Importance of organic Fe complexing ligands in the Mississippi River plume

As part of a larger program focused on understanding the biogeochemistry of large river plumes, we participated in two expeditions during 2000 to sample the Mississippi River plume. Surface water samples were collected using a trace metal clean towed fish and analyzed for total dissolved Fe, organic Fe complexing ligands and their associated conditional stability constants. The ligands in the river plume have conditional stability constants (log K′_FeL between 10.5 and 12.3 with an average of 11.2 and standard deviation of 0.6) very similar to ligands found in the open ocean. Comparison of high flow and low flow regimes indicates that variability in flow may be the main cause of the variability in Fe concentrations in the plume. The organic Fe complexing ligands are in greatest excess during a time of higher flow. These ligands are responsible for maintaining very high (5 nM) Fe concentrations throughout the plume. Due to complexation with these organic ligands, the concentration of Fe remains above the Fe-hydroxide solubility level until a salinity above 35 is reached where there appears to be a sink for Fe in the less productive waters. Therefore, Fe is transported a great distance from the river source and is available for biological utilization in the coastal zone.     

Thermodynamic characterization of the partitioning of iron between soluble and colloidal species in the Atlantic Ocean

Recent electrochemical measurements have shown that iron (Fe) speciation in seawater is dominated by complexation with strong organic ligands throughout the water column and have provided important thermodynamic information about these compounds. Independent work has shown that iron exists in both soluble and colloidal fractions in the Atlantic Ocean. Here we have combined these approaches in samples collected from a variety of regimes within the Atlantic Ocean. We measured the partitioning of Fe between soluble (< 0.02 μm) and colloidal (0.02 to 0.4 μm) size classes and characterized the concentrations and conditional stability constants of Fe ligands within these size classes. Results suggest that equilibrium partitioning of Fe between soluble and colloidal ligands is partially responsible for the distribution of Fe between soluble and colloidal size classes. However, a significant fraction of the colloidal Fe was inert to ligand exchange as soluble Fe concentrations were generally lower than values predicted by a simple equilibrium partitioning model.In surface waters, strong ligands with conditional stability constants of 10¹³ relative to total inorganic Fe appeared to dominate speciation in both the soluble and colloidal fractions. In deep waters these ligands were absent, and instead we found ligands with stability constants 12–15 fold smaller that were predominantly in the soluble pool. Nevertheless, significant levels of colloidal Fe were found in these samples, which we inferred must be inert to coordination exchange.     

Complexation of mercury by dissolved organic matter in surface waters of Galveston Bay, Texas

The chemical speciation of dissolved mercury in surface waters of Galveston Bay was determined using the concentrations of mercury-complexing ligands and conditional stability constants of mercury-ligand complexes. Two classes of natural ligands associated with dissolved organic matter were determined by a competitive ligand exchange-solvent solvent extraction (CLE-SSE) method: a strong class (L_s), ranging from 19 to 93 pM with an average conditional stability constant (K_HgLs) of 10²⁸, and a weak class (L_w) ranging from 1.4 to 9.8 nM with an average K_HgLs of 10²³. The range of conditional stability constants between mercury and natural ligands suggested that sulfides and thiolates are important binding sites for dissolved mercury in estuarine waters. A positive correlation between the estuarine distribution of dissolved glutathione and that of mercury-complexing ligands supported this suggestion. Thermodynamic equilibrium modeling using stability constants for HgL, HgCl_x, Hg(OH)_x, and HgCl(OH) and concentrations of each ligand demonstrated that almost all of the dissolved mercury (> 99%) in Galveston Bay was complexed by natural ligands associated with dissolved organic matter. The importance of low concentrations of high-affinity ligands that may originate in the biological system (i.e., glutathione and phytochelatin) suggests that the greater portion of bulk dissolved organic matter may not be important for mercury complexation in estuarine surface waters.     

Investigations of iron coordination and redox reactions in seawater using Fe radiometry and ion-pair solvent extraction of amphiphilic iron complexes

Iron coordination and redox reactions in synthetic and coastal seawater were investigated at nanomolar concentrations using ⁵⁹Fe radiometry and ion-pair solvent extraction of iron chelated by sulfoxine (8-hydroxyquinoline-5-sulfonate) and BPDS (bathophenanthroline disulfonate). Using sulfoxine, we determined the rate at which the monomeric Fe(III) hydroxide species present in seawater of pH 8 are complexed by the microbial siderophore deferriferrioxamine B and the synthetic chelator EDTA (ethylenediaminetetraacetic acid). Forward rate constants of 2 × 10⁶M⁻¹s⁻¹ and 20 M⁻¹s⁻¹, respectively, were obtained. The kinetics of these reactions have not been measured previously at pH values near that of seawater. Conditional equilibrium constants measured for the Fe(III)-EDTA system are consistent with published stability constants for EDTA complexes and for Fe(III) hydrolytic equilibria minus the neutral Fe(OH)₃^o species, suggesting it is not quantitatively significant near pH 8. Commercial humic acid was found to have sufficient affinity for iron to compete with Fe(III) hydrolysis in seawater, and limited evidence was obtained for an interaction with dissolved organic matter in coastal seawater.In our investigations of redox reactions using BPDS to trap Fe(II) produced in the medium, we observed enhanced photoreduction of Fe(III) by humic acid as well as reduction induced by solutes released from phytoplankton in seawater of pH 8. Although the method is sensitive enough to work at near-oceanic levels of iron, the difficulty in distinguishing Fe(II) generated by Fe(III)-BPDS interactions from Fe(II) produced by other means limits its utility. This analytical ambiguity may be generalizable to other methods which measure ferrous iron in seawater using Fe(II)-specific ligands.     

Metal partitioning between colloidal and dissolved phases and its relation with bioavailability to American oysters

Kinetics and the extent of metal partitioning between colloidal and dissolved phases and coagulation of metals associated with colloids were examined to determine their effects on the bioavailability of selected metals (Cd, Co, Hg, Ag, Fe, and Zn) to American oysters (Crassostrea virginica) using radiotracer and short term exposure experiments. After dispersion of radiolabeled colloids into low molecular weight (LMW, < 1 kDa) seawater, metal partitioning between dissolved (<1 kDa) and colloidal (1 kDa-0.2 microm) phases resulted in a consistent pattern, with a relatively constant percentage in the colloidal phase for each metal. On average, about 90% of Hg and Fe, approximately 60% of Ag and approximately 40% of Zn, Co, and Cd were measured in the colloidal fraction during a short term exposure experiment, consistent with their partitioning in natural waters. Controlled laboratory experiments carried out in parallel using radioactively tagged colloids showed that coagulation of colloidal species, quantified as the fraction retained by a 0.2 microm filter, was insignificant for most metals under the conditions and time periods of the uptake experiments. The bioavailability of colloidally complexed metals, measured in terms of dry weight concentration factor (DCF, ml g(-1)) and uptake rate constant (ml g(-1) h(-1)), was somewhat depressed compared with their counterpart in the LMW treatment, but could be well predicted from the results of the LMW treatment and metal partitioning. Both DCF values and uptake rate constants were higher in the LMW treatment than in the colloidal treatment. In addition, B-type metals, such as Ag, Hg, and Zn, all had higher values of DCF and uptake rate constants, regardless of treatments, except for Cd which had a lower DCF and uptake rate constant. In contrast, Co and Fe had significantly lower DCF values and uptake rate constants. Most of Hg and Ag (60-80%) were measured in the soft tissue of oysters in both LMW and colloidal treatments. In contrast, 80% of Fe, 75% of Co, and approximately 60% of Cd were observed on the shell, while Zn was found evenly distributed between shell and soft tissue of oysters. These results agree well with the variation pattern of both DCF value and uptake rate constant for these two groups of metals.     

Dissolved iron(III) speciation in the high latitude North Atlantic Ocean

On voyages in the Iceland Basin in 2007 and 2009, we observed low (ca. 0.1 nM) total dissolved iron concentrations [dFe] in surface waters (<150 m), which increased with depth to ca. 0.2–0.9 nM. The surface water [dFe] was low due to low atmospheric Fe inputs combined with biological uptake, with Fe regeneration from microbial degradation of settling biogenic particles supplying dFe at depth. The organic ligand concentrations [L_T] in the surface waters ranged between 0.4 and 0.5 nM, with conditional stability constants (log K′_FeL) between 22.6 and 22.7. Furthermore, [L_T] was in excess of [dFe] throughout the water column, and dFe was therefore largely complexed by organic ligands (>99%). The ratio of [L_T]/[dFe] was used to analyse trends in Fe speciation. Enhanced and variable [L_T]/[dFe] ratios ranging between 1.6 and 5.8 were observed in surface waters; the ratio decreased with depth to a more constant [L_T]/[dFe] ratio in deep waters. In the Iceland Basin and Rockall Trough, enhanced [L_T]/[dFe] ratios in surface waters resulted from decreases in [dFe], likely reflecting the conditions of Fe limitation of the phytoplankton community in the surface waters of the Iceland Basin and the high productivity in the Rockall Trough.Below the surface mixed layer, the observed increase in [dFe] resulted in a decrease of the [L_T]/[dFe] ratios (1.2–2.6) with depth. This indicated that the Fe binding ligand sites became occupied and even almost saturated at enhanced [dFe] in the deeper waters. Furthermore, our results showed a quasi-steady state in deep waters between dissolved organic Fe ligands and dFe, reflecting a balance between Fe removal by scavenging and Fe supply by remineralisation of biogenic particles with stabilisation through ligands.     

Strong ligands for thorium complexation in marine bacteria

The interaction between thorium and marine organisms (cultured heterotrophic bacteria) was experimentally examined by using chemical equilibrium techniques. Thorium (Th) quantitatively reacts with a binding site on bacteria (Alteromonas, Vibrio, Pseudomonas and Flavobacterium) in 0.1 M HCl solution. According to mass balance analysis of adsorption experiment data, Th forms a 1:1 complex with a binding site similar in reactivity among bacteria used in this study, whose conditional stability constants are in the range from 10(6.63) to 10(7.07) M-1 under the experimental conditions of a 0.1 M HCl solution. The mole ratio of the strong ligand to organic carbon in bacteria ranged from 2.3 to 4.3 mmol/mol C. The strong ligand/carbon ratios in bacteria were more than one order of magnitude greater than in phytoplankton, zooplankton or other organic ligands in surface waters. The results suggest that the strong organic ligand reacting with Th is one of the functional groups commonly existing in oceanic microorganisms. The conditional stability constants of the Th complexes with the binding site in marine microorganisms are in the same order of magnitude as that with the strong ligand found in particulate and dissolved organic matter. These findings strongly suggest that the strong ligand in particulate and dissolved organic matter, reacting with trace metals under the conditions of seawater, originates from marine organisms.     

Speciation of metals in the oceans. I. Inorganic complexes in seawater,and influence of added chelating agents

The speciation of the metal ions in the surface waters of the open ocean has been computed by using critical stability constants describing all of the inorganic complexes that can be formed by the metal ions and inorganic ligands present. The stability constants used were adjusted to change the literature values from 0.10 M ionic strength to the 0.70 M ionic strength of seawater by using a new empirical technique involving experimentally observed variations of stability constants with ionic strength for complexes of similar charge types. For stability constants not listed in Critical Stability Constant compilations, values were estimated by using known trends of stability constants of similar complexes. The speciation determinations made include equilibria with suspended solid phases such as hydroxides, carbonates, and basic carbonates of the metal ions present. Perturbation of metal speciation resulting from the addition of some common chelating agents is determined and the results obtained are discussed in terms of the effects on seawater by the introduction of these chelating agents into the environment. In the 10⁻⁹ to 10⁻⁷ range of added chelant, EDTA (ethylenediaminetetraacetate), NTA (nitrilotriacetate), TPP (tripolyphosphate) and CIT (citrate) exist in the ocean primarily as their Ca(II) and Mg(II) chelates. EDTA and NTA are the dominant ligands for Cu(II) and Ni(II), while TPP is extensively complexed to Fe(III). CIT strongly influences Cu(II) speciation, while SSA (sulfosalicylate, a well-known Fe(III) reagent) has no influence on any of these metal ions or their complexes.     

Biological effects on viviparous blenny exposed to chrysene and held in synthetic as well as in natural brackish water

Viviparous blenny (Zoarces viviparous) were exposed to chrysene and held in natural brackish water as well as in two different synthetic brackish waters, all with a salinity of 6.8%. After 10 weeks of exposure a number of different biomarkers was analysed. The result indicated increased ethoxyresorufin-O-deethylase activity and the formation of two different DNA adduct spots in the liver tissue, as a result of the chrysene exposure. Very similar results of the chrysene exposure were obtained in blenny held in the two synthetic brackish waters. No influence of the synthetically made waters compared to the natural brackish water could be observed on the biomarkers investigated in the study.     

A study of metal ion adsorption at low suspended-solid concentrations

A procedure for conducting adsorption studies at low suspended solid concentrations in natural waters (<50 mg l⁻¹) is described. Methodological complications previously associated with such experiments have been overcome. Adsorption of zinc ion onto synthetic colloidal titania (TiO₂) was studied as a function of pH, supporting electrolyte (NaCl) concentration (0·1-0·002 m) and particle concentration (2–50 mg l⁻¹). The lack of success of the Davis Leckie site bonding model in describing Zn(II) adsorption emphasizes the need for further studies of adsorption at low suspended-solid concentrations.     

The rate of oxidation of ferrous iron in seawater and the partition of elements between iron oxides and seawater

At the Minamichita Beach Land (Mihama-cho, Aichi, Japan), seawater is pumped up from underground and is supplied to aquaria. The underground seawater containsca. 2 ppm of Fe (II), 0.1 ppm of Mn (II) and a little dissolved oxygen. Iron oxide is formed in the seawater when aerated. The oxidation rate of Fe (II) was measured to be 1.4×10¹⁴ mol^–3 l ³ min^–1, which is comparable to the lowest values in the literature. The slow rate of Fe (II) oxidation obtained here can be attributed to the presence of organically bound iron in the seawater. The distribution coefficient of cations between seawater and iron oxide phase was in the order of Cu>Ni>Co>Cd>Mn, which is consistent with that predicted from their hydrolysis constants. The adsorption affinity sequence of oxyanions was phosphate >vanadate> molybdate. The difference in phosphate from the prediction of the adsorption theory was attributed to the formation of ferriphosphate on the oxide surface. On the basis of these data, the limitation and usefulness in the application of the distribution coefficients to marine environments were discussed.     

Trace metal adsorption on inorganic solid phases under estuarine conditions

The adsorption of trace metals (Pb, Zn, Cu, Cd and Hg) on calcite, kaolinite and bentonite has been studied in Krka river water of various salinities (S = 3, 20 and 38‰).Model experiments were performed at pH 8 in natural water and in UV-irradiated water samples to determine the importance of natural soluble organic matter. Cold vapour atomic absorption spectrometry was used for the Hg determination, and a differential pulse anodic stripping voltammetry (DPASV) method for determination of other trace metals, using either a hanging mercury drop electrode or a mercury-coated glassy carbon working electrode. The following results have been obtained on calcite:

1. (1) a powerful absorption of Pb and Zn was observed for both natural and UV-irradiated waters at S=20‰;

2. (2) a strong adsorption of Hg was observed in natural water, and significantly lower adsorption in UV-irradiated water at all three salinities, particularly at low salinity;

3. (3) Cd adsorption was of medium intensity in both waters at S=20‰;

4. (4) Cu was practically not adsorbable in both waters at S=20‰ if the total Cu ion concentration was lower than the copper complexing capacity value;

5. (5) Cu adsorption was of a medium intensity at S=38‰. It has not influenced by the EDTA model ligand.

Trace metals Pb, Zn and Cu are adsorbable on kaolinite and bentonite at S=38‰, whereas Cd is practically not adsorbed. The Krka River is a calcareous river, which supplies the sea with calcite and aluminosilicates. In the light of our model experiments, we suggest that the self-purification of the Krka River is remarkable in respect to Pb, Zn and Hg; this occurs to a lesser extent for Cu, and is negligible in the case of Cd.     

Calibration of a chalcogenide glass membrane ion-selective electrode for the determination of free Fe in seawater: I. Measurements in UV photooxidised seawater

Calibration of a chalcogenide glass membrane, Fe(III)ISE [Fe_2.5(Ge₂₈Sb₁₂Se₆₀)_97.5], in buffered saline media has been undertaken in order to assess the suitability of this ISE for seawater analyses. The electrode slopes in saline citrate and salicylate buffers were 26.3 and 28.2 mV/decade, respectively, for Fe³⁺ concentrations ranging from 10⁻¹⁰ M to less than 10⁻²⁵ M Fe³⁺. The calibration lines in the citrate and salicylate buffers were essentially collinear with the response in unbuffered chloride-free standards containing >10⁻⁵ M Fe³⁺, demonstrating that the response of the FeISE is unaffected by chloride ions. A mechanism involving a combination of charge transfer and ion-exchange of Fe(III), at the electrode diffusion layer, can be used to explain the ≈30 mV/decade slope of the FeISE. The response of the FeISE in UV photooxidised seawater containing 8 nM total Fe was measured as the pH was changed from 8.27 to 3.51. The slope of the response was 24.2 mV/decade [Fe³⁺] calculated as a function of pH using Fe(III) hydrolysis constants for seawater. Moreover, the response was essentially collinear with that in citrate buffers and in unbuffered solutions containing >10⁻⁵ M Fe³⁺ and the slope for the combined data was 26.2 mV/decade. This study was restricted to organic-free seawater because the certainty in Fe(III)–ligand stability constants is insufficient to warrant the selection of an ideal calibration buffer system, and there is evidence that powerful chelating ligands (e.g., EDTA along with humic and fulvic acids) may alter the response of the Fe(III)ISE. The Fe dissolution rate of the FeISE in UV photooxidised seawater was found to be 1.6×10⁻² nmol Fe/min, as measured by cathodic stripping voltammetry (CSV). This would contaminate a 100-ml sample by 0.8–1.6 nM Fe over a typical measurement period of 5–10 min obtained using a stability criterion of 0.5 mV/min. Various methods are proposed for reducing the level of contamination in open ocean samples that contain sub-nanomolar concentrations of iron. The FeISE has the potential to detect free Fe³⁺ at concentrations typically found in natural seawater.     

Cellular iron contents of plankton during the Southern Ocean Iron Experiment (SOFeX)

Iron (Fe) availability limits phytoplankton biomass and production in large regions of the Southern Ocean and influences community composition and size structure, which may affect C export and other system-level functions. To improve our understanding of Fe partitioning within communities and the response of different components to fertilization, we assessed the cellular Fe contents of individual diatoms, autotrophic flagellates, and heterotrophic flagellates during the recent Southern Ocean Fe Experiment using synchrotron-based X-ray fluorescence (SXRF). Dual ⁵⁵Fe/¹⁴C radioisotope incubations were also conducted to estimate Fe:C ratios in size-fractionated plankton. Cellular Fe quotas determined by the two techniques were in close agreement when low amounts of ⁵⁵Fe (0.2 nM) were added, but ⁵⁵Fe additions of 2 nM resulted in 2–3-fold higher quotas. SXRF assessments of cellular Fe quotas (normalized to C) were generally in good agreement with prior bulk analyses of natural assemblages, but revealed compositional differences among protistan taxa not previously detected. Mean Fe:C ratios for diatoms, autotrophic flagellates, and heterotrophic flagellates from unfertilized waters were 6.0, 8.7, and 14.1 μmol mol C⁻¹, respectively. Smaller cells had higher Fe:C ratios than larger cells. Fertilization enhanced Fe quotas in all cell types, with mean Fe:C ratios increasing approximately 4-fold (from about 10 to about 40 μmol mol C⁻¹) after two Fe additions. This study provides some of the first measurements of Fe quotas in phytoplankton cells from natural communities and the first measurements of Fe quotas in natural protozoa.     

Determination of conditional stability constants of organic copper and zinc complexes dissolved in seawater using ligand exchange method with EDTA

To clarify the nature of organic metal complexes dissolved in seawater, a ligand exchange reaction between ligands of natural origin and an aminopolycarboxylic acid (EDTA) was used to determine the conditional stability constants of organic metal complexes. The results indicate that more than two organic molecules complexed with copper and zinc exist in surface seawater. It is found that the conditional stability constants of these naturally-occurring organic metal complexes are 1–3 orders of magnitude higher than those of EDTA-Cu and EDTA-Zn complexes. These estimates of the conditional stability constants for the dominant species of organic copper and zinc complexes are 10^11.8 and 10^9.3, respectively, at pH 8.1. The results indicate that these naturally-occurring organic metal complexes are stable species and not easily dissociated or displaced with others in the marine environment.     

Zinc speciation in the Northeastern Atlantic Ocean

Measurements of zinc and zinc complexation by natural organic ligands in the northeastern part of the Atlantic Ocean were made using cathodic stripping voltammetry with ligand competition. Total zinc concentrations ranged from 0.3 nM in surface waters to 2 nM at 2000 m for open-ocean waters, whilst nearer the English coast, zinc concentrations reached 1.5 nM in the upper water column. In open-ocean waters zinc speciation was dominated by complexation to a natural organic ligand with conditional stability constant (log K_ZnL′) ranging between 10.0 and 10.5 and with ligand concentrations ranging between 0.4 and 2.5 nM. The ligand was found to be uniformly distributed throughout the water column even though zinc concentrations increased with depth. Organic ligand concentrations measured in this study are similar to those published for the North Pacific. However the log K_ZnL′ values for the North Atlantic are almost and order of magnitude lower than those reported by Bruland [Bruland, K.W., 1989. Complexation of zinc by natural organic-ligands in the central North Pacific. Limnol. Oceanogr., 34, 269–285.] using anodic stripping voltammetry for the North Pacific. Free zinc ion concentrations were low in open-ocean waters (6–20 pM) but are not low enough to limit growth of a typical oceanic species of phytoplankton.     

Rare earth elements in seawater during an iron-induced phytoplankton bloom of the western subarctic Pacific (SEEDS-II)

During an iron-enrichment experiment in the western subarctic Pacific (SEEDS-II), concentrations of dissolved and acid-soluble rare earth elements (REEs) were determined in shallow waters. Detailed vertical profiles of dissolved REEs were compared with those of nutrients in a preliminary survey. The results showed good correlation, except for Ce and Y. Along with phytoplankton growth within the iron-enriched patch, dissolved REEs were decreased in surface waters. The changes in REE concentrations were small but systematic: light REEs were adsorbed preferentially over heavy REEs. This systematic trend is consistent with results of earlier studies of REE adsorption onto biogenic particle surfaces. However, we were unable to detect a decrease of acid-soluble REEs in surface waters. During the phytoplankton bloom decline phase, dissolved REEs were elevated to concentration levels at the preliminary survey, although nutrients were decreased. The liberation of REEs from the suspended particles might be related to disaggregation from large particles to colloid particles, desorption from the particulate phase, and remineralization from organic matter. Outside the Fe-patch, time variations of REEs were also observed during SEEDS-II, indicating that both artificially induced phytoplankton blooms and natural variation of biogenic particles can affect the biogeochemical cycles of REEs in shallow waters within a short time.     

Cu complexation by organic ligands in the sub-arctic NW Pacific and Bering Sea

Cu speciation was characterized at three stations in the sub arctic NW Pacific and Bering Sea using cathodic stripping voltammetry with the competing ligands benzoylacetone and salicylaldoxime. A single ligand model was fit to the titration data, yielding concentrations throughout the water column of ∼3–4 nM, and conditional stability constants ranging from 10^12.7 to 10^14.1, this range being partly due to the choice of competing ligand. Free Cu²⁺ in surface waters was 2–4×10⁻¹⁴ M, in close agreement with values reported by previous workers in the NE Pacific using anodic stripping voltammetry (ASV). However, those results showed that complexation by strong organic ligands becomes unimportant below 200–300 m, while our data indicated Cu is strongly complexed to depths as great as 3000 m. Free Cu²⁺ concentrations in surface waters reported here and in previous work are close to the threshold value where Cu can limit the acquisition of Fe by phytoplankton.     

Collection and detection of natural iron-binding ligands from seawater

Iron (Fe) is an essential element for the biochemical and physiological functioning of terrestrial and oceanic organisms, including phytoplankton, which are responsible for the primary productivity in the world's oceans. However, due to the low solubility of Fe in seawater, phytoplankton are often limited by their inability to incorporate enough Fe to allow for optimal growth rates in regions with dissolved Fe concentrations below 1 nM. It has been postulated that certain phytoplankton may produce compounds to facilitate the uptake of Fe from seawater to overcome this limitation. Dissolved Fe in the oceans is overwhelmingly complexed (>99%) by strong organic ligands that may control the uptake of Fe by microbiota; however, the identity, origin, and chemical characteristics of these organic chelates are largely unknown. Although it has been implied that some components of natural Fe-binding ligands are siderophores, no direct analyses of such compounds from natural seawater have been conducted. Here, we present a simple solid-phase extraction technique employing Biobeads SM-2 and Amberlite XAD-16 resins for concentrating naturally occurring dissolved iron-binding compounds from large volumes (>200 l) of seawater. Additionally, we report on the first successful determination of molecular weight size classes and preliminary iron-binding functional group characterization within those size classes for isolates collected from the surface and below the photic zone (150 m) in the central California coastal upwelling system. Electrochemical analyses using competitive ligand equilibration/adsorptive cathodic stripping voltammetry (CLE-ACSV) showed that isolated compounds had conditional Fe-binding affinities (with respect to inorganic iron—Fe′) of K_FeL,Fe′^cond=10^11.5–10^11.9 M⁻¹, similar to purified marine siderophores produced in laboratory cultures and to the ambient Fe-binding ligands observed in seawater. In addition, 63% of the extracted compounds from surface-collected samples fall within the defined size range of siderophores (300–1000 Da). Hydroxamate or catecholate Fe-binding functional groups were present in each compound for which Fe binding was detected. These results illustrate that the functional groups previously shown to be present in marine and terrestrial siderophores extracted and purified from laboratory cultures are also present in the natural marine environment. These data provide evidence that a significant fraction of the organic Fe-binding compounds we collected contain Fe-binding functional groups consistent with biologically produced siderophores. These results provide further insight into characteristics of the Fe-binding ligands that are thought to be important in controlling the biological availability of Fe in the oceans.