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Clinical Chemistry 28: 508-511, 1982;
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Clinical Chemistry, Vol 28, 508-511, Copyright © 1982 by American Association for Clinical Chemistry

Methemoglobin in blood as determined by double-wavelength spectrophotometry

M Sakata, A Yoshida and M Haga

We describe spectrophotometric determination of methemoglobin (Met-Hb) in blood by absorbance differences at two wavelengths (characteristic of the instrument used) in the 500-600 nm region. Blood is diluted 100- to 200-fold with a solution containing KCN and carbon monoxide to convert hemoglobin to carboxyhemoglobin and Met-Hb to cyanomethemoglobin (CN-Met-Hb). Carboxyhemoglobin has the same absorbance at the two wavelengths used, so that the differences in absorbance reflect only the CN-Met-Hb component. After measurement of absorbance differences, potassium ferricyanide is added to convert all hemoglobin derivatives to CN-Met-Hb; samples are then remeasured at the same wavelengths. The percentage of Met-Hb is determined from the ratio of the absorbance differences. Results agree satisfactorily with currently accepted procedures and with calculated theoretical results for mixtures of oxyhemoglobin, carboxyhemoglobin, and Met-Hb.




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Copyright © 1982 by the American Association for Clinical Chemistry.