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Interference of Methylene Blue with CO-Oximetry of Hemoglobin Derivatives

¹ Lab. de Toxicol., and
² Réanimation Toxicol., Hôpital Fernand Widal, 200 rue du Fg-St-Denis, 75475 Paris Cedex 10, France;
³ C.N.E.H., 9 rue Antoine Chantin, 75014 Paris, France;
^a author for correspondence: fax +33 1 40 05 48 78

Methylene blue (MB) is frequently used as an antidote in treating methemoglobinemia (1) because it facilitates the reducing activity of the NADPH-dependent methemoglobin reductase system in erythrocytes (2). However, MB absorbs strongly between 550 and 700 nm (Fig. 1 ), the same spectrophotometric region as that of the various hemoglobin derivatives: oxyhemoglobin (O₂Hb), deoxyhemoglobin (HHb), methemoglobin (MetHb), and carboxyhemoglobin (COHb). To evaluate the potential magnitude and direction of errors linked to the presence of MB for the results for total hemoglobin (tHb) and its derivatives, we evaluated six CO-Oximeters. The wavelengths used by each instrument for these determinations are as follows: IL 482 (Instrumentation Laboratory, Lexington, MA), 535, 585.2, 594.5, and 626.6 nm; CCD 270 (Chiron Diagnostics, Medfield, MA), 557, 577, 597, 605, 624, 635, and 650 nm; CCD 835 (Chiron; wavelengths not communicated); OSM3 (Radiometer, Copenhagen, Denmark), 535, 560, 577, 622, 636, and 670 nm; ABL 520 (Radiometer; same wavelengths as OSM3); AVL 912 (AVL Scientific Corp., Roswell, GA), 530, 536, 542, 548, 554, 560, 566, 572, 578, 584, 590, 604, 612, 622, 630, 640, and 648 nm.

View larger version (12K): [in this window] [in a new window]   Figure 1. Spectra of hemoglobin derivatives and methylene blue. The absorbance of MB in physiological (isotonic) saline solution peaks at 663 nm.

Blood was collected from five healthy volunteers with informed consent. Because the study involved only blood sampling, Institutional Ethics Committee Review was not required in France. The five samples were combined to obtain 120 mL of pooled blood, which were then separated into three 40-mL fractions:

–Fraction N, which had no enrichment in CO or MetHb.

–Fraction CO, which was enriched in CO by tonometry with use of an IL 237 tonometer (Instrumentation Laboratory) and CO in nitrogen, 10 mL/L (Société Cosma, Igny 91430, France). Because . . . [Full Text of this Article]

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				B. H. A. Maas, A. Buursma, R. A. J. Ernst, A. H. J. Maas, and W. G. Zijlstra Lyophilized bovine hemoglobin as a possible reference material for the determination of hemoglobin derivatives in human blood Clin. Chem., November 1, 1998; 44(11): 2331 - 2339. [Abstract] [Full Text] [PDF]