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Food Additives & Contaminants: Part A Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/tfac20 Determination of atrazine and degradation products in Luxembourgish drinking water: origin and fate of potential endocrine-disrupting pesticides T. Bohn a , E. Cocco a , L. Gourdol a , C. Guignard a & L. Hoffmann a a Centre de Recherche Public – Gabriel Lippmann, Department of Environment and Agro- Biotechnologies, 41, rue du Brill, L-4422 Belvaux, Luxembourg Available online: 27 Jun 2011
To cite this article: T. Bohn, E. Cocco, L. Gourdol, C. Guignard & L. Hoffmann (2011): Determination of atrazine and degradation products in Luxembourgish drinking water: origin and fate of potential endocrine-disrupting pesticides, Food Additives & Contaminants: Part A, 28:8, 1041-1054 To link to this article: http://dx.doi.org/10.1080/19440049.2011.580012
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Determination of atrazine and degradation products in Luxembourgish drinking water: origin and fate of potential endocrine-disrupting pesticides T. Bohn*, E. Cocco, L. Gourdol, C. Guignard and L. Hoffmann Centre de Recherche Public – Gabriel Lippmann, Department of Environment and Agro-Biotechnologies, 41, rue du Brill, L-4422 Belvaux, Luxembourg (Received 5 January 2011; final version received 5 April 2011)
Several pesticides have been hypothesized to act as endocrine-disrupting compounds, exhibiting hormonal activity and perturbing normal physiological functions. Among these, especially s-triazine herbicides have received increased attention. Despite being banned in many countries, including the European Union, atrazine is still the world’s most widely used herbicide. Despite its discontinued use, considerable concentrations of atrazine and its degradation products, mainly desethylatrazine (DEA) and deisopropylatrazine (DIA), are still found in the environment, including drinking water sources. The aim of this investigation was to study concentrations of especially s-triazine herbicides and major degradation products in drinking water, including spring water, tap water and bottled water in Luxembourg. Spring water (2007/2008/2009, n ¼ 69/69/69), tap water (2008/2009, n ¼ 19/26), and bottled water (2007/2008/2009, n ¼ 5/13/7) were sampled at locations in Luxembourg and investigated for pesticides by LC-ESI-MS/MS. Atrazine was the predominant triazine, detectable in many spring water locations, tap and bottled water, ranging (mean) from 0–57 (9), 0–44 (4), and 0–4 (1) ng l 1, respectively. DEA and DIA in spring water ranged (mean) from 0–120 (19) and 0–27 (3) ng l 1, with higher concentrations from agricultural areas and low molar ratios of DEA:atrazine 50.5 and high ratios of atrazine:nitrate suggesting point-source contamination. Levels (mean) of DEA and DIA in tap water were 0–62 (14) and 0–6 (51) ng l 1 and in bottled water 0–11 (2) and 0–7 (2) ng l 1. Simazine and other triazines were detected in traces (55ngl 1). Thus, the conducted monitoring suggested the presence of low concentrations of s-triazines in raw and finished water, presumably partly due to non-agricultural contamination, with concentrations being below thresholds advocated by the European Union Directive 98/83/EC. Keywords: s-triazine herbicides; atrazine degradation products; tap and spring water; DEA:atrazine ratio; nitrate; 2,6-dichlorobenzamide
Introduction Among pesticides, especially the s-triazine herbicides Endocrine-disrupting compounds (EDC) are molecules such as terbutylazine, cyanazine, simazine and atra- exhibiting hormonal or antihormonal activities, thus zine, have achieved much attention in recent years, interacting and potentially perturbing normal physio- even though they have been used since the 1950 s logical functions. These include natural compounds, (Ribaudo and Bouzaher 2010). Although the strict such as mycotoxins, e.g. zearalenone (Mantovani et al. classification of atrazine (2-chloro-4-ethylamino-6-iso- 2009; Giraud et al. 2010) and soy isoflavonoids propylamine-s-triazine) as an EDC has been discussed (Cederroth et al. 2010), and man-made chemicals (Solomon 2009), it has been suggested to inflict such as pharmaceuticals (Pailler et al. 2009) and damage to the central nervous system (Rodriguez
Downloaded by [CRP Gabriel Lippmann (Centre de Recherch] at 04:44 05 January 2012 polychlorinated biphenyls (Lasserre et al. 2009), and et al. 2005; Coban and Filipov 2007), the endocrine chemicals employed in agriculture. The latter group system (Rayner et al. 2004; Hayes et al. 2006) and the comprises a large number of pesticides, including, for immune system (Brodkin et al. 2007; Rowe et al. 2008), example, herbicides, fungicides and insecticides. affecting the reproductive health of rats (Kniewald As these compounds have been released on purpose et al. 2000), pigs (Betancourt et al. 2006) and amphib- and are often in direct contact with the future food ians (Hayes et al. 2006), fostering feminization in the product, great prudence in their choice, way of latter (Hayes et al. 2010). However, data on the impact application and concentration has to be maintained. on mammals and particularly humans are scant and Nevertheless, with the increased research on EDC, the effects remain unclear (Solomon 2009). including pesticides, several compounds have been Nevertheless, atrazine is still the most used herbi- hypothesized to impact the human hormonal system. cide in many countries and possibly worldwide
*Corresponding author. Email: [email protected]
ISSN 1944–0049 print/ISSN 1944–0057 online ß 2011 Taylor & Francis DOI: 10.1080/19440049.2011.580012 http://www.informaworld.com 1042 T. Bohn et al.
(Thurman and Scribner 2008). It is used for weed including the s-triazine herbicides atrazine, simazine, removal, especially against broadleaf and grassy weeds, terbuthylazine, sebuthylazine, cyanazine and hexazi- employed in agricultural but also in urban areas such as non, and the atrazine degradation products desethyla- in private gardens, on railway tracks or other weed- trazine and deisopropylatrazine, the urea herbicides control programmes. Atrazine possesses a high ground- isoproturon, chlortoluron, monolinuron, metha- water ubiquity score (GUS) of approximately 3.6 benzthiazuron, metoxuron, diuron, linuron and meto- (Vogue et al. 1994) and thus a high potential for bromuron, the chloroacetanilide herbicides groundwater contamination. In fact, the major way of metazachlor and metolachlor, and 2,6-dichlorobenza- human intake for this pesticide is via water (Bray et al. mide (BAM; Figure 1), a breakdown product of the 2008). In the European Union usage of atrazine was herbicide dichlobenil, were obtained from LGC abandoned in 2004 following general precautionary Promochem (Molsheim, France). Methanol, acetone principles; however, significant concentrations can still and acetonitrile were procured from Biosolve be found in the environment such as in groundwater (Valkenswaard, the Netherlands). and surface water (Administration de la Gestion de l’Eau 2005; Planas et al. 2006; Kuster et al. 2010; Loos et al. 2010), often in concentrations above the 0.1 mgl 1 Sampling campaigns limit set by the European Union Directive 98/83/EC Several campaigns were carried out in the framework (European Commission 1998). In addition to raw water, of this study: atrazine has been detected in finished (tap) water, up to 60 mgl 1 in the United States (Wu et al. 2010), while . Spring water: 69 springs in the Luxembourg concentrations in bottled water are generally assumed City area from sandstone aquifer, the major to be much lower due to filtration steps through aquifer in Luxembourg (for a detailed descrip- activated carbon during production. tion, see Gourdol et al. 2010), were monitored Despite its persistence, atrazine has been shown to between summer 2007 and spring 2009, with a undergo degradation in the soil due to bacterial total of ten collection campaigns (July/ activities. These breakdown products include mainly August, November 2007; January, March, the comparatively stable desethylatrazine and deiso- May, June/July, September, October/ propylatrazine (Papiernik and Spalding 1998; Jason November 2008; January, March 2009; et al. 2010), which have likewise been detected in Figure 2). All these springs are involved in groundwater (Shomar et al. 2006; Loos et al. 2010), the water drinking supply of Luxembourg and tap water (Gfrerer et al. 2002). However, the s- City (capital and largest city) and were triazine ring can be further hydroxylated (Krutz et al. revisited during the entire period to compare 2010) and finally cleaved (Scribner et al. 2005). changes in individual springs. These springs Whether and to what extent these breakdown products could be grouped into nine distinct hydro- possess endocrine-disrupting functions is not entirely geological zones whose recharge areas covered clear, albeit it is being assumed for DEA and DIA a range of different surface usages including (Ralston-Hooper et al. 2009). forest, cropland, urban areas and grassland While data on s-triazines, especially atrazine content (Figure 2). For each individual sampling, in spring water and tap water, have been reported for approximately 2 500 ml of water were sam- some European countries, such as for France (Mirgain pled in pre-washed glass bottles (analyses of et al. 1993; Ministe` re de la Sante´ et des Solidarite´ s 2005; pesticides) or polyethylene bottles (analysis of Brignon 2007) and Germany (Leistra and Boesten 1989; nitrate), and stored at 4 C until analysis Downloaded by [CRP Gabriel Lippmann (Centre de Recherch] at 04:44 05 January 2012 Tappe et al. 2002; Bundesministerium fu¨ r Umwelt (conducted within 48 h of collection). During 2005), no data have recently been published for the sampling campaigns, spring discharges Luxembourg. Thus, the aim of this study was to were also evaluated. investigate pesticide, especially s-triazine, contamina- . Tap water: Luxembourgish tap water from tion of individual spring water locations in Luxembourg various locations in Luxembourg (n ¼ 34, to verify whether contamination is related to agricul- from n ¼ 28 different locations, analysed in tural activities, and finally to determine concentrations duplicate) in spring 2008 and spring 2009 were in finished drinking water (tap water) and bottled water analysed (Figure 3). For this purpose, tap in Luxembourg. water from private households was collected after having let the water tap open for at least 2 min, and then approximately 1.5 L were Material and methods collected in glass bottles and stored at 4 C Reagents and standards until analysis. The campaign focused on Unless otherwise stated, all chemicals were of analyt- southern Luxembourg with its higher ical grade or superior. All herbicide standards population and population density. Food Additives and Contaminants 1043
Cl Cl Cl
N N N N N N
N N N N N NH H2N N N H H H 2 H atrazine desethylatrazine (DEA) deisopropylatrazine (DIA)
Cl Cl Cl
N N N N N N
N N N N N N N N N H H H H H H simazine terbuthylazine sebuthylazine
N H N O N N N N N N N O cyanazine Cl hexaxinon
Cl Cl O O O O N N N N N N H H H isoproturon chlortoluron monolinuron
Cl S O H O Cl N N N N N N N O O H Cl
metabenzthiazuron metoxuron diuron
Cl Cl H O N N O N N O H Br O linuron metobromuron
O Cl NH2 N Cl O O Cl
metazachlor metolachlor 2,6-dichlorobenzamide
Downloaded by [CRP Gabriel Lippmann (Centre de Recherch] at 04:44 05 January 2012 Figure 1. Pesticides and degradation products monitored during sampling campaigns in Luxembourg between summer 2007 and spring 2009.
. Bottled water: a sampling of various bottled Pesticide extraction and concentration water varieties sold in Luxembourg in summer The water pre-concentration was realized similar as 2007, 2008 and 2009, with a focus on locally described previously (Trenholm et al. 2006) using an produced water varieties (n ¼ 25 in total, with automatic SPE workstation (Autotrace SPE worksta- n ¼ 15 from Luxembourg), analysed in dupli- tion, Caliper, Hopkinton, MA, USA), and cartridges cate, was procured. Both sparkling (n ¼ 4) and containing a vinylpyrolidone–divinylbenzene copoly- plain water (n ¼ 21) were included; a total of mer (oasis HLB, 200 mg, Waters, Elstree, UK). For ten different brands was investigated. Samples this purpose, SPE cartridges were preconditioned with were procured in local supermarkets 10 ml methanol followed by 10 ml of water. Then, (CACTUS S.A., CORA), which constitute 500 ml of water were loaded at 10 ml min 1. Pesticides the most visited chains in Luxembourg. were then eluted twice with 5 ml of methanol. Samples 1044 T. Bohn et al.
Figure 2. Locations of spring water (n ¼ 69, circles) included for pesticide analyses from Luxembourg City between summer 2007 and spring 2009. All spring water locations used for drinking water supply in Luxembourg City were included.
were then evaporated to near dryness with a Speedvac with a mobile phase consisting of water containing concentrator (Heto, Denmark) and reconstituted in 0.1% formic acid (FA) (solvent A) and acetonitrile Downloaded by [CRP Gabriel Lippmann (Centre de Recherch] at 04:44 05 January 2012 approximately 0.5 ml of water:acetonitrile 80/20 (v/v) containing 0.1% FA (solvent B). The gradient started and stored at 4 C until analysis. at 20% of B for 3 min and was changed to 40% B until 12 min, to 90% B until 15 min, to 100% B until 15.5 min, remaining at 100% until 18 min, decreased to 20% B at 18.5 min and stayed at 20% B until 20 min. HPLC-MS-MS detection The flow rate was 0.25 ml min 1; injection volume was Detection and quantification was achieved by HPLC 25 ml. Quantification was achieved by using external coupled to a tandem mass spectrometer (LC-MS/MS, standards and external calibration curves, typically in Dionex Ultimate 3000, Sunnyvale, CA, USA, together the range of 1 ng l 1, and deemed acceptable if with an AB Sciex API 3200 mass spectrometer, R 4 0.98. For the MS-MS parameters, vaporizer Concord, ON, Canada), using electrospray ionization temperature was set to 600 C. The capillary voltage (ESI) in positive mode and multiple reaction monitor- was 5 kV. Nebulizer and turbo gas were set at 55 and ing (MRM), similar as described earlier (Baker et al. 50 psig; and curtain and collision gas at 15 and 5 psig, 2000; Panuwet et al. 2008). An Acclaim 120 C-18 respectively. Collision offset voltages were individually column (3 mm, 2.1 100 mm; Dionex) was used at 40 C optimized for each component (Table 1). The retention Food Additives and Contaminants 1045
Figure 3. Locations of tap water sampling (n ¼ 28) for the determination of pesticides in 2008 (stars) and 2009 (triangles) in Luxembourg. Downloaded by [CRP Gabriel Lippmann (Centre de Recherch] at 04:44 05 January 2012
time, parent mass and quantifier ion for each pesticide were filtered through an IC acrodisc Gelman LC13 are displayed in Table 1. 0.45 mm filter (Pall Gelman, Dreieich, Germany) and separated by ion chromatography employing a DX 500 (Dionex) and a AS12A-4 mm column, using a micro- membrane suppressor (ASRS-Ultra, Dionex) and a Nitrate gradient eluent of 2.7 mM Na2CO3/3.0 mM NaHCO3. Nitrate was assessed from water originating from Quantification was achieved using external standards. Luxembourg City springs in order to assess agricul- tural usage of the land (i.e. coming from fertilizer usage) and to compare results with pesticide concentrations. Nitrate in spring water was measured Data analyses by ion chromatography, following the manufacturer’s Unless otherwise reported, all values represent suggestion (Dionex Corp. 1991). In short, samples mean standard deviation (SD). For correlation 1046 T. Bohn et al.
Table 1. Retention time, parent mass, transition ions, collision energy and declustering potential for measuring pesticides following extraction of 500 ml of water samples for each pesticide and SRM conditions for the LC-MS-MS analyses in positive mode.
Retention Parent Transition 1 Transition 2 Pesticide time (min) ion (m/z) DP (V)a CE (V)b CE in (V)b
Atrazine 12.39 216.1 49 174.2 (22) 104.0 (40) Desethylatrazine 4.71 188.1 42 146.1 (22) 104.1 (35) Deisopropylatrazine 2.60 174.1 70 68.0 (42) 104.0 (30) Simazine 9.01 202.1 50 104.1 (33) 124.1 (24) Terbuthylazine 14.76 230.1 52 174.1 (24) 104.0 (43) Sebuthylazine 14.41 230.1 52 174.1 (24) 104.0 (43) Cyanazine 9.67 241.1 50 214.1 (24) 104.1 (40) Hexaxinon 8.40 253.1 50 171.1 (50) 71.0 (50) Isoproturon 13.15 207.2 44 72.0 (40) 165.1 (19) Chlortoluron 11.95 213.1 38 72.0 (37) 140.0 (32) Monolinuron 12.91 215.0 38 125.9 (25) 99.0 (47) Methabenzthiazuron 11.50 222.1 39 165.1 (22) 150.1 (44) Metoxuron 8.04 229.1 46 72.0 (8) 229.1 (22) Diuron 13.20 233.1 38 72.0 (37) 233.1 (34) Linuron 14.87 249.0 35 160.0 (24) 132.9 (47) Metobromuron 13.77 259.0 36 170.1 (23) 90.9 (47) Metazachlor 14.12 278.1 29 134.1 (31) 210.1 (15) Metolachlor 15.59 284.2 39 252.1 (21) 176.2 (33) 2,6-Dichlorobenzamide 3.30 190.0 44 109.0 (50) 172.9 (23) Atrazine-d5 (IS)c 12.26 221.2 40 179.2 (24) –
Notes: aDP, declustering potential. bCE, collision energy. cIS, internal standard (monitoring purpose).
between land usage and concentration of pesticides, sebuthylazine and hexaxinon, were not detected in Pearson correlation coefficients were used to evaluate any of the samples. Only four springs were completely the relation between nitrate and pesticide concentra- free of detectable pesticides. tion, relation of atrazine and degradation products The range of land usage varied from 1.7 to 38.8 (using their molar ratios), and pesticide and nitrate cropland (average ¼ 16.1%; Ministe` re de concentration and land use. For the latter purpose, the l’Environnement Luxembourg 1999), and from 3.2% mean total discharge of the respective pesticides (and to 26.6% (mean ¼ 15.4%) for urban usage, with a clear nitrate) from springs of the same geographical area was correlation between nitrate and cropland (R ¼ 0.86, calculated and plotted against the respective land usage p 5 0.05). A low but significant correlation between as a percentage. For all correlations, both slope and the usage of land as cropland and concentration of intercept were considered. A p 5 0.05 (two-sided) was DEA was found (R ¼ 0.44, p 5 0.001), with lower considered to be statistically significant. positive correlations for atrazine and DIA. Interestingly, a high correlation between BAM and urban land use was found (R ¼ 0.81, p 5 0.05). When Downloaded by [CRP Gabriel Lippmann (Centre de Recherch] at 04:44 05 January 2012 investigating the ratio between DEA and atrazine Results (expressed on a molar basis; Figure 5), a ratio that has Spring water locations been used to discriminate between point-source con- The most common pesticide derived compound was tamination and non-point source contamination 2,6-dichlorobenzamide, a degradation product of (Scribner et al. 2005), most springs plotted on a dichlobenil (Table 2), and detectable in the majority regression line had a mean slope of approximately 2.8 of springs. Atrazine and/or degradation products (R ¼ 0.94). However, a second group of measurements (DEA and DIA) were also detected in most springs, was determined with a slope of approximately 0.3 and concentrations were similar during the entire (R ¼ 0.95), belonging to a group of six springs located 2-year period and the ten campaigns (Figure 4), with together, and associated with low cropland usage no detectable global trend. The major molecule of the (3.4%), but a comparatively high urban coverage s-triazine herbicides and their degradation products (16.8%). When plotting atrazine versus nitrate, a found was DEA, followed by atrazine and DIA similar pattern with the same two groups of springs (Table 2). Simazine was detected in traces while was revealed, with a ratio of atrazine (pmol l 1):nitrate other s-triazine herbicides, including cyanazine, (mg l 1) of approximately 43.4, indicative for the Food Additives and Contaminants 1047
Table 2. Pesticides monitored in Luxembourgish spring water (n ¼ 69), number of findings above the limit of detectiona, and concentrations between summer 2007 and spring 2009.
Locations with Analyses with Minimum Maximum Mean detectable detectable concentration concentration concentration Pesticide concentration (n)a concentrations (%) (ng l 1) (ng l 1) (ng l 1)
Atrazine 58 78.3 0 57 9 Desethylatrazine 59 82.6 0 120 19 Deisopropylatrazine 47 49.6 0 27 3 Simazine 27 24.9 0 5 1 Terbuthylazine 1 0.2 0 1 0 Sebuthylazine 0 0 0 0 0 Cyanazine 0 0 0 0 0 Hexaxinon 0 0 0 0 0 Isoproturon 11 2.0 0 5 0 Chlortoluron 11 10.9 0 6 0 Monolinuron 1 0.2 0 4 0 Methabenzthiazuron 0 0 0 0 0 Metoxuron 0 0 0 0 0 Diuron 7 4.5 0 11 0 Linuron 0 0 0 0 0 Metobromuron 0 0 0 0 0 Metazachlor 0 0 0 0 0 Metolachlor 0 0 0 0 0 P2,6-Dichlorobenzamide 58 79.9 0 346 30 b PATZ þ DEA þ DIA 61 83.5 0 202 32 All pesticides 65 89.4 0 352 63
Notes: aNumber of locations with at least one finding above the limit of detection (1 ng l 1). bATZ, atrazine; DEA, desethylatrazine; DIA, deisopropylatrazine.
Atrazine 50 DEA
45 DIA Simazine 40 Chlortoluron
) 35 BAM –1 30
25
20
15 Concentration (ngl
10
Downloaded by [CRP Gabriel Lippmann (Centre de Recherch] at 04:44 05 January 2012 5
0
Nov-07 Jan-08 Mar-08 May-08 Sep-08 Jan-09 Mar-09 Oct-Nov-08 June-July-08 July/August 07
Figure 4. Concentrations of major herbicides and degradation products detected in spring water from Luxembourg City (n ¼ 69) between summer 2007 and spring 2009. Values are the mean SE. BAM, 2,6 dichlorobenzamide; DEA, desethylatrazine; DIA, deisopropylatrazine.
six springs. BAM and nitrate were only very weakly to indicate that other DIA precursors such as correlated (R ¼ 0.11). Nitrate range was between 3.8 cyanazine and simazine were present (Scribner et al. and 78.1 (mean ¼ 31.5) mg l 1 for all springs. 2005). In the present investigation, the DIA:DEA ratio In addition, a high ratio of DIA:DEA was reported was generally below 1, mostly below 0.5. 1048 T. Bohn et al.
90 700
80 600 70 ) ) 500 –1 60 –1 l 400
mg l 50
40 300 DEA (pmol Nitrate ( Nitrate 30 200 20 100 10 0 0 0 50 100 150 200 250 300 0 50 100 150 200 250 300 Atrazine (pmol l–1) Atrazine (pmol l–1)
90 180 80 160
70 140 ) )
–1 60 120 –1 l
mg l 50 100
40 80 DIA (pmol DIA
Nitrate ( Nitrate 30 60
20 40
10 20
0 0 0 200 400 600 800 0 50 100 150 200 250 300 DEA (pmol l–1) Atrazine (pmol l–1)
90 180
80 160
70 140 ) )
–1 60 120 –1 l
mg l 50 100
40 80 DIA (pmol DIA
Nitrate ( Nitrate 30 60 20 40
10 20
0 0 0 50 100 150 200 0 200 400 600 800 DIA (pmol l–1) DEA (pmol l–1)
Downloaded by [CRP Gabriel Lippmann (Centre de Recherch] at 04:44 05 January 2012 Figure 5. Relation between atrazine, degradation products desethylatrazine (DEA), deisopropylatrazine (DIA) and nitrate in Luxembourgish spring water (n ¼ 69) during 2007–2009 and ten measurement campaigns.
Tap water Tap water concentrations for southern Luxembourg Tap water was collected at a number of locations (n ¼ 15 (defined here as all locations and including those south villages or cities, with each one to five sampling of Luxembourg City) were lower in atrazine and 1 locations) in Luxembourg. Again, DEA was the degradation products DEA and DIA (13 ng l ) com- predominant s-triazine herbicide derivative, found at pared with those north of Luxembourg City (37 ng l 1); concentrations between 0 and 62 (mean ¼ 14) ng l 1, however, results were not significantly different. followed by atrazine with 0–44 (mean ¼ 4) ng l 1 and BAM (five from 15 sampling locations negative) DIA, ranging from 0 to 6 ng l 1, with a sum of atrazine, was the only other herbicide detected, with DEA and DIA of 0–111 (mean ¼ 18) ng l 1 (Figure 6). concentrations of 0 96 (mean ¼ 13) ng l 1, except for Only two out of 15 sampling locations were negatively terbuthylazine, which was found in one occasion at tested for atrazine and degradation products. approximately 1 ng l 1. Food Additives and Contaminants 1049
DIA 100 DEA Atrazine ) –1
l 80
60
40
Herbicide concentration (ng 20
0
Itzig Attert Bissen Belvaux Howald Schoos Dalheim Keispelt OberkornPetange Rodange SoleuvreSteinselTutange Welfrange DifferdangeDudelange Pontpierre Schifflange LeudelangeLuxembourg Rumelange Wellenstein Bettembourg Esch-AlzetteHupperdange Remerschen Mean overall Grevenmacher
Figure 6. Concentration of atrazine and degradation products desethylatrazine (DEA) and deisopropylatrazine (DIA) in tap water from various locations in Luxembourg, collected in spring 2008 and spring 2009. Concentrations shown are means of n ¼ 1–5 locations, depending on the number of tap water locations investigated per location.
Bottled water values for Luxembourg from unfinished and finished Due to the limited number of brands available in drinking water. It also is suggested that the ratio of Luxembourg, seven Luxembourgish water varieties, atrazine:nitrate, in addition to the ratio of with (n ¼ 2) and without carbon dioxide, from two DEA:atrazine, could be used to differentiate between major companies and areas, were investigated. non-point-source agricultural contamination of atra- In addition, three French brands commonly traded zine and non-agricultural contamination, also detected and consumed in Luxembourg were investigated in the present study. Other s-triazine herbicide con- (Table 3). From the n ¼ 14 Luxembourgish samples tamination was negligible by comparison. investigated, only one contained measurable amounts Usage of atrazine had been officially declared of atrazine (no degradation products), with a content illegal by individual states, as in Germany (1991) or of approximately 1.5 ng l 1. Two of the French brands France (2002), before being eventually banned were also negative, but the third (n ¼ 3 samples) con- European Union wide and thus also in Luxembourg tained measurable amounts of atrazine. Overall mean in 2004. However, also in countries where atrazine was concentrations for atrazine were 1 (range ¼ 0–4) ng l 1, banned for a comparably long time, still significant and degradation products DEA 2 (range ¼0–11) ng l 1 concentrations of the parent compound and its main and DIA 2 (range ¼ 0–7) ng l 1. In addition to these, bacterial breakdown products, DEA and DIA, can be Downloaded by [CRP Gabriel Lippmann (Centre de Recherch] at 04:44 05 January 2012 only BAM could be detected in four out of 25 samples detected, with concentrations of DEA typically sur- (all originating from Luxembourg), with average passing that of atrazine. For example, in Bavaria, 1 concentrations of 2 (range ¼ 0–14) ng l . Germany, approximately 5% of sampling points had concentrations 40.1 mgl 1 atrazine and DEA, and in almost 50% of the sampling points either DEA, Discussion atrazine or both could be detected in 1999, 8 years In the present study we have demonstrated that despite after the atrazine ban (Tappe et al. 2002). In countries its ban in 2004, low but measurable concentrations of where atrazine is still in use, such as the United States, the s-triazine herbicides atrazine, and its degradation concentrations are even higher, with atrazine peak 1 products DEA and DIA, were detectable in the concentrations in raw water of up to 230 mgl (Wu majority of spring waters from Luxembourg City, et al. 2010). The long persistence of these compounds is and at similar concentrations at most tap water related to the long half-life of atrazine and their de- locations, but were almost absent in bottled water alkylated metabolites DEA and DIA in deeper soil from Luxembourg, being the first study reporting layers or water, with half-lives of atrazine and DEA 1050 T. Bohn et al.
Table 3. Investigation of several bottled water types frequently sold in Luxembourg procured from local supermarkets during spring 2008 and spring 2009.
Atrazine DEAf Others Number Origin na Type (ng l 1) (ng l 1) (ng l 1)
1 Luxembourg, north (Canton Echternach) 1 Sparkling n.d.b n.d. n.d. 2 Luxembourg, north (Canton Echternach) 3 Weakly sparkling n.d. n.d. n.d. 3 Luxembourg, north (Canton Echternach) 3 Non-sparkling n.d. n.d. 11.9 2.4 c 4 Luxembourg, west (Canton Redingen) 4 Non-sparkling 0.4 0.7 e n.d. 3.3 3.7 c 5 Luxembourg, west (Canton Redingen) 1 Non-sparkling n.d. n.d. n.d. 6 Luxembourg, west, Canton Redingen 1 Non-sparkling n.d. n.d. n.d. 7 Luxembourg, west (Canton Redingen) 2 Non-sparkling n.d. n.d. n.d. 8 France (Haute-Savoie) 3 Non-sparkling n.d. n.d. n.d. 9 France (Alsace-Lorraine) 2 Non-sparkling n.d. n.d. n.d. 10 France (Auvergne) 5 Non-sparkling 2.7 0.9 9.2 1.4 6.2 1.1 d 0.1 0.1 c Mean s-triazinesg 0.3 0.9 1.8
Notes: aNumber of samples from each variety. Each purchased variety was investigated in duplicate. bn.d., Not detected. c2,6,-dichlorobenzamide. dDIA, deisopropylatrazine. eConcentration below the detection limit (1 ng l 1) results from a mean of four samples measured with only one being positive. fDEA, desethylatrazine. gMean of atrazine, DEA and DIA over the ten investigated water brands, respectively.
around 28–178 days (Krutz et al. 2010), with shorter The only other herbicide or derivative with notable half-lives in surface and especially adapted soils, i.e. concentrations detected was BAM, a degradation soils regularly treated with s-triazines (Krutz et al. product of dichlobenil, with concentrations of up to 1 2010). As DEA is more polar and about 20 times more 346 ng l . Dichlobenil has been applied mainly for water soluble (Scribner et al. 2005), it was suggested lawns and gardens. As it was also found to be that it binds less strongly to the soil than atrazine, persistent in the groundwater, it has meanwhile also therefore resulting in higher concentration in ground- been banned in European Union countries, including water, in which degradation is impeded due to lower Luxembourg in 2010 (European Commission 2008). Its bacteria counts that could result in degradation endocrine-disrupting potential is currently not under- stood, but it has been found to affect reproduction in (Scribner et al. 2005). The DIA conversion rate from animals (Cox 1997), and it constitutes the major atrazine is low compared with DEA, explaining its groundwater contaminant in other European Union lower abundance in the environment (Krutz et al. countries (Holtze et al. 2008). In the present investi- 2010). gation, BAM was found in 51 spring water samples The spring water concentrations in the present 1 (out of approximately 700) to surpass slightly the study, ranging from 0 to 57 ng l for atrazine and 0.1 mgl 1 European Union limit. In addition, these from 0 to 120 ng l 1 for its degradation product DEA, 1 high concentrations were only found in few individual were generally, but not always (seven cases 40.1 mgl springs, which are usually not consumed unpooled. DEA), in agreement with the European Union After pooling of various water sources, including Downloaded by [CRP Gabriel Lippmann (Centre de Recherch] at 04:44 05 January 2012 Directive 98/83/EC (European Commission 1998) additional water from northern Luxembourg as is (even though defined for finished water), demanding typical for Luxembourg City, concentrations might be 1 a concentration of a single pesticide below 0.1 mgl . more likely to fall below 0.1 mgl 1. 1 The sum parameter of 50.5 mgl required by the While DEA, the major s-triazine degradation same European Union directive was not violated. compound detected, can be formed from atrazine and Older data from Luxembourgish groundwater from propazine (a less frequently used herbicide in Europe), various aquifers (n ¼ 72 locations from various DIA can be formed from atrazine, simazine or regions) from 2005 and older (Administration de la cyanazine. Thus, a high ratio (40.6; Scribner et al. Gestion de l’Eau 2005) indicated similar, even slightly 2005) of DIA:DEA would indicate several parental higher, concentrations for atrazine (range ¼ sources of DIA. However, this ratio was almost never 0–550 ng l 1, mean ¼ 16 ng l 1) and DEA (range ¼ reached in the present investigation (only in three out 0–158 ng l 1, mean ¼ 16 ng l 1), although DEA was of approximately 700 point measurements), indicating, not detected in the majority of groundwater sampling in addition to the actual low measured concentrations, points by then (47 negative out of 72), nevertheless that simazine and cyanazine were much less frequently highlighting their long persistence. used herbicides compared with atrazine. Food Additives and Contaminants 1051
Another ratio that has been employed is periods of times. Processes used for preparing drinking DEA:atrazine, with a low ratio (approximately 0.5) (tap) water typically include chlorination, flocculation, suggesting point-source contamination, as otherwise, sedimentation, filtration, followed by a second disin- due to its increased formation in soil over time from fection stage such as chlorination, UV or ozonization. atrazine (due to microbiological activity), DEA con- It has been shown in earlier reports that atrazine and centrations would tend to be higher. Thus, the degradation products are quite resistant to these DEA:atrazine ratio was used as an indicator of processes and are hardly removed (Benotti et al. point-source contamination (Scribner et al. 2005), 2009). In our study, when comparing mean spring perhaps suggesting non-agricultural usage of atrazine, water contamination (unfinished water) of atrazine even though additional methods, including several and degradation products (sum of 31 ng l 1) with mean geostatistic approaches (Leterme et al. 2006), have tap water concentrations in Luxembourg City aimed to differentiate between point and non-point (23 ng l 1), only a slight reduction in s-triazine content contamination. In the present study we recognized that could be assumed. However, the comparison is by using this cut-off ratio (0.5), six springs located impeded by the fact that different locations were together possessed such a remarkably lower ratio, investigated, and that tap water for Luxembourg City indicating point-source contamination. In addition, the is diluted with drinking water from northern same springs showed also lower nitrate concentrations 1 1 Luxembourg. and a ratio of atrazine (pmol l ):nitrate (mg l ) 43.4, Bottled water exhibited the lowest concentrations also suggesting that the atrazine source was not linked compared with individual spring water locations and to general agricultural farming practices, and that the tap water, and does not seem to contribute consider- ratio of atrazine to nitrate could be an additional ably to the intake of potential EDC. The one positive suitable marker to differentiate between non-point- sample produced in Luxembourg contained very low source agricultural use and other applications. A amounts of s-triazine herbicides, below 1.5 ng l 1, correlation between nitrate and atrazine has already interestingly only atrazine. The absence or low pesti- been noted (Umweltbundesamt Oesterreich 2006). cide concentrations measured in those waters may be Interestingly, the poor correlation of BAM and explained by the fact that they were obtained from nitrate and the high correlation between BAM and deeper or more remote groundwater sources, and that urban land usage suggested that dichlobenil was not bottled water is typically undergoing filtration through necessarily originating from agricultural use. However, charcoal filters, a process which is known to bind and despite the indication of local atrazine contamination remove pesticides, including s-triazine herbicides (Wu sources, the positive correlation of crop land use and et al. 2010), a step which has not generally been put DEA concentration indicated that agriculture was, in into practice for tap water, partly due to its compar- general, the major source of contamination for atrazine. atively high costs. Corresponding with spring water contamination When estimating a mean consumption of 2.1 L with atrazine, tap water also clearly showed the water per day and capita from various beverage presence of atrazine and its metabolites DEA and sources (Walti et al. 2005), with approximately 35% DIA. Tap water contamination was also reported for coming from tap water, approximately 15% bottled other countries, including the United States (up to water and the remainder from various other sources 60 mgl 1; Wu et al. 2010), China (Gfrerer et al. 2002), (tea, coffee, fruit juices, sodas) that are assumed to be and France, where atrazine and DEA surpassed in low in s-triazine herbicides, i.e. comparable with approximately 1% of the investigated drinking water bottled water, and combining this consumption pattern Downloaded by [CRP Gabriel Lippmann (Centre de Recherch] at 04:44 05 January 2012 0.1 mgl 1 (Ministe` re de la Sante´ et des Solidarite´ s with mean triazine and degradation product concen- 2005), in some areas such as Alsace, with even trations obtained in this study (Æatrazine þ DEA þ 1 413% (atrazine) and 417% (DEA; see http:// DIA in tap and bottled water: 18 ng l and approx- 1 vorort.bund.net/suedlicher-oberrhein/wasser-oberrhein. imately 2 ng l , respectively), it can be estimated that 1 html). Concentrations in the present study were all via this route approximately 16 ng day s-triazine below this concentration; however, individual concen- herbicides are consumed in Luxembourg by its trations varied considerably, with some places showing approximate 0.5 million inhabitants. Even though concentrations just below the European Union require- other food items may contribute to atrazine intake ment, with up to 208 ng l 1 for the sum of detected (United States Department of Public Health and pesticides including their degradation products and up Human Services 2003), water is assumed to be by far to 96 ng l 1 for individual pesticides or their deriva- the most contributing source. An acceptable daily tives, i.e. BAM, emphasizing the importance of intake (ADI) of 5 mgkg 1 body weight for atrazine has continuous efforts further to lower their concentration been suggested (National Registration Authority in drinking water and monitor drinking water locally 1997), thus the estimated intake as determined in this to observe changes of these contaminants over longer study would be far lower. 1052 T. Bohn et al.
In conclusion, the large majority of pesticides and Cederroth CR, Auger J, Zimmermann C, Eustache F, Nef S. their degradation products measured in water samples 2010. Soy, phyto-oestrogens and male reproductive func- in Luxembourg were in accordance with European tion: a review. Int J Androl. 33:304–316. regulations, emphasizing the fact that these safeguards Coban A, Filipov NM. 2007. Dopaminergic toxicity associ- were generally observed. Given that the environment is ated with oral exposure to the herbicide atrazine in juvenile male C57BL/6 mice. J Neurochem. 100:1177–1187. most often polluted by several pollutants that could act Cox C. 1997. Herbicide factsheet – dichlobenil. J Pest Ref. in synergy, it would be prudent in the future to 17:14–20. continue measuring pesticides, their degradation prod- Dionex Corp. 1991. Determination of nitrite and nitrate in ucts and their intake, which is especially important for drinking water using ion chromatography with direct UV pregnant women and infants, who are most prone to detection. Dionex Application Update AU 132. Sunnyvale adverse effects such as those caused by endocrine (CA): Dionex Corp. disruptor activity. European Commission. 1998. Council directive 98/83/EC of 3 November 1998 on the quality of water intended for human consumption. Off J Eur Union L. 330:32–54. Acknowledgements European Commission. 2008. Commission Decision of 18 September 2008 concerning the non-inclusion of dichlobe- The authors thank Mr Jean-Franc¸ ois Iffly and Mr Franc¸ ois nil in Annex I to Council Directive 91/414/EEC and the Barnich for aiding in the water collection, Ms Johanna Ziebel withdrawal of authorisations for plant protection products for helping in the pesticide measurements, Ms Sonia Heitz for cartographic assistance, and Mr N. 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