Clinical Chemistry
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Clinical Chemistry 33: 1767-1770, 1987;
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Clinical Chemistry, Vol 33, 1767-1770, Copyright © 1987 by American Association for Clinical Chemistry

Determining methemoglobin in blood by zero-crossing-point first- derivative spectrophotometry

A Taulier, P Levillain and A Lemonnier
Laboratoire Central de Biochimie, Centre Hospitalier de Bicetre, Kremlin-Bicetre, France.

We determined methemoglobin in blood by zero-crossing-point first- derivative spectrophotometry. After lysis of erythrocytes, hemoglobin was converted into oxyhemoglobin and the first derivative spectrum was recorded between 405 and 425 nm. At the exact point where the first- derivative spectrum of oxyhemoglobin was zero ("zero-crossing point"), the first-derivative value of oxyhemoglobin and methemoglobin mixture was proportional to the methemoglobin concentration. The standard curve was linear for all proportions of methemoglobin. Within-assay precision (CV) was 3.4% for a 20% methemoglobin content. Correlation with results by the Evelyn and Malloy method was very good for high proportions of methemoglobin (greater than 10%), but the proposed technique was far better for low methemoglobin percentages because of its linearity, its high sensitivity, and its low detection limit.




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Copyright © 1987 by the American Association for Clinical Chemistry.