Mechanism of interference of a polymerized hemoglobin blood substitute in an alkaline phosphatase method

J. J. Chance, E. J. Norris, M. H. Kroll

Research output: Contribution to journalArticle

Abstract

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. (C) 2000 American Association for Clinical Chemistry.

Original languageEnglish (US)
Pages (from-to)1331-1337
Number of pages7
JournalClinical Chemistry
Volume46
Issue number9
StatePublished - 2000
Externally publishedYes

Fingerprint

Blood Substitutes
Alkaline Phosphatase
Hemoglobins
Assays
Alkalies
Denaturation
Wavelength
Ultraviolet Spectrophotometry
Oxygen
Hemolysis
Polymerization
Spectrophotometry

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Mechanism of interference of a polymerized hemoglobin blood substitute in an alkaline phosphatase method. / Chance, J. J.; Norris, E. J.; Kroll, M. H.

In: Clinical Chemistry, Vol. 46, No. 9, 2000, p. 1331-1337.

Research output: Contribution to journalArticle

Chance, J. J. ; Norris, E. J. ; Kroll, M. H. / Mechanism of interference of a polymerized hemoglobin blood substitute in an alkaline phosphatase method. In: Clinical Chemistry. 2000 ; Vol. 46, No. 9. pp. 1331-1337.
@article{d9945ec5ccca47b890d8b77d7291b979,
title = "Mechanism of interference of a polymerized hemoglobin blood substitute in an alkaline phosphatase method",
abstract = "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. (C) 2000 American Association for Clinical Chemistry.",
author = "Chance, {J. J.} and Norris, {E. J.} and Kroll, {M. H.}",
year = "2000",
language = "English (US)",
volume = "46",
pages = "1331--1337",
journal = "Clinical Chemistry",
issn = "0009-9147",
publisher = "American Association for Clinical Chemistry Inc.",
number = "9",

}

TY - JOUR

T1 - Mechanism of interference of a polymerized hemoglobin blood substitute in an alkaline phosphatase method

AU - Chance, J. J.

AU - Norris, E. J.

AU - Kroll, M. H.

PY - 2000

Y1 - 2000

N2 - 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. (C) 2000 American Association for Clinical Chemistry.

AB - 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. (C) 2000 American Association for Clinical Chemistry.

UR - http://www.scopus.com/inward/record.url?scp=0033817334&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033817334&partnerID=8YFLogxK

M3 - Article

VL - 46

SP - 1331

EP - 1337

JO - Clinical Chemistry

JF - Clinical Chemistry

SN - 0009-9147

IS - 9

ER -