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Fast and sensitive chemiluminescence determination of H2O2 concentration in stimulated human neutrophils

✍ Scribed by Mueller, S. ;Arnhold, J.

Publisher
John Wiley and Sons
Year
1995
Weight
773 KB
Volume
10
Category
Article
ISSN
0884-3996

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✦ Synopsis

A fast and sensitive chemiluminescence assay for the determination of H 2 0 2 in stimulated neutrophils without the use of enzymes was developed. The method is based on the oxidation of luminol by hypochlorous acid. The chemiluminescence of this reaction is highly dependent on the concentration of hydrogen peroxide.

Changes in H202 concentration in PMA-stimulated neutrophils were followed by injection of NaOCl t o cell suspension at different times after cell stimulation. The short integration time of 2 s permits calculation of actual concentrations of H202 without influence of H202 decomposition by cellular enzymes or newly produced HzOz due t o dismutation of superoxide anion radicals. Concentrations of H202 were diminished by catalase and enhanced by sodium azide owing t o inhibition of cellular catalase and myeloperoxidase. Changes in H2O2 concentration upon stimulation could be observed at 3000 cells/mL.

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