Milne, Angela, Davey, Margaret S., Worsfold, Paul J., Achterberg, Eric P. and Taylor, Alison R.
Real-time detection of reactive oxygen species generation by marine phytoplankton using flow -injection chemiluminescence
Limnology and Oceanography: Methods, 7, .
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Little is known about the biological production of reactive oxygen species (ROS) such as H2O2 and O2
– in the
surface waters of the oceans. This article describes two flow injection–chemiluminescence methods to measure
H2O2 and O2
– production by marine diatoms. These methods each incorporated a polycarbonate filter unit in
which live cells were immobilized, followed by downstream determination of H2O2 and O2
– in the cell filtrate. The
sample analysis rate was ~1 min–1 for H2O2 and continuous for O2
– during assays with diatom cells. The respective
detection limits for each system were 1.9 nM (H2O2) and 1.6 nM (O2
–). An initial examination of the effect of
changing light intensity showed that a rapid light-induced production of both O2
– and H2O2 by T. weissflogii cells
could be readily detected. Moreover, this production was proportional to the biomass present on the flushed filter.
These methods enable the monitoring of real-time fluctuations of biological ROS production in response to
changing environmental conditions, and therefore facilitate analysis of the biotic component of ROS production
and the subsequent impacts on chemical speciation of nutrients and trace metals in aquatic ecosystems.
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