REV. CHIM. (Bucharest) 63 No. 8 2012 http://www.revistadechimie.

ro 759
Copper Speciation Assessment in Aquatic Ecosystem Affected by
Historical Mining Activities
ELENA BIRSAN
1,2
, ELENA DIACU
1
*
1
University Politehnica of Bucharest, Faculty of Applied Chemistry and Materials Science, 1-7 Polizu Str., 011061, Bucharest,
Romania
2
National Research-Development Institute for Industrial Ecology Bucharest- ECOIND, Romania
Abrud river is an aquatic ecosystem exposed to anthropogenic influences exerted by long-term mining. This
study examines the level of copper concentrations by assessing both total copper content and ionic forms in
surface waters and sediments. These components were studied in 10 control sites located in a section of
river that has been exposed to acidic mine water spills and leaks. The partition coefficient (k
d
) of copper
between the liquid and solid phase suspension was calculated, and filtered and unfiltered liquid samples
were subjected to analytical laboratory determinations required to create a complete image of the chemical
composition of the matrix. The results were integrated into the mathematical model PHREEQC to determine
the weight of chemical species of copper. Speciation of copper in sediment was performed using sequential
extraction procedure. We conclude that the sites most affected by pollution are those with the highest levels
of the bioavailable mobile fraction.
Keywords: aquatic environment, acid mine drainage,copper, speciation, sequential extraction
* email: elena_diacu@yahoo.co.uk
Heavy metal pollution captures more attention to the
whole world because of the dramatic increase of its
presence in air, water and soil. This type of pollution in
aquatic environment is a long term and irreversible process.
Pollution may have natural origin, contamination being due
to the dissolution of minerals and rocks in an aqueous
medium [1], or anthropogenic origin [2, 3], especially the
mining industry, which produced in time large amounts of
waste with high metal content [4].
Environmental contamination due to historical mining
activities has been observed in many regions, the most
common effects are acidification and metal contamination
of soil, sediment, surface water and groundwater.
Production of acidic mine waters is due to exposure of
pyrite (FeS
2
) and other sulfide minerals to air or water. So
takes place sulfide oxidation and that produces a high acid
concentration due to sulfates, iron and other metals,
because between pH and heavy metals concentrations
there is a strong negative correlation [5]. The production of
acid mine water is a quick and self-supportive process
that continues as long as pyrite, air and water are available.
Similarly and more or less simultaneously are occurring
reactions which are releasing large amounts of other
metals such as Mn, Cu, Zn, etc., even long after the mining
activity was stopped [6, 7].
Chemical composition of surface water will influence
and be influenced by the composition of river sediments.
Their old deposits can be displaced in the outer curves of
the river or, during floods along the entire river course. During
floods, particles will be transported from the shores and
banks in the rivers water. Considerable quantities of heavy
metals can then be transported by river water and can be
found again in particulate form [8-10]. As these particles
are larger, their ability to remain in water decreases, so
that particles containing metals are deposited and released
several times from the source. Such processes occur
especially during floods, by two different mechanisms: (i)
the hanging of particles containing metals, (ii) the
adsorption of dissolved species on the surface of particles.
It is widely accepted that only the total metal content
determination into an environmental component is not a
sufficient method for evaluating the potential of
environmental contamination, and is thus required an
assessment of possible mobile bioavailable forms of the
total content. Determination of partition coefficient k
D
[11]
and analysis of chemical speciation of metals in aquatic
ecosystems are tools used to predict environmental risks.
For this purpose it is used for surface water samples the
relatively simple partitioning of metal between the bound
fraction and the dissolved one. This approach assumes
that dissolved metals are mobile and can be taken over by
biota.
Metals are found in surface water mainly in two phases:
dissolved in the water column and in particulate phase -
adsorbed on suspended particles and sediment. Dissolved
phase can be transported through the water column
advection and dispersion processes, while the particulate
fraction can be transported with sediment.
Partition of metals between dissolved and particulate
phases is based on the characteristics of suspended
particles as well as different environmental conditions: pH,
salinity, dissolved organic matter. Fine sediments, acting
as a reservoir for organic compounds and heavy metals
are a potential threat to the aquatic environment.
Contaminated bed sediment may be carried by water
currents, thereby enabling release significant amounts of
metals in water. This desorption of contaminants in the
particle phase may have a pronounced impact on the
environment and therefore on the aquatic ecosystem.
Numerical models which are used provide valuable tools
for predicting the evolution and transport of heavy metals
and are becoming increasingly used in studies of the
surface water hydro management.
Investigations on the content of heavy metals in
sediments are extremely numerous in recent years [12,
13]. If the sediments were originally regarded as deposits
for various chemical species, are now considerate as
active aquatic components which are playing an important
REV. CHIM. (Bucharest) 63 No. 8 2012 http://www.revistadechimie.ro 760
Fig. 1. Location of control sections
and sampling
role in the redistribution of these chemical species to
aquatic biota.
In the absence of anthropogenic influences metals in
sediment are mainly associated with silicates and primary
minerals, so have limited mobility, instead of chemical
elements introduced by human activities that have a higher
mobility and are associated with other phases of the
sediment such as carbon, oxides, hydroxides and sulfides.
Sequential extraction is a technique that provides
detailed information on the origin, propagation mode,
physico-chemical and biological availability, mobility and
transport of metals. This procedure simulates the mobility
and retention of these species in the natural environment
using the changes in environmental conditions for pH,
oxido-reduction potential and organic matter degradation.
It uses a series of reagents, increasing extraction capacity
with each step in order to dissolve the metals present in
different phases of the sediment. Extractors are inert
electrolytes, weak acids, reducing agents, oxidizing agents
and strong acids.
In order to standardize the different extraction schemes
described in literature, a sequential extraction procedure
in three steps was developed by BCR (Community Bureau
of Reference). This procedure has been unanimously
accepted by different authors and applied to different types
of solid samples: river sediments, marine, soil, sludge,
particles in suspension. The idea of this method is that
each extractor dissolves a different component, so that
the final distribution can provide information about the
mobility and bioavailability of an element or potential
environmental risk [14, 15].
The purpose of this study is to assess the distribution of
copper in a river system considered representative for this
type of historical pollution in a region of the Apuseni
Mountains, western Romania, a region that have held
permanent activities of ore extraction gold for 2,000 years.
Achieving this goal was made by measuring the
concentration of copper in the dissolved phase (<0.45 m),
suspended matter and surface sediments and examined
the geochemical composition of surface water and
sediments near the pollution sources.
Experimental part
Sampling, digestion and selective extraction of water and
sediment samples.
The study was conducted in Abrud basin located in the
Western Carpathians, Romania, which is an aquatic
ecosystem exposed to anthropogenic influences exerted
by long-term mining. Surface water and sediment samples
were taken from 10 sites control (May 2010) located in a
section of river about 20 km length that has been exposed
to acidic mine water spills and leaks from dumps for many
decades. The samples were taken from the lower depths
of water courses. (fig.1).
Three sets of liquid samples of 2 L for each sampling
point were collected: The first set was for the complete
characterization of the site (pH, electrical conductivity,
redox potential, temperature); the second one was for the
determination of major ions present in aqueous matrix (in
cold storage) and the third, acidified, was for analysis of
copper and other metallic ions. Homogeneous sample was
used to determine the total copper content and dissolved
copper was determined in the sample filtered through
membranes with porosity of 0.45 m. Sediment samples
were collected from the upper layer of sediment and
collected with the water surrounding, in order to estimate
the natural background of the analytes of interest. To
determine the total content of copper, sediment samples
were subject to laboratory operations: air drying, grinding,
sieve to 63 m grain size fractions, after the removal of
interstitial water. Sediment samples were mineralized
using the method of aqua regia digestion (HCl: HNO
3
v: v
3:1). For each type of sample digestion control solutions
were prepared.
Instrumentation and reagents
The instruments used for the experiments were: atomic
absorption spectrometer Nova A300 (Analytik Jena,
Germany) for total copper determination from water and
sediments, Dionex 3000 ion chromatograph for sulphates
and chlorides determination, SevenGo pH meter (Mettler
Toledo) equipped with 413 Inlab SG glass electrode based
on polymer electrolyte, mechanical shaker - GFL 3018,
Germany for sample preparation; Rotofix 32 (Hettich,
Germany) centrifuge, digestion oven Speedwave MWS-2
(Berghof, Germany), water purification system (Millipore),
REV. CHIM. (Bucharest) 63 No. 8 2012 http://www.revistadechimie.ro 761
Tabel 1
VALUES THAT CHARACTERIZE THE CHEMICAL COMPOSITION OF WATER SAMPLES
water bath DC 10-W19 / B (Thermo Haake, Germany),
analytical balance-AB204-S (Mettler Toledo) and water
bottom dredge sampler Van Veen (Wagtech, UK).
All the reagents used for sample processing and for the
preparation of the extraction solutions have been of
Suprapur (Merck) quality; the water used for solutions was
prepared by a Millipore system.
Determination of partition coefficient of copper in surface
water samples
Partition coefficient was calculated according to
equation based on total concentrations of metals [M]
T
,
concentrations of dissolved phase metals [M]
D
,
concentrations of total suspended solids (TSS). K
d
is
expressed in L/kg, [M]
T
and [M]
D
in mg/L and TSS in mg/L
[16].
(1)
Copper speciation in surface water samples
Since the total metal content does not provide enough
clear information about processes that alter the
characteristics of surface water, for their geochemistry
interpretation is necessary to know the actual distribution
of dissolved species. To this end, it was used a special
computer that can calculate and determine the distribution
and activity of dissolved species. Water chemistry data
were analyzed using geochemical modeling program
PHREEQC [17] provided with a thermodynamic database
that allows interpretation of chemical processes and
reactions occurring in aqueous matrix. This modeling was
done in order to estimate the chemical forms in which
copper is found in surface water. The experimental data
input determined for these calculations were: pH,
temperature, oxidation-reduction potential and data on the
chemical composition of water (dissolved metal
concentrations and major ions).
Copper speciation in river sediment samples
On the 9 sediment samples and a mud sample collected
in May 2010 was applied BCR optimized extraction
procedure. This procedure developed by Quevauviller et al.
[17] has been modified, so in the second stage of
extraction it was used hydroxyl ammonium chloride in a
concentration of 0.5 mol/L, at pH = 2, unlike the original
BCR procedure, which requires the use of hydrochloride
concentration of 0.1 mol/L. Were separated the following
four fractions:
- easily soluble fraction (exchangeable) F1- contains
the metals bounded of carbonates and in the easily soluble
fraction;
- Fe-Mn hydrated oxides fraction (reducible fraction) F2
- contains metals associated with iron and manganese
oxides;
- fraction bound with organic matter and sulfides F3
(oxidable fraction)
- residual fraction, R - containing metals strongly related
to the crystalline structure of minerals, primary and
secondary minerals derived from geological formations.
Sequential extraction of copper from sediment was
carried out in four steps, in the centrifuge tubes, using for
determination 0.5 g sediment sieved by the 63 m sieve.
For the first extraction stage to sediment sample were
added 20 mL acetic acid 0.11 mol/L. The tubes were
agitated for 16 h with 30 rpm with a mechanical shaker,
then were centrifuged at 2,200 rpm for 20 min. Supernatant
- easily soluble fraction copper was analyzed by flame
atomic absorption spectrometry. The sediment in this phase
was washed with 10 mL of distilled water; aqueous phase
was removed after separation by centrifugation for 20 min
at 2,200 rpm.
For the second step of extraction in tube were added 20
mL of hydroxylammonium chloride 0.5 mol/L, pH = 2.
Then, the processes of shaking, centrifugation and washing
were repeated, and the supernatantcontaining copper
associated with iron and manganese oxides, the copper
from the resulting solution being analyzed as described in
first step. As in the previous stage, the sediment was washed
with double distilled water which was removed by
centrifugation.
In the third step, in each tube with the sediment, 5 mL
of hydrogen peroxide were added; then, the tubes were
covered and left at room temperature for one hour and in
water bath at 85 C for another hour. The tube caps were
removed and hydrogen peroxide was evaporated, until the
volume was reduced to about 1 mL. An additional quantity
of 5 mL hydrogen peroxide was added to each tube; tubes
was closed and kept in water bath for 1 hour, following
evaporation to dryness. After cooling, 25 mL ammonium
acetate (acidified to pH 2 with HNO
3
) were added, the
operations of shaking and centrifugation were repeated,
and the supernatant solution representing the fraction
bound to organic matter and sulfides was analyzed from
the copper content.
In the fourth step, the copper from the residual fraction
R was determined, performing solid sample digestion with
28 mL mixture of 1:3 (v / v) nitric acid 65% - 37%
hydrochloric acid.
Results and discussions
Partition coefficient of copper in surface water
Analytical results are presented in table.1. As seen, water
samples have strong acid character and reducing potential
in sections 5 (acid mine water) and 6 (surface water after
discharge of wastewater), as in other sections pH in the
neutral range and oxidizing potential.
REV. CHIM. (Bucharest) 63 No. 8 2012 http://www.revistadechimie.ro 762
Fig.2. Partition coefficient variation
Tabel. 2
COPPER SPECIES DISTRIBUTION (%
OF TOTAL CONTENT) IN SURFACE
WATER SAMPLES
Fig. 3. Total copper content in
river sediment samples
The values obtained for the partition coefficient and for
dissolved copper show that in the investigated area
suspended materials are contaminant transporters. Also,
can be observed the dependence of the k
d
values versus
pH, which shows that at acidic pH values, k
d
are lower,
increasing when the pH samples approach to the neutral
range. Thus we can say that the release of the metals
related to particles is controlled by pH, and the retention
and accumulation of metals in those particles are taking
place at alkaline pH. The chart presented in figure 1shows
the distribution coefficient dependence on concentration
of suspended materials.
Copper speciation in surface water samples
To assess the chemical forms of copper were used
analytical data and PHREEQC program. Thus was found
that the oxidation state I of copper is prevalent, more than
98% of dissolved copper being in this form, the rest exist in
the oxidation state II. Among the divalent copper ionic
forms are distinguished forms of free ionic copper Cu
+2
,
Cu(OH)
2
and CuSO
4
(tabel 2).
Copper speciation in river sediments
Values obtained for total copper concentration and each
extracted fraction concentrations are graphically
represented in figure 3 and 4.
Figure 3 indicates copper level contamination of
watercourses investigated, namely river stream Abrud and
Foies brook, and this can be explained due to inputs
represented primarily by acid mine water discharges 5 into
Foies brook, and leakages from careers. The sediments
have a high content of copper, for the entire watercourse,
in most locations well above the limit allowed by legislation
(Ord.161/2006 CMA 40 mg/kg d.w.), except the value for
locations 5 and 9. Lowest value of 18.51 mg/kg d.w. in
location 5 can be explained because the deposited mud in
mine is due to waste water drain (with a particularly very
acidic pH) and in this condition the copper cannot
precipitates. The location 9, corresponding to Aries River
upstream from the confluence with the Abrud river is also
less polluted, the water being clean. In other locations
investigated, copper concentration values are considerable,
the highest being in section 1, which demonstrates the
significant contribution of water leakages from hillsides
and Rosia Poieni career.
Graphic diagram for copper content of the four fractions
of sediment samples from different locations is as follows
(fig.4).
As it is evident from this figure, copper is found in many
locations: 2, 3, 4, 5, 6, 7, 8. The highest copper content is in
the easily soluble fraction F1 of the sediment, with values
ranging in the area of 29.74% to 47.13%, and in the oxidable
fraction F3. The lower copper content was recorded in the
reducible fraction F2, and in the residual fraction, with
values between 5.73% to 16.86%, and 5.73% to 25.74%,
respectively. The largest share of the residual fraction was
determined for locations 1 and 3, and it shows age deposit
of copper and copper tendency to accumulate in those
sections of the river with large bed and small flow rate. In
REV. CHIM. (Bucharest) 63 No. 8 2012 http://www.revistadechimie.ro 763
Fig.4. Distribution of copper fractions
the locations 1, 3 and 4, copper exist mainly in oxidable
fraction F3, and the fraction R (less accessible for the
solubilisation), which prove the importance of the
contribution of metal to the sediment composition in long
time. For the other locations (2, 7 - 10) the residual fraction
is less represented.
Conclusion
The results of this study show the variability of copper
content in surface water and sediment in rivers located
near sources of pollution from a mining area. Investigation
of the copper content along investigated watercourses,
from Foies brook, as receiver of Acid Mine Water to Abrud
river and Aries river, demonstrates that the effects of copper
pollution are not localized. The conclusion is that in surface
water and sediment, copper is mobile and can be found in
significant amounts away from the pollution source,
phenomenon explained by the values of distribution
coefficient k
d
of copper between dissolved phase and
suspended particles. In addition, the study demonstrated
that copper in the aqueous medium could generate
different chemical species, which can exist in different
quantities. Fractions of the sediment in which copper was
found, show that long-term pollution of mining activities
has two major consequences: long-distance mobility and
the highest bioavailability, but also a tendency for
accumulation of copper in the sediment.
The heavy metal pollution has been studied also by
other researchers [19].
Acknowledgement: The work has been funded by the Sectorial
Operational Programme Human Resources Development 2007-2013
of the Romanian Ministry of Labor, Family and Social Protection
through the Financial Agreement POSDRU/6/1.5/S/16.
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Manuscript received: 15.02.2012