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Determination of the Components in Erythrocytes using an Automated Analyzer by Shinobu Ttmaka, Honami Takada, Chiknra Kurata, Yoshimasa Haga, Yasuyuki Okamoto

Determination of the biochemical components in erythrocytes should provide unique pathophysiological information. We optimized a simple alcohol binding method for the selective removal of hemoglobins from hemolysates, and enabled simultaneous determination of several components in erythrocytes using commercially available assay kits in an automated analyzer. Venous blood was collected in a vacutainer containing lithium heparin. The washed cells were hemolyzed with distilled water, frozen, and then thawed. Nine volumes of the hemolysates were mixed with one volume of Tris-HCl buffer. One volume of n-butanol was then added to nine volumes of the buffered hemolysates. After vigorous mixing, the mixture of n-butanol and hemolysates was left to stand. The butanol-bound hemoglobins were precipitated by centrifugation, and the clear supernatant below the butanol layer was applied directly to an automated analyzer. Using sera, we determined the effects of the hemoglobin removal procedures on the chemical analytes. Sufficient recovery was noted in most analytes, except for several enzyme activities and lipids. Accordingly, we determined five components present in erythrocytes: creatine, potassium, magnesium, and aspartate aminotransferase as well as superoxide dismutase activities in healthy subjects. We suggest that our simple method is applicable to the simultaneous determination of erythrocyte components in routine laboratory tests.