assessment of mobility and bioavailability of contaminants in msw incineration ash with aquatic and...

Waste Management xxx (2014) xxx–xxx

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Waste Management

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Assessment of mobility and bioavailability of contaminants in MSWincineration ash with aquatic and terrestrial bioassays

0956-053X/$ - see front matter � 2014 Elsevier Ltd. All rights reserved.http://dx.doi.org/10.1016/j.wasman.2013.12.024

⇑ Corresponding author. Tel.: +46 709 923 733.E-mail address: [email protected] (V. Ribé).

Please cite this article in press as: Ribé, V., et al. Assessment of mobility and bioavailability of contaminants in MSW incineration ash with aquaterrestrial bioassays. Waste Management (2014), http://dx.doi.org/10.1016/j.wasman.2013.12.024

V. Ribé ⇑, E. Nehrenheim, M. OdlareFuture Energy Research Group, School of Business, Society and Engineering, Mälardalen University, SE-721 23 Västerås, Sweden

a r t i c l e i n f o a b s t r a c t

Article history:Received 13 June 2013Accepted 31 December 2013Available online xxxx

Keywords:MSWWTE ashMobilityBioavailabilityToxicityLeachingCr

Incineration of municipal solid waste (MSW) is a waste treatment method which can be sustainable interms of waste volume reduction as well as a source of renewable energy. In the process fly and bottomash is generated as a waste material. The ash residue may vary greatly in composition depending on thetype of waste incinerated and it can contain elevated levels of harmful contaminants such as heavy met-als. In this study, the ecotoxicity of a weathered, untreated incineration bottom ash was characterized asdefined by the H14 criterion of the EU Waste Framework Directive by means of an elemental analysis,leaching tests followed by a chemical analysis and a combination of aquatic and solid-phase bioassays.The experiments were conducted to assess the mobility and bioavailability of ash contaminants. A com-bination of aquatic and terrestrial bioassays was used to determine potentially adverse acute effects ofexposure to the solid ash and aqueous ash leachates. The results from the study showed that the bottomash from a municipal waste incineration plant in mid-Sweden contained levels of metals such as Cu, Pband Zn, which exceeded the Swedish EPA limit values for inert wastes. The chemical analysis of the ashleachates showed high concentrations of particularly Cr. The leachate concentration of Cr exceeded thelimit value for L/S 10 leaching for inert wastes. Filtration of leachates prior to analysis may have under-estimated the leachability of complex-forming metals such as Cu and Pb. The germination test of solidash and ash leachates using T. repens showed a higher inhibition of seedling emergence of seeds exposedto the solid ash than the seeds exposed to ash leachates. This indicated a relatively low mobility of tox-icants from the solid ash into the leachates, although some metals exceeded the L/S 10 leaching limit val-ues for inert wastes. The Microtox� toxicity test showed only a very low toxic response to the ashleachate exposure, while the D. magna immobility test showed a moderately high toxic effect of theash leachates. Overall, the results from this study showed an ecotoxic effect of the solid MSW bottomash and the corresponding ash leachates. The material may therefore pose an environmental risk if usedin construction applications. However, as the testing of the solid ash was rather limited and the ash leach-ate showed an unusually high leaching of Cr, further assessments are required in order to conclusivelycharacterize the bottom ash studied herein as hazardous according to the H14 criterion.

� 2014 Elsevier Ltd. All rights reserved.

1. Introduction frequently contains elevated levels of environmentally hazardous

In today’s consumer society the need for efficient and environ-mentally friendly waste management methods is greater than ever.Incineration of municipal solid waste (MSW) is a waste treatmentmethod which can be sustainable both in terms of waste volumereduction as well as a source of renewable energy. In the incinera-tion process fly and bottom ash is generated as a waste. Thebottom ash material may be used as a fill material for road con-struction (OECD, 1977; Forteza et al., 2004), a stabilizer for liquidwastes such as sewage sludge (Bednarik et al., 2004) or as a landfillcover (Wiles, 1996). The MSW ash residue may vary greatly incomposition depending on the type of waste incinerated and it

contaminants such as heavy metals (Hasselriis and Licata, 1996).Potentially adverse environmental effects of MSW bottom ash uti-lization mainly stems from leaching of heavy metals (Ecke andAberg, 2004) and organic compounds such as PAH́s (Wheatleyand Sadhra, 2004). However, some previous studies have shownthat only a small amount of the heavy metal content of the ashis leached (Wiles, 1996).

The EU waste directive 2008/98/EC (EC, 2008) prescribes hazardcharacterization of waste materials according to 15 assessmentcriteria. The H14 criterion, which characterizes a waste as‘‘ecotoxic’’, should preferably be assessed through the use ofbiotests. A wide range of bioassays are available for ecotoxicitytesting, but when selecting an appropriate test method for hazardcharacterization considerations regarding material properties andexposure conditions need to be made. Several researchers have

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2 V. Ribé et al. / Waste Management xxx (2014) xxx–xxx

investigated suitable ecotoxicity test batteries for the assessmentof the H14 criterion (Pandard et al., 2006; Wilke et al., 2008) anda European ring test, initiated by the German Environmental Pro-tection Agency, UBA, was performed to evaluate available stan-dardized test protocols for the assessment of waste hazard(Moser et al., 2010). Leaching tests are commonly used to assessthe mobility of contaminant substances from a solid waste intoan aqueous solution (Kosson et al., 2002). There are different typesof leaching tests, using leachants such as deionised water, calciumchloride or acidic solutions. The ISO 12457 standard protocol, usingdeionised water as leachant, is considered a less aggressive leach-ing method in comparison to the US EPA Toxicity CharacterizationLeaching Procedure (TCLP) (US EPA, 1992) using an acetic acid/so-dium hydroxide solution as leachant (Mantis et al., 2005). The ISO14735 leaching protocol for the preparation of waste leachates forecotoxicity testing is analogous to the ISO 12457 protocol but uti-lizes a weak calcium chloride solution as an artificial rain leachant.If the leachates from a waste can be classified as ‘‘ecotoxic’’ accord-ing to the H14 criterion, the material should also be classified ashazardous according to the H15 criterion of the waste directive;i.e. a waste capable by any means, after disposal, of yieldinganother substance, e.g. a leachate, which possesses any of theprevious fourteen hazard properties, H1-H14.

In this study, the ecotoxicity of a matured, untreated incinera-tion bottom ash was characterized as defined by the EU WasteFramework Directive H14 criterion by means of an elemental anal-ysis, leaching tests and a combination of aquatic and solid-phasebioassays. The aim of the study was to assess the mobility and bio-availability of ash contaminants to determine the ecotoxicity of theash according to the H14 criterion and to assess whether the mate-rial may be used in a construction application. A chemical analysisof the ash leachates was performed to study the extent of metalleachability. Aquatic and terrestrial bioassays were used to deter-mine potentially adverse acute effects of exposure to the solidash and to aqueous ash leachates.

2. Materials and methods

2.1. Experimental outline

The mobility and bioavailability of contaminants in the solidash were assessed in leaching and toxicity tests according to theexperimental outline of Fig. 1.

Fig. 1. Experimental outline of the ash contam

Please cite this article in press as: Ribé, V., et al. Assessment of mobility and bterrestrial bioassays. Waste Management (2014), http://dx.doi.org/10.1016/j.w

2.2. Ash

The bottom ash of the study originated from a municipal wasteincineration plant in Sweden. Metal parts were separated from theash after incineration. The ash was allowed to mature, i.e. weatherin order to stabilize the metal content through carbonation reac-tions with water and carbon dioxide. This reduces future leachingof metals from the ash (ISWA, 2006). Elemental analysis of thewhole ash was performed with ICP–MS/IC-AES within one month(Table 1) by a commercial accredited laboratory. Particles with agrain size >4 mm were separated from the ash sample prior tothe germination test.

2.3. Control soil for germination test

The control garden soil was a commercial seedling and cactussoil. The soil was a 95% peat and 5% sand mixture, with 0.8 kg/m3 manure supplied. Nutrients were added at the following con-centrations: 112 g/m3 N; 56 g/m3 P; and 120 g/m3 K. The soil pHwas 5.5–6.5 and the soil density was 400 kg/m3.

2.4. Ash leachate

Three sets of two parallel batch leaching tests were performedaccording to ISO standard protocol 12 457-2 using deionised wateras the leachant in a 10:1 L/S ratio. Large particles (>4 mm) wereseparated from the whole ash sample through sieving. Followingcentrifugation and filtration over a 0.45 lm membrane filter, theash leachates were assessed chemically, (see Section 2.4) and withterrestrial and aquatic bioassays (Section 2.5).

2.5. Chemical characterization of leachate

The metal concentrations of the leachate replicates L-A and L-Bfrom the third batch leaching were determined by ICP–MS and ICP-AES at the accredited laboratory Analytica, according to US EPAstandard procedures (US EPA, 1994a,b) on filtered leachate sam-ples. No acid digestion was carried out on the leachate samples.pH and conductivity was measured using standard procedures.

2.6. Ecotoxicity assays

The toxicity of sieved ash samples (see Section 2.2 Ash) was as-sessed in a germination test using Trifolium repens (White clover).

inant mobility and bioavailability study.

ioavailability of contaminants in MSW incineration ash with aquatic andasman.2013.12.024


Table 1Elemental composition of the bottom ash.

Element mg/kg dry mass

Al 87,300As 37.5B 487Ba 2110Ca 107,000Cd 1.14Co 62.5Cr 731Cu 17,100Fe 67,300Hg 0.013K 8160La 17.5Mg 14,400Mn 1050Mo 74.7Na 32,300Ni 200P 2100Fb 1250Pb 77.8SP 0.572Se 232,000Si 9850Ti 94.1Zn 4050

Table 2Chemical characteristics of the ash leachates.

Element Conc. Sample

L-A L-B

Al lg/l 10,200 10,800As lg/l <1 <1Ba lg/l 27.4 28.5Ca mg/l 321 302Cd lg/l 0.294 0.315Co lg/l 0.598 0.496Cr lg/l 134 121Cu lg/l 307 266Fe lg/l 0.0129 0.0358Hg lg/l <0.02 <0.02K mg/l 54.3 50.5Mg mg/l 0.429 0.482Mn lg/l 2.27 74.9Mo lg/l 221 202Na mg/l 360 333Ni lg/l 7.05 6.3Pb lg/l 1.01 1.72V lg/l 3.43 3.11Zn lg/l 25.9 14.4Cond. mS/m 315 300PH 7.7 7.7

V. Ribé et al. / Waste Management xxx (2014) xxx–xxx 3

The toxicity of the leachates were assessed in a germinationstudy using T. repens (first batch leaching); in the acute bacterialluminescent bioassay Microtox� using Vibrio fischeri (second batchleaching) and in an acute crustacean toxicity test using Daphniamagna (third leaching round).

2.6.1. Germination test with T. repensThe bioavailability and mobility of the ash contaminants in the

solid ash and the ash leachates were assessed in a germination testusing the trifoliate, eudicot white clover (T. repens).

The toxicity assays were performed in plastic Petri dishes (9 cm£) with triplicate samples at solid ash or ash leachate concentra-tions of 6.25%, 12.5%, 25.0%, 50.0% and 100% (Fig. 2a and b). The di-rect-contact test using solid ash sample was performed by dilutionof the sample with commercial garden soil (Fig. 2a). Pure gardensoil was used as the test control. Petri dishes were half-filled withsample (38 ml) and subsequently soil, ash/soil and ash sampleswere watered. The leachate test was performed by dilution of theash leachate sample with doubly deionised water (Fig. 2b). Doublydeionised water was also used as test control. Aliquots of the sam-ples (38 ml) were applied to Petri dishes containing cellulose filterpaper (Whatman). Ten seeds of white clover were placed on the

Fig. 2. a and b. Germination test using T. repens. The direct-contact test


surface of solid and leachate samples. Petri dishes were coveredwith lids and incubated in darkness at 20 �C for six days. Seedlingemergence was measured after the six days of incubation.

2.6.2. 30-min acute Microtox� test (V. fischeri)The toxicity of ash leachates to V. fischeri was assessed in the

Microtox� test according to the ISO 11348-3 protocol using Micro-toxOmni™ Software (Azur Environmentals, USA), as previously de-scribed by Ribé et al. (2012). One leachate sample exceeded the pHlimits for the test (pH 6.5–8.0). However, the upper pH limit valuewas exceeded by less than 5% (pH: 8.3) and the sample pH wastherefore not adjusted. The sample concentrations tested were;80.0%, 50.0%, 33.3%, 25.0%, 16.7%, 12.5%, 8.33% and 6.25%.

2.6.3. Immobility of D. magnaThe acute aquatic toxicity of the ash leachate samples was as-

sessed in a 48 h D. magna test according to ISO standard 6341,using Daphtoxkit™ (MicroBioTests Inc., Belgium).

Each leachate sample (L-A and L-B) was diluted to give finalleachate concentrations of 6.25%, 12.5%, 25.0%, 50.0% and 100%.Each concentration of the dilution series was tested with four rep-licate sample wells, each containing five test organisms, togetherwith a control for each dilution series. The validity of the testswas assessed according to ISO 6341. A reference test withK2Cr2O7 was performed in parallel with the ash leachate assay.

using solid ash sample (a). The leachate test using ash leachate (b).



Fig. 3. Germination of T. repens after exposure to solid ash (mean ± S.D.). Three replicates per sample concentration (incl. control), each containing 10 seeds of T. repens, wereexposed to ash samples for six days. Error bars represent standard deviation around the mean.


2.7. Data analysis

Dose–response relationships between exposure concentrationsand toxic effects from the T. repens germination tests were ex-plored graphically. No effect concentrations were established forthe germination tests.

Inhibition values and EC50 (30-min) values for the Microtox�

test were calculated for all test concentrations from duplicatesamples, where possible. If no dose–response was observed, orthe extrapolated EC50 value exceeded 100% sample concentration,the inhibition effect at the highest tested concentration (80%sample) was given.

Fig. 4. Germination of T. repens after exposure to ash leachate (mean ± S.D.). Three repliwere exposed to ash leachates for six days. Error bars represent standard deviation arou


EC50 values for immobility after 48 h exposure were calculatedfrom the sigmoidal concentration–response curves fitted by theleast-squares method using the Sigma Plot 4.0� software (byWindows).

Significant effects between the different exposure concentra-tions of the D. Magna test were evaluated by one-way analysis ofvariance (ANOVA) followed by Tukey’s HSD multiple comparisontest. The evaluation was performed within and between allconcentrations, where the sample dilution series from leachatesL-A and L-B were treated as separate samples. The data analysiswas performed using the software package SPSS 19 (SPSS Inc.,Chicago, IL.).

cates per sample concentration (incl. control), each containing 10 seeds of T. repens,nd the mean.



Fig. 5. a and b. Immobility of D. magna after exposure to ash leachates L-A (left) andL-B (right).Four replicates per sample concentration (incl. control), each containing5 neonates were exposed to ash leachate samples for 24 h (dash-dot-dot lines) and48 h (solid lines).

V. Ribé et al. / Waste Management xxx (2014) xxx–xxx 5

3. Results and discussion

3.1. Physicochemical character of the ash leachates

The leachate chemical parameters of the third batch leaching(L-A and L-B) are shown in Table 2. The leachates from all threebatch leachings were slightly alkaline (7.7–8.3), but in comparisonto other studies pH was relatively low (Stiernstrom et al., 2011).The conductivity of the ash leachates was quite low (Bagchi,1990; Hjelmar, 1996; Lundtorp et al., 2003). The levels of mineralsand metals, particularly Cu, Cr, Pb and Zn, were very high in thewhole ash sample (Barna et al., 2000; Van Gerven et al., 2005),and this was reflected in the elevated concentrations of Cr in theash leachate. The Swedish EPA has published limit value concen-trations for a selection of metals, sulfate, chloride and three PAHfractions in solid materials and leachates for wastes intended forrecycling in road filling and other construction applications (TheSwedish EPA, 2010). The concentrations of the aforementionedmetals all exceeded the Swedish EPA limit values for solid inertwaste as well as the limit for Cr in waste leachings. As the leachateswere centrifuged and filtered prior to the chemical analysis, mainlythe free ion concentrations were measured in the ICP–MS/AESanalysis. The Pb and Cu ions complex readily with organic ligands,e.g. humic acids and only a small fraction of the total metal contentis normally available as the free ion (Christensen et al., 1999). Mei-man and Comans (1999) showed in their study of the leachingbehavior of MSW bottom ash at different stages of weathering, thatthe leaching of Cd, Pb, Cu, Zn and Mo was generally reduced after aweathering period in excess of 1.5 years. It is hypothesized that asthe pH of weathered ash decreases, less soluble metal complexspecies, such as aluminium hydroxides with Cu and Pb are formed(Meiman and Comans, 1998, 1999). Although dissolved metal com-plexes with organic ligands or sorbent minerals pass through a0.45 lm membrane filter, the bioavailability of the complexed me-tal is likely to be reduced.

3.2. Ecotoxicity assays

3.2.1. Germination test with T. repensThe results from the germination tests with solid ash showed a

dose–response with increasing level of inhibition of seedling emer-gence with higher ash concentrations (Fig. 3). At ash concentra-tions of 50% and higher there was no emergence of seedlings.The high concentrations of metals such as Cu, Pb and Zn were prob-ably the main cause of the inhibition of seedling emergence but thelow organic content of the ash material may also have contributedto the inhibited germination (Weng et al., 2002). As this study wasa rather limited, preliminary study no EC50 concentrations wereestablished from the dose–response curves.

The results from the germination tests with ash leachates didnot show a clear dose–response. Average seedling emergence forthe different ash leachate concentrations varied between 8.3 forthe 12.5% ash leachate samples and 4.7 for the 100% ash leachatesamples (Fig. 4). Noteworthy is the marked variation in seedlingemergence between ash leachate replicates at ash leachate concen-tration of 25%. Although no clear dose–response was shown for theash leachate germination test, the rather high deviation from themean at 25% leachate concentration, may have indicated the toxic-ity threshold level.

The marked difference between inhibition of germination be-tween the direct-contact test with solid ash and the ash leachatetest indicates that the mobility of contaminants from the solid ashinto the leachate is low and the dilution of contaminants in theleachate is high due to the L/S ratio. This is in line with the resultsfrom the chemical analysis and also with an earlier study performedby Barna et al. (2000), which demonstrated a low leaching level of


ash contaminants such as Pb and Zn. Ferrari et al. (1999) reportedsimilar results, where only 1% of the total metal content of the solidMSW bottom ash was detected in the aqueous leachate and a corre-sponding difference in toxicity response between the solid-phaseand liquid-phase phytotoxicity tests was shown.

The concentrations of metals were very high in the ash of thisstudy, and although the bioavailability of these metals may be gener-ally reduced by an alkaline pH of the solid matrix (Barna et al., 2000),the seeds were nevertheless exposed to elevated levels of metals.

3.2.2. 30-min acute Microtox� test (V. fischeri)The results from the acute bacterial Microtox� test of the ash

leachates showed a similar toxic response to the T. repens germina-tion tests. Ash leachate A showed no toxic response; the extrapo-lated EC50 concentration exceeded 1000% leachate. Ash leachateB did not show any dose–response and the highest toxic effect incomparison to the control was 10% at 80% leachate concentration.

3.2.3. Immobility of D. magnaThe D. magna immobility test was valid according to the proto-

col validity criteria. The K2Cr2O7 reference test showed a 24 h-EC50

value of 1.3 mg/l which was within the specified limits of the ISO6341 protocol. The D. magna immobility test with leachatereplicates L-A and L-B from the third leaching batch showed amoderately high toxic effect of the leachates, see Fig. 5a and b.The 48 h half maximal effective concentrations, 48 h-EC50, were




29% leachate concentration and 18% leachate concentration for L-Aand L-B, respectively. There were no results for the L-A 25% leach-ate sample concentration.

The ANOVA analysis showed no significant differences betweenthe same dilution levels of samples L-A and L-B. The NOEC value forsample L-B was 12.5% leachate concentration and the LOEC was25% leachate concentration. For sample L-A, where there were noresults for the 25% leachate concentration, there was a significantdifference between the 12.5% sample concentration and the 50%sample concentration. There were no significant differences shownwithin groups.

The results showed a higher sensitivity of D. magna to the leach-ate contaminants in comparison to V. fischeri and T. repens. In aleaching and ecotoxicity study performed by Tsiridis et al. (2012)with fly ash, the sensitivity of D. magna to metals such as Cu wasdemonstrated to be greater than that of V. fischeri.

There are, however, additional physicochemical factors thatmay modify the toxic effects of contaminants to the V. fischeri bio-luminescence. Berglind et al. (2010) showed that the tolerance ofthe bioluminescent bacterium to contaminants such as phenolwas enhanced by increased concentrations of Ca and K.

4. Conclusions

The results from this study showed that the bottom ash origi-nating from a municipal waste incineration plant in mid-Swedencontained high levels of metals such as Cr, Cu, Pb and Zn, which ex-ceeded the Swedish EPA limit values for inert wastes.

The chemical analysis of the ash leachates showed high concen-trations of particularly Cr. The leachate concentration of Cr ex-ceeded the Swedish EPA limit value for L/S 10 leaching for inertwastes. As the leachates were filtered over a fine membrane filterprior to analysis, the leachability of complex-forming metals suchas Cu and Pb may have been underestimated.

The germination test of solid ash and ash leachates using T. re-pens showed a higher inhibition of seedling emergence of seeds ex-posed to the solid ash than the seeds exposed to ash leachates. Thisindicated a low mobility of contaminants from the solid ash intothe leachates.

The Microtox� toxicity test showed only a very low toxic re-sponse to the ash leachate exposure, while the D. magna immobil-ity test showed a moderately high toxic effect of the ash leachates.

While the leaching of metals from the bottom ash is relativelylimited, in terms of the solid ash metal content, there is still a sig-nificant leaching of Cr from the material and the leachates show amoderately high toxicity to D. magna.

Overall, the results from this study showed an ecotoxic effect ofthe solid MSW bottom ash and the corresponding ash leachates.The material may, hence, pose an environmental risk if used inconstruction applications. However, as the ecotoxicity testing ofsolid ash was rather limited, the bottom ash herein studied couldnot be conclusively characterized as ecotoxic according to theH14 criterion. Similarly, additional assessment of the leachingproperties of the ash ought to be performed over a wider pH range,in order to provide further information about the Cr leaching for anaccurate characterization of the bottom ash according to the H15criterion.

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