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HUNGARIAN JOURNAL
OF INDUSTRIAL CHEMISTRY
VESZPRÉM
Vol. 39(1) pp. 95-99 (2011)
ENVIRONMENTAL SIGNIFICANCE AND IDENTIFICATION OF
METAL-CHELATE COMPLEXES USING ION CHROMATOGRAPHY
R. TÓFALVI , A. SEPSEY, K. HORVÁTH, P. HAJÓS
University of Pannonia, Department of Analytical Chemistry, 8200 Veszprém, Egyetem u. 10., HUNGARY
E-mail: tofalvir@freemail.hu, hajosp@almos.uni-pannon.hu
The trace analysis of metal-complexes has long been an area of interest for analytical chemists and environmental
researchers due to the biological and toxic properties of these compounds. The method for the simultaneous separation of
the metal cations and organic and inorganic anions is based on the use of strong chelating anion with high charge. When
basic solution contains an excess of strong complexing anion of high charge, such as ethylenediaminetetraacetate (EDTA) or
trans-1,2-diamine-cyclohexane-tetraacetic acid (DCTA) ion, most heavy and transition metal ions will occur as anionic
complexes. Hence this method provides simultaneous metal and anion separation. The EDTA and DCTA chelating
agents exhibit strong complexing power. These aminopolycarboxylic acids can remobilize metals in nature. Because
aminopolycarboxylic acids are a potential risk to the environment, it is important to develop an effective analytical
technique for their determination. Several factors affect the retention in the separation of the complex anions: complex
formation reactions, ion-exchange equilibria and protolysis depending on pH. The aim of this work is the optimization of
a simultaneous chromatographic separation and identification of metal ions complexed by the ligand EDTA or DCTA.
The method was utilized to separate CuEDTA2-, CuDCTA2-, ZnEDTA2-, ZnDCTA2-, AlEDTA-, AlDCTA-, Cl-, piruvate
and maleate anions. An advantage of the developed method is that the same basic pH-range is favourable to the stability
of the metal complexes and to the elution.
Keywords: transition metal complexes, EDTA, DCTA ligands, ion exchange chromatography
Introduction
Metal ion speciation and the environment
The presence of transition and heavy metals in the
environmental and biological materials justifies the
importance of high performance environmental qualitative
and quantitative analysis of these species [1]. The
transition metals exist in different oxidation states
possessing different physical and chemical properties
and different toxicity. The main sources of metal-
contamination of the environment are the industrial
emission, vehicle exhaustion, corrosion processes,
households, agriculture, hazardous storage tanks, and
waste disposal sites. The presence of inorganic pollutants,
especially toxic metal ions, is a serious issue, as metal ions
may often be carcinogenic in nature. The identification
of pollutants in environmental matrices is a difficult task
because of strong interference from other components of
the sample. The extended use of palladium in automotive
catalytic converters and in the chemical industry has
also led to increasing concentrations of this metal in
environmental compartments. Platinum group and
heavy metals may enter the environment and interact
with complexing materials, such as humic substances.
Determination of palladium by ion chromatography with
ICP-MS detection was developed by M. C. Bruzzoniti
et al. [2]
Aluminium plays probably a role in the development
of Alzheimer’s disease [3]. The route of these toxic
metal ions to the human body is through water and other
foodstuffs. Therefore, the monitoring of metal ions with
different oxidation states in water bodies and foodstuffs is
essential and important. Some toxic metal ions are also
present in the atmosphere and indirectly affect our health.
Some metal ion in oxoanion forms (AsO4
3-, CrO4
2-) are
transported across cell membranes.
Copper and zinc within those metals that are essential
to life although inherently toxic. The characteristic
oxidation forms of copper are: Cu(I) and Cu(II). In case
of zinc the most frequent forms are Zn(I) and Zn(II).
The change of oxidation state of an element affects the
degree of its bio-availability and toxicity. The different
oxidation states of a particular metal ion possess different
physical and chemical properties. These oxidation states
differ in their redox potential, complexation, and
hydration properties. Therefore the speciation analysis
can differentiate the complexed and free forms of metal
ions. Measuring the total concentration of metal ions
gives no information about the actual chemical forms it
exist, that is important to understand its toxicity and
biotransformation. Therefore the speciation of elements
can not be omitted.
96
Aminopolycarboxylic acids as ligands can remobilize
heavy and transition metals and their release in nature
may cause release of metals into ground water and their
uptake by plants. Degradation of chelating agents is
controversial. While excessive uptake of heavy metals
was viewed as a deterrent for the use of EDTA in
agriculture, the same process is now being researched
because of the possibility that it could be applied to the
phytoremediation of heavy-metal contaminated soils.
However, due to the lack of selective analytical
techniques, the mechanisms of metal uptake by plants in
the presence of EDTA still remain largely speculative.
Since aminopolycarboxylic acids are a potential risk to
the environment, it is also important to develop a
selective analytical technique for their determination.
Chelate chromatography
Chelate chromatography is a special type of ion-
chromatography in which chelating agents as eluent
additives are employed. The ion chromatography is a
suitable speciation technique and it offers reproducible
results.
Simultaneous separation of metals and anions is
based on the use of a strong complexing anion of high
charge [4]. Ethylenediaminetetraacetic acid (EDTA) and
trans-1,2-diamincyclohexanetetraacetic acid (DCTA)
are excellent chelating agents that are able to form
sufficiently stable chelates with different metal ions.
The strong complex-forming anions with high charge
react with most of the di- and trivalent metal cations and
they form complexes with one or two negative charge
that makes the simultaneous separation of metal cations,
organic and inorganic anions possible.
Several factors affect the chromatographic retention
of complex anions. These are the (1) complex formation
reactions, (2) the ion-exchange equilibria, and (3) the
protolysis that depends on the pH of elution.
Retention models have been developed by Hajos et al.
In order to study the retention behavior of metal-complexes
in anion exchange chromatography [4, 5]. The theory [5]
is based on the generalized ion-exchange-, protonation-
and complex-formation equilibria. The unknown ion-
exchange equilibrium constants for the sample and the
eluent species can be determined from experimental
retention data [6] by iterative minimization, using a
non-linear regression algorithm. The model was utilized
to predict the retention behavior of CdEDTA2-, CoEDTA2-,
MnEDTA2- and NiEDTA2- ions. It was concluded that the
chromatographic separation of these species are strongly
influenced by the size of ion, the type, concentration, and
pH of eluent, and the stability of complex.
Experimental
Instrumentation
A Dionex DX-300 ion chromatograph (Sunnyvale, CA,
USA) equipped with a conductivity detector and a Dionex
AMMS-I cation micromembrane suppressor was used
during the work. The separations were carried out by
AS9-HC and AS4A-SC separator columns (250 x 4 mm
i.d.) packed with anion-exchangers functionalized with
alkyl/alkanol quaternary ammonium ions. The
recommended pH-range for the columns was 2–13. All
chromatograms were obtained at room temperature at a
flow rate of 1.2 mL min-1. The injection volume was
50 μl. The micromembrane suppressor was regenerated
with sulphuric acid (0.025 M) at flow rate 3,5 mL min-1.
Reagents and solutions
Eluents were prepared by using analytical grade Na2CO3
and NaHCO3 (Fluka, Switzerland). High purity water
was obtained by using a Milli-Q system (Millipore,
Bedford, MA). The specific resistance of the water was
18.2 MΩ cm-1. Sample solutions of metals, organic and
inorganic anions and the chelating agents (EDTA and
DCTA) were prepared by dilution of a concentrated stock
solution of analytical-grade salts (Fluka). The sample
solutions contained chloride salts of metal cations and
complex forming ligands. Before analysis, all eluents
were treated in an ultrasonic bath for 5 mins in order to
remove air.
Basic components and practice of chelate-
chromatography
The basic components of chelate chromatography are
presented in Fig. 1. The delivery system consists of an
eluent container (Na2CO3, NaHCO3, pH 8–11), liquid
transfer lines, eluent and sample selection valves and a
pump. The sample components (metal-halogenides,
oxoanions) together with complexing agents (EDTA,
DCTA) are injected into the separation system via a
valve injector. Plastic valves made of chemically inert
materials are used. Typical injection volumes are between
10–100 μL. The separator columns are packed with
pellicular anion exchanger in order to obtain optimum
separation condition for ionic components (MEDTA2-,
A-) with an adequately short analysis time. After leaving
the separator column, the separated species pass into the
conductivity detector. The suppressor-type ion
chromatography systems has a unique detection system
in which an ion-exchange membrane enhances the
sensitivity of analysis. The main function of the suppressor
is to chemically reduce the high background conductivity
97
of the electrolyte used as eluent (NaOH → H2O;
Na2CO3 → H2CO3), and to convert the sample anions
into a much more conductive form (NaCl → HCl). A
major advantage of chelate-chromatography – in
contrast to other instrumental analysis such as atomic
spectroscopy – is its ability to detect different species of
anions and cations simultaneously.
Figure 1: Schematic flow diagram of chelate
chromatography
Results and discussion
By adding negatively charged EDTA or DCTA ligand
to positively charged metal ions complex anions with
negative charge form in the solution according to the
following equilibria:
EDTA4- + M2+ ↔ [MEDTA]2- (1)
DCTA4- + M2+ ↔ [MDCTA]2- (2)
The conjugate bases of EDTA and DCTA are 6-dentate
ligands. In case of complex formation, the 6 donoratoms
of the ligand (4 oxygen and 2 nitrogen atoms) are located
at octahedron vertices around the central metal ion. The
high stability of the metal chelates is due to the fact that
the ligand surrounds fully the metal ion and isolates it
from molecules of the solvent. The stability of the
complexes depends on the pH. When the pH increases,
the chelating agents are more and more deprotonated and
exhibit their complexing power. During the formation of
metal chelates pH-dependent side-reactions occur. At the
eluent pH range investigated EDTA and DCTA exist in
two forms: HY3- and Y4-.
In this work, carbonate/hydrogencarbonate electrolyte
was used as eluent at various concentrations and pHs.
The separation system contains three ionic species in the
eluent (CO3
2-, HCO3
- and OH-) and various forms of
organic, inorganic and complex ions in the sample.
During elution, the following simultaneous equilibria
take place in the separator column:
2R-E + MEDTA2- ↔ R2-MEDTA + 2E
- (3)
R-E + MHEDTA- ↔ R-MHEDTA + E- (4)
2R-E + MDCTA2- ↔ R2-MDCTA + 2E
- (5)
R-E + MHDCTA- ↔ R-MHDCTA + E- (6)
where:
R – the charged functional group of the ion-
exchanger
E – the anion of the eluent.
In the suppressor reaction carbonic acid is formed
from the eluent anions:
CO3
2- + 2 H+ ↔ H2CO3 (7)
The retention factors (log k’) of the investigated
anions at different eluent concentrations and pHs can be
seen in Table 1. The result shows clearly that the
increasing eluent concentration decreases the retention
of anions. At the same time, the changing eluent pH
affects the sample composition by changing the fractions
of differently protonated species.
It can be seen in Fig. 2 (pH vs. Φ) that at the pH of
elution the EDTA can exist in two distinct forms with
tri- and four negative charges. It is important to note that
the changing pH of the eluent does not affect the order
of elution of the metal complexes.
An advantage of the applied method is that the same
basic pH-range is favorable to the stability of the metal
complexes and also to the elution.
Figure 2: Partial molar fractions of EDTA in the eluent
at different pHs. The cross-hatched area represents
the pH region of the eluents used
With this separation method, complexes with different
ligands and the complexes of different metal ions can be
separated in the same run (Fig. 3-5). The simultaneous
separation of negatively charged metal chelates and
carboxylate anions can be performed as well (Fig. 4).
The results indicate that the retention is influenced
by the pH of eluent (Table 1). Increasing eluent pH leads
to decrease in retention (k’) because the predominant
form of eluent species is the divalent carbonate above
pH 10 that has higher elution strength than the monovalent
hydrogen carbonate anion. This important factor has to
be considered during the optimization of separation.
98
Table 1: The effect of pH and eluent concentration on the retention of complexes and ligands
k’
Celu [mM] 5.0 6.5 8.0 9.0
pH 10.27 10.86 11.03 9.90 10.27 10.50 10.86 9.44 10.27 10.86 11.03
Cl- 3.88 3.45 3.60 4.20 3.56 3.42 3.08 5.34 3.53 3.14 2.85
EDTA4- 12.00 8.06 9.00 11.89 8.16 6.93 5.95 12.27 6.29 5.38 3.92
EDTA3- 26.09 19.38 20.16 25.18 17.78 15.81 14.63 17.29 14.73 12.68 10.58
DCTA4- 12.80 9.28 9.81 13.54 10.04 8.73 7.11 n.r. 7.96 6.32 5.07
DCTA3- 15.86 11.23 11.93 14.36 12.10 10.55 8.62 21.01 16.96 13.27 10.84
CuEDTA2- 30.27 25.98 25.95 22.53 21.67 19.92 18.87 n.r. 17.55 15.86 14.28
ZnEDTA2- 22.87 19.30 19.95 n.r. 17.84 16.12 14.54 18.20 14.62 12.77 10.72
CuDCTA2- 27.60 19.14 20.84 n.r. 20.41 17.08 14.66 20.22 16.96 13.27 10.65
ZnDCTA2- 28.58 20.01 21.31 n.r. 21.45 17.84 15.30 21.90 17.60 13.72 10.50
n.r.: no retention data
Figure 3: Simultaneous separation of Al and Zn
complexes with EDTA and DCTA chelating agents.
Peaks: 1. Cl-, 2. EDTA4-, 3. DCTA4-, 4. [AlEDTA]-,
5. [AlDCTA]-, 6. [ZnEDTA]2- and [ZnDCTA]2-. Eluent:
9.0 mM Na2CO3, pH = 11.027.
Column: AS9-HC anion exchanger.
Calibration data (Table 2) demonstrate that the
simultaneous IC analysis of metal-chelate complexes
and ligands is sensitive and can be used for quantitation
as well.
Figure 4: Chromatogram of simultaneous separation of
aliphatic carboxyl acids and copper-EDTA complex.
Peaks: 1. Piruvate, 2. Cl-, 3. EDTA4-, 4. Maleate,
5. [CuEDTA].
Eluent: 0.7 mM Na2CO3 + 1.8 mM NaHCO3; pH=9.66.
Column: AS4A-SC.
Figure 5: Chromatogram of EDTA-metal complexes.
Peaks: 1. Cl-, 2. EDTA4-, 3. [CuEDTA], 4. [ZnEDTA].
Eluent: 8.0 mM Na2CO3+ NaHCO3, pH = 10.27.
Column: AS9-HC anion exchanger.
Table 2: Calibration data of EDTA and DCTA chelating
agents and their complexes.
Sample Sensitivity
(μS sec L mg-1)
Linearity (r2)
EDTA4- 3×108 0.8645
EDTA3- 3×108 0.8442
DCTA4- 4×108 0.8771
DCTA3- 2×107 0.7558
[Cu-EDTA]2- 109 0.9931
[Cu-DCTA]2- 109 0.9894
Conclusion
Our experiments verified that the simultaneous analysis
of anions and metal cations can be achieved and the
change of concentration of components can be detected,
the metal complexes and their ligands can be identified.
The retention data of chelate-complexes (Cu2+, Zn2+,
EDTA, DCTA) were given by the use of anion exchange
column packed with pellicular stationary phase, and by the
use of carbonate-hydrogencarbonate eluent and suppressed
conductivity detection. The effective parameters of the
separation were determined considering the composition
of eluent.
99
ACKNOWLEDGEMENTS
Present article was published in the frame of the
projects TÁMOP-4.2.1/B-09/1/KONV-2010-0003 and
TÁMOP-4.2.2/B-10/1-2010-0025. The projects are
realized with the support of the Hungarian Government
and the European Union, with the co-funding of the
European Social Fund. Financial and infrastructural
support of the Hungarian Scientific Research Fund
(OTKA K 81843), and the NTP_OKA_VIII_A_85
Student Grant is also gratefully acknowledged.
REFERENCES
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/NLD (Gebruik deze instellingen om Adobe PDF-documenten te maken die zijn geoptimaliseerd voor prepress-afdrukken van hoge kwaliteit. De gemaakte PDF-documenten kunnen worden geopend met Acrobat en Adobe Reader 5.0 en hoger.)
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/UKR <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>
/ENU (Use these settings to create Adobe PDF documents best suited for high-quality prepress printing. Created PDF documents can be opened with Acrobat and Adobe Reader 5.0 and later.)
>>
/Namespace [
(Adobe)
(Common)
(1.0)
]
/OtherNamespaces [
<<
/AsReaderSpreads false
/CropImagesToFrames true
/ErrorControl /WarnAndContinue
/FlattenerIgnoreSpreadOverrides false
/IncludeGuidesGrids false
/IncludeNonPrinting false
/IncludeSlug false
/Namespace [
(Adobe)
(InDesign)
(4.0)
]
/OmitPlacedBitmaps false
/OmitPlacedEPS false
/OmitPlacedPDF false
/SimulateOverprint /Legacy
>>
<<
/AddBleedMarks false
/AddColorBars false
/AddCropMarks false
/AddPageInfo false
/AddRegMarks false
/ConvertColors /ConvertToCMYK
/DestinationProfileName ()
/DestinationProfileSelector /DocumentCMYK
/Downsample16BitImages true
/FlattenerPreset <<
/PresetSelector /MediumResolution
>>
/FormElements false
/GenerateStructure false
/IncludeBookmarks false
/IncludeHyperlinks false
/IncludeInteractive false
/IncludeLayers false
/IncludeProfiles false
/MultimediaHandling /UseObjectSettings
/Namespace [
(Adobe)
(CreativeSuite)
(2.0)
]
/PDFXOutputIntentProfileSelector /DocumentCMYK
/PreserveEditing true
/UntaggedCMYKHandling /LeaveUntagged
/UntaggedRGBHandling /UseDocumentProfile
/UseDocumentBleed false
>>
]
>> setdistillerparams
<<
/HWResolution [2400 2400]
/PageSize [612.000 792.000]
>> setpagedevice