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Anal Biochem. 2013 May 15;436(2):66-8. doi: 10.1016/j.ab.2013.01.024. Epub 2013 Jan 31.

Overcoming matrix effects in the chemiluminescence determination of extracellular adenosine triphosphate in erythrocyte suspensions.

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  • 1Department of Chemistry and Biochemistry, Concordia University, Montreal, Quebec, Canada H4B 1R6.

Abstract

As erythrocyte-derived extracellular adenosine triphosphate (ATP) gains recognition as a key vasodilator, its accurate determination is critical. Erythrocytes' high hemoglobin content can act as an inner filter when measuring ATP concentrations by chemiluminescence. We evaluated two approaches to correct for this matrix effect: addition of cell-free hemoglobin to the ATP standards and standard addition of ATP to erythrocyte suspensions. In addition, we reduced sample hematocrit to minimize the absorbance. We conclude that extracellular ATP should be determined in erythrocyte suspensions at 0.06 to 0.004% hematocrit. This gives robust signals without matrix effects and requires only microliters of blood.

Crown Copyright © 2013. Published by Elsevier Inc. All rights reserved.

[PubMed - indexed for MEDLINE]
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