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Water Research 38 (2004) 38453858 www.elsevier.com/locate/watres

Xenobiotic organic compounds in leachates from ten Danish MSW landllschemical analysis and toxicity tests
A. Baun, A. Ledin, L.A. Reitzel, P.L. Bjerg, T.H. Christensen
Environment and Resources DTU, Technical University of Denmark, Bygningstorvet, Building 115, DK-2800 Kongens Lyngby, Denmark Received 5 March 2003; received in revised form 1 July 2004; accepted 12 July 2004

Abstract A monitoring program comprising chemical analysis and biological toxicity testing of leachate samples from 10 Danish landlls (six engineered and four uncontrolled) revealed the presence of 55 different xenobiotic organic compounds (XOCs) and 10 degradation products of XOCs. The compounds belong to the following groups: BTEX, C3-benzenes, bicyclo compounds, napthalenes, chlorinated aliphatics, phenols (chloro-, methyl-, dimethyl, nonyl-), pesticides, and phthalates. Concentrations of single XOCs ranged from o0.1 to 2220 mg/L. A pesticide screening including 101 different compounds resulted in detection of 18 pesticides and three degradation products. The ndings of degradation products of toluene, phenols, phthalates, pesticides, and nonylphenol ethoxylates show that degradation occurred inside the landlls. In biotests with bacteria and algae it was found that the non-volatile organic compounds were toxic as the samples only needed to be pre-concentrated from 1.3 to 9.4 times to give 50% inhibition of the test organisms. One of the ten samples proved to be genotoxic in the umuC test after 141 times pre-concentration. A major part of the organic chemicals causing toxicity remains unknown and it is recommended to combine chemical analyses and biotests in future monitoring programs. r 2004 Elsevier Ltd. All rights reserved.
Keywords: Chemicals; Degradation products; Ecotoxicity; Genotoxicity; Landlls; Leachate

1. Introduction Household waste and non-hazardous industrial waste disposed of in municipal solid waste (MSW) landlls contain a mixture of many chemical compounds originating from the various discarded products. A number of these chemicals are released during the lifetime of the landll and result in emission of xenobiotic organic compounds (XOCs; i.e. organic chemicals with an anthropogenic source) to the envirCorresponding author. Tel: +45-45-25-15-67; fax: +45-4593-28-50. E-mail address: anb@er.dtu.dk (A. Baun).

onment. Consequently, an unknown number of XOCs will be present in the landll leachates, where they constitute a potential risk to the quality of receiving water bodies (e.g. surface water or groundwater), when the leachates are released into the environment. In a number of studies the issue of XOCs in landll leachates have been addressed (for overview see Kjeldsen et al., 2002), but there is still a limited database and an urgent need for more information about the compounds present in leachate, both with respect to their identity and to the concentrations in MSW leachates. This kind of information forms the minimum requirements for the evaluation of fate and effects of leachates and the information is needed for selection of treatment methods

0043-1354/$ - see front matter r 2004 Elsevier Ltd. All rights reserved. doi:10.1016/j.watres.2004.07.006

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3846 A. Baun et al. / Water Research 38 (2004) 38453858

as well as for the risk assessments of landll emissions. In many leachate studies, the information available on organic compounds is, however, in terms of collective parameters like total organic carbon (TOC), dissolved organic carbon (DOC), non-volatile organic carbon (NVOC), chemical oxygen demand (COD), or biological oxygen demand (BOD). These parameters are relatively easy to measure and give some valuable information about the oxygen consuming potential in the recipient. However, they do not give any information about the identity of XOCs and thereby assessment of toxicity, bioaccumulation, or persistence of XOCs in leachate is often not possible. With regard to the individual organic compounds or groups of compounds the focus has mainly been on hydrophobic, volatile, aromatic and aliphatic organic substances, e.g. benzene, toluene, ethylbenzene and xylenes (BTEX), polyaromatic hydrocarbons, and chlorinated aliphatic and aromatic compounds (see e.g. man, Christensen et al., 2001; Kjeldsen et al., 2002; O 1998; and references herein). Only a limited number of investigations published in the literature have focused on groups of compounds containing polar/ionic groups (i.e. more hydrophilic substances) and these studies have shown that phthalates, phenols and pesticides as well as aliphatic, alicyclic and aromatic carboxylic acids can be present in leachates from MSW landlls (Kjeldsen et al., 2002; Paxeus, 2000). In this context also degradation products xenobiotic organic chemicals should be mentioned as these are important indicators of on-going degradation; however, degradation products may be more polar than the parent compounds, making detection via traditional chemical analysis difcult. Examples of degradation products that may be found in MSW landll leachates are benzylsuccinic acid (BSA) related to toluene degradation (see e.g. Baun et al., 2003), nonylphenol-ethoxycarboxylates (NPECs) generated when the non-ionic detergents nonylphenol-ethoxylates (NPEs) are degraded (Talmage, 1994), and monoesters of o-phthalic acid originating from degradation of diesters of o-phathalic acid (Jonsson et al., 2003). A limitation to traditional chemical analyses of organic compounds is that not all the organic compounds present in leachates from landlls can be identied and therefore potential biological effects of leachates may be underestimated. Testing of landll leachates in biological toxicity tests (biotests) provides a direct functional response that relates to the overall toxic properties of the mixture of compounds present in the leachate sample. Direct biotesting of landll leachates has also been carried out previously (e.g. ment et al., 1996), but a Plotkin and Ram, 1984; Cle study by Baun et al. (1999a) showed that pre-concentration and fractionation of leachate and leachate-polluted groundwater is essential when the toxicity of XOCs is in focus. Solid phase extraction has proven to be a suitable

method for pre-concentration and fractionation in combination with biotests and the approach has for example been applied to leachate-polluted groundwater (Baun et al., 2000). The main objective for this study was to evaluate the occurrence and toxicity of XOCs in leachates from both old landlls (uncontrolled landlls) and landlls still in use (engineered landlls with liners and leachate collection). The monitoring program included sampling at ten Danish landlls and the analytical program included both the chemicalanalytical methods and biotests covering acute, subchronic, and genotoxic effects. The study also focussed on whether degradation products of pesticides, toluene, NPEs, and phthalates could be found in landll leachate and thereby provide evidence of in situ of degradation of XOCs in landlls.

2. Materials and methods 2.1. Landll description Leachates samples were collected from the ten Danish landlls listed in Table 1. Six of the landlls were engineered (i.e. with liners and leachate collection systems), while the other four were old uncontrolled landlls. It can be seen from Table 1, that the landlls, beside the differences in age, also have received different types of wastes. All ten landlls have, however, primarily received MSW and the quantities of waste vary from 1.5 105 to 50 105 m3 of waste. 2.2. Sampling procedure Leachate samples were collected in the spring of 1999 from existing wells. Leachate collection wells covering the whole landll were sampled at all the engineered landlls (Landlls Nos. 16). At the uncontrolled landlls no such collection wells were installed and hence samples were collected in the groundwater below the Landlls Nos. 7 and 8 and in wells located inside the landll just above the groundwater table at Landlls Nos. 9 and 10. At Landlls Nos. 1, 36 and 910 samples were collected using a Grundfos MP1 pump (Grundfos, Bjerringbro, Denmark) tted with Teon tubing. At Landll No. 2, the sampling bottles were lowered into the well, and at landlls 7 and 8, a vacuum system, consisting of Teon tubes and glass asks, was used to collect leachate from Teon multi-level wells (see e.g. Jakobsen et al., 1998). Prior to sampling, the well was purged until steady values in pH, temperature and electrical conductivity were reached. All samples were collected in glass bottles. The prewashing procedure for the bottles depended on the compound that should be analysed for, but 5 L acid

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A. Baun et al. / Water Research 38 (2004) 38453858 Table 1 General characteristics of the MSW landlls included in the monitoring program Landll No. Location Type of landll Waste volume/ landll area (103 m3/104 m2) 1050/7 4950/33 700/7 330/2 500/6 1060/8.5 250/10 450/75 500/4.5 150/2.2 Period of operation Waste type (besides MSW) 3847

1 2 3 4 5 6 7 8 9 10

Forlev Sandholt-Lyndelse Hjer Skovsted Lgstr Esbjerg Grindsted Vejen Srup Arnitlund

Eng. Eng. Eng. Eng. Eng. Eng. Unctrl. Unctrl. Unctrl. Unctrl.

1978 1977 1983 1987 1978 1986 19301977 19621981 19571972 19711980

WWTP sludge, non-hazardous industrial, demolition, combustion cinders WWTP sludge, oil- and creosote contaminated soil WWTP sludge, non-hazardous industrial, demolition, bulky WWTP sludge, asbestos waste, shredder, demolition, contaminated soil Non-hazardous industrial, garden, bulky WWTP sludge, bulky, asbestos, contaminated soil WWTP sludge, demolition, industrial, chemical, bulky Demolition, industrial, chemical WWTP sludge, industrial, demolition, bulky, WWTP sludge, industrial, chemical

Eng.: Engineered landll with liners and leachate collection system; Unctrl.: Uncontrolled old landlls without liners and leachate collection system; MSW: Municipal solid waste; WWTP: Waste water treatment plant.

washed blue cap bottles tted with Teon lined caps were used for most of the parameters. In total, 25 L of leachate were collected at each site. The bottles were lled with nitrogen prior to sampling. During and after sampling, the headspaces in the bottles were ushed with nitrogen. These precautions were taken to avoid aeration of the samples, hereby minimizing the risk for sorption/co-precipitation of XOCs to/with Fe- or Mnprecipitates formed due to changes in the pressure of CO2 and O2 during sampling, transportation, and storage of the samples. Preventing the introduction of oxygen also limited the risk for initiating aerobic degradation processes in the samples during storage. The samples were transported to the laboratory where they were kept at 4 1C until further preparations were carried out.

nonylphenol ethoxycarboxylate (NPEC1), nonylphenol diethoxycarboxylate (NPEC2), naphthalenes (3), pesticides (101), and phenols (28). Phthalate diesters, phthalate monoesters, and phthalic acid were also included in the study as described by Jonsson et al. (2003). The analyses were performed at our laboratory according to Table 2, with the exception of the pesticides, NPEs, and nonylphenol, which were carried out by commercial laboratories by means of GC/MS and LC/MS (SGAB-Analytica, Stockholm, Sweden and MILJ-KEMI, Dansk Milj Center A/S, Albertslund, Denmark).

2.4. Solid phase extraction for biotest application Solid phase extractions (SPE) of leachate samples were carried out using Isolutes ENV+ (International Sorbent Technology Ltd., Hengoed, UK) as SPE resin. Pre-packed columns holding 200 mg of SPE-sorbent were cleaned by successive solvent washes (acetone, methanol, n-hexane). The leachate samples (typically 0.251.0 L) were adjusted to pH 7.0 and the samples were pumped through the SPE columns at ow rates of 3040 mL/min. The column efuents were adjusted to pH 2.0 using conc. HNO3 and passed through precleaned SPE columns conditioned to pH 2.0 by acidied Milli-Q water. The loaded columns were dried with N2 and then eluted using 9 mL of n-hexane/acetone (85/ 15 v/v) followed by 6 mL of methanol. For each water sample, the extracts obtained from the two columns (pH

2.3. Chemical analyses The hydrochemical characterization covered the following parameters: pH, alkalinity, electrical conductivity, temperature, ammonium, chloride, oxygen and dissolved NVOC. Temperature, pH, electrical conductivity, and oxygen were measured in the eld using electrodes, and all other parameters were measured using laboratory standard methods. The following XOCs were selected for analyses (No. of compounds in parentheses): BTEX (5) and the toluene metabolite BSA, C3-benzenes (7), bicyclo compounds (2), chlorinated aliphatics (3), NPEs (2),

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3848 A. Baun et al. / Water Research 38 (2004) 38453858 Table 2 Sample preparations and analytical methods used in the GC/MS analyses of XOCs in leachate samples Compounds Aromatics, chlorinated aliphatics Benzylsuccinic acid NPEC Phenols Phthalates Sample preparation Liquidliquid extraction SPE with in-vial elution and derivatization SPE with in-vial elution and derivatization SPE and derivatization Liquidliquid extraction, derivatization Method 1 2 3 4 5

References: 1. Ru gge et al. (1995), 2. Baun et al. (2003), 3. Field and Reed (1996), 4. Reitzel and Ledin (2002), 5. Jonsson et al. (2003) NPEC: Nonylphenolethoxycarboxylates; SPE: solid phase extraction

2.0 and pH 7.0) were combined. The volume was reduced using gentle aeration with atmospheric air. Extract volumes were reduced and pre-concentration factors from about 25150 times were obtained. 2.5. Biotests A biotest battery consisting of an acute toxicity test (Biotox, Vibrio scheri), a sub-chronic toxicity test (algae, Pseudokirchneriella subcapitata), and a genotoxicity test (umuC, Salmonella typhimurium) was applied. The extracts pre-concentrated by SPE were diluted with the test medium of each test and the acetone was removed by gentle aeration for 18 h using N2 (algal and umuC-tests) or atmospheric air (luminescent bacteria). Samples tested in umuC and luminescent bacteria tests were adjusted to pH 7.0. Extracts tested in algal tests were adjusted to pH 8.1. Algal bioassays were carried out according to the ISO-standard for algal toxicity testing (ISO, 1989) applying a mini-scale test version, described by Arensberg et al. (1995). Ten concentrations without replicates and six controls were included in each test setup. Acid washed 20 mL glass scintillation vials containing 4 mL of test solution were inoculated with a log-phase culture of P. subcapitata (formerly know as Selenastrum capricornutum) to 104 cells/mL. The vials were placed on shaker under continuous light (90710 mE/m/s) and at a temperature of 2172 1C. Algal growth rates were determined from acetone extractions as described by Mayer et al. (1997). The tests were conducted at pH 8.170.2 with typical control growth rates of 1.82.0 d1 during the 48 h of incubation. Concentrationresponse curves were described by the Weibull equation which was tted to data using nonlinear regression applying a computer program (Andersen et al., 1998). The biotests with luminescent bacteria were carried out according to the ISO-standard (ISO, 1996) using lyophilized Vibrio scheri supplied in the BioToxTM Kit (Bio-Orbit, Turku, Finland) as test organisms. Tests were carried out in 1 mL glass vials using ve

concentrations in duplicate and two controls in each test. The reduction in luminescence after 15 min was used as endpoint. Concentrationresponse curves and EC-values were estimated as described for the algal test. The umuC tests for genotoxicity were carried out using four concentrations in triplicate and 12 controls (ISO, 1997). Tests were carried out both with and without metabolic activation by S9 liver homogenate (ISO, 1997). Test results were quantied as described by Baun et al. (1999b) in order to minimize the inuence of growth inhibition on the genotoxicity response.

3. Results and discussion 3.1. General hydrochemical parameters The characterization of the landll leachates in terms of general hydrochemical parameters showed that there were large differences between the landlls (Table 3). For instance, the electrical conductivity varied from 2.6 to 54.1 mS/cm and correspondingly the observed chloride concentrations were from 126 mg/L up to the extremely high concentration of 18,400 mg/L in the leachate from Landll No. 1. The alkalinity was 21.779.4 meq/L and the pH-values were 6.98.0. Leachates from the uncontrolled landlls (Nos. 710) generally had lower values for all these parameters, except for the pH-values, for which no signicant differences between the two groups could be observed (Table 3). These results are within the ranges generally observed in landlls (Kjeldsen et al., 2002), with the exception for the high chloride concentration and electrical conductivity measured in Landll No. 1. The concentration of NVOC was 66363 mg/L (Table 3), which is in the lower range of what has been observed in other landlls (range 3029,000 mg/L; Kjeldsen et al., 2002). It should be noted that all samples were anaerobic when sampled and after storage (evidenced by measurements of oxygen, data not shown), but no further effort was made to establish the redox conditions of the leachates.

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A. Baun et al. / Water Research 38 (2004) 38453858 Table 3 Characteristics of landll leachates with respect to general hydrochemical parameters Parameter Landll No 1 PH Electr. cond. (mS/cm) Alkalinity (meq/L) Chloride (mg/L) Ammonium (mgN/L) NVOC (mg/L) 7.0 54.1 55.7 18,400 860 363 2 8.0 15.3 79.4 2730 546 218 3 7.1 10.1 79.0 2560 623 361 4 7.7 14.5 35.7 3770 340 110 5 7.7 16.2 47.1 4210 340 170 6 7.0 14.3 64.6 3470 546 178 7 6.7 2.57 25.8 126 104 114 8 6.7 3.76 25.5 472 154 121 9 7.1 4.67 28.7 606 205 209 10 6.9 2.57 21.7 195 110 66 3849

b.d.=Below detection limit.

3.2. Occurrence of xenobiotic organic compounds (XOCs) The results of the analyticalchemical investigations of XOCs in landll leachates are shown in Tables 46. It should be mentioned that the complex composition of leachate samples caused severe analytical problems in the present study resulting in varying detection limits and reproducibility for most parameters and especially for the XOCs. The complex composition of the samples may be illustrated by a comparison of the NVOC content with the total carbon content of all the XOCs quantied in the leachate samples. The result of this comparison was that the XOCs accounted for 0.05% to 0.92 % (mean: 0.25%, st. dev.: 0.26) of the total concentration of NVOC in the leachate samples. 3.3. Aromatic compounds The aromatic compounds such as toluene and the alkyltoluenes, alkylbenzenes, the xylenes, naphthalene, and methylnapthalenes were present in all ten landlls. In general the concentrations were in the range 1200 mg/L, with exception of m/p-xylene, which was found in a very high concentration (2220 mg/L) in the leachate from Landll No. 10. Benzene was observed in concentrations above the detection limit in nine of the ten landlls with a maximum concentration of 38.9 mg/L (Table 4). The results are well in accordance with other ndings regarding the BTEXs and naphthalene, although these ndings are in the lower part of the previously reported concentration ranges (cf. Kjeldsen et al., 2002). The present study provides the rst analytical evidence of BSA present in actual landll leachates; however, it has previously been found in a landll leachate plume (Baun et al., 2003). BSA is one of the anaerobic degradation products of toluene and the compound was found in all landll leachates in concentrations ranging from 0.6 to 19.3 mg/L. Therefore,

it is concluded that anaerobic degradation of toluene in the landlls is occurring, though a correlation between the concentration of toluene and the concentration of BSA could not be established. 3.4. Chlorinated aliphatic compounds The chlorinated aliphatic compounds (1,1,1-trichlorethane, trichloroethylene and tetrachloroethylene) have previously been found in a relative large number of landlls (Kjeldsen et al., 2002). Thus, it was somewhat surprising that they not were found in the ten Danish landlls; however, the three compounds may have been degraded due to microbial dechlorination yieding e.g. dichloroethylenes or vinylchloride. This can, however, not be veried, as the degradation products were not included in the present study due to analytical problems with the lower chlorinated compounds. 3.5. Bicyclo-compounds The bicyclo-compounds camphor and fenchone were found in high concentrations (7.3255 mg/L) in Landll Nos. 1, 410 (not included in analyses carried out at Landll Nos. 2 and 3). These compounds may originate from degradation of general organic matter in the waste, but the presence of camphor could also be related to disposal of lacquers, varnishes, and plastics (Paxeus, 2000). 3.6. Phenols and nonylphenol ethoxylates The most frequently found phenols were o/p-cresol and the dimethylated phenols (xylenols), which were observed in all landlls in concentrations reaching a maximum of 61 mg/L (o/p-cresol in Landll No. 3). The chlorinated phenols 3,5 dichlorophenol (3,5 DCP) and 4-chloro-o/mcresol was found in all leachate samples, whereas the mono-substituted chlorophenols (2/3- and 4-chlorophenol) were found less frequently. Other dichlorophenols as

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3850 A. Baun et al. / Water Research 38 (2004) 38453858 Table 4 Concentrations (mg/L) of xenobiotic organic compounds identied in landll leachates Compounds Landll No. 1 2 3 4 5 6

Aromatic hydrocarbons and degradation products Benzene 8.5 b.d. Toluene 15.4 16.6 Ethylbenzene 30.0 10.4 o-Xylene 12.8 18.5 M/p-Xylene 33.5 51.4 n-Propylbenzene 3.9 Isopropylbenzene 92.9 1,2,3-Trimethylbenzene 7.9 1,2,4-Trimethylbenzene 13.8 1,3,5-Trimethylbenzene 5.5 2-Ethyltoluene 6.5 4-Ethyltoluene 9.3 Fenchone 59.3 Camphor 255 Benzyl succinic acid 15.5 19.3 Naphthalene 8.6 0.6 1-Methylnaphthalene 2.7 2-Methylnaphthalene 4.5 Chlorinated hydrocarbons 1,1,1-Trichlorethane Trichlorethylene Tetrachlorethylene NPEs/degradation products Nonylphenol NPE1 NPE2 NPEC1 NPEC2 Phenols Phenol m-Cresol O/p-Cresol 3,5-Dimethylphenol 2,5-Dimethylphenol 2,4-dimethylphenol 3,4-Dimethylphenol 2,6/2,3-Dimethylphenol 2-Methoxyphenol 2/3-Chlorphenol 4-Chlorphenol 4-Chlor-o/m-cresol 3,5-Dichlorphenol Compound b.d. b.d b.d b.d. (25) b.d. (20) b.d. (20) 0.5 b.d. 2.2 17.0 29.0 27.3 4.5 12.5 10.4 1.9 b.d. 1.6 1.3 4.6 0.37 Landll No. 7 8 b.d b.d b.d. b.d. (20) b.d. (15) b.d. (15) 3.0 b.d. 0.61.9 19.2 13.6 17.3 3.2 2.1 5.9 1.2 b.d. 1.1 0.86 8.7 0.63

6.9 241 66.1 40.9 118 11.6 1.8 b.d b.d b.d. b.d. (3.0) b.d. (6.0) b.d. (6.0) b.d. b.d. 0.61.9 20.9 60.9 14.7 3.1 10.3 9.3 1.8 b.d. 1.3 0.44 10.2 0.38

3.0 4.7 16.5 9.5 23.2 3.9 4.0 71 15.4 4.8 5.4 6.8 83.0 56.2 4.5 8.8 3.1 4.7 b.d b.d b.d. b.d. (2.0) b.d. (5.0) b.d. (5.0) b.d. b.d. b.d. b.d. 0.50 2.4 1.0 0.22 0.07 1.1 0.020.06 b.d. b.d. 5.6 0.29

2.3 5.9 7.8 5.5 19.4 3.3 3.7 6.2 9.1 4.3 5.1 6.6 7.3 20.6 3.8 4.9 2.6 4.2 5.48 8.61 2.19 6.3 b.d. (5.0) b.d. (5.0) b.d. b.d. b.d. b.d. 0.36 0.71 0.41 0.12 0.03 1.1 0.020.06 b.d. b.d. 3.3 0.09

14.1 19.1 150 53.2 201 7.5 7.5 19.6 47.0 11.6 16.2 21.7 70.3 168 3.5 16.6 4.3 5.9 b.d b.d b.d b.d. (3.0) b.d. (5.0) b.d. (5.0) b.d. b.d. b.d. 1.0 2.5 6.2 1.1 4.3 3.6 0.47 b.d. 1.0 1.2 6.7 0.40

10

LOD/LOQa

Aromatic hydrocarbons and degradation products Benzene 38.9 5.8 Toluene 1.4 4.1 Ethylbenzene 12.4 10.1 o-Xylene 11.8 8.8 M/p-Xylene 25.7 34.1

14.4 4.1 2.5 2.0 5.4

6.1 1.9 32.1 6.4 2220

0.13/0.42 0.04/0.13 0.10/0.35 0.05/0.17 0.16/0.53

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A. Baun et al. / Water Research 38 (2004) 38453858 Table 4 (continued) Compound 7 n-Propylbenzene Isopropylbenzene 1,2,3-Trimethylbenzene 1,2,4-Trimethylbenzene 1,3,5-Trimethylbenzene 2-Ethyltoluene 4-Ethyltoluene Fenchone Camphor Benzyl succinic acid Naphthalene 1-Methylnaphthalene 2-Methylnaphthalene Chlorinated hydrocarbons 1,1,1-Trichlorethane Trichlorethylene Tetrachlorethylene NPEs/degradation products Nonylphenol NPE1 NPE2 NPEC1 NPEC2 Phenols Phenol m-Cresol O/p-Cresol 3,5-Dimethylphenol 2,5-Dimethylphenol 2,4-Dimethylphenol 3,4-Dimethylphenol 2,6/2,3-Dimethylphenol 2-Methoxyphenol 2/3-Chlorphenol 4-Chlorphenol 4-Chlor-o/m-cresol 3,5-Dichlorphenol 3.6 3.3 12.0 13.5 6.7 8.5 7.6 17.1 85.2 0.9 8.7 2.6 4.4 b.d b.d 6.92 b.d. (3.0) b.d. (5.0) b.d. (25) b.d. b.d. 0.61.9 b.d. 1.6 4.0 1.9 1.5 0.93 1.5 0.020.06 0.52 0.030.11 3.1 0.18 Landll No. 8 6.6 5.6 33.3 51.6 14.5 26.1 20.0 58.8 131 0.6 91.2 9.0 9.1 b.d b.d b.d b.d. (2.0) b.d. (4.0) b.d. (4.0) b.d. b.d. b.d. b.d. 0.12 1.0 b.d. 1.0 0.12 0.32 b.d. b.d. b.d. 3.3 0.010.04 9 2.9 3.3 4.8 5.3 3.9 4.5 5.1 9.8 73.0 11.4 9.7 2.4 4.2 b.d. b.d. b.d. b.d. (1.0) b.d. (1.0) b.d. (2.0) b.d. b.d. b.d. 1.0 6.8 3.1 1.6 3.7 1.6 1.5 b.d. b.d. 0.030.11 1.2 0.26 10 15.6 18.7 67.7 152 36.8 34.0 69.8 8.8 65.3 6.9 114.7 12.2 12.2 b.d. b.d b.d. 7.0 b.d. (3.0) b.d. (5.0) b.d. b.d. b.d. b.d. 0.24 3.6 1.4 8.7 0.69 0.52 b.d. 0.03 0.18 3.6 0.08 LOD/LOQa 0.44/1.47 0.09/0.30 0.09/0.30 0.14/0.48 0.25/0.85 0.10/0.34 1.38/4.59 1.74/5.80 1.83/3.43 0.1/0.1 0.08/0.28 0.09/0.3 0.23/0.78 0.09/0.09 0.14/0.45 0.09/0.09
b b b

3851

0.5/0.5 2.0/2.0 0.60/1.9 0.16/0.52 0.02/0.06 0.006/0.02 0.006/0.02 0.02/0.08 0.006/0.02 0.05/0.15 0.02/0.06 0.004/0.01 0.03/0.11 0.01/0.04 0.01/0.04

NPE1: nonylphenol ethoxylate; NPE2: nonylphenol diethoxylate; NPEC1: nonylphenol ethoxycarboxylate; NPEC2: nonylphenol diethoxycarboxylate. a LOD=limit of detection; LOQ=limit of quantication. b Value in parenthesis is the detection limit, as LOD for these compounds varied due to the samples matrixnot included in the analyses.

well as tri-, tetra- and pentachlorophenols were not detected in any of the leachates (limit of detection 0.0040.073 mg/L). The occurrence of 3,5 DCP in all of the ten landlls is striking, as this compound has no commercial use. The ndings of 3,5 DCP may be related to the presence of higher chlorinated phenols like pentachlorphenol (PCP), either as an impurity in technical grade phenols (identied in PCP-products by Lanouette et al. (1984) or due to anaerobic degradation of PCP (e.g. Hendriksen et al., 1992).

Relatively high concentrations of 4-chloro-o/m-cresol was found (range 1.210.2 mg/L) indicating degradation of pesticide compounds (MCPP/MCPA) or disposal of the pure compound/products containing 4-chloro-o/mcresol. Nonylphenol was found in quantiable amounts in landlls Nos. 6 and 10, i.e. concentrations of 6.3 and 7 mg/L, respectively. None of the different forms of the detergent NPE was found in concentrations above the detection limit in any of the landlls. However, the

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3852 A. Baun et al. / Water Research 38 (2004) 38453858 Table 5 Concentrations of mono- and diesters of o-phthalic acid found in landll leachates. Landll No. 1 Compounds Concentration (mg/L) 260 2 5 6 14 2 2 3 21 1 60 340 4 2 1

2 3 7

9 10

Phthalic acid (PA) Monoethylphthalate (MEP) Monobutylphthalate (MbutP) Monobenzylphthalate (MbenzP) Monoethylhexylphthalate (MEHP) Phthalic acid (PA) Butylbenzylphthalate (BBP) Diethylhexylphthalate (DEHP) Phthalic acid (PA) Monomethylphthalate (MMP) Diethylphthalate (DEP) Monoethylphthalate (MEP) Monobutylphthalate (MbutP) Diethylhexylphthalate (DEHP) Diethylhexylphthalate (DEHP)

phthalate contents of leachate from Landll No. 1 is interesting as several degradation products were found (PA, MEP, MbutP, MbenzP, MEHP), but none of the parent compounds. Especially interesting is the relatively high concentration of MEHP (14 mg/L), which is related to degradation of DEHP in the landll. The overall ndings of phthalate monoesters indicate that degradation of phthalate diesters have occurred inside the landlls and as a consequence hereof the monoester degradation products should be included in future monitoring programs and risk assessments of MSW landll leachates. 3.8. Pesticides and their degradation products Twenty-one different pesticides were identied through the screening program, which covered 101 different pesticides. The pesticides found in the landll leachates are listed in Table 6 and the compounds not found are listed in Appendix A, which also shows the detection limits of compounds not found. The most frequently found pesticides were the phenoxyacid herbicides: MCPP (or Mecoprop; present in nine landlls), Dichlorprop (eight landlls), and MCPA (three landlls). These dissociated acids are expected to be easily transported through the landll matrix due to man, their inherent physicalchemical properties (O 2001). The phenoxyacids have also been identied in leachates from other landlls and especially MCPP has frequently been found (Schultz and Kjeldsen, 1986; man and Gintautas et al., 1992; Kjeldsen, 1993; O man, 1999). A number of pesticide Hynning, 1993; O compounds not previously reported as constituents of MSW landll leachates were also identied in the present study, e.g. Ametryn, Atrazine, Bentazon, Chlorpropham, and Dichlobenil (Table 6). An extended pesticide screening program for leachates from Landlls Nos. 2 and 3 resulted in detection of an array of pesticides not previously found in MSW leachates (i.e. Chloridazon, 4-CPP, 2,6-DCPP, Glyphosate, Hexazinon, Isoproturon, and Propoxuron). Furthermore, degradation products of Glyphosate (AMPA) and Simazine (hydroxysimazine) were found (Table 6). The degradation product hydroxyatrazine was also found in leachates from Landlls Nos. 2 and 3, whereas the other known degradation products of Atrazine (desethylatrazine and desisopropylatrazine) could not be detected (cf. Appendix A). It is noteworthy that the triazine parent compounds were not found in concentrations above the detection limits (0.05 mg/L for Atrazine and 0.1 mg/L for Simazine) in leachates from Landlls Nos. 2 and 3, but their degradation products were found in concentrations from 0.6 to 1.7 mg/L. Only three pesticide chemicals (i.e. Dichlobenil, Dichlorprop, MCPP) were found in leachates from the old landlls (Nos. 710) and concentrations were low

Note: Results from landlls Nos. 13 have previously been published (Jonsson et al., 2003).

degradation product nonylphenol ethoxycarboxylate (NPEC1) was found in Landll Nos. 1 and 2. The reason for nding the degradation products and not the parent compounds could be that the NPEs were degraded in the landll, but it could also be due the detection limits of these compounds. The detection limits for the NPEs varied from 1 to 20 mg/L depending on the sample matrix, while the detection limit for the NPEC1 and NPEC2 were 0.5 and 2 mg/L, respectively. 3.7. Mono- and diester of o-phthalic acid The occurrence of mono- and diesters of o-phthalic acid in landll leachates included in the present study has been described by Jonsson et al. (2003) and a summary is given in Table 5. Jonsson et al. (2003) found that diesters of o-phthalic acid were present in leachates from landlls Nos. 2, 3, 7, 9, and 10 (DEP, BBP, DEHP), whereas the corresponding degradation products, the monoesters (MEP, MbutP/MbenzP, MEHP), were found only in Landlls Nos. 1 and 7. In this respect, the concomitant presence of high concentrations of DEP (60 mg/L) and MEP (340 mg/L) is interesting and unprecedented in eld studies on landll leachate. The monoester MMP was found in leachate from Landll No. 7, but the parent compound DMP could not be detected. Phthalic acid, which may be either a degradation product of the diesters or could have been disposed of as a chemical, was found in leachates from landlls Nos. 1, 2, and 7 in concentrations up to 260 mg/L. The

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A. Baun et al. / Water Research 38 (2004) 38453858 Table 6 Concentrations of pesticides found in leachate samples Landll No. 1 Glycines and degradation products AMPA Glyphosate Phenoxy alkanoic acids 4-CPP 2,6-DCPP Dichlorprop MCPA Mechlorprop (MCPP) Triazines and degradation Ametryn Atrazine Hexazinon Hydroxyatrazine Hydroxysimazine Simazine Other pesticides Bentazon Chloridazon Chlorpropham Dichlobenil Isoproturon Fenpropimorf Propoxur Tridemorph 0.87 23 products 1.1 26 0.26 2.1 2 3 4 5 6 7 8 9 10 LOD 3853

4.3 1.7 15 1.3 0.93 0.22 6.7 1.3 1.7 1.7 1.0 0.12

3.8 27 19 0.74 2.8 0.46 28 0.66 0.61 4.0 1.6 1.3 2.6

5.2 9.1 15 0.16 1.4 0.23

1.3 5.2 2.3 2.2 0.12

0.66 16 0.12 0.27

0.27

0.38 150

13

0.84 0.29

0.05 0.05 0.10 0.10 0.15 0.20 0.25 0.1 0.10 0.10 0.40 0.40 0.1 0.10 1.0 0.10 0.10 0.50 0.10 1.0 0.1

Concentrations in mg/L. 4-CPP: 4-chlorophenoxypropionic acid; 2,6-DCPP: 2,6-dichlorophenoxypropionic acid. : Below detection limit; : not included in analyses.

with the exception of MCPP in leachate from Landll Nos. 8 and 9 (Table 6). 3.9. Genotoxicity of landll leachates Genotoxicity tests of the SPE-extract from Landll No. 1 revealed genotoxic effects, but all other samples did not shown any signs of genotoxicity in the range of concentrations tested (from 1 to 150 times pre-concentrated, see Table 7). At the highest concentration also clear cytotoxic effects occurred when testing leachate from Landll No. 1 with a 36% inhibition of the biomass. Taking this general toxic response into account, by use of the procedure suggested by Baun et al. (1999b), the induction ratio was still 1.9 times higher than the induction ratio of the control group in the test with metabolic activation (i.e. with addition of S9 liver homogenate) when the sample was preconcentrated 141 times. According to the ISO-standard of the umuC-test, an induction ratio above 1.5 should be interpreted as a positive result (ISO, 1997). Therefore, it is concluded that non-volatile organic compounds, which can be

converted to genotoxic compounds by liver enzymes, were present in the leachate collected at Landll No. 1. 3.10. Toxicity of landll leachates The non-volatile organic compounds present in the leachates gave toxic responses in both algal and bacterial test systems. As an example of results obtained in these tests, Fig. 1 shows the concentrationresponse curves obtained in algal tests with SPE-extracts of leachates from all the ten landlls. From these curves the effect concentrations (EC-values) can be estimated and the results of tests with algae and bacteria are listed in Table 7 in terms of EC50-values. Judged from these results the most toxic sample was collected at Landll No. 3 followed by leachates of Landlls Nos. 1, 2, and 7. Leachates from Landlls Nos. 810 were the least toxic ones. In general the bacterial test was more sensitive to the constituents of the sample than the algal test. The EC50-values of the two tests were, however, all in the same order of magnitude. Thus, pre-concentration

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3854 A. Baun et al. / Water Research 38 (2004) 38453858 Table 7 Summary of results of toxicity tests of SPE-extracts of landll leachates Landll No. EC50 (P. subcapitata) EC50 (V. scheri) Genotoxicity test (S. typhimurium)a With S9b 1 2 3 4 5 6 7 8 9 10 6.1 3.8 2.5 3.8 2.9 6.5 2.2 9.4 6.8 3.3 [4.8;8.1] [3.0;4.6] [2.2;2.8] [2.9;4.9] [2.7;3.1] [5.4;7.8] [1.8;2.6] [6.7;13] [5.4;9.9] [2.0;5.4] 1.9 1.6 1.3 3.0 5.4 1.8 2.5 3.1 6.1 5.9 [1.9;2.0] [1.1;2.0] [1.2;1.5] [2.8;3.4] [4.4;7.9] [1.3;2.3] [2.2;2.8] [2.5;3.7] [4.3;13.1] [5.5;8.6] + Without S9b

EC50-values are listed as pre-concentration factors (i.e. number of times the samples needed to be concentrated to yield 50% response towards the test organisms). 95% condence intervals are shown in brackets. : Not genotoxic (Induction ratioo1.5 at all concentrations). +: Genotoxic (Induction ratio41.5 and o50% inhibition of bacterial biomass). a Maximum pre-concentration factor: 100150 times. b S9 is the metabolic activation system (liver homogenate).

factors ranging from 1.3 times to 9.4 times were sufcient to give 50% effect in both test system for all of the tested leachate samples. In a few cases the algal test was more sensitive than the bacterial test and this emphasizes the need of using more than one toxicity test for assessing toxicity of samples of an unknown and complex chemical composition. The SPE-treatment of leachates results in both a fractionation and a pre-concentration of the constituents in the leachate (Baun et al., 1999a). Although the SPE-resins are capable of retaining a large variety of both polar and non-polar organic xenobiotic compounds, it should be emphasized that SPE methods to some extent are selective with respect to the kind of organic compounds that will be retained. It is also important to recognize that volatile compounds will be lost during the change of solvent prior to biotests. Therefore, the biotest responses recorded in Table 7 reect the toxicity of non-volatile organic compounds present in the landll leachates. Analysis of content of NVOC before and after SPE-treatments showed that 1236% of the total amount of NVOC was retained in the columns (in average 26%). The major part of the NVOC was retained at pH 2.0 and this indicates that a dominating part of the NVOC retained in the columns was organic acids. 3.11. Relationship between toxicity and non-volatile organic chemicals Concentration addition and comparison with literature values of inhibition of algal growth due to single chemicals was used in an attempt to evaluate how many

percent of the measured toxicity could be attributed to identied chemicals. In order to use this kind of additivity model the chemicals must have similar mode of action (van Leeuwen and Hermens, 1995) and therefore pesticides (which are known to have specic targeted modes of action) were not included in the comparison, but were evaluated separately. The calculations were carried out by comparison of the results obtained in algal tests with leachate extract with literature algal toxicity results from ECOTOX database (US-EPA, 2002). It was not possible for all compounds to nd data from tests with P. subcapitata. For these compounds results of tests with Chlorella vulgaris or Scenedesmus subspicatus were used. It was found that from 2% (Landll No. 5) to 37% (Landll No. 8) of the observed toxicity could be explained by the compounds identied. Examples of the results obtained by this analysis are shown in Fig. 2 for leachates Nos. 6 and 8. For all samples it was found that naphthalene and 4chloro-o/m-cresol contributed signicantly to the toxicity (129%), but also compounds like camphor (Landlls Nos. 1, 6, and 8), 4-chlorophenol (Landll Nos.1, 2, and 6), 3,5-dichlorophenol (Landll Nos. 1), o/p cresol (Landll No. 1), and methylnaphthalenes (Landll No. 8) accounted for more than 1% of the toxicity observed. It should be noted that these results are valid for algal toxicity only and cannot be extrapolated to other species with different sensitivities towards the compounds found in the leachates. The very phytotoxic pesticides Ametryn (EC50=6 mg/ L), Chloridazon (EC50=170 mg/L), Hexazinon (EC50=7 mg/L), Isoproturon (EC50=77 mg/L), and Simazine (EC50=100 mg/L) were found in leachates

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A. Baun et al. / Water Research 38 (2004) 38453858 3855

Fig. 2. Percentage of toxicity accounted for by single chemicals assuming concentration additivity (pesticides were not included in the comparison).

3.12. Toxicity ranking and correlation with identied XOCs

Fig. 1. Example of observed inhibition and estimated concentrationresponse curves in a 48-h algal growth inhibition tests of SPE treated leachates from Landlls Nos. 110. Test organism: P. subcapitata.

from Landlls Nos. 2, 3, 5, and 6 (Table 6) in relatively high concentrations compared to the EC50values reported in the literature. A calculation shows that each of these pesticides can account for 2.4% (Chloridazon, Landll No. 3) to 73% (Hexazinon, Landll No. 2) of the algal toxicity observed in the present study. The pesticide atrazine is also very toxic towards algae (EC50=50180 mg/L), but the low concentration (0.16 mg/L) found in leachate from Landll No. 4 do not contribute signicantly to the toxicity found in this leachate sample.

In another attempt to explain the toxicity observed, a statistical ranking analysis was carried out to evaluate whether any of the general hydro-chemical parameters (e.g. chloride, ammonium, or NVOC) or the measured xenobiotic organic compounds correlated with the toxicity measured in SPE-extracts. The toxicity ranking was carried out based on the lowest EC50-values measured in tests with algae and bacteria (Table 7). The Spearman ranking procedure (Lehmann and DAbrera, 1998) was used to investigate the correlation between the toxicity ranking and the ranking of leachate pollution based on each of the following chemical descriptors: Chloride concentration, ammonium concentration, TAL, NVOC concentration, total concentration of XOCs, total concentration of phenols, and total detected number of XOCs including only the pesticides that were analyzed for in all leachate samples. The Spearman rank correlation coefcient (rs) was

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3856 A. Baun et al. / Water Research 38 (2004) 38453858

calculated and used to test the hypothesis of no correlation between toxicity and each of the chemical descriptors (Lehmann and DAbrera, 1998). Only the sum of concentrations of phenols (rs=0.6485, p=0.0245) and total number of detected XOCs (rs=0.8282, p=0.0024) correlated with the toxicity, whereas no signicant correlations were found for any of the other chemical descriptors at a 5% signicance level. Thus, neither the total concentrations of XOCs nor the concentrations of NVOC could be correlated with toxicity. As a result of the ranking analysis it can be concluded that the most polluted samples, in terms of number of detected organic chemicals, also were found to be the most toxic samples. On the other hand the summed concentrations of chemicals (or other hydro-chemical parameters) could not be used to point out the most toxic samples. Thus, the application of the biotest approach provides valuable information that cannot be deduced from chemical analyses alone. However, the identity of potential problematic chemicals cannot be revealed by biotests and therefore chemical analyses and biotests should be used in combination in future monitoring programs.

4. Conclusion The varying and very complex composition of the ten landll leachates included in the present study (e.g. electrical conductivity ranging from 2.6 to 54 mS/cm, NVOC from 66 to 363 mg/L, and chloride concentrations from 195 to 18,400 mg/L) affected the reproducibility of chemical analysis and also resulted in varying detection and quantication limits for several of the XOCs. When the occurrence and toxicity of XOCs is in focus fractionation and pre-concentration techniques are viable options for chemical as well as for biological analyses. For the latter this is particularly pertinent as e.g. the high chloride and ammonium concentrations found in some of the landlls in itself would be toxic to some aquatic organisms. The complexity of leachates analyzed in the present study may be illustrated by the observation that in no case the XOCs found accounted for more than 1% of the total organic carbon content. It should however be emphasized that the major part of the organic carbon content may well be harmless compounds, such as humic acids and volatile organic acids. Six of the landlls were engineered landlls that started landlling in 19771987 and were still receiving waste at the time of sampling (1999). Since 1976 Danish regulation has regarded discarded pesticide products as hazardous chemical waste that is illegal to deposit at ordinary MSW landlls. Special collection systems and treatment facilities exist for this kind of waste and therefore these compounds should not be present in

leachates from any of the new landlls. The study demonstrated that pesticides indeed were present in all the engineered landlls (18 pesticide compounds and three degradation products) with concentrations of single pesticides reaching a maximum of 28 mg/L. This nding points to the fact that extensive pesticide screening programs are necessary in the environmental risk assessment of leachates from engineered landlls eventhough source control already have been implemented. The 55 XOCs and 10 known degradation products identied in the present study belonged to the following groups: Aromatic compounds (18 compounds, including one degradation product: BSA), chlorinated aliphatics (three compounds), phenols (14 compounds, including one possible degradation product: 3,5 DCP), nonylphenol ethoxycarboxylate (one compound), phthalates (eight compounds, including ve degradation products: monoesters of o-phthalic acid), and pesticide (21 compounds, including three degradation products of glyphosate, atrazine, and simazine). It is concluded that inclusion of degradation products in future monitoring programs may provide additional insight in the fate of chemicals in landlls, as the present study provided several examples of concomitant presence of parent compounds and degradation products (e.g. di/ monophthalates, toluene/BSA, Glyphosate/AMPA), but also presence of degradation products in samples where the parent compounds were not found e.g. phthalate monoesters in leachate No. 1, nonylphenol ethoxycarboxylate in leachates Nos. 1 and 2, and hydroxyatrazine and -simazine in leachates Nos. 2 and 3. The use of toxicity biotests showed that the nonvolatile organic compounds in leachates have toxic potentials and that assessments of leachate toxicity must include results from biotest batteries. Genotoxicity tests revealed that only leachate from Landll No. 1 had the ability to cause genetic damage after 141 times preconcentration. A ranking correlation showed that the most toxic samples also were the most polluted ones in terms of the number of XOCs present, but the toxic potential of the samples could not be predicted by chemical analysis alone. In future monitoring programs aimed at risk assessments of XOCs in landll leachates, it is therefore recommended to combine chemical analyses covering a wide range of XOCs (including degradation products and pesticides) with a battery of biotests.

Acknowledgements Dr. Susanne Jonsson, Linko ping University, Sweden, is thanked for performing the phthalate analyses. Assistance with sampling and analyses by Mrs. Susanne Kruse, Mrs. Lena D. Troelsen, and Mr. Jens S. Srensen is gratefully acknowledged.

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A. Baun et al. / Water Research 38 (2004) 38453858 3857

Appendix A. Limit of detection for pesticides analyzed for, but not found in the ten leachates LOD (mg/L) 0.1 Compounds Alachlor, Ametryn, Bifentrin, Bromopropylate, Bupririmaat, Chlorfenvinfos, Chlorpyrifos-methyl, Chloroxynil, Chlorthalonil, Desmetryn, Diazinone, Dichlouanide, 2,6dichlorbenzamid (BAM), Difenoconazole, Dimethoaat, Ethylparation, Etridiazole, Fenamifos, Fenithothion, Fenvalerat, Fluazifopbutyl, Fonofos, Furalaxyl, Heptenofos, Hexachlorbenzene, HTI, Ioxynil, Lindan, Metalaxyl, Methyl-parathion, Metolachlor, Pendimethalin, Penconazole, Pentachlorphenol, Permethrin, Pirimifos-methyl, Prochloraz, Procimidon, Propachlor, Prosulfocarb, Pyrazofos, Pyrimethanil, Tebuconazole, Terbutryn, Terbutylazine, Tetrachloorvinfos, Tetramethrine, Tolclofos-methyl, Triallaat, Vinclozolin Azinphos-ethyl, Cypermethrin, Deltamethrin, Demeton-s-methyl, Desethylatrazine, Desisopropylatrazin, Disulfoton, Fenarimol, Fenpropathrin, Fention, Fluazinam, Fosfamidon, Metamitron, Mevinfos, Pentachlorbenzene, Pirimicarb, Propham, Propyzamide, Tri-allaat, Triazofos Bromoxynil, Chlorpyrifos, Chlorsulfuron, Cyanazine, 2,4-D, Demeton-o/s, Dichloorvos, Dinoterb, Ethoprofos, Hydroxycarbofuran, Koumafos, Malathion, Metasulfuronmethyl, Metazachlor, Metribuzine, Propiconazole, Pyrifenox, 2,4,5-T, 2,4,5-TP, Tolyuanide, Triazofos Dinoseb, Diuron, DNOC, Fenoxycarb, Methidothion Alletrin, Azinphos-methyl, Dodemorf

0.10.2*

0.10.5*

0.11.0* 0.11.5*
*

Minimum and maxium values of detection limits within the group. Detection limits were varying due to sample matrix.

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