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Clinical Chemistry 46: 1331-1337, 2000;
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(Clinical Chemistry. 2000;46:1331-1337.)
© 2000 American Association for Clinical Chemistry, Inc.

Articles

Mechanism of Interference of a Polymerized Hemoglobin Blood Substitute in an Alkaline Phosphatase Method

Jeffrey J. Chance1,1, Edward J. Norris2 and Martin H. Kroll1,a

1 Division of Clinical Chemistry, Department of Pathology, and
2 Department of Anesthesiology and Critical Care Medicine, Johns Hopkins University School of Medicine, 600 N. Wolfe St., Baltimore, MD 21287-7065.
a Address correspondence to this author at: Dallas Veterans Affairs Medical Center, 4500 Lancaster Road, 113, Dallas, TX 75216. Fax 214-857-0739; e-mail Martin.Kroll{at}med.va.gov

Background: Hemoglobin-based oxygen carriers can cause profound interferences in many analytical procedures. We determined the mechanism of interference in the assay of alkaline phosphatase activity and identified approaches that might be used to correct for this interference.

Methods: Interference of a polymerized hemoglobin blood substitute with the assay of alkaline phosphatase was examined with a Hitachi 917 analyzer and ultraviolet-visible spectrophotometry.

Results: Hemoglobin-based oxygen carrier solutions had substantial absorbance at 415 nm, the wavelength of analysis used to measure the formation of 4-nitrophenol. In addition to offsetting the initial absorbance at the analytical wavelength, polymerized hemoglobin gave rise to a strong negative interference plot because of alkali denaturation of the substitute. The same interference mechanism was also observed for native hemoglobin (hemolysate), indicating that the interference was not derived from the polymerization process. The interference can be corrected by implementing a rate-correction procedure, or the interference can be avoided by measurement at 450 nm.

Conclusions: The interference of polymerized hemoglobin in the alkaline phosphatase assay is a result of an absorbance offset caused by alkali denaturation of hemoglobin. The interference can be corrected or avoided by modifying the calculation or the analytical wavelength. The correction strategy may also be applicable to improving the hemolysis index for this method.




The following articles in journals at HighWire Press have cited this article:


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 Anesth. Analg.Home page
J. S. Jahr, S. Osgood, S. J. Rothenberg, Q.-L. Li, A. W. Butch, R. Gunther, A. Cheung, and B. Driessen
Lactate Measurement Interference by Hemoglobin-Based Oxygen Carriers (Oxyglobin(R), Hemopure(R), and HemolinkTM)
Anesth. Analg., February 1, 2005; 100(2): 431 - 436.
[Abstract] [Full Text] [PDF]


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