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Viollier Et Al - 2000 - The Ferrozine Method Revisited
Viollier Et Al - 2000 - The Ferrozine Method Revisited
Abstract
The original ferrozine method has been modied to sequentially determine the Fe(II)/Fe(III) speciation in small
volumes of fresh and marine water samples, at the submicromolar level. Spectrophotometric analyses of the Fe(II)
ferrozine complex are performed on a single aliquot before and after a reduction step with hydroxylamine. The
procedure is calibrated using Fe(III) standards stable under normal conditions of analysis. It is shown also that the
presence of high concentrations of dissolved NOM (natural organic matter) do not create any signicant artifacts.
The method was used to measure Fe(II) and Fe(III) depth distribution in salt marsh pore waters and in a stratied
marine basin. # 2000 Elsevier Science Ltd. All rights reserved.
0883-2927/00/$ - see front matter # 2000 Elsevier Science Ltd. All rights reserved.
PII: S 0 8 8 3 - 2 9 2 7 ( 9 9 ) 0 0 0 9 7 - 9
786 E. Viollier et al. / Applied Geochemistry 15 (2000) 785790
Fe(III) species) are generally performed on two separ- A1 eFeII lCFeII eFeIII lCFeIII 1
ate sample fractions (Fadrus and Maly, 1975). Electro-
active Fe(II) concentration is also measured by volta- where A1 is the measured absorbance before the re-
metric methods complemented by an independent de- duction step, eFe(II) and eFe(III) are molar absorption
termination of total Fe (Bue et al., 1987). More coecients, l is the optic path length and CFe(II) and
recently, Elrod et al. (1991) developed a method of CFe(III) are the Fe species concentrations.
detection based on the work of Yamada et al. (1985) After addition of a reducing agent and a buer,
to measure subnanomolar levels of Fe(II) and Fe(III) Fe(III) reduces to Fe(II), thereby increasing the con-
(after reduction to Fe(II) with ascorbic acid) in ocean centration of Fe(II)ferrozine complex in solution. The
water by ow injection analysis, coupled to a chemilu- additive property of the LambertBeer law yields the
minescence detector. A major drawback of these absorbance:
approaches is the requirement for a separate analysis
of dissolved Fe(II) and total dissolved Fe (Fe(III) is A2 eFeII lCFeII CFeIII a 2
calculated by dierence).
The concentrations of total Fe measured on ltered where A2 is the measured absorbance after the re-
samples (0.45 mm pore size) from anoxic environments duction step and a is the dilution factor due to ad-
are generally interpreted as representing the concen- dition of the reducing agent and buer. The simple
trations of Fe(II) species (e.g. Albrechtensen and linear system of Eqs. (1) and (2) is solved for the
Christensen, 1994). However, O2 contamination during Fe(II) and Fe(III) concentrations:
sampling and ltration may induce fast precipitation
of Fe(III) oxyhydroxides and decrease the dissolved A1 eFeII la A2 eFeIII l
Fe(II) concentration. Moreover, when collecting natu- CFeII 3
eFeII laeFeII l eFeIII l
ral waters, contamination by colloidal particles may
occur. The preservation of samples by acidication
may cause the dissolution of these particles, and hence
A2 A1 a
increase the total dissolved Fe concentration. These CFeIII 4
aeFeII l eFeIII l
two eects are not reproducible and emphasize the
need for an immediate speciation measurement, prefer-
ably on a single sample fraction. Calibration curves with Fe(III) standards allow one
The authors have designed an experimental protocol to derive the values of eFe(II)l at A1 and eFe(II)la at A2
to work with sample volumes of 1 ml or less in order (Fig. 1). Due to the successive additions of reagents
to determine the dissolved concentrations of Fe(II) and during the procedure, the uncertainty on the calculated
Fe(III) at the submicromolar level. The method, suit- value of a may become important. However, an accu-
able for eld work, consists of recording the absor- rate value of a can be obtained directly during the pro-
bances of an Fe(II) colored complex before and after cedure: one has to repeat the reduction step for the
an Fe(III) reduction step. standards and record A2' . a is the coecient of propor-
tionality between A'2 and A2.
2. Theory
3. Experimental
3.1. Reagents
Fig. 4. (a) Standard addition of Fe(III) to an organic-rich water sample (experimentally derived salt marsh pore water). (b) Stan-
dard addition of Fe(III) to an organic-rich water sample (25 mg C/l from Suwannee River, GA).
A potential problem with the classical ferrozine the reduction step and the absence of NOM-bound Fe
method is the incomplete reduction of organically com- precipitation. Though reduction of the ferric ferrozine
plexed Fe(III), as pointed out by Luther et al. (1996). complex in the presence of humic substances could po-
These authors also attribute a poor recovery of total tentially lead to an artefact, no signicant change in
Fe to the precipitation of Fe(III) and Fe(II) humic absorbance with time was observed in the experiment.
complexes upon acidication of the sample by hydro- Thus, the proposed method can be used to measure
xylamine hydrochloride. In order to test the reliability Fe(II)/Fe(III) speciation, even in organic-rich natural
of the proposed analytical protocol in the presence of waters. The method detection limit (MDL) is estimated
organically complexed Fe(III), standard additions of to 0.3 mmol/l with a 1 cm cell. Tests on a 20 mmol/l Fe
Fe(III) were performed to two water samples rich in (III) solution prepared in NaCl 0.7 M lead to 20.1 2
dissolved organic matter (NOM). 0.2 mmol/l Fe(III) and 0.1 2 0.1 mmol/l Fe(II) n 5).
A pore water sample was extracted under anaerobic Only slight absorbance dierences occur for salinity
conditions from a ow-through reactor containing an above 5-. Salinity, when above this latter value,
undisturbed core section of a salt marsh sediment should be known within 220%. Interferences with
(Roychoudhury et al., 1998). The presence of relatively major inorganic ions are described in Stookey (1970).
high levels of dissolved NOM was apparent from the
yellow to light brown color of the pore waters. Dis-
solved organic C (DOC) concentrations in marshes are 4.2. Applications
among the highest found in natural environments,
typically in the range 15 to 30 mg C/l (Leenheer, The proposed method was used to measure the
1994). A second water sample was prepared by adding depth distributions of Fe(II) in anoxic pore waters
NOM extracted by reverse osmosis from the Suwannee retrieved by dialysis from a salt marsh (Skidaway
River (Georgia, USA) (SR) to deionized water (1409 Island, Georgia), and in the water column of the Orca
mg C/l SR-NOM sample provided by E.M. Perdue). Basin (Gulf of Mexico) whose deeper part contains an
The original total dissolved Fe content (363226 mmol/ anoxic brine (salinity 250-) (Shokes et al., 1977).
l) was determined using graphite furnace atomic Iron(II) concentrations show the expected depth dis-
absorption (GFFAS) with 800 times sample dilution. tribution in the salt marsh pore waters (Fig. 5). Iro-
The nal DOC concentration of the solution was 25 n(II) produced by reductive dissolution right below the
mg/l. The SR-NOM was chosen because of its high sedimentwater interface is removed from the pore
abundance of strong cation-binding functional groups waters by formation of FeS in the lower part of the
(Cantrell et al., 1990; Gu et al., 1996; Leenheer, 1998). prole. The amounts of Fe(III) are negligible except
In both NOM-rich solutions, excellent linearity of for 4 samples in the top of the prole. These anoma-
the standard additions was observed (Fig. 4). The lous concentrations are explained by contamination of
extrapolation of the calibration curve to X axis leads the samples by sediment particles sticking to the dialy-
to 337 2 18 mmol/l for the original SR-NOM Fe con- sis membrane. These particles dissolve after addition of
tent. These observations demonstrate the reliability of the acidic reagent leading to a release of Fe(III) in the
E. Viollier et al. / Applied Geochemistry 15 (2000) 785790 789
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