Andrew L. Rose

Kinetics of iron complexation by dissolved natural organic matter in coastal waters

Abstract In coastal waters, where conditions are variable and can change rapidly, the kinetics of iron complexation by NOM is a major factor affecting its speciation. In this study we have examined the kinetics of complexation of ferrous and ferric iron by terrigenous NOM in terms of formation rate constants and dissociation rate constants by employing competitive ligand methods with visible spectrophotometry for determination of the complexed iron. Rate constants for organic ferrous complex formation ranged from 500 to 7.5×10 4 M −1 s −1 , and rate constants for complex dissociation ranged from 1×10 −6 to 3.6×10 −3 s −1 . Formation rate constants for organic ferric complexes were comparable to those determined for strong iron binding ligands found in the open oceans, and from 2.1×10 5 to 9.6×10 7 M −1 s −1 . Ferric complex dissociation data were fitted by a two ligand model, with rate constants for both ligand classes typically higher than those for the strong ligands in the open ocean. Rate constants varied from 2×10 −4 to 4×10 −3 s −1 for the ‘weak’ ligand class, and from 1.0×10 −6 to 1.3×10 −4 s −1 for the ‘strong’ class. All rate constants varied by several orders of magnitude between NOM samples of different origin, reflecting the highly variable composition of these substances. Calculated conditional stability constants for organic complexes with ferric species in seawater, Fe′, were generally similar to those measured in the open oceans. Conditional stability constants for the ferrous complexes were less than those for the ferric complexes by two to four orders of magnitude. There was a weak positive correlation between the conditional stability constants for ferrous and ferric complexes, suggesting that similar functional groups are involved in binding each of the two forms of iron. One of the samples of NOM had a conditional stability constant with Fe′ that was comparable with those for siderophores and similar strong iron binding compounds. These results suggest that in the absence of oxidants, complexes between ferrous iron and NOM may be relatively long-lived. Our work also suggests that organic complexes between iron and terrigenous NOM may be quite strong, and will have a major effect on iron solubility in coastal waters.

The influence of extracellular superoxide on iron redox chemistry and bioavailability to aquatic microorganisms.

Superoxide, the one-electron reduced form of dioxygen, is produced in the extracellular milieu of aquatic microbes through a range of abiotic chemical processes and also by microbes themselves. Due to its ability to promote both oxidative and reductive reactions, superoxide may have a profound impact on the redox state of iron, potentially influencing iron solubility, complex speciation, and bioavailability. The interplay between iron, superoxide, and oxygen may also produce a cascade of other highly reactive transients in oxygenated natural waters. For microbes, the overall effect of reactions between superoxide and iron may be deleterious or beneficial, depending on the organism and its chemical environment. Here I critically discuss recent advances in understanding: (i) sources of extracellular superoxide in natural waters, with a particular emphasis on microbial generation; (ii) the chemistry of reactions between superoxide and iron; and (iii) the influence of these processes on iron bioavailability and microbial iron nutrition.

Methods for reactive oxygen species (ROS) detection in aqueous environments

This review summarizes direct and indirect analytical methods for the detection and quantification of the reactive oxygen species (ROS): 1O2, O2·−/HOO·, H2O2, HO·, and CO3·− in aqueous solution. Each section briefly describes the chemical properties of a specific ROS followed by a table (organized alphabetically by detection method, i.e., absorbance, chemiluminescence, etc.) summarizing the nature of the observable (associated analytical signal) for each method, limit of detection, application notes, and reaction of the probe molecule with the particular ROS.

Determination of superoxide in seawater using 2-methyl-6-(4-methoxyphenyl)-3,7- dihydroimidazo[1,2-a]pyrazin-3(7H)-one chemiluminescence

Superoxide, the one-electron reduced form of dioxygen, is known to be generated in marine environments by photochemical and biological processes. Because of its selective reaction with only a few commonly occurring compounds, superoxide is expected to approach concentrations in the high picomolar or low nanomolar range in seawater. Most currently existing methods do not have both the necessary sensitivity and selectivity to measure naturally occurring concentrations. In contrast, we demonstrate here that the chemiluminescence reagent 2-methyl-6-(4-methoxyphenyl)-3,7-dihydroimidazo[l,2-a]pyrazin-3(7H)-one (MCLA) is selective for superoxide in seawater and can be used with a detection limit of around 50 pM. Although a wide range of potential interferences were shown not to react with MCLA directly, some care must be taken when analyzing samples containing nanomolar concentrations of Fe(II), Cu(I), Mo(V), V(III), or V(IV), since these compounds can react with oxygen to produce superoxide during analysis that is subsequently detected. We describe two methods for calibrating the system, one employing photochemically generated superoxide standards and the other employing the superoxide-generating xanthine/xanthine oxidase system and discuss limitations on the use of each. The method was successfully used in the field to determine steady-state superoxide concentrations in the water column in the eastern equatorial Pacific Ocean.

Predicting iron speciation in coastal waters from the kinetics of sunlight-mediated iron redox cycling

Abstract.Iron is a critical nutrient in marine systems, whose solubility is strongly influenced by the presence of natural organic ligands and sunlight. Because these parameters are never constant, and because they initiate processes that occur on a variety of timescales, equilibrium models are inadequate at predicting iron speciation in the presence of natural organic matter (NOM) and light. Instead, a dynamic redox cycle based on kinetically governed transformations of iron is proposed as a tool for predicting iron speciation. The presence of NOM is a critical inhibitor of precipitation of iron as insoluble oxyhydroxides, and dissociation of organic ferric complexes represents a limiting step in loss of iron from the soluble pool. Sunlight causes photochemical reduction of organic ferric complexes, producing transient amounts of ferrous iron. Although the process drives iron away from the ferric state, photo-oxidation of ligands allows some organically complexed iron to “escape” into the inorganic part of the cycle, and thus increases the rate at which soluble iron is lost from the system. Model predictions of spatial and temporal variations in ferrous iron and hydrogen peroxide concentrations are in excellent agreement with field measurements. These results indicate that detailed kinetic modelling of fundamental chemical processes is an extremely useful approach to prediction of iron speciation at an ecosystem scale.

Effects of pH, chloride, and bicarbonate on Cu(I) oxidation kinetics at circumneutral pH.

The oxidation kinetics of nanomolar concentrations of Cu(I) in NaCl solutions have been investigated over the pH range 6.5-8.0. The overall apparent oxidation rate constant was strongly affected by chloride, moderately by bicarbonate, and to a lesser extent by pH. In the absence of bicarbonate, an equilibrium-based speciation model indicated that Cu(+) and CuClOH(-) were the most kinetically reactive species, while the contribution of other Cu(I) species to the overall oxidation rate was minor. A kinetic model based on recognized key redox reactions for these two species further indicated that oxidation of Cu(I) by oxygen and superoxide were important reactions at all pH values and chloride concentrations considered, but back reduction of Cu(II) by superoxide only became important at relatively low chloride concentrations. Bicarbonate concentrations from 2 to 5 mM substantially accelerated Cu(I) oxidation. Kinetic analysis over a range of bicarbonate concentrations revealed that this was due to formation of CuCO(3)(-), which reacts relatively rapidly with oxygen, and not due to inhibition of the back reduction of Cu(II) by formation of Cu(II)-carbonate complexes. We conclude that the simultaneous oxygenation of Cu(+), CuClOH(-), and CuCO(3)(-) is the rate-limiting step in the overall oxidation of Cu(I) under these conditions.

Hydroxyl radical production by H2O2-mediated oxidation of Fe(II) complexed by Suwannee river Fulvic Acid under circumneutral freshwater conditions

The Fenton reaction, the oxidation of ferrous iron by hydrogen peroxide (H(2)O(2)), is typically assumed to be a source of hydroxyl radical (HO(•)) in natural systems, however, formation of HO(•) in this process is strongly dependent upon solution pH and the ligand environment, with HO(•) only formed when Fe(II) is organically complexed. In this study we examine the formation of HO(•) when Fe(II)-NOM complexes are oxidized by H(2)O(2) using phthalhydrazide as a probe for HO(•). We demonstrate that HO(•) formation can be quantitatively described using a kinetic model that assumes HO(•) formation occurs solely from the reaction of Fe(II)-NOM complexes with H(2)O(2), even though this reaction is sufficiently slow to play only a negligible role in the overall oxidation rate of total Fe(II). As such, NOM is seen to play a dual role in circumneutral natural systems in stabilizing Fe(II) toward oxidation by H(2)O(2) while enabling the formation of HO(•) through this oxidation process.

Effect of Light on Iron Uptake by the Freshwater Cyanobacterium Microcystis aeruginosa

Visible light was observed to induce reductive dissociation of organically complexed Fe and dramatically increase the short-term uptake rate of radiolabeled Fe by Microcystis aeruginosa PCC7806 in Fraquil* medium buffered by a single metal chelator, ethylenediaminetetraacetic acid (EDTA). Only wavelengths <500 nm activated Fe uptake indicating that Fe photochemistry rather than biological factors is responsible for the facilitated uptake. The measured rate of photochemical Fe(II) production combined with a significant decrease in (55)Fe uptake rate in the presence of ferrozine (a strong ferrous iron chelator) confirmed that photogenerated unchelated Fe(II) was the major form of Fe taken up by M. aeruginosa under the conditions examined. Mathematical modeling based on unchelated Fe(II) uptake by concentration gradient dependent passive diffusion of Fe(II) through nonspecific transmembrane channels (porins) could account for the magnitude of Fe uptake and a variety of other observations such as the effect of competing ligands on Fe uptake. Steady-state uptake rates indicated that M. aeruginosa acquires Fe predominantly during the light cycle. This study confirms that Fe photochemistry has a dominant impact on Fe acquisition and growth by M. aeruginosa in EDTA-buffered culture medium.

Effect of Fe(II) and Fe(III) transformation kinetics on iron acquisition by a toxic strain of microcystis aeruginosa

We have investigated the mechanism of Fe uptake by a toxic strain of the freshwater cyanobacterium Microcystis aeruginosa (PCC7806) with particular attention given to the effect of Fe(II) and Fe(III) transformation kinetics on Fe uptake. Chemiluminescence analysis revealed that M. aeruginosa produces extracellular superoxide (a moderate Fe reducing agent) at rates of 0.4-1.2 amol cell(-1) h(-1) depending on initial Fe concentration in the culture medium. Short-term assimilation assays using (55)Fe showed that reduction of Fe(III) in both organic and inorganic forms by cell-generated superoxide or ascorbate facilitated Fe uptake via formation of unchelated Fe(II), when Fe availability was low because of the use of the strong Fe chelator ethylenediaminetetraacetate (EDTA) as a ligand. In contrast, Fe reduction was unimportant for Fe uptake in the presence of low concentrations (< or =100 microM) of the weak Fe-binding ligand citrate because of a high concentration of unchelated Fe(III), indicating that the contribution of reduction to Fe uptake depends on the nature of Fe binding and availability of unchelated Fe(III) in the external medium. A kinetic model incorporating uptake of both unchelated Fe(II) and Fe(III) and based on similar models developed for marine microalgae successfully described Fe uptake rates by M. aeruginosa PCC7806.

Phthalhydrazide chemiluminescence method for determination of hydroxyl radical production: modifications and adaptations for use in natural systems

The hydroxyl radical is formed through a variety of processes pertinent to natural and anthropogenic systems. Here we report development of a simple and sensitive trap-and-trigger chemiluminescence method based upon the hydroxylation of phthalhydrazide to 5-hydroxy-2,3-dihydro-1,4-phthalazinedione, which emits chemiluminescence when oxidized under alkaline conditions. Cu(III) is employed as an oxidant and is shown to be relatively insensitive to a range of interferences likely to be encountered. The method has been standardized by use of γ-radiolysis of water as a primary source of hydroxyl radical, with a convenient secondary calibration procedure developed that uses Fenton chemistry. Detection limits varied from 7.4 nM (at pH 3) and 6.2 nM (at pH 8.1) of accumulated hydroxyl radical production in a simple 10 mM NaCl matrix to around 30 nM in an artificial seawater medium, due to competition for hydroxyl between the phthalhydrazide probe and bromide. The method has been used to characterize the kinetics of the Fenton system employed for calibration and is shown to be consistent with published models of this process over time scales of several hours. The application of this method to a range of matrices and for photochemical studies is also described.