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Microchemical Journal
journal homepage: www.elsevier.com/locate/microc
Geomicrobiology Laboratory, School of Earth Sciences, University of Melbourne, Parkville, VIC 3010, Australia
Ecochemistry Laboratory, Institute for Applied Ecology, University of Canberra, ACT 2601, Australia
a r t i c l e
i n f o
Article history:
Received 9 August 2016
Accepted 15 August 2016
Available online 28 August 2016
Keywords:
Arsenic speciation
Thioarsenate
HPLC
ICP-MS
Preservation
a b s t r a c t
Recently developed analytical techniques allowed for the detection of a range of dissolved arsenic-sulphur species in sulphur-rich environments. These so called thioarsenates are unstable, however, and can degrade upon
handling and storage. An experiment evaluating the effect of exposure to air on arsenic and sulphur-enriched
geothermal waters demonstrated a near to complete loss of thioarsenate species to arsenite or arsenate during
short oxidation times. In contrast, thioarsenic standards were stable for the duration of analysis in spite of exposure to air. For samples containing thio-methylated arsenic species, the extent of oxidation varied for different
methylated arsenic species. This study recommends ash freezing of samples in liquid nitrogen immediately
after recovery and further storage under anaerobic conditions at 80 C. A second experiment to test the efciencies of different HPLC columns for separating arsenic species resulted in the preference for an IonPac column
with NaOH as the mobile phase when analysing arsenic thioanions, over the commonly used PEEK PRP-X100
anion exchange and Atlantis C18 reverse phase column with ammonium phosphate mobile phases. Distinct separation of thio-methylated arsenic species with the IonPac column, however, was not successful potentially due
to matrix components. Acceptable detection, separation and quantication of thio-methylated arsenic species
were only achieved with the Atlantis C18 column. This study shows that preservation and analysis of samples
is matrix dependent, which holds important implications for efforts to interpret arsenic speciation in geothermal
waters, especially those of low pH (23), low oxygen (49% saturation), low iron (5 mg L1) and high sulphur
concentrations (91 mg L1).
2016 Elsevier B.V. All rights reserved.
1. Introduction
Accurate measurement of arsenic speciation is critical for understanding the arsenic biogeochemical cycle in hydrothermal systems.
Until recently the inorganic oxyanions arsenate and arsenite were believed to be the only dissolved arsenic species in hydrothermal waters
[1,8,22,25]. Arsenic bonds with free oxygen and forms arsenic
oxyanions in different oxidation states: arsenite (AsIIIO33 ) is stable
under reduced geochemical conditions and arsenate (AsVO3
4 ) under
more oxidising conditions [20]. An increase in redox potential promotes
the oxidation of arsenite to arsenate, while an increase in pH results in
deprotonation of protonated arsenic species.
H3 AsO3 HS H2 AsO3 S H2
H3 AsO3 S0 H2 AsO3 S H
Corresponding authors.
E-mail addresses: katrin.hug@gmail.com (K. Hug), bill.maher@canberra.edu.au
(W.A. Maher).
1
Present address: Institute of Groundwater Ecology, Helmholtz Center Munich,
German Research Center for Environmental Health, 85764 Neuherberg, Germany.
http://dx.doi.org/10.1016/j.microc.2016.08.008
0026-265X/ 2016 Elsevier B.V. All rights reserved.
163
critical. Acidication and storage at 4 C is a commonly used preservation technique [25]. In sulphide- or sulphur-rich waters, however, acidication can potentially lead to total dissolved arsenic loss via induced
arsenic-sulphur precipitation under low pH [12] and storage of samples
at 4 C could lead to thioarsenate degradation, due to the rapid oxidation
of arsenic thioanions compared to oxyanions [12].
For analysis, high-performance liquid chromatography coupled to inductively coupled plasma mass spectrometry (HPLCICPMS) using an
IonPac column and a sodium hydroxide gradient is a well-established laboratory method for thioarsenate speciation [13,21,23], while an Atlantis
C18 column with phosphate buffer is commonly used to separate thiomethylated arsenic species [9]. These separation techniques, however, require an efcacy evaluation with different sample types. In this paper we
describe our experiences in preserving geothermal samples and the analysis of thioarsenates and methylated thioarsenic species in arsenicenriched hot spring waters by HPLCICPMS.
Fig. 1. EhpH diagrams for the AsOHS system (S = 1 104 M and As =
2 105 M) (modied from [2]). The diagram is based on the arsenic thioanions and
thermodynamic data is provided in [4].
3
1
222:2 kJ mol
2. Experimental methods
2.1. Field site and water chemistry
The sampling sites are situated in the Taupo Volcanic Zone (TVZ) at
Waiotapu (Fig. 2) and were selected on the basis of distinctive physical
and chemical characteristics such as pH, temperature, redox potential
and sulphur concentration, all factors known to inuence arsenic speciation [12,14,21,26]. In close vicinity of Champagne Pool (CPr), the biggest and most prominent feature in the area, Alum Cliffs (AC), stream
to Frying Pan (FPs) and pool at Frying Pan (FPp) are characterised
by low pH and compared to Champagne Pool higher dissolved oxygen
saturation and redox potential, which indicates mixing of deep hydrothermal uids with oxygenated water close to the surface. Temperature,
pH, redox potential and dissolved oxygen saturation (DO%) were measured in situ using a Professional Plus multimeter (YSI, USA). The
pools in this area contain signicant sulphur but low iron concentrations, which were measured via inductively coupled plasma mass spectrometry (ICP-MS).
2.2. Sampling and storage
Water samples for arsenic speciation and quantication were collected in February 2012. Each suite of samples was stored in opaque
125 mL high-density polyethylene bottles (Nalgene, USA) washed
Oxic conditions deplete thioarsenate species very quickly via conversion to less thiolated arsenic species and eventually arsenite [12].
Planer-Friedrich et al. [14] showed that the [SH]:[OH] activity ratio
affects the stability of thioarsenic species. When [SH] N [OH] under
anaerobic conditions, the most stable arsenic species is thioarsenite,
whereas under aerobic conditions thioarsenate is more stable. When
[SH] b [OH] under either aerobic or anaerobic conditions, the most
stable arsenic species is arsenite, as thiolated arsenic species would be
unstable at excess OH. Arsenite oxidation, however, only starts to
occur in signicant levels when the thioarsenate species are almost
completely transformed to arsenite.
2
H2 AsOS3 2:5 O2 H2 AsO2 S
2 SO4
2
H2 AsO2 S
2 2:5 O2 H2 AsO3 S SO4
9
10
H2 AsO3 S H H3 AsO3 S0
11
H3 AsO3 O2 H2 AsO4
12
Fig. 2. Sampling sites at Waiotapu in the Taupo Volcanic Zone on the North Island of New
Zealand (photo credit: Google Earth). Alum Cliffs (AC), stream to Frying Pan (FPs), Frying
Pan pool (FPp).
164
Table 1
Temperature, pH, redox potential and dissolved oxygen saturation (DO%) at Waiotapu sites, New Zealand.
Temperature (C)
(0.2 C)
pH (0.2)
DO (%) (2%)
Alum Cliffs
85
2.5
242
34
1.02 0.02
FPs
33
2.2
249
49
1.97 0.02
FPp
41
2.6
228
35
5 0.04
CPr
68
5.5
75
20
b0.008
Site ID
Site description
AC
Image
165
Table 2
MTA spiked environmental Champagne Pool sample CPr held at room temperature for the
duration of 4 days with measurements on the rst and fourth day over a period of 4 h.
MTA: monothioarsenate, DTA: dithioarsenate, TriTA: trithioarsenate, TetraTA:
tetrathioarsenate.
As(V)
MTA
DTA
TriTA
TetraTA
% Total
peak area
Day 1
CPr + MTA
CPr + MTA
CPr + MTA
Day 4
CPr + MTA
CPr + MTA
CPr + MTA
Mean SD
% peak area (n = 6)
Retention time (n = 6)
Fig. 3. Arsenic speciation in water samples from Waiotapu, New Zealand before
and after oxygen exposure. MTA: monothioarsenate, DTA: dithioarsenate, MA:
monomethylarsenate, DMA: dimethylarsenate, MTMA: monomethylmonothioarsenate,
MTDMA: dimethylmonothioarsenate, DTDMA: dimethyldithioarsenate. AC: Alum Cliffs,
FPs: stream to Frying Pan.
59.2
57.1
57.5
20.1
22.5
21.4
3.4
3.4
3.5
17.4
17.0
17.6
0.62
0.53
0.64
58.0
58.5
58.3
20.3
20.1
19.9
5.1
5.0
4.3
15.9
19.5
17.1
0.46
0.74
0.65
58 1
21 1
4.1 0.8
12.6 0.1 14.2 0.1 16 0.1
17 1
0.6 0.1
17.7 0.1 19.1 0.3
Fig. 4. HPLCICPMS chromatogram of arsenic species standards using the IonPac column.
Dimethylarsenate (DMA), dimethylmonothioarsenate (MTDMA), monomethylarsenate
(MA), monomethylmonothioarsenate (MTMA), monothioarsenate (MTA), dithioarseante
(DTA), and trithioarsenate (TriTA). cps: counts per second.
166
Fig. 5. HPLCICPMS chromatogram of arsenic species at Frying Pan pool (FPp) using the
IonPac column. Dimethylarsenate (DMA), dimethylmonothioarsenate (MTDMA),
monomethylmonothioarsenate (MTMA), monothioarsenate (MTA), dithioarsenate
(DTA), and trithioarsenate (TriTA). cps: counts per second.
167
Fig. 8. HPLCICPMS chromatogram of a spiked water sample using the Atlantis C18 column. Dimethylarsenate (DMA), dimethylmonothioarsenate, monomethylarsenate (MA),
dimethylmonothioarsenate (MTDMA), monomethylmonothioarsenate (MTMA). cps: counts per second.
Albert Shimmins Memorial Fund (UTR7.220) to K.H. and Dyason Fellowship to J.W.M.
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