Chemical Speciation & Bioavailability

ISSN: 0954-2299 (Print) 2047-6523 (Online) Journal homepage: https://www.tandfonline.com/loi/tcsb20

Optimisation of analytical parameters in inorganic

arsenic (III and V) speciation by hydride generation
using L-cysteine as prereducing agent in diluted
HCl medium

Emil A. Cordos, Tiberiu Frentiua, Michaela Pontaa, Bela Abrahamb & loan
Margineana

To cite this article: Emil A. Cordos, Tiberiu Frentiua, Michaela Pontaa, Bela Abrahamb & loan
Margineana (2006) Optimisation of analytical parameters in inorganic arsenic (III and V) speciation
by hydride generation using L-cysteine as prereducing agent in diluted HCl medium, Chemical
Speciation & Bioavailability, 18:1, 1-9, DOI: 10.3184/095422906782146276

To link to this article: https://doi.org/10.3184/095422906782146276

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Chemical Speciation and Bioavailability (2006), 18(1) 1

Optimisation of analytical parameters in inorganic

arsenic (III and V) speciation by hydride generation using
L-cysteine as prereducing agent in diluted HCl medium
Emil A. Cordosa*, Tiberiu Frentiua, Michaela Pontaa, Bela Abrahamb
and Ioan Margineana
a
Babes Bolyai University, Department of Chemistry, 400028 Cluj-Napoca, Arany Janos 11, Romania
b
Research Institute for Analytical Instrumentation, 400293 Cluj-Napoca, Donath 67, Romania

ABSTRACT
The effects on the arsenic spectrophotometric absorption signal and hydride generation of the reaction
medium (HCl: 0 – 0.05 M), reducing agent (NaBH4: 0.2 – 1% stabilised in NaOH: 0.2 – 1%), prereducing
agent (L-cysteine: 0 – 2%) and contact time between L-cysteine and arsenic (III) and arsenic (V) solutions
were investigated using FIA – HG – QFAAS. The speciation procedure involved three steps: (i) arsenic (III)
determination immediately after the sample was mixed with L-cysteine; (ii) total arsenic determination
after a 10 min-contact time between sample and L-cysteine by boiling in water bath at 90
5 C and (iii)
arsenic (V) determination as difference between total arsenic and arsenic (III). Under optimised
conditions to generate arsine from arsenic (III) (0.01 M HCl, 0.2% L-cysteine, 0.6% NaBH4 in 0.5%
NaOH), the arsenic speciation was possible with good recovery (98 – 105%) for arsenic (III), arsenic (V)
and total arsenic. A rigorous pH control at 2.00
0.01 prior to arsine generation was necessary. The
detection limit based on 3s criteria and parameters of calibration curve was 0.1 ng mL  1 arsenic. The
simple and inexpensive optimised method is used for the determination of leachable inorganic arsenic
species within a distribution study of arsenic in the Baia-Mare area, Romania, which is the subject of
another paper.

Keywords: inorganic arsenic speciation; hydride generation; quartz furnace atomic absorption
spectrometry; L-cysteine reducing agent

INTRODUCTION Numerous sensitive techniques have been devel-

Arsenic speciation in nature is important from a
number of perspectives: relation between toxicity
and species nature, bioavailability of species and
relation between sample nature and the majority
species. Different forms of arsenic have vastly
different toxicity on humans due to their different
bioavailability. For example, inorganic arsenic
[As(III) and As(V)] are carcinogenic and can cause
neurological, cardiovascular and haematological
disorders (Dannan et al., 1984), while monomethy-
larsonic acid (MMA), dimethylarsinic acid (DMA)
are believed to be far less toxic (Tezuka et al., 1993;
Vahter and Marafante, 1983).
*To whom correspondence should be addressed:
E-mail: cordos@icia.ro
www.scilet.com
oped to speciate traces of arsenic species, but most
are too laborious for routine analysis and require
expensive analytical equipment that is generally
not available in most laboratories. For environ-
mental studies, speciation of arsenic involves the
use of separation techniques coupled with a sensi-
tive atomic detector. High performance liquid
chromatography (HPLC) has been successfully
directly coupled to inductively coupled plasma –
optical emission spectrometry (HPLC – ICP – OES)
(Amran et al., 1997) and inductively coupled
plasma mass spectrometry (HPLC – ICP – MS)
(Kavanagh et al., 1998; Le et al., 1994a;
Milstein et al., 2002; Moldovan et al., 1998;
Wrobel et al., 2002). In order to increase the
sensitivity of hyphenated techniques a post-
column hydride generation (HG) to convert the
2 Optimisation of analytical parameters in inorganic arsenic (III and V) speciation by hydride generation
arsenic compounds into volatile arsine prior to the
detection is used, such as in atomic fluorescence
spectrometry using a hydrogen diffusion flame
(HPLC – HG – AFS) (Coehlo et al., 2003) or
HPLC – HG – ICP – MS (Magnuson et al., 1996).
However, arsenobetaine (AsB), arsenocholine
(AsC) and arsenosugars (AsS) do not form volatile
hydrides and destruction of the organic chain of
the molecule before the hydride generation is
required. This has been achieved by using both
on-line microwave digestion (MW) and photo-
oxidation with UV radiation in the presence of
potassium persulfate and detection by atomic
absorption spectrometry (HPLC – MW – HG – AAS)
(Le et al., 1994b; Moldovan et al., 1998; Villa-
Lojo et al., 2002) or mass spectrometry (HPLC –
(UV) – HG – ICP – MS) (Wei et al., 2000; Caruso
et al., 2001; Pongratz, 1998; Wrobel et al., 2002;
Yehl et al., 2001). Although hyphenated-techni-
ques are the most sensitive, they are not always
accessible. A more economical option is to employ
flow injection analysis-hydride generation-atomic
absorption spectrometry, usually in a quartz
atomiser cell (FIA – HG – QFAAS) (Chappel et al.,
1995; Nielsen et al., 1996; Shraim et al., 1999,
2000), hydride generation atomic fluorescence
spectrometry (FIA – HG – AFS) (Cava-Montesinos
et al., 2003) flow injection analysis-hydride
generation-inductively coupled plasma-atomic
emission spectrometry (FIA – HG – ICP – AES)
(Huang et al., 1988) or FIA – HG – ICP – MS
(Anderson and Pergantis, 2003), which offer a
good sensitivity, accuracy and low matrix effects.
The technique is based on the addition of a
reducing agent (usually a solution of sodium
tetrahydroborate stabilised with sodium hydroxide)
to an acidified sample solution. The analysed
elements form volatile covalent hydrides, which
are separated from the reaction solution and
transported to the spectral detector. The technique
has two main advantages over conventional nebu-
lization: a more efficient sample introduction,
which enables a better sensitivity and the separa-
tion of the hydride-forming elements from poten-
tial interferents prior to measurement.
In HG techniques, arsenic speciation could be
carried out by using two procedures: (i) separation
of arsenic (III) from arsenic (V) by extraction in
an organic reagent (Balasoiu et al., 2001;
Chappell, 1995) or (ii) selective reduction of
arsenic species by carefully controlling the pH
(Shraim et al., 1999, 2000). It has been shown
that in 10 M HCl solution AsCl3 is a covalent
molecule and can be extracted into an organic
phase (chloroform), while AsCl5 exists as
complex ions [AsCl6]
and [AsCl4] þ not extrac-
table in chloroform. For selective reduction an
attractive method is that based on L-cysteine as
a prereducing agent. The benefits of undertaking
L-cysteine in arsenic speciation are well docu-
mented in the literature (Chen et al., 1992; Welz
and Sucmanova, 1993a,b). Thus, in HCl or HClO4
medium, by controlling the concentration of the
acid, pre-reducing (L-cysteine) and reducing
(NaBH4) reagents, the speciation of the four
arsenic species in water sample (arsenite, arsenate,
MMA and DMA) is possible (Shraim et al., 1999,
2000). However, the experimental results from the
above mentioned papers, emphasise a diversity of
the optimum conditions to generate arsine
depending not only on the nature of species but
also on the concentration of acids, reducing agents
and mixture ratio of solutions.
The aim of this work was to optimise a simple,
rapid, inexpensive, accurate and precise technique
for the arsenic speciation (arsenite, arsenate) and
the quantification of total arsenic based on FIA –
HG – QFAAS. The effect of diluted HCl as reac-
tion medium on the arsine generation from the two
arsenic inorganic species has been investigated.
Effects of the concentration of the reducing and
hydride-generating agent (NaBH4), of the preredu-
cing agent (L-cysteine) and of the contact time
between sample and L-cysteine were also studied.
Accuracy was checked by determining the content
of As(III) and As(V) in synthetic solutions
prepared in the laboratory. The simple and inex-
pensive optimised method is used for the determi-
nation of leachable inorganic arsenic species
within a distribution study of arsenic in the Baia-
Mare area, Romania, which is the subject of
another paper (Cordos et al., 2005).
METHODOLOGY
Apparatus
A Perkin-Elmer, model 5000, atomic absorption
spectrometer was used in combination with a
vapour generation accessory (VGA-77 Varian). A
quartz atomiser cell (170 mm length, 12 mm i.d.)
heated at controlled temperature (900 + 10 C) by
700 W power supply using a Nichrome resistance
heaters (Research Institute for Analytical
Instrumentation, Cluj-Napoca, Romania) was
employed. The VGA-77 system was equipped
with a three-channel peristaltic pump, with PTFE
(polytetrafluoroethylene) tubing, and a reaction coil
connected to a gas – liquid separator. The FIA – HG
unit was operated according to the Manufacturer’s
instructions; the instrument parameters are
 Emil A. Cordos, Tiberiu Frentiu, Michaela Ponta, Bela Abraham and Joan Marginean 3

Figure 1 Experimental set-up of FIA – HG – QFAAS.

summarised in Table 1, while the experimental set-
up in Figure 1.
An arsenic hollow cathode lamp (S.&J. Juniper,
Harlow, UK) operated at 12 mA was used at the
wavelength of 193.7 nm with a band-pass of 0.7 nm.
Data processing was performed by a house-software
(Research Institute for Analytical Instrumentation,
Cluj-Napoca, Romania). The acidified sample with
diluted HCl containing L-cysteine was pumped and
mixed with the carrier flow (diluted HCl) and the
reducing agent (NaBH4 in NaOH solution) at the
flow-rates indicated in Table 1. The developed arsine
was purged from the liquid phase using an Ar stream
introduced in the coil where the sample and reducing
reagent were mixed. The gaseous phase containing
arsine is separated and then introduced into the T-
shaped quartz cell using a supplemental Ar stream.
The out-put signal was processed in steady-state
mode using a 60 s-time delay after the sample was
introduced into the HG and was recorded and
displayed in absorbance units as average for a
measuring time of 10 s.
The accurate pH adjustment of the samples to
2.00 + 0.01 prior to hydride generation was

Table 1 Operating conditions for FIA – HG – QFAAS

Parameter Condition
FIA – HG (Varian VGA-77)
1
Sample flow rateymL min 7
HCl carrier flow rateymL min  1 1
NaBH4 flow rateymL min  1 1
Ar carrier flow rateymL min  1 100
Time delay ys 60
Quartz cell atomiser (Research Institute for Analytical Instrumentation, Cluj-Napoca, Romania)
Lengthymm 170
Inner diameterymm 12
Heating temperaturey C 900
10
Perkin Elmer Model 5000 atomic absorption spectrometer
Hollow cathode lamp currentymA 12
Wavelengthynm 193.7
Bandpassynm 0.7
Calibration mode steady-stateyconcentration
Replicates 5
Time constantys 0.5
Measurement timeys 10
Background correction (deuterium lamp) On
4 Optimisation of analytical parameters in inorganic arsenic (III and V) speciation by hydride generation
performed with a pH-meter 540 GLP (WTW GmbH,
Weilheim, Germany).
Reagents and standard solution
Chemicals of puriss p.a. or puriss p.a. plus grade
(Fluka) and double distilled water were used
throughout. Arsenic trioxide ACS (499.95%),
As2O55 H2O (99.99%) plus, NaBH4 for the
determination of hydride formers by AAS
(496%), NaOH ACS (499%) and L-cysteine
BioChemika Mikroselekt (499.5%), hydrochloric
acid 10 M (32% mym) for the determination of As
(55610
7 % As) and HNO3 65% were employed.
Percentages of all NaBH4, NaOH and L-cysteine
solutions are expressed as myv.
All glassware was soaked in 5 M HNO3 for a
minimum of 12 h and washed with distilled water
and rinsed with double distilled water before use.
Solutions were stored refrigerated at 4 C.
Stock solution of 1000 mg mL
1 As (III) in HCl
1 M media was prepared by dissolution of the
appropriate mass of As2O3 in 25 ml 20% NaOH,
followed by neutralisation with 1 M HCl using
phenolphthalein as indicator. Stock solution of
1000 mg mL
1 arsenic (V) was prepared by dissol-
ving As2O5 5 H2O in 20% NaOH followed by
neutralisation with 1 M HCl. The 1 L final volume
of As(III) was adjusted to 1 M HCl, while that of
As(V) to 1 M HCl and 0.5 M HNO3. These solutions
were used to prepare daily the stock solutions of
100 ng mL
1 arsenic (III) in 0.005 – 0.05 M HCl
media and 100 ng mL
1 arsenic (V) in 0.01 M and
0.02 M HCl, respectively.
Stock solutions in the range of 0.005 – 0.05 and
1 M HCl prepared by dilution of HCl 10 M with
double distillate water were used to prepare stan-
dard arsenic solutions, blank, stock L-cysteine
solution and carrier fluid to the hydride generator.
A solution of 5 M HNO3 obtained from that of
65% was used to prepare the stock solution of
1000 mg mL
1 arsenic (V).
Stock solutions of 10% L-cysteine were prepared
by dissolving 10 g L-cysteine in 100 mL water and
0.005 – 0.05 M HCl medium, respectively. L-cysteine
was used as both pre-reducing agent of arsenic (V) to
arsenic (III) and hydride generation reagent together
with NaBH4 in NaOH medium.
To optimise the concentration of L-cysteine in
arsine generation, standard solution containing
10 ng mL
1 arsenic and 0 – 2% L-cysteine in
0.01 M HCl and 0.02 M HCl media were used.
Also, various concentrations of NaBH4 solutions in
the range of 0.2 – 1% stabilised with 0.5% NaOH and
solutions containing 0.6% NaBH4 stabilised with
NaOH in the range 0.2 – 1%, respectively, were
daily prepared and then filtered.
Analytical procedure and calibration
The speciation procedure involved three steps: (i)
arsenic (III) determination by selective hydride
generation with NaBH4 immediately after the
sample was mixed with L-cysteine; (ii) total
arsenic determination by prereducing arsenic (V) to
arsenic (III) with L-cysteine by boiling in water bath
at 90 + 5 C and contact time of 10 min and (iii)
arsenic (V) determination as difference between total
arsenic and arsenic (III). Throughout this study the
contact time refers to the time necessary to prereduce
arsenic (V) to arsenic (III) with L-cysteine before
determination by FIA – HG – QFAAS.
Calibration curves were plotted in the range of
0.5 – 25 ng mL
1 arsenic (III) and arsenic (V)
respectively, in the optimum conditions to generate
arsine. In the pre-reducing step of arsenic (V) to
arsenic (III), 5 mL of 10% L-cysteine in 0.01 M HCl
were added to appropriate aliquot volumes from the
100 ng mL
1 arsenic (V) stock solution. The volu-
metric flasks were placed in a boiling water bath for
10 min, the solutions were then cooled to room
temperature. Prior to analysis, the solutions were
diluted so that the final concentration of L-cysteine
reached 0.2%. The pH in solution was strictly
adjusted to 2.00 + 0.01. For each calibration stan-
dard the absorption was considered as average of five
replicate measurements. The limit of detection was
calculated according to 3s criteria using the slope of
the calibration curve (m) and residuals (sd):
LOD ¼ 3 sd ym
To assess the accuracy of the proposed method
standard deviations and recovery degrees for
different mixtures of arsenic (III) and arsenic (V)
were evaluated from five replicate measurements.
Analytical results were checked using the t-test and
Q-test.
RESULTS AND DISCUSSION
Optimisation of arsine generation from arsenic
(III)
The effects of HCl as reaction media and concentra-
tion of L-cysteine, NaBH4 in NaOH solution as well
as the contact time on arsine generation were studied
at the constant reagents flow rates in the VGA77-
Varian FIA-HG system shown in Table 1.
 Emil A. Cordos, Tiberiu Frentiu, Michaela Ponta, Bela Abraham and Joan Marginean
Figure 2 Influence of HCl concentration in sample with 0.2%
L-cysteine on the absorbance signal of 10 ng mL  1 arsenic (III);
reducing agent: 0.6% NaBH4 in 0.5% NaOH; carrier: 0 – 0.05 M
HCl; appropriate blank.
a. Effect of HCl concentration
The influence of HCl concentration within the range
of 0 – 0.05 M in the presence of 0.2% L-cysteine
when using 0.6% NaBH4 in 0.5% NaOH on the
absorption signals of 10 ng mL
1 arsenic (III) is
presented in Figure 2. The carrier was HCl solution
of identical concentration to that of the arsenic (III)
sample. The zero concentration of HCl corresponded
to the arsenic sample containing only L-cysteine in
water as reaction medium, while a water stream was
used as carrier.
Data in Figure 2 show that in the presence of a
concentration of 0.2% L-cysteine in the sample, the
absorption signal increased over a narrow range of
HCl concentration (0.005 – 0.02 M). The maximum
absorption signal occurred for a concentration of
0.01 M HCl both for the sample solution and the
carrier.
There is a large amount of experimental results in
the literature about the influence of HCl concentra-
tion on the arsine generation from different arsenic
species. According to Shraim et al. (1999), arsenite,
arsenate, MMA and DMA have generated approxi-
mately equal absorption signals in HCl medium
within 0.03 – 0.06 M when using 0.6% NaBH4 in
0.5% NaOH after a contact time of 2 h with 0.4%
L-cysteine. When the concentration of L-cysteine
was increased to 5% in 0.01 M HCl, and that of
NaBH4 to 2%, the time to reduce the arsenic (V)
species to arsenic (III) decreased to 5 min. Thus, the
optimisation of HCl and L-cysteine concentrations
and contact time respectively, allows the speciation
as arsenic (III) and arsenic (V).
Our results are very similar to those previously
reported by Chen et al. (1992) obtained in a batch
system. They showed that in the presence of 0.02 M
HCl or HNO3 and 0.33% L-cysteine the absorption
5
signal measured immediately after the solutions
were mixed without prereduction was due only to
arsenic (III) even when arsenic (V) was present.
Under these conditions, the authors reported an
optimum concentration of 0.02 M HCl. Brindle
et al. (1992) reported very little influence on the
absorbance of arsenic (III) of HCl concentration
within 0.01 – 0.06 M when using FIA – HG. By
using a FIA – HG – AAS system, Welz and
Sucmanova (1993a,b) found an optimum concentra-
tion of 0.1 M HCl as carrier in the presence of 1% L-
cysteine to develop arsine from arsenic (III). Le et al.
(1994b) investigated the influence of HCl concentra-
tion up to 4 M on arsine generation from arsenic
(III), arsenic (V), MMA and DMA in the presence
and absence of L-cysteine with FIA – HG – QFAAS
system when using 2.5% NaBH4. They obtained
identical responses for these arsenic species when
using 0.3 – 0.7 M HCl, after 10 – 20 min contact time
of sample with the 2.5% L-cysteine solution. These
findings are different compared to ours and those of
Anderson et al. (1986), who reported almost similar
responses for the four arsenic species when using a
much lower and narrower acid concentration range
of 0.01 – 0.03 M HCl. Thus, there is similar beha-
viour in the hydride generation in the presence of L-
cysteine within the concentration range investigated
by us and that reported by Anderson et al. (1986).
The reason of the disagreement between our results
and those of Welz and Sucmanova (1993a,b) and Le
et al. (1994b) are unclear but the use of higher
concentrations of L-cysteine could be responsible.
Also, our results demonstrated very clearly that the
use of L-cysteine required the careful optimisation of
all the analytical parameters, especially that of pH.
Brindle and Le (1990) have previously shown that
there is a compound formed between the tetrahy-
droborate and the thyol group of L-cysteine whose
reactivity is strongly influenced by the pH of the
solution. Unlike Welz and Sucmanova (1993a,b)
who reported a little influence of acid concentration
within 0.05 – 0.15 M in the presence of 1% L-cyst-
eine on arsine generation, we found that pH was a
critical parameter. This could be due to the concen-
tration of HCl 10 times more diluted and a lower L-
cysteine concentration in our samples. We proceeded
further to study the effect of L-cysteine concentra-
tion in 0.01 M and 0.02 M HCl.
b. Effect of L-cysteine
The effect of L-cysteine in a concentration range of
0 – 2% in the sample on arsine generation was
evaluated from the absorption signals of solutions
containing 10 ng mL
1 arsenic (III) in 0.01 M and
0.02 M HCl, respectively. The concentration of
6 Optimisation of analytical parameters in inorganic arsenic (III and V) speciation by hydride generation
Figure 3 Influence of L-cysteine concentration on the absor-
bance signal of 10 ng mL  1 arsenic (III). A. 0.01 M HCl in
sample solution and carrier; reducing agent: 0.6% NaBH4 in
0.5% NaOH. B. 0.02 M HCl in sample solution and carrier;
reducing agent: 0.6% NaBH4 in 0.5% NaOH.
NaBH4 stabilised in basic media was kept constant at
0.6% NaBH4 in 0.5% NaOH. Flow rates of the
reagents are shown in Table 1, while experimental
results in Figure 3. In each case a solution containing
appropriate levels of L-cysteine and HCl was used as
blank. As shown in Figure 3, the absorbance
increased steeply when the L-cysteine concentration
was increased up to 0.2%, which suggested a kinetic
effect on the arsine development due to the inter-
mediate species formed between the tetrahydrobo-
rate and the thyol group of L-cysteine. Hence, the
absorption was higher up to two orders of magnitude
in the presence of L-cysteine compared to that in its
absence.
For further increase of L-cysteine concentration,
the absorbance decreased slowly. The absorbance
signals were always higher in 0.01 M HCl as
compared to those in 0.02 M HCl. Data in the
literature have mentioned a maximum absorption
signal corresponding to arsenic (III) when using L-
cysteine usually in the range 0.1 – 0.5%. However, in
order to decrease the contact time and minimise the
interference due to transitional metals, Chen et al.
(1992) reported a concentration of 0.33% L-cysteine
instead of 0.1% when using a batch system (0.02 M
HCl). For the same reasons, Welz and Sucmanova
(1993a,b) used 1% L-cysteine instead of 0.5% in a
FIA – HG system (0.1 M HCl). In agreement with
results reported by Chen et al. (1992), we have also
remarked the little influence of L-cysteine concen-
tration in the range of 0.1 – 0.3% on the absorption
signal of arsine ( + 10%). Unlike the results of Welz
and Sucmanova (1993a,b), who reported an insignif-
icant influence on the arsenic (III) absorption signal
when using L-cysteine in the range of 0.5 – 4% in
HCl 0.1 M without a rigorous control of pH, we
Figure 4 Influence of NaBH4 concentration in 0.5% NaOH on
the absorption signal of 10 ng mL  1 arsenic (III) in 0.01 M HCl
with 0.2% L-cysteine; carrier: 0.01 M HCl; blank: 0.2% L-
cysteine in 0.01 M HCl.
found a decrease of the absorption up to half when
using L-cysteine in the range of 0.3 – 2%. Finally, we
used an L-cysteine concentration of 0.2% in 0.01 M
HCl and a rigorous adjustment of pH to 2.00 + 0.01
prior to hydride generation throughout the study. In
these conditions, the interference of arsenic (V) on
arsenic (III) in arsine generation is substantially
reduced since those due to transitional metals are
overcome.
c. Effect of NaBH4 concentration
The influence of NaBH4 concentration in the range
of 0.2 – 1% in the presence of 0.2% L-cysteine on the
absorption signals of 10 ng mL
1 arsenic (III) is
presented in Figure 4.
As shown in Figure 4, for a concentration of
0.01 M HCl both in sample solution and carrier and
in the presence of 0.2% L-cysteine the maximum
absorption signal corresponds to a concentration of
0.6% NaBH4 in 0.5% NaOH. Also, the absorption
increases sharply for NaBH4 concentrations up to
0.6% suggesting a kinetic influence on the hydride
development. For further increases in NaBH4
concentration a limitation of the absorbance signal
is observed.
d. Effect of NaOH concentration
The influence of NaOH concentration in the range of
0 – 1% as stabiliser for the solution of 0.6% NaBH4
on the absorption signal of 10 ng mL
1 arsenic (III)
is presented in Figure 5.
Unlike NaBH4, which seemed to have a kinetic
effect on arsine development, the NaOH concentra-
tion had no significant influence between 0.2 and 1%
on the absorption signal and hence no significance in
the kinetics of hydride generation. The concentration
 Emil A. Cordos, Tiberiu Frentiu, Michaela Ponta, Bela Abraham and Joan Marginean
Figure 5 Influence of NaOH concentration as stabiliser for
0.6% NaBH4 on the absorption signal of 10 ng mL  1 arsenic
(III) in 0.01 M HCl with 0.2% L-cysteine; carrier: 0.01 M HCl;
blank: 0.2% L-cysteine in 0.01 M HCl.
of 0.5% NaOH was chosen as stabiliser for the 0.6%
NaBH4 solution.
The main analytical performances for the determi-
nations of arsenic (III), arsenic (V) and total arsenic
by FIA – HG – QFAAS system were established in
the optimum conditions to generate hydride
presented in Table 2. The detection limit of 0.1 ng
mL
1 arsenic was calculated using the slope of the
calibration curve in the range of 0.5 – 25 ng mL
1
arsenic (III) or prereduced arsenic (V) and the 3s
criteria. This detection limit allowed quantification
upward from 0.5 ng mL
1 arsenic.
Optimisation of arsine generation from arsenic
(V)
To study the generation of arsine from the pentava-
lent arsenic, the absorption signal of 10 ng mL
1
arsenic (V) in 0.01 M HCl by using 0.2% and 0.3%
L-cysteine was measured. Solutions of 0.01 M HCl
and 0.6% NaBH4 in 0.5% NaOH were used as carrier
and reducing agent, respectively. Change in absor-
bance signal as function of contact time is shown in
Figure 6.
It was found that in acid media of 0.01 M and
0.02 M HCl respectively and in the absence of L-
Table 2 Optimum conditions to generate arsine from arsenic (III) in FIA – HG – QFAAS (VGA77-Varian) system
Parameter
Sample flow rate
HCl concentration in sample solution
L-cysteine concentration in sample solution
Carrier flow rate (HCl)
HCl concentration in carrier
NaBH4 flow rate
NaBH4 concentration
Blank
7
Figure 6 Effect of contact time on the absorption signal of
10 ng mL  1 arsenic (V) in 0.01 M HCl when using: A. 0.2% L-
cysteine; B. 0.3% L-cysteine; carrier: 0.01 M HCl; reducing
agent: NaBH4 0.6% in NaOH 0.5%; blank: 0.2% L-cysteine in
0.01 M HCl; room temperature.
cysteine, arsenic (V) in solution gave an absorption
signal not significantly different from that of blank.
Obviously, the ground is that arsenic (V) develops
arsine in different conditions as compared to those of
arsenic (III) and an intermediate prereduction step to
arsenic (III) is necessary. When using 0.2% L-cyst-
eine in the arsenic (V) sample solution (curve A), no
significant absorbance signal was observed at room
temperature for approximately 10 minutes after L-
cysteine had been added. After a contact time of 10
minutes, a small absorption signal corresponding to
arsine became visible, due to prereduction of arsenic
(V) to arsenic (III) by L-cysteine (Figure 6, curve A).
The absorption signal gradually increases with
contact time, which demonstrates that the reducing
process occurs slowly at room temperature. The use
of 0.3% L-cysteine in 0.01 M HCl shortens to 5 min
the minimum contact time with arsenic (V) after that
a small absorption signal could be measured. Curves
A and B show that the pre-reduction of arsenic (V) at
room temperature in 0.01 M HCl needs more than
60 min. For the arsenic (V) solution in 0.01 M HCl in
the presence of 0.2% L-cysteine heated in boiling
water bath at 90 + 5 C for 10 min, the absorption
signal of arsine was similar to that of an arsenic (III)
Optimum value
7 mL min  1
0.01 M (pH ¼ 2.00
0.01)
0.2% myv
1 mL min  1
0.01 M HCl
1 mL min  1
0.6% myv in 0.5% NaOH myv
0.2% L-cysteine in 0.01 M HCl
8 Optimisation of analytical parameters in inorganic arsenic (III and V) speciation by hydride generation
Table 3 Recovery degree in synthetic solutions of arsenic (III) and arsenic (V) (n ¼ 5)
As(III) As(V) Unboiled sample solution
Determination of As (III)
ng mL  1 Found Recovery
(ng mL  1) (%)
mean
SD*
3.00 3.00 2.95
0.10 98
3
8.00 1.00 8.00
0.07 100
1
1.00 8.00 1.05
0.05 105
5
*Standard deviation
solution of identical concentration, suggesting a
complete reduction of arsenic (V) to arsenic (III).
A mechanism of the prereduction step of arsenic (V)
to arsenic (III) in the presence of L-cysteine
followed by the generation of arsine has been
proposed by Howard (1997). As a result of our
studies, it was concluded that to determine arsenic
(V) by hydride generation in 0.01 M HCl a 10 minute
contact time in boiling water bath is necessary. In
order to achieve the complete reduction of arsenic
(V) to arsenic (III), 1% L-cysteine in 0.01 M HCl
(pH ¼ 2.00 + 0.01) was used followed by the sample
dilution to 0.2% L-cysteine to generate arsine.
Validation of arsenic speciation as arsenic (III)
and arsenic (V)
To evaluate the possibility to speciate arsenic (III)
and arsenic (V) as well as arsenic losses during
heating in a boiling water bath, the recovery degree
for mixtures of the two oxidation states in synthetic
solutions was considered. Table 3 presents the
concentration levels of arsenic (III), arsenic (V) and
total arsenic found experimentally in three synthetic
solutions and the expected values. All measurements
were carried out under the optimum conditions to
generate arsine for the two arsenic species.
As shown in Table 3, the recovery of arsenic (III)
in synthetic solution samples was in the range of
98 – 104% while that of total arsenic was within 98 –
101%, which has demonstrated that no significant
arsenic losses occurred through evaporation during
the prereduction step of arsenic (V) to arsenic (III).
Arsenic (V) calculated as difference of the total
arsenic and arsenic (III) show a recovery degree
within 98 – 105%. Also, there was no significant
difference for a 95% confidence interval between
the recovery degree and that theoretically considered
(100%).
CONCLUSIONS
The results reported in this paper show that the
analysis and speciation of arsenic (III) and arsenic
Boiled sample solution Determination of As (V)
Determination of total As as difference of total As and As (III)
Found Recovery Found Recovery
(ng mL  1) (%) (ng mL  1) (%)
mean
SD mean
SD
5.95
0.14 99
2 3.00
0.12 100
4
9.05
0.10 101
1 1.05
0.08 105
4
8.85
0.18 98
2 7.80
0.13 98
2
(V) using a method based on the selective reduc-
tion and FIA – HG – QFAAS technique is rapid and
provides accurate results. The determination of
arsenic (V) as the difference of total arsenic and
arsenic (III) required a prereduction step for
10 min in a boiling water bath. Also, a rigorous
pH control at 2.00 + 0.01 prior to hydride genera-
tion was necessary for a concentration of 0.2% L-
cysteine in the sample. The developed method
could be used to determine leachable arsenic (III)
and arsenic (V) species and total arsenic in soil
samples.
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