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Rhodazine D O2/H2O2 assay. O2 and H2O2 concentrations were
determined by the Rhodazine D assay (RDA) (CHEMetrics, Calverton,
Virginia, United States). All Quick-Seal centrifuge tubes containing
cells were centrifuged at 4 8C (2,000 3 g, 10 min) after irradiation.
Supernatants were tested by the RDA. The RDA is suitable for
measuring 0.1¨C1.0 mg/l O2, and 1¨C10 3 105 M H2O2. Mn(III,IV) are
oxidants that will also cause a positive RDA test result. However,
Mn(III,IV) dioxides are insoluble in water (circa-neutral pH) and
readily removed from suspension by centrifugation (2,0003g, 5 min)
or standing. RDA results indicating 1 mg/l or more O2 or 103105 M
or greater H2O2 were also tested by the Indigo Carmine assay
(CHEMetrics), which yields a blue color suitable for measuring higher
concentrations of O2 and H2O2. Once a test solution had filled an
assay vial, the open end was sealed anaerobically with vacuum grease,
and the vial was stored in the dark.
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3Â¥2008-09-27 23:17:04
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