobin. The median potassium/ hemoglobin (mmol/g) ratio was 28.0, with a mean of 28.4, and the 95% population limits were 21.0 and 34.5 (Fig. 1). Although the population range included published individual experimental values (1), the median and mean were lower than other individual estimates (1-4, 6, 7). There was also a substantial difference between the 2.5 and 97.5 percentiles, which would create difficulty in the choice of ratio when correcting potassium values for hemolysis in a given case. Thus, because of the variability in the potassium/hemoglobin ratio demonstrated here, the correction of potassium increase in specimens exhibiting in vitro hemolysis is not advocated.


(1.) Jay D, Provasek D. Characterization and mathematical correction of hemolysis interference in selected Hitachi 717 assays. Clin Chem 1993; 39:1804-10.

(2.) Sonntag O. Haemolysis as an interference factor in clinical chemistry. J Clin Chem Clin Biochem 1986;24:127-39.

(3.) Brydon W, Roberts L. The effect of haemolysis on the determination of plasma constituents. Clin Chim Acta 1972;41:435-8.

(4.) Frank J, Bermes E, Bickel M, Watkins B. Effect of in vitro hemolysis on chemical values for serum. Clin Chem 1978;24:1966-70.

(5.) Meites S. Reproducibly simulating hemolysis, for evaluating its interference with chemical methods [Letter]. Clin Chem 1973;19:1319.

(6.) Caraway W. Chemical and diagnostic specificity of laboratory tests. Am J Clin Pathol 1962;37: 445-64.

(7.) Mather A, Mackie N. Effects of hemolysis on serum electrolyte values. Clin Chem 1960;6: 223-27.

Robert Hawkins

Department of Pathology and Laboratory Medicine

Tan Tock Seng Hospital

11 Jalan Tan Tock Seng

Singapore 308433

Fax 65-253-6507; e-mail Robert
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Title Annotation:Letters
Author:Hawkins, Robert
Publication:Clinical Chemistry
Article Type:Letter to the editor
Date:May 1, 2002
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