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Chemosphere 70 (2007) 237–247

www.elsevier.com/locate/chemosphere

Effects of organic matter on the distribution of uranium

in soil and plant matrices
a,*
A.J. Bednar , V.F. Medina a, D.S. Ulmer-Scholle b, B.A. Frey c, B.L. Johnson a,
W.N. Brostoff d, S.L. Larson a
a
US Army Corps of Engineers, Engineer Research and Development Center, 3909 Halls Ferry Road, Vicksburg, MS 39180, United States
b
Department of Earth and Environmental Sciences, New Mexico Institute of Mining and Technology, Socorro, NM 87801, United States
c
New Mexico Bureau of Geology and Mineral Resources, New Mexico Institute of Mining and Technology, Socorro, NM 87801, United States
d
US Army Corps of Engineers, San Francisco District, 1455 Market Street, San Francisco, CA 94103-1298, United States

Received 22 September 2006; received in revised form 11 June 2007; accepted 14 June 2007
Available online 20 August 2007

Abstract

This work studied interactions of uranium with pure organic compounds, such as glutathione, and more complex mixtures, such as
humic acid and aqueous plant extracts. High performance liquid chromatography with UV absorption interfaced to inductively coupled
plasma mass spectrometry sequential detection was used to detect organouranium complexes in a variety of soils and plant materials,
indicating that nearly 100% of the uranium extracted from certain plant tissues was bound to organic ligands. In addition, soil sorption
experiments indicated that humic acid generally decreased uranium sorption to soils and promoted subsequent desorption of uranium
because of uranium partitioning to the organic phase. These experiments demonstrate that organic compounds influence the mobility
and chemistry of uranium in the environment.
Published by Elsevier Ltd.

Keywords: Uranium; Complexation; Organic matter; Speciation

1. Introduction 1997; Casas et al., 1998). Uranium (IV) generally exists

Natural organic matter (NOM) contains functional
groups that can form complexes with metals, and these
complexes can affect the physical and chemical properties
of the affected metal (Langmuir, 1997; Christensen et al.,
1999; Nierop et al., 2002; Schmitt et al., 2002) or radionu-
clide (Czerwinski et al., 1994; Zhang et al., 1997; Lenhart
et al., 2000; Pacheco and Havel, 2001). Complexation
affects metal and radioisotope mobility, sorption to soils,
and bioavailability (Zhang et al., 1997; Morton et al.,
2002). Inorganic reactions on natural uranium ore deposits
can produce redox states of +4 and +6, which have dra-
matically different geochemical properties (Langmuir,
*
Corresponding author. Tel.: +1 601 634 3652.
E-mail address: Anthony.J.Bednar@erdc.usace.army.mil (A.J. Bed-
nar).
as an insoluble oxide, although it can be complexed with
various ligands including NOM. Uranium (VI), however,
is soluble in natural waters as the uranyl ion ðUO2þ2 Þ and
has an extensive complexation chemistry with inorganic
and organic moieties (Czerwinski et al., 1994; Lenhart
et al., 2000; Pacheco and Havel, 2001; Jackson et al.,
2005), which affects mobility in soils and uptake by plants
(Zhang et al., 1997; Ebbs et al., 1998; Read et al., 1998).
The uranyl ion also has the ability to complex with simple
inorganic ligands, such as carbonate (Langmuir, 1997;
Elless and Lee, 1998), and is generally mobile in the envi-
ronment as UO2þ 2 and its complexes, although sorption
to soil constituents is common (Morrison et al., 1995;
Langmuir, 1997; Ebbs et al., 1998; Read et al., 1998). These
complexes are more or less mobile than the parent ion
depending on solution chemistry and their interaction with
the solid phase (Chisholm-Brause et al., 2001).

0045-6535/$ - see front matter Published by Elsevier Ltd.

doi:10.1016/j.chemosphere.2007.06.032
238 A.J. Bednar et al. / Chemosphere 70 (2007) 237–247
Uranium can affect reproduction and fetal development
(Arfsten et al., 2001; Sztajnkrycer and Otten, 2004). Conse-
quently, the US Environmental Protection Agency has
established a maximum contaminant level of 30 lg l1 for
uranium in water (Federal Register, 2004). Anthropogenic
activities, such as mining, milling, and enrichment proce-
dures for nuclear fuel, can increase the potential mobility
of uranium (Abdelouas et al., 1998). Additionally, the
waste material from the enrichment process can be intro-
duced directly into the terrestrial environment through mil-
itary activities. This isotopically depleted uranium (DU) is
used in armor and armor piercing munitions (Chen and
Yiacoumi, 2002; Sztajnkrycer and Otten, 2004). The Uni-
ted States Army conducts testing of DU munitions at var-
ious facilities and DU has historically been used in military
conflicts, these both being sources of environmental release
(Sztajnkrycer and Otten, 2004). Although metallic uranium
is essentially immobile, corrosion reactions with air and
water can yield oxidized products, such as schoepite and
metaschoepite [hydrated uranium (VI) oxides] (Chen and
Yiacoumi, 2002; Johnson et al., 2006). Other minerals have
also been identified from uranium corrosion in soil, includ-
ing becquerelite, fourmuilerite, and sodium zippeite,
among other trace phases (Buck et al., 2004).
Schoepite and other uranium minerals can dissolve to sol-
uble U(VI), as UO2þ 2 , and therefore may migrate away from
the source area (Chen and Yiacoumi, 2002; Sztajnkrycer and
Otten, 2004). Furthermore, complexation of UO2þ 2 with nat-
ural ligands (organic and inorganic) will further alter mobil-
ity depending on how these secondary compounds interact
with soil (Abdelouas et al., 1998; Elless and Lee, 1998; Len-
hart and Honeyman, 1999; Jackson et al., 2005).
Natural organic matter and its interaction with UO2þ 2 acetic acid (EDTA) and sodium nitrate (for ionic strength
was the central focus of this research. The work addressed
the extent to which organic matter complexes with uranium
and how these complexes affect sorption to soil. Findings
from various sorption/desorption, complexation and sepa-
ration experiments are reported confirming the occurrence
of uranium-organic matter interactions in the environment.
Humic acid and model organic compounds were used in
complexation studies herein to simulate natural organic
matter and its interactions with dissolved uranium.
2. Methods and materials
2.1. Elemental analysis
Inductively coupled plasma mass spectrometry (ICP-MS)
(Perkin-Elmer Elan 6000) was used for elemental analysis of
all aqueous samples (Pawlak and Rabiega, 2002). The
plasma power was 1200 W with a nebulizer flow of
0.85 ml min1. Samples were prepared for analysis by filtra-
tion (0.45 lm unless otherwise specified) and acidification to
pH  2 with nitric acid, unless otherwise specified. Because
of its rarity in most samples and its similar mass to 238U, bis-
muth (209Bi as bismuth nitrate, J.T. Baker, Phillipsburg, NJ)
was used as the internal standard for all analyses. ICP-MS
was used for determination of uranium in samples because
of its higher sensitivity and faster analysis speed than tradi-
tional sodium iodide gamma spectroscopy. A detection limit
of 0.1 lg U l1 in aqueous solution was determined for
direct nebulization ICP-MS analysis, and is in agreement
with the literature (Pawlak and Rabiega, 2002).
2.2. Chemicals and standards
All chemicals used were of reagent grade or higher pur-
ity and used without further purification; the DI water used
was from a Milli-QÒ ultrapure water system (resistivity of
18.3 mX cm). The DI water system included UV-steriliza-
tion and the final water product was free of organic com-
pounds and filtered to 0.2 lm.
A humic acid standard (HA, CAS 1415-93-6) in the
sodium form, which was used to represent the acid insolu-
ble fraction of NOM, was purchased from Aldrich Chem-
ical Company (St. Louis, MO). NOM, and the HA
fraction, consists of a heterogeneous blend of various high
molecular weight, multi-functional organic moieties that
contain phenolic and carboxylic groups, among others.
These functional groups are known to complex metals
and contribute to the pH-dependant operational defini-
tions of humic and fulvic acid fractions of NOM (Lenhart
et al., 2000; Nierop et al., 2002). Cysteine and glutathione
were also purchased from Aldrich. Dissolved uranium
(973 mg l1) was also purchased from Aldrich for use as
a spike solution for sorption and complexation experi-
ments. An additional uranium calibration standard
(10 mg l1) was purchased from SPEX Certiprep for use
as an analytical standard. Disodium ethylenediaminetetra-
adjustments) were purchased from J.T. Baker.
2.3. Chromatography equipment and supplies
Molecular weight cut off (MWCO) centrifuge filter
devices (Millipore Amicon Ultra, Billerica, MA), with
nominal pore filter sizes of 5, 10, 30, 50, and 100 kilodal-
tons (kDa), were used to separate organouranium com-
plexes based on relative molecular size. The filter devices
have volume capacities of 4 or 15 ml, depending on the part
purchased. Generally a 4 ml aliquot was filtered because
aliquots were diluted for ICP-MS analysis. The solution
was added to the filter devices and centrifuged at
2000 rpm until the entire volume had passed through the
filter membrane, and was collected for analysis.
An Agilent series 1100 high pressure liquid chromato-
graph (HPLC) with an Agilent G1314A UV absorbance
detector, set at 230 nm, was used for all chromatographic
separations and was interfaced to the ICP-MS. Details of
the coupling of HPLC to ICP-MS have been described else-
where (Bednar et al., 2004; Bednar et al., 2007). The col-
umn compartment was maintained at 25 °C, and samples
were kept at 10 °C in the thermostat-controlled autosam-
pler prior to analysis to inhibit degradation. Typically
 A.J. Bednar et al. / Chemosphere 70 (2007) 237–247 239

50 ll were injected for analysis. Reagent grade chemicals
were used for preparation of mobile phases and all solu-
tions were filtered (0.45 lm) prior to introduction into
the instrument. Mobile phases were degassed with an in-
line vacuum degasser during operation. Two specific forms
of HPLC separation were used in this work: reverse phase
and molecular size exclusion chromatography.
Reverse phase (RP) chromatography coupled to ICP-
MS as the element specific detector was used to investigate
specific organic compound binding to uranium. An Agilent
Zorbax Stablebond Aqueous column, using an isocratic
mobile phase of 10 mM ammonium acetate (pH 5.5) in
0.1% acetonitrile at 1.0 ml min1 flow rate, was used for
separation. Selection of the column and eluent configura-
tions were based on compatibility with the column manu-
facturer’s suggestion, and the ICP-MS’s low tolerance for
organic solvent introduction.
Molecular size exclusion chromatography (SEC) sepa-
rates compounds by molecular weight, or more precisely,
hydrodynamic size. A Shodex KW-803 SEC column was
used to separate organouranium complexes based on
molecular weight. The mobile phase used in all experiments
was 21 mM Tris(hydroxymethyl)aminomethane–HCl buf-
fer (TRIS buffer, pH 7) because of the known protein
molecular weight standards used to correlate elution time
with molecular size (Miyazawa and Murayama, 2007).
The ICP-MS was used as the elemental specific detector,
however, an Ultraviolet–Visible (UV) absorbance detector
was added in-line prior to the ICP-MS to determine when
the organic moiety eluted from the column by monitoring
absorbance at 230 nm. UV absorbance has been demon-
strated to correlate well with organic matter (Benedetti
et al., 2003). This dual detector capability allowed the
simultaneous determination of organic moieties and
uranium.
2.4. Soils
A silty clay loam (loess) soil from Vicksburg, MS
(Waterways Experiment Station – ‘WES’ soil, Tardy
et al., 2003; Larson et al., 2005) was used as the comparison
soil in the sorption studies. This soil contained 52 g Fe
kg1, 26 g Al kg1, and 4 g Mn kg1 (Perkin–Elmer 3000
DV ICP-AES analysis by US EPA method 6010B). Two
silty clay loam soils from Yuma, AZ containing 23, 23,
and 0.4 g kg1 of iron, aluminum, and manganese, respec-
tively, were also used for sorption, extraction, and leach
experiments. The first Yuma soil (‘Yuma Clean’) and the
WES soil each contained 1 mg U kg1; whereas, the second
Yuma soil contained 87 mg U kg1 (‘Yuma DU’) from the
historical testing of DU munitions. All soils were sieved
using a #10 ASTM sieve (<2 mm) to produce a more
homogeneous sample for laboratory experiments. Subsam-
ples of the soils were heated in a muffle furnace at 400 °C
for 1 h to remove native NOM (Gustafsson et al., 1997).
The humic acid standard material was then added to the
soil subsamples at 1 wt.% by dissolving it in DI water, mix-
ing the solution with the soil, and allowing the water to
evaporate at ambient conditions to create ‘NOM normal-
ized’ soils for comparative studies.
2.5. Plant materials
Two sources of naturally occurring organic materials
were collected to investigate the natural occurrence of
organouranium complexes (Table 1). The first source con-
sisted of three plant species harvested from a phytoremedi-
ation experiment in which the plants were grown in soil
that contained up to 100 mg U kg1 from the historic test-
ing of DU munitions. Two plant standards (CLV-1 and
CLV-2) from the Canadian Certified Reference Materials
Project also were included (CANMET Mining and Mineral
Science Laboratory, Ontario, CA).
The second source of uranium-containing organic mate-
rial was cryptobiotic crusts. Cryptobiotic crusts are water-
stable soil aggregates held together by algae, bacteria,
fungi, lichens, or mosses, and the substances they produce
(Johansen, 1993). Within the crusts is an intimate associa-
tion of soil particles with the biotic component, a muco-
polysaccharide gel, which has the ability to bind and

Table 1
Plant samples used in the speciation study and the uranium concentration (mg U kg1 plant determined by ICP-MS) in acid digests and aqueous
extractions
Lab ID Scientific name Sample information Total [U] DI extract [U] 1% MeOH extract [U]
CLV-1a Picea sp. Spruce twigs 87 0.72 1.0
CLV-2a Picea sp. Spruce needles 3.6 0.02 0.03
IRM 9824 Kochia scoparia Kochia leaves, seeds 35 0.52 0.40
IRM 8891 Kochia scoparia Kochia leaves, seeds 11 0.08 0.11
IRM 9634 Kochia scoparia Kochia leaves, seeds 60 1.1 1.1
F1-Brju Brassica juncea Indian mustard 2 0.06 0.05
F3-Brju Brassica juncea Indian mustard 17 1.4 0.94
F5-Brju Brassica juncea Indian mustard 33 0.21 2.6
20-NE Amarathus cruentus Purple amaranth 20 0.17 0.31
3PO51 Machaeranthera tanacetifolia Tansy aster 37 0.26 2.5
Soil crust ‘A’ Microcoleus vaginatus Soil crust 0.5 0.005 0.003
Soil crust ‘B’ Microcoleus vaginatus Soil crust 2.7 0.22 0.45
a
Standard reference materials, certified values listed.
240 A.J. Bednar et al. / Chemosphere 70 (2007) 237–247
possibly to allow uptake of soil constituents by the bacteria
and algae. These crust samples were collected from a dry
lakebed in the southwestern United States on which
Microcoleus vaginatus was the dominant microorganism,
and had no known exposure to anthropogenic uranium
sources (Brostoff, 2002). Uranium concentrations in the
soil crust samples are listed in Table 1.
3. Experimental
3.1. MWCO separation
The formation of organouranium complexes under var-
ious ionic strength and pH conditions was investigated
using MWCO filtration (5, 10, 30, 50, and 100 kDa).
Humic acid solutions ranging from 0 to 100 mg l1 (0,
10, 50, and 100 mg/l) were spiked with 100 lg U l1 and
allowed to react and equilibrate for 1 h prior to size sepa-
ration filtration. These solutions were made in either DI
water or 10 mM sodium nitrate. Additionally, experiments
with the same matrix constituents were performed in neu-
tral and acidic (pH  4) solutions, with pH adjusted with
addition of trace nitric acid. After the 1-h equilibration
time, the samples were size filtered, diluted, acidified, and
analyzed by ICP-MS for dissolved uranium in the filtrate.
Dissolved inorganic uranium passed through the filters
used. As organic moieties complexed with uranium, the
apparent molecular weight (size) of the uranium complex
increased (due to the attached organic portion). These lar-
ger complexes were then removed more effectively by filtra-
tion as the filter pore size decreased.
3.2. Reverse-phase and size exclusion chromatography
Chromatography was used to separate organouranium
complexes based on their chemical properties (i.e., charge,
hydrophobicity, or molecular size). Cysteine, glutathione,
and EDTA were chosen as model compounds with the
potential to bind uranium. EDTA is a common metal com-
plexation agent (Snoeyink and Jenkins, 1980), while cys-
teine and glutathione are important in sorption and plant
uptake of metals (Grill et al., 1987; Ebbs et al., 1998; Read
et al., 1998). Complexes of uranium with model organic
ligands were prepared by spiking a 0.1% solution of the
organic compound (except EDTA, which was at 1% of
0.25 M [saturated]) with 100 lg U l1. The solutions were
allowed to react and equilibrate for 1 h before analysis.
Plant extracts for chromatographic analysis were prepared
as described below. All solutions were centrifuged or fil-
tered (0.45 lm) prior to HPLC-ICP-MS analysis.
3.3. Adsorption, desorption, and extraction procedures
Uranium sorption to soils was conducted in the presence
and absence of humic acid and natural organic matter. DI
water extraction of soil was used to determine the most
mobile and bioavailable fraction of uranium in a sample.
All adsorption, desorption, and extraction solutions were
agitated on a reciprocal shaker. Adsorption solutions con-
tained uranium from a standard solution dissolved in DI
water, and were added to a soil sample at a 10:1 solu-
tion:solid ratio and allowed to equilibrate for 24 h. The
desorption solutions used were DI water and 100 mg l1
humic acid. Desorption solutions were added to a soil sam-
ple in the same 10:1 ratio, and mixed for 1 h before aliquots
were filtered and diluted for analysis. In addition to DI
water and humic acid solution extractions, 0.1% nitric acid
(pH < 2) or sodium bicarbonate (pH  9) was used to
determine if pH would affect uranium extraction from soil.
Solutions were retrieved for analysis by centrifugation and
filtration (0.45 lm), prior to acidification and analysis by
ICP-MS.
A sequential extraction experiment was performed on
Yuma clean and DU soils to investigate possible leaching
of uranium after successive exposures to mildly acidic
leaching solutions (pH 5, 60:40 sulfuric:nitric acid [syn-
thetic precipitation leaching procedure (SPLP), US EPA,
1994]). The extraction procedure was performed nine times
with centrifugation between each stage for separation of
the aqueous and solid phases. These results were compared
to multiple DI water leaches of the same soil. In all soil
extraction experiments, a constant 1 g soil to 10 ml of
extraction fluid ratio was maintained. Extraction times
for the sequential extraction experiments were 24 h. Solu-
tions were prepared for total uranium analysis by ICP-
MS as previously described.
For uranium speciation of plant materials, the possibil-
ity exists of finding organouranium complexes (Grill et al.,
1987; Ebbs et al., 1998). Therefore, DI water and DI water
with 1% methanol were used to determine the effect of
organic solvent on the extraction efficiency of organoura-
nium complexes from plant materials. The same 1 g to
10 ml ratio was maintained, although extraction times were
increased to 7 days to allow for the extraction from the bio-
logical material. Samples were prepared for dissolved ura-
nium analysis by ICP-MS or speciation by HPLC-ICP-MS
as previously described.
4. Results and discussion
4.1. MWCO separation study
Initial work focused on determining the extent of HA
complexation (0 to 100 mg HA l1) with dissolved ura-
nium. Measurable complexation occurred even at the low-
est (non-zero) HA concentration (10 mg l1) (Fig. 1). For
the control sample (no HA added), uranium recovery was
roughly the same (about 80%) for all MWCO filter sizes,
although there did appear to be some interaction between
dissolved uranium and the 10 kDa filter material (particu-
larly in the DI water solution), leading to a lower recovery.
The filter material is regenerated cellulose, which could
potentially sorb inorganic and organo-complexes. Ura-
nium recoveries subsequently decreased as organic matter
 A.J. Bednar et al. / Chemosphere 70 (2007) 237–247
Fig. 1. Results of MWCO filter experiments in DI water and 10 mM sodium nitrate under neutral and acidic conditions. A general reduction of uranium
concentration in the filtrate is seen as a function of filter pore size and HA concentration.
was added to the solutions, indicating that the organic
material was binding with the uranium, creating a larger
apparent molecular size that was removed by filtration. A
decreased concentration of uranium is observed to be both
a function of increased humic acid concentration and
decreased filter pore size.
Ionic strength had little effect over the range tested,
although the 10 kDa filter had a higher uranium recov-
ery from the 10 mM ionic strength solution than from
the DI water sample (see 0 mg l1 HA samples, Fig. 1).
This suggests competition for sorption sites on the
10 kDa filter material with the electrolyte ions, resulting
in better recovery of uranium in the control samples if
electrolyte ions are present. Selected solutions in Fig. 1
were performed in triplicate, and subjected to all size fil-
trations; resultant standard deviations for triplicate
analyses ranged from <1 to 4.4 lg l1, showing excellent
reproducibility.
The effect of pH was minimal for the range tested. The
neutral pH solutions (Fig. 1) had similar results to the acid
studies: less uranium in the filtrate with decreasing pore
sizes in the presence of humic acid. The anomaly in the
10 kDa filter pore size sample attributed to ionic strength
was observed again, further suggesting a sorption interac-
tion of dissolved uranium with the filter material that is
reduced by the presence of competing ions (e.g., Na+ or
H+).
241
4.2. Reverse-phase chromatography
The MWCO filter study indicated that uranium binds
significantly to HA. The next experiment was designed to
investigate uranium binding to model organic compounds.
Fig. 2 presents RP-HPLC–ICP-MS chromatograms of
organouranium complexes with ‘standard’ organic com-
pounds (glutathione, cysteine, humic acid, EDTA) and
dissolved inorganic uranium. RP-HPLC separated the ura-
nium species according to the organic ligand. Column-
mobile phase compatibility is critical for this type of chro-
matography, in that equilibrium partitioning of the analyte
between the two phases must be maintained, while keeping
the organic solvent constituent low enough for the ICP-MS
to tolerate.
Using a buffered ammonium acetate mobile phase with
0.1% acetonitrile on an Agilent Zorbax SB-Aq column,
separation of EDTA and glutathione complexes of ura-
nium was accomplished (Fig. 2). The results for HA and
cysteine were not definitive as these potential complexes
were unresolved from the dissolved inorganic uranium
peak, even with varied amounts of acetonitrile and acetate
buffer.
Experiments were then conducted to determine if
organic complexes with uranium could be found in ‘natu-
ral’ samples. Fig. 3 shows selected expanded RP chromato-
grams of various plant extracts. The relative peak heights
242 A.J. Bednar et al. / Chemosphere 70 (2007) 237–247
Fig. 2. Reverse phase HPLC-ICP-MS chromatogram of various orga-
nouranium complexes using the 10 mM ammonium acetate buffer + 0.1%
acetonitrile mobile phase. ‘R’ represents an organic moiety, specifically
EDTA and glutathione species are resolved.
Fig. 3. Reverse phase HPLC-ICP-MS chromatogram of organouranium
complexes extracted from plant material with DI water and 1% methanol.
At least two forms of uranium are separated under these chromatographic
conditions for the plant species shown.
between the two chromatograms indicate DI water and 1%
methanol extract different amounts of uranium in each
plant species tested. The plant names and the laboratory
reference identification numbers, along with the aqueous
extractable uranium and the total uranium in the plant
material, are listed in Table 1. Some plant samples, such
as 3PO51 and F5-Brju, had higher methanol-labile ura-
nium concentrations, while others, such as IRM9824 and
F3-Brju, had higher DI water extractable uranium concen-
trations. The differences in uranium concentrations
between the two extraction solutions were up to an order
of magnitude for some plant species (e.g., 3PO51 and F5-
Brju), while others had very little difference. Although the
peaks in Fig. 3 overlap and are not baseline resolved, reten-
tion times varied for the organouranium complexes
observed depending on the plant sample. The plant stan-
dard CLV-1 (spruce twigs) had two resolvable forms of
uranium. Because of the complex nature of these plant
extracts, these peaks still represent multiple organoura-
nium species rather than the model compounds shown in
Fig. 2.
4.3. Size exclusion chromatography
Fig. 4 is a size exclusion HPLC-UV-ICP-MS chromato-
gram (SEC) of uranium-containing plant material, simulta-
neously using ICP-MS and UV absorbance detection
(230 nm). The addition of methanol to the extraction fluid
generally resulted in more complex UV absorbance (e.g.,
Fig. 4), indicating that slightly different organic moieties
were extracted from the plant material. Multiple forms of
uranium are seen with these chromatographic conditions,
and substantially more uranium is leached with the 1%
methanol solution.
The SEC–UV-ICP-MS chromatograms of cryptobiotic
soil crust DI water extracts show the complex nature of
the organouranium species extracted by DI water
(Fig. 5). The ‘crust A’ sample contained very little dissolved
uranium, and had very simple UV and ICP-MS chromato-
grams. However, the ‘crust B’ sample contained a higher
level of uranium with multiple inorganic and organoura-
nium species. The uranium peak observed at 6 min
appears to be an inorganic colloidal form (e.g., uranium
oxide), as it did not have a significant UV signature,
whereas the peaks observed within a range of 10–15 min
consisted of uranium associated with the organic moieties.
The most notable information gained from the SEC
data in Figs. 4 and 5 was the distribution of uranium in
these samples. In SEC, large molecules elute earlier than
smaller ones. This separation assumes, however, that there
are no hydrophobic or electrostatic interactions of the ana-
lyte with the packing material due to the ionic strength of
the mobile phase. The data for plant extract 3PO51
(Fig. 4) and the soil crust extracts (Fig. 5) indicate that
the inorganic uranium peaks (those without a large UV
absorbance) were in particulate form and were of a larger
size than the organouranium complexes because they elute
first. Analysis of protein MW standards on this column
with this mobile phase suggest the organouranium com-
plexes eluting at 10–15 min have a hydrodynamic size sim-
ilar to 3 kDa MW proteins.
4.4. Soil sorption and desorption
The experiments presented above indicated that ura-
nium and organic compounds readily form complexes
 A.J. Bednar et al. / Chemosphere 70 (2007) 237–247 243

Fig. 4. Size exclusion HPLC–UV-ICP-MS chromatograms of organouranium complexes extracted from tansy aster plant material (sample 3PO51) with
DI water and 1% methanol.

Fig. 5. Size exclusion HPLC–UV-ICP-MS chromatograms of organouranium complexes extracted from soil crust materials with DI water.
244 A.J. Bednar et al. / Chemosphere 70 (2007) 237–247

and are found in naturally occurring matrices (cryptobiotic
crusts and plant materials). The next goal was to investi-
gate whether these complexes could measurably influence
uranium sorption to soil and thus its mobility in the envi-
ronment. Both Yuma soils and the WES soil contained
native organic matter: the WES soil had approximately
4.8% mass loss on ignition, whereas the Yuma soils were
about 3.2%. Some of this weight loss can be attributed to
loss of residual soil moisture and chemical waters of hydra-
tion, however, the bulk of it is likely attributable to organic
carbon.
Fig. 6 shows the extent of uranium sorption to the native
soils before and after they were combusted in a muffle fur-
nace to remove native organic matter. Soil samples marked
with ‘+HA’ were amended with 1 wt.% HA. The HA was
added to the soil as a solution and the water was allowed
to evaporate under ambient conditions. The adsorption
spike solution was dissolved U(VI) in DI water at 10 and
100 lg l1. DI water was used as the subsequent desorption
solution.
The combusted WES soil was remarkably efficient at
sorptive removal of uranium from solution, with nearly
100% removal at both spike levels. The addition of HA
to the combusted and native WES soils prior to the ura-
nium spike resulted in approximately 75% and 90% of
the 100-lg l1 uranium spike being sorbed, respectively.
This indicates that HA hindered the sorption of uranium
to the WES soils, likely because of competition for binding
sites on the soil particles. The sorbed uranium in the HA-
treated WES samples was only slightly desorbed (6–9%)
compared to the non-HA amended soils.
Unlike the WES soils, the Yuma clean soil did not have
complete sorptive removal of the uranium from solution,
with only about 60–70% sorption observed. The addition
of HA to the combusted and native soils reduced sorption
to about 25% or less. However, the subsequent desorption
experiments indicated that the HA applied to the Yuma
soils slightly increased the release of sorbed uranium. This
effect could indicate differences in sorption mechanisms of
the two soils. The WES soil contains approximately 2 and
10 times more iron and manganese, respectively, than the
Yuma soils suggesting that oxides may be the most impor-
tant sorbing phases in this soil (Morrison et al., 1995; Gab-
riel et al., 1998; Lenhart and Honeyman, 1999; Schmitt
et al., 2002). Similar competitive adsorption effects in other
soils have been observed between NOM and other metals,
such as arsenic (Redman et al., 2002).
The experiments shown in Fig. 6 indicate that adsorp-
tion and desorption of uranium are highly dependant upon
soil and solution chemistry. Fig. 7 shows the effect of

Fig. 6. Adsorption of uranium to native WES and Yuma soils before and after being heated in a muffle furnace (MF), with and without amendment with
1% (weight) HA. The WES soil is more effective in sorbing uranium, although HA inhibits sorption and promotes subsequent desorption. The error bars
shown are the standard deviation of duplicate experiments. The original spike concentrations were 10 and 100 lg U l1.
 A.J. Bednar et al. / Chemosphere 70 (2007) 237–247 245

Table 2
Results of sequential extraction of the two Yuma soils using SPLP and DI
water
Extraction Yuma DU Yuma DU Yuma clean Yuma clean
number [U] DI [U] SPLP [U] DI [U] SPLP [U]
1 82 61 0.4 0.4
2 58 0.3 0.2 <0.1
3 27 17 <0.1 0.2
4 51 19 <0.1 <0.1
5 28 10 <0.1 <0.1
6 20 9.0 <0.1 <0.1
7 16 7.8 <0.1 <0.1
8 66 17 0.3 0.3
9 42 14 <0.1 <0.1
Concentrations given are in the leach solution (lg U l1).

To further investigate the extent of sequential uranium

Fig. 7. Effects of leach solution chemistry on extraction of uranium from
three soils studied. Almost two orders of magnitude more uranium is
leached from the Yuma DU soil by 100 mg l1 HA solution than by DI
water, comparable to that extracted by 0.1% acid or base solutions.
extractant solution composition on uranium desorption
from Clean Yuma and WES soils that have had no ura-
nium added to them. Also shown in the figure is desorption
of uranium from the Yuma DU soil that ‘naturally’ con-
tained uranium from the historical testing of DU muni-
tions (i.e., not a laboratory spike). Four extraction
solutions were investigated: DI water (DI), 0.1% nitric acid
(acid), 0.1% sodium bicarbonate (base), and 10 mg l1 HA.
Each soil was extracted sequentially with each extraction
fluid.
The Yuma DU soil released much higher concentrations
of U, compared to the two soils (Yuma Clean and WES)
that did not contain much uranium. Additionally, pH
effects, and the presence of HA, increased uranium
extracted from the Yuma DU soil by at least an order of
magnitude compared to DI water extraction. Both 0.1%
nitric acid and sodium bicarbonate extracted similar
amounts of uranium as the HA solution. The second
desorption removed approximately 50% as much uranium
as the first desorption, suggesting that the supply of readily
soluble uranium may become exhausted with subsequent
leaching. The total uranium in the 1 g Yuma DU soil sam-
ple was 0.087 mg, where approximately 0.012 mg was
removed by the two extraction experiments.
desorption from soil, a sequential extraction experiment
was performed on the two Yuma soils. Both the SPLP
solution and DI water were used for the nine sequential
extractions. DI water generally extracted 2–3 times more
uranium than the pH 5 SPLP solution (Table 2). Virtually
no uranium was leached from the Yuma clean soil, as
expected.
The difference in uranium concentration between the
two extraction fluids is possibly explained by the solubility
of UO2þ2 in the presence of inorganic ligands, such as car-
bonate (Ebbs et al., 1998; Chen and Yiacoumi, 2002). At
pH 5 of the SPLP solution, dissolved carbonate will be
minimal, but in the DI water extraction, the solution will
acquire the native pH of the soil (Yuma soil pH  8.5).
Water extractable alkalinity from the Yuma soil is
750 mg CaCO3 kg1, whereas the WES soil is about
75 mg CaCO3 kg1. Because the Yuma soils naturally con-
tain carbonate minerals, this will allow for carbonate disso-
lution and complexation of UO2þ 2 , thereby increasing its
solubility compared to the mildly acidic SPLP solution.
5. Conclusions
From the experiments presented, it is clear that NOM
interactions must be considered if the chemistry and mobil-
ity of uranium is to be understood in natural systems. Both
MWCO filters and on-line HPLC-ICP-MS were used to
demonstrate the binding of uranium to organic moieties,
including HA. Furthermore, HPLC-UV-ICP-MS was used
to speciate inorganic and organouranium forms in aqueous
extractions of plant materials.
The RP approach proved to be effective at resolving ura-
nium complexes with glutathione and EDTA, but HA- and
cysteine-uranium complexes produced similar chromato-
grams as ionic uranium, suggesting further method devel-
opment is needed. The challenge that lies in RP is mobile
phase compatibility with the ICP-MS. Typically, RP meth-
ods employ 50% or greater organic solvent mobile phases
(e.g., 50% methanol in water); however, mobile phase solu-
tions with greater than 5% organic solvent can cause ICP
246 A.J. Bednar et al. / Chemosphere 70 (2007) 237–247
instability or even extinguish the plasma. Serious matrix
incompatibilities may result when RP columns designed
for high organic mobile phases must still separate com-
pounds in nearly 100% aqueous solutions because of ICP
sample introduction limitations.
SEC was used to investigate uranium binding with
organic moieties in plant extracts. The analyses showed
multiple inorganic and organouranium forms in natural
plant samples, and indicates that SEC is useful for studying
the binding of uranium to complex organic moieties found
in plants.
Soil sorption characteristics were perhaps the most tan-
gible aspects of uranium-organic matter interactions. In
general, it was assumed that organic matter would increase
the sorptive properties of soil. However, this assumption
focuses on stationary organic matter in soils (e.g., attached
to soil particles), which can supply additional sorption
sites. Soil particles can interact with dissolved organic mat-
ter, such that the soluble organic material competes with
other dissolved constituents for sorption sites on soil par-
ticles. The results presented focused on dissolved organic
matter, and yielded results different than would be
expected with stationary organic matter bound to soil
particles.
The results of the sorption studies indicated that HA did
not uniformly promote sorption, but could hinder it. It is
likely that the dissolved HA competed with uranium for
sorption sites on oxide and clay particles. In all soils, the
addition of HA decreased sorption; additionally, HA pro-
moted subsequent desorption. Heating the soils in a muffle
furnace to remove native organic matter caused a slight
decrease in initial sorption of uranium compared to the
native soil, but also caused a slight decrease in subsequent
desorption of uranium.
Uranium chemistry in soils, particularly as it relates to
mobility, is complex, including both inorganic and organic
ligands. However, the investigation of its geophysiochemi-
cal speciation is a step to understanding its behavior. These
experiments provide a glimpse into the impact of organic
matter interactions in natural matrices on uranium
chemistry.
Acknowledgements
The use of trade, product, or firm names in this report is
for descriptive purposes only and does not imply endorse-
ment by the US Government. The tests described and the
resulting data presented herein, unless otherwise noted,
were obtained from research conducted under the Range-
Safe program, and the Army Research, Development,
and Engineering Command – Armaments Research, Devel-
opment, and Engineering Center. Permission was granted
by the Chief of Engineers to publish this information.
The findings of this report are not to be construed as an
official Department of the Army position unless so desig-
nated by other authorized documents. The authors also
thank Scott Waisner and Burton Suedel of the USACE
for their editorial comments and suggestions.
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