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Department of Natural Resources and Environmental Sciences, University of Illinois, W-321 Turner Hall, 1102 South Goodwin Avenue,
Urbana, IL 61801, USA
b
Department of Chemistry, Federal University of Minas Gerais, Campus Pampulha, Belo Horizonte, MG, Brazil
Received 22 April 2008; accepted in revised form 10 July 2008; available online 24 July 2008
Abstract
The quantitative assay of clay minerals, soils, and sediments for Fe(II) and total Fe is fundamental to understanding biogeochemical cycles occurring therein. The commonly used ferrozine method was originally designed to assay extracted forms
of Fe(II) from non-silicate aqueous systems. It is becoming, however, increasingly the method of choice to report the total
reduced state of Fe in soils and sediments. Because Fe in soils and sediments commonly exists in the structural framework
of silicates, extraction by HCl, as used in the ferrozine method, fails to dissolve all of the Fe. The phenanthroline (phen) method, on the other hand, was designed to assay silicate minerals for Fe(II) and total Fe and has been proven to be highly reliable.
In the present study potential sources of error in the ferrozine method were evaluated by comparing its results to those
obtained by the phen method. Both methods were used to analyze clay mineral and soil samples for Fe(II) and total Fe.
Results revealed that the conventional ferrozine method under reports total Fe in samples containing Fe in silicates and gives
erratic results for Fe(II). The sources of error in the ferrozine method are: (1) HCl fails to dissolve silicates and (2) if the analyte solution contains Fe3+, the analysis for Fe2+ will be photosensitive, and reported Fe(II) values will likely be greater than
the actual amount in solution. Another diculty with the ferrozine method is that it is tedious and much more labor intensive
than the phen method. For these reasons, the phen method is preferred and recommended. Its procedure is simpler, takes less
time, and avoids the errors found in the ferrozine method.
2008 Elsevier Ltd. All rights reserved.
1. INTRODUCTION
Quantitative chemical analysis of clay minerals to reveal
the oxidation state of structural Fe is an essential aspect of
characterizing the eects of Fe oxidation state on important
properties of soils, sediments, and clay minerals. A variety
of methods have been employed and examined (Amonette
et al., 1994, 1998), but the colorimetric methods have become the most widely used because of their simplicity,
accuracy, and precision. Among colorimetric methods, the
phenanthroline method (phen method, Stucki and
Anderson, 1981; Stucki, 1981; Amonette and Templeton,
*
Corresponding author.
E-mail address: jstucki@illinois.edu (J.W. Stucki).
0016-7037/$ - see front matter 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.gca.2008.07.009
5002
and Luther (1994) calibrated the methods needed to selectively extract Fe(III) from amorphous and crystalline
matrices, non-structural Fe(II) in silicates, and acid-volatile
sulde (AVS) forms of Fe from salt marsh sediments and
mineral samples. In general, the more crystalline the Fe
mineral phase, the greater the concentrations of HCl required to eect Fe extraction, leading to a number of studies recommending 0.5, 1, 2, or even 3 N HCl (Caneld,
1988; Leventhal and Taylor, 1990; Jolivet et al., 1992; Wallmann et al., 1993; Zachara et al., 1998).
With regard to determining Fe(II) in silicate minerals,
Kostka and Luther (1994) successfully extracted Fe(II)
from chlorite (a phyllosilicate clay mineral) using 0.5 M
HCl, followed by ferrozine complexation. They recognized,
however, that total mineral dissolution by HFH2SO4 was
necessary for the measurement of total Fe by ferrozine; but,
unfortunately, they measured only total Fe in the HF
H2SO4 extract, leaving open the possibility that some
non-HCl-extractable Fe(II) could have been present in the
chlorite.
Over time the ferrozine method, with HCl as the extracting agent, has become commonly used to assess the reduced
state of Fe in microbiologically active soils and sediments,
including the level of Fe(II) in the silicates. Since Kostka
and Luther (1994) made no claim of complete Fe(II) extraction from chlorite by this method and current practice lacks
the use of HFH2SO4 to determine total Fe(II) and total
Fe, its reliability as a quantitative tool for assessing the redox state of Fe in soils and sediments containing silicate
minerals is in doubt. Such uncertainty has signicant consequences because of the current widespread use of the method without critical assessment of its relevance for the
system(s) to which it is being applied.
The objectives of the present study were to document the
eects of photoreactivity and choice of extraction acid on
results from the ferrozine method applied to the measurement of Fe(II) and total Fe in silicate minerals. Results will
be compared with the phen method under controlled, but
varied, lighting conditions and with or without the use of
HFH2SO4 in the extraction medium.
2. MATERIALS AND METHODS
The materials used in this study were standard reference
clay minerals of varying Fe(II) and total Fe contents and
soils with a mixture of clay minerals and Fe (oxyhydr)oxides. The clay minerals were: Upton, Wyoming,
montmorillonite (American Colloid Company), designated
UP; ferruginous smectite (sample SWa-1, Source Clays
Repository of The Clay Minerals Society), designated S1;
and Uley green nontronite (sample NAu-1, Source Clays
Repository of The Clay Minerals Society), designated N1.
The clay samples were homoionically saturated with Na+,
fractionated to <2-lm particle size, dialyzed by centrifuge
washing to a NaCl concentration of 5 104 M, and freeze
dried prior to use.
Two of these reference clay minerals, namely, UP and
S1, were also prepared in their chemically reduced state,
using the procedure of Stucki et al. (1984), which included
the addition of sodium dithionite to a citratebicarbonate-
5003
5004
dierence between HCl and HFH2SO4 digestion, but revealed a detectable degradation eect on the Fe(II)ferrozine complex. When carried out in the dark (CFHF-D),
results from the conventional ferrozine method were virtually indistinguishable from those carried out under normal
room light (CFHF); however, when the Fe(II)ferrozine
complex was exposed to light for extended periods of time
(424 h), it appeared to degrade increasingly as the total Fe
content increased. The extent of degradation was greater in
room light than in UV light. A condent explanation for
this phenomenon will require further understanding of the
exact photoreactive nature of Feferrozine complexes.
3.2. Fe(II)
The most important use of the conventional ferrozine
method (CFHCl) is to measure the Fe(II) content of a
mixed mineral system. In the present study, however, values
for Fe(II) found by this method were erratic (Fig. 3, Table
2), both surpassing and falling short of results from the phen
method. Total Fe analyses found that HCl produces incomplete dissolution of the sample, which could explain why
some of the Fe(II) values obtained by CFHCl were less
than those obtained by phen. It fails, however, to explain
why the Fe(II) results would be greater than found by phen.
A plausible hypothesis is that some of the Fe dissolved by
HCl is present as Fe(III), which then complexes with ferrozine and is subsequently reduced photochemically to
Feferrozine3 2 if exposed to light. This hypothesis was
tested by carrying out the CFHCl analysis away from normal light, using the same red-lamp system as in the phen
method. Soil samples R4A and R7A were among those that
Table 1
Results for Total Fe analysis of soil and clay mineral samples by the phenanthroline (phen) method and variations of the ferrozine methoda
Sample
CFHCl
(mmol/g)
CFHCl-D
(mmol/g)
CFHF
(mmol/g)
Mean
SD
Mean
Mean
SD
Mean
SD
2.85
0.46
3.1
0.72
0.75
0.769
0.799
1.05
0.432
3.94
2.08
0.9
0.78
0.79
0.84
0.9
2.84
0.50
0.08
0.008
0.2
0.03
0.03
0.004
0.007
0.04
0.006
0.08
0.02
0.2
0.02
0.01
0.02
0.2
0.04
0.03
0.5
0.06
0.06
0.01
0.33
0.01
0.3
0.29
0.01
0.27
0.01
0.03
0.13
0.05
0.02
0.02
0.02
0.03
1.14
0.2
0.02
0.03
0.07
0.04
0.02
0.02
0.01
0.01
0
0.08
CFHF-D
(mmol/g)
CFHF-4R
(mmol/g)
CFHF-24U
(mmol/g)
CFHF-24R
(mmol/g)
Mean
Mean
Mean
Mean
SD
0.19
0.6
0.33
0.36
0.32
0.33
0.30
0.31
0.02
0.3
0.01
0.02
0.002
0.03
0.03
0.09
Mean
SD
0.005
0.72
0.02
0.73
0.02
0.02
0.756
0.003
0.761
0.004
0.34
0.002
0.75
0.65
0.63
0.66
0.08
0.04
0.01
0.05
0.39
3.5
1.72
0.62
0.64
0.62
0.69
1.01
SD
CFHF-4U
(mmol/g)
0.08
0.1
0.06
0.02
0.06
0.03
0.01
0.03
0.35
2.7
0.99
0.65
0.66
0.66
0.64
0.77
SD
0.01
0.2
0.06
0.06
0.05
0.01
0.09
0.04
0.12
0.5
0.69
0.24
0.201
0.20
0.17
0.21
SD
0.02
0.3
0.03
0.02
0.005
0.01
0.02
0.04
0.40
3.2
1.2
0.66
0.66
0.66
0.7
0.70
SD
0.01
0.2
0.06
0.05
0.04
0.06
0.1
0.05
Total Fe
S1A
UPA
N1A
R1A
R4A
R5A
R7A
R2A
UPB
N1B
CAB
R1B
R4B
R5B
R7B
R2B
S1R
UPR
Phen
a
CFHCl: conventional ferrozine; CFHCl-D: CFHCl method carried out in the dark, under red lamps; CFHF: CFHCL method carried out with HFH2SO4 replacing HCl as the digesting
solution; CFHF-D: the same as CFHF, except carried out in the dark, under red lamps; CFHF-4U: the same as CFHF-D, except subsequently exposed to UV light for 4 h; CFHF-4R: the
same as CFHF-D, except subsequently exposed to room light for 4 h; CFHF-24U: the same as CFHF-D, except subsequently exposed to UV light for 24 h; CFHF-24R: the same as CFHFD, except subsequently exposed to room light for 24 h.
5005
5006
Table 2
Results for Fe(II) analysis of soil and clay mineral samples by the phenanthroline (phen) method and variations of the ferrozine method (see
footnote to Table 1 for abbreviations)
Sample
Fe(II)
S1A
UPA
N1A
R1A
R4A
R5A
R7A
R2A
UPB
N1B
CAB
R1B
R4B
R5B
R7B
R2B
S1R
UPR
Phen (mmol/g)
CFHCl
(mmol/g)
CFHCl-D
(mmol/g)
Mean
SD
Mean
SD
0.018
0.063
0.016
0.159
0.161
0.148
0.092
0.134
0.0351
0.005
0.049
0.13
0.14
0.13
0.093
0.122
1.0
0.31
0.004
0.002
0.003
0.006
0.005
0
0.001
0.003
0.0005
0.001
0.002
0.02
0.01
0.02
0.003
0.002
0.2
0.04
0.26
0.050
0.08
0.005
0.20
0.03
0.16
0.1
0.022
0.01
0.04
0.017
0.017
0.010
0.02
0.011
0.46
0.03
0.005
0.01
0.02
0.006
0.007
0.005
0.04
0.008
0
0.02
Mean
SD
0.098
0.086
0.082
0.046
0.005
0.006
0.009
0.007
CFHF (mmol/g)
CFHF-D
(mmol/g)
Mean
Mean
SD
0.152
0.007
0.116
0.008
0.0300
0.0009
0.12
0.110
0.090
0.08
0.02
0.005
0.006
0.01
0.063
0.057
0.061
0.16
0.136
0.11
0.10
0.11
SD
0.006
0.001
0.004
0.03
0.009
0.01
0.03
0.02
5007
Fig. 4. Eect of light exposure on Fe(II) analysis. Phen, phenanthroline method; CFHCl, conventional ferrozine method dissolving with HCl; CFHCl-D, same as CFHCl except samples kept in
the dark.
5008
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