Determination of Low-Level Ethylenediaminetetraacetic Acid in Water Samples by Ion Chromatography with Ultraviolet Detection

2007, 65, 229232

T. Kemmei1,&, S. Kodama1, A. Yamamoto2, Y. Inoue3, K. Hayakawa4

1 2

Toyama Institute of Health, 17-1 Nakataikoyama, Imizu, Toyama 939-0363, Japan; E-Mail: tomoko.kenmei@pref.toyama.lg.jp Department of Biological Chemistry, College of Bioscience and Biotechnology, Chubu University, Matsumoto-cho 1200, Kasugai 487-8501, Japan Department of Preventive Medicine and Environmental Health, Osaka City University Medical School, Asahi-machi, Abeno, Osaka 545-8585, Japan Graduate School of Natural Science and Technology, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan

Received: 25 August 2006 / Revised: 17 November 2006 / Accepted: 21 November 2006 Online publication: 14 December 2006

Abstract
A convenient and sensitive ion chromatographic (IC) method for the analysis of ethylenediaminetetraacetic acid (EDTA) in water samples was proposed. Using a fast reversible reaction of free EDTA and metalEDTA complexes into Fe(III)EDTA complex in the presence of Fe(III) ions, sample solutions were applied to an ion-exchange column using a mobile phase (pH 2.3), which was composed of 100 lM Fe(III) chloride and 5 mM methanesulfonic acid. The addition of Fe(III) solution (100 lL) containing 10 mM Fe(III) chloride and 0.5 M methanesulfonic acid to the sample solution (10 mL) permitted the injection of a large volume (400 lL) of sample, which allowed for greater sensitivity. The proposed IC method gave a highly linear (r2 > 0.999) calibration curve ranging 0.0051.0 lM EDTA and had a limit of detection of 1.5 nM. High repeatability (RSD < 2.1%) and recoveries (88108%) were also obtained. With this method, total EDTA level in raw and drinking waters were analyzed successfully.

Keywords
Ion chromatography Drinking water Ethylenediaminetetraacetic acid

Introduction
Ethylenediaminetetraacetic acid (EDTA), the most widely employed aminopolycarboxylic acid, is a synthetic complexing agent that is used in industrial and commercial processes. The hexadentate

ligand of EDTA is a very powerful chelating agent that forms a stable complex with polyvalent metal ions in a 1/1 proportion [13]. EDTA is provided 4,5005,500 ton year)1 in Japan. Huge amounts of EDTA from many sources are discharged to the

aquatic environment. EDTA is not removed during wastewater treatment [46] and is hard to biologically degrade [5, 7, 8]. It has been reported that only the Fe(III)EDTA complex was quickly degraded by photolysis and other metal EDTA complexes were very slowly transformed [6]. While EDTA may not pose a signicant risk to human health or ecological systems [9], it increases the levels of certain heavy metals dissolved even at very low concentrations [10] by both releasing adsorbed metals from sediments and preventing their removal through precipitation. Thus, EDTA could be considered as a critical environmental chemical with regard to contamination of surface water and groundwater [5]. Due to the potential effects of EDTA on metal fate during treatment and after discharge, sensitive analytical techniques are needed for determining metalEDTA complexes in environmental samples as well as in drinking water. A variety of chromatographic methods have been developed for the analysis of EDTA. Gas chromatographic methods using a nitrogen phosphorus-specic detector [6, 11, 12] or mass spectrometer (MS) [13] have been used. These methods, however, require a preliminary derivatization step, which is tedious and time-consuming. The complexation con-

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Other chemicals (analytical grade) were purchased from Wako (Osaka, Japan).

Apparatus for IC

(b)

The IC system consisted of a Toyo Soda CCPD pump (Tokyo, Japan), a Rheodyne 7125 injector with a 400 lL sample loop, a Shimadzu SPD-10AV UV detector, a Shimadzu CTO-10AC column oven (Kyoto, Japan), a Shodex DEGAS degasser (Tokyo, Japan).

column (Sigma-Aldrich Japan, Tokyo, Japan) were done usuing 300 mM methanesulfonate as a mobile phase (pH 0.7). This column was made of poly(divinylbenzene/methacryrate) and packed with 5 lm particles and could be used a below pH 2.

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Results and Discussion

Conversion of EDTA and MetalEDTA Complexes into Fe(III)EDTA Complex Through an IC Column
In the rst experiment, it was tried to analyze EDTA as free EDTA by reversed phase HPLC with a mobile phase at very low pH. That is, 1 mM of EDTA solution was applied to a SUPELCOGEL TPR100 column with 300 mM methanesulfonate (pH 0.7) as a mobile phase. Two peaks were detected at 260 nm (Fig. 1a). When 0.2 mM Fe(III)EDTA complex was applied to the column, only one peak was detected (Fig. 1b), whose retention time was the same as that of the rst peak detected by applying EDTA. Therefore, the rst and second peaks in Fig. 1a appeared to correspond to Fe(III)EDTA complex and free EDTA, respectively. Free EDTA or metalEDTA complexes owing through an HPLC column were previously found to react with Fe(III) in the column, forming Fe(III)EDTA complex, and free EDTA and many metalEDTA complexes could be easily converted into Fe(III)EDTA complex by addition of Fe(III) ions [15]. Moreover, the peak height of Fe(III)EDTA at 260 nm was signicantly higher than that of free EDTA. Thus, on the basis of the reaction of free EDTA and metalEDTA complexes into Fe(III)EDTA complex in the presence of Fe(III) ions, 0.1 mM EDTA solution was analyzed as Fe(III) EDTA complex using a Hitachi gel #2740 IC column and a mobile phase containing 5 mM methanesulfonate (pH 2.3). The conversion rate of free EDTA into Fe(III)EDTA complex increased with increasing Fe(III) chloride concentration in the mobile phase, reaching 95% at 50100 lM Fe(III) chloride (Fig. 2). Similarly, the conversion rates of metalEDTA complexes into Fe(III) EDTA complex were analyzed. Metal EDTA complexes such as Ca(II), Cu(II), Mg(II), Mn(II), Pb(II), Zn(II), Co(II), and Ni(II)EDTA were commercially Full Short Communication

(a)
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Standard and Sample Preparation

0 2 4

Retention time (min)

Fig. 1. Chromatograms of (a) 1 mM free EDTA and (b) 0.2 mM Fe(III)EDTA complex applied to a SUPELCOGEL TPR)100 column with 300 mM methanesulfonate (pH 0.7) as the mobile phase

stant of EDTA with Fe(III) ion is particularly high (pKc 25.1) [3], and Fe(III) EDTA complex has a UV absorption peak. So, UV detection of Fe(III) EDTA complex has been used in high performance liquid chromatography (HPLC) [1419], ion chromatography (IC) [20] and capillary electrophoresis [21]. However, these methods require a concentration step for EDTA concentrations less than 10 lg L)1. Recently, a low level of EDTA has been analyzed by HPLCMS [22], ICMS [23, 24] and ICinductively coupled plasma MS [25]. However, these apparatus are not used widely. In this paper, we propose a conventional IC method using UV detection for the analysis of EDTA at 1.5 nM concentration (about 0.5 lg L)1).

A stock standard of EDTA (1 mM) prepared by dissolving disodium EDTA dihydrate in puried water. The stock solution was stored at 4 C and diluted daily. Environmental water samples (15 river waters, 8 riverbed waters, 37 ground waters, and 12 spring waters) were collected in Toyama prefecture, placed in polyethylene bottles and stored in the dark at 4 C until analysis. Nine mineral water samples were purchased from a local market. Fe(III) solution containing 10 mM Fe(III) chloride and 0.5 M methanesulfonic acid was prepared by dissolving 135 mg of Fe(III) chloride hexahydrate and 2.4 g of methanesulfonic acid in 50 mL puried water. Unless stated otherwise, 100 lL of the Fe(III) solution was added to 10 mL of the diluted standard solutions or samples before they were applied to IC.

Chromatographic Conditions
Separations by IC were attained with a 4.6 mm i.d. 150 mm Hitachi gel IC column (#2740; Hitachi, Tokyo, Japan) thermostated at 40 C. This column was made of polymethacryrate bonded with alkyl quarternary ammonium and packed with 10 lm particles. The mobile phase (pH 2.3), unless stated otherwise, was composed of 100 lM Fe(III)Cl3 and 5 mM methanesulfonic acid, and the ow rate was 1 mL min)1. Analytes were detected at 260 nm. Calculations of EDTA concentrations were based on peak heights. Primary investigations with a 4.6 mm i.d. 150 mm SUPELCOGEL TPR-100

Experimental
Chemicals
Disodium EDTA dihydrate (Na2H2 EDTA2H2O), ferric monosodium EDTA tri-hydrate (FeNaEDTA3H2O), calcium disodium EDTA dihydrate (CaNa2EDTA 2H2O) and other metal EDTA hydrates were obtained from Dojindo (Kumamoto, Japan). Water was puried with a Milli-Q SP.TOC. (Millipore, Tokyo, Japan).

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available as their hydrates and were dissolved to 0.1 mM solution. Al(III) and Cr(III)EDTA complexes could not be purchased, so we prepared Al(III) and Cr(III)EDTA solutions(0.1 mM) by mixing Aluminium(III) nitrate solution or Chromium chloride solution with EDTA solution and boiled for 1 h. As a result, Ca(II), Cu(II), Mg(II), Mn(II), Pb(II), Zn(II) and Al(III) complexes were converted into Fe(III) EDTA complex at rates more than 90% (Fig. 3). However, Co(II)EDTA complex showed only 71% conversion, and Cr(III) and Ni(II)EDTA complexes were not almost converted to Fe(III)EDTA complex. Nowack et al. [15] also reported that Cr(III) and Ni(II)EDTA complexes were hard to react with Fe(III) ions. EDTA exists mainly in the form of Ca(II)EDTA or Zn(II)EDTA complexes in natural waters [11]. So, it seems that, in the case of drinking waters, the low conversion rates of Co(II), Cr(III) and Ni(II) EDTA complexes into Fe(III)EDTA complex have little effect on measurements of the total EDTA concentrations.

100

Conversion rates (%)

50

50

100

FeCl3(M)
Fig. 2. Eect of the concentration of Fe(III) chloride in the mobile phase on the conversion rate of free EDTA into Fe(III)EDTA complex. A Hitachi gel #2740 column with 5 mM methanesulfonate (pH 2.3) as a mobile phase was used

100

Conversion rates (%)

80 60 40 20 0 Ca Cu Mg Mn Pb Zn Al Co Cr Ni

Metal species of metal-EDTA complexes

Fig. 3. Conversion rates of metalEDTA complexes into Fe(III)EDTA complex in a Hitachi gel #2740 column using 100 lM Fe(III)Cl3 and 5 mM methanesulfonate (pH 2.3) as the mobile phase

Analysis of EDTA
In order to analyze with high sensitivity, the eect of sample volume injected on the analysis of EDTA was investigated. When a stock standard solution of EDTA was diluted in water, an increase in injection volume resulted in broader system peaks. In IC, system peaks are generally derived from dierences of the composition between injection solution and mobile phase. When water was applied to the IC system, system peaks were appeared before EDTA peak and a larger volume injection of water broadened the width of them. As a result, with an injection volume of 100 lL, system peaks overlapped with the peak of EDTA. On the other hand, when a mixture of 10 mL of EDTA solution and 100 lL of Fe(III) solution(in which the concentrations of Fe(III) chloride and methanesulfonic acid were the same as those in the mobile phase) was applied to the column, EDTA was successfully analyzed with injection volume ranging from 10 to 400 lL. Therefore, a 400 lL injection volume composed of 100 lM Fe(III)Cl3 and 5 mM methanesulfonic acid was used for further experiments.

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(b)

(a)

Retention time (min)

Fig. 4. Chromatograms of (a) steel-canned mineral water sample and (b) 0.1 lM EDTA standard solution. Chromatograms were obtained with a Hitachi gel #2740 column with 100 lM Fe(III)Cl3 and 5 mM methanesulfonate (pH 2.3) as the mobile phase

Linearity (r2 > 0.999) was demonstrated in the range 0.0051.0 lM by a standard curve for EDTA. The detection limit, dened as a signal-to-noise ratio of 3, was 1.5 nM (approximately 0.5 lg L)1). The large volume injection allowed low

detection limit without a separate preconcentration step before injection. The precision of ve consecutive determinations was evaluated at 0.1 lM of EDTA. High repeatability of peak height (RSD 2.1%) was obtained. To measure

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recoveries, a standard solution of EDTA was added to 15 water samples to a nal concentration of 0.1 lM. Good recoveries (88108%) were obtained. Using the proposed IC method, EDTA in 75 water samples containing 15 river waters, 8 riverbed waters, 37 ground waters, 12 spring waters, and 9 mineral waters were analyzed. EDTA was detected in only one mineral water sample (Fig. 4a). The peak corresponded to a concentration of 0.01 lM (3.0 lg L)1).

Acknowledgements
This work was supported by a Grant-inAid for Scientic Research (C)18580337 from the Japan Society for the Promotion of Science.

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Conclusion
Since free EDTA and its metal complexes, except Co(II), Cr(III) and Ni(II)EDTA, were easily converted to Fe(III)EDTA in the presence of Fe(III) ions, EDTA was detected as Fe(III) EDTA at 260 nm. Total EDTA in raw and drinking water samples, which exists mainly in the form of Ca(II)EDTA or Zn(II)EDTA, was successfully analyzed at low level (1.5 nM) by using the large volume injection without a tedious preconcentration step.

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