Science of the Total Environment 442 (2013) 302–309
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Science of the Total Environment
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Chlorine disinfection of dye wastewater: Implications for a commercial azodye mixture
Francine Inforçato Vacchi a, Anjaina Fernandes Albuquerque a, Josiane Aparecida Vendemiatti a,Daniel Alexandre Morales a, Alexandra B. Ormond b, Harold S. Freeman b, Guilherme Julião Zocolo c,Maria Valnice Boldrin Zanoni c, Gisela Umbuzeiro a,⁎a Faculdade de Tecnologia, Universidade Estadual de Campinas, Limeira, SP, 13484-332, Brazilb Department of Textile Engineering, Chemistry, and Science, North Carolina State University, Raleigh, NC 27695-8301, USAc Departamento de Química Analítica, Universidade Estadual Paulista Júlio de Mesquita Filho, Instituto de Química de Araraquara, Araraquara, SP 14801-970, Brazil
H I G H L I G H T S
► Aqueous solutions of Disperse Red 1 were treated with chlorine.► The chlorination products of Disperse Red 1 were identified using LC–ESI-MS/MS.► Daphnia and Salmonella/microsome were employed for eco/genotoxicity testing.► The chlorinated dye was more mutagenic than the dye itself.► Chlorination should be avoided in effluents containing azo-dyes.
⁎ Corresponding author.E-mail address: [email protected] (G. Umbuzeir
Article history:Received 5 August 2012Received in revised form 3 October 2012Accepted 3 October 2012Available online 22 November 2012
Keywords:C.I. Disperse Red 1ChlorinationCommercial dye mixtureLC–MSDaphniaSalmonella/microsome assay
Azo dyes, themost widely used family of synthetic dyes, are often employed as colorants in areas such as textiles,plastics, foods/drugs/cosmetics, and electronics. Following their use in industrial applications, azo dyes havebeen found in effluents and various receiving waters. Chemical treatment of effluents containing azo dyesincludes disinfection using chlorine, which can generate compounds of varying eco/genotoxicity. Among thewidely known commercial azo dyes for synthetic fibers is C.I. Disperse Red 1. While this dye is known to existas a complexmixture, reports of eco/genotoxicity involve the purified form. Bearing inmind the potential for ad-verse synergistic effects arising from exposures to chemical mixtures, the aim of the present study was to char-acterize the components of commercial Disperse Red 1 and its chlorine-mediated decoloration products and toevaluate their ecotoxicity and mutagenicity. In conducting the present study, Disperse Red 1 was treated withchlorine gas, and the solution obtained was analyzed with the aid of LC–ESI-MS/MS to identify the componentspresent, and then evaluated for ecotoxicity and mutagenicity, using Daphnia similis and Salmonella/microsomeassays, respectively. The results of this study indicated that chlorination of Disperse Red 1 produced four chlori-nated aromatic compounds as themain products and that the degradation productsweremore ecotoxic than theparent dye. These results suggest that a disinfection process using chlorine should be avoided for effluentscontaining hydrophobic azo dyes such commercial Disperse Red 1.
Of the annual worldwide production of synthetic dyes, more than60% are azo dyes andaround2000of themare used in the textile, leather,plastics, paper, cosmetic and food industries (Stolz, 2001). Azo dyes in-clude a very important family of hydrophobic dyes known as dispersedyes and have been identified as aquatic contaminants (Bafana et al.,2011; Oliveira et al., 2006; Osugi et al., 2009; Umbuzeiro et al., 2005).
o).
rights reserved.
For industrial application, azo disperse dyes are often employed as com-mercial products containing subsidiary colorants arising from the meth-od of synthesis and chemical auxiliaries such as dispersing agents toimprove water solubility and color transfer (Reife and Freeman, 1995;Venkataraman, 1978).
Although the ecotoxicity and genotoxicity of certain azo dyes havebeen evaluated (Bae and Freeman, 2007; Gottlieb et al., 2003; Novotnýet al., 2006; Osugi et al., 2009; Sponza, 2006; Umbuzeiro et al., 2005;Wang et al., 2009), invariably the tests were performed on pure forms.It is also known that conventional biological treatment of effluentsfrom dyeing processes in the textile industry often does not efficientlyremove the dyes employed (Oliveira et al., 2006; Osugi et al., 2009;
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Umbuzeiro et al., 2004, 2005); therefore, dyes and their auxiliaries canbe found in raw and treated effluents and the adjacent aquaticenvironment.
In several countries, effluents from biological treatment aredisinfected (decolorized) using chlorine for pathogen removal. Ithas been found, however, that the chlorination of colored wastewatercan generate genotoxic compounds known as phenylbenzotriazoles(PBTAs) (Kummrow and Umbuzeiro, 2008; Nukaya et al., 1997).These chlorinated compounds were also found in drinking water(Oliveira et al., 2006). In a study involving the chlorination of themonoazo dye methyl orange, para-hydroxybenzene sulfonic acidand para-chloro-N,N-dimethylaniline were identified as reactionproducts (Laitinen and Boyer, 1972).
Monoazo dye C.I. Disperse Red 1 is available worldwide in the formof 57 different commercial products (Colour Index, 2011). This dye hasbeen extensively used for dyeing polyester fabrics in industrial plantssuch as those located in São Paulo State, Brazil. Its wide use has led tostudies such as the one conducted by Oliveira et al. (2010)who showedthat pure Disperse Red 1 was genotoxic before and after chlorination.Also, pure Disperse Red 1 was studied for acute toxicity using Daphniasimilis and an EC50 of 130 μg/L was reported (Ferraz et al., 2011).
Bearing inmind that commercial Disperse Red 1 is sold and used as amixture of colored components rather than a pure dye, the aim of thepresent study was to establish the contribution of the components ofthe commercial dye and its chlorination products to the observedecotoxicity and mutagenicity. As would be anticipated, it has beenreported that the presence of multiple toxic compounds in a mixturecan lead to individual toxicities that are additive, less than additive, orgreater than additive (Neal, 1987). The behavior of commercial DisperseRed 1 within this range of possibilities was determined.
2. Material and methods
2.1. Analysis of commercial dye
Disperse Red 1, was purchased as a commercial product containinga dispersing agent. Preparative thick layer chromatography platesemployed for isolating the colored components of the commercial dyewere 20×20 cm Whatman brand Partisil® PLK5F plates from FisherScientific, coated with 1000 μm thick silica gel 150 A. Commercial Dis-perse Red 1 (5 mg) was stirred with methylene chloride (0.5 mL), andthe solution was spotted along a horizontal line, 1″ from the bottom ofthe thick layer plate and 1″ from both sides of the plate, using with aTLC capillary. Spotting was repeated until the entire sample was trans-ferred to the plate and produced a continuous band no more than3 mm thick. The resultant plate was allowed to dry for 1 h, and then de-veloped in a 29×9.5×27 cm TLC chamber containing 2 toluene/1 ethylacetate (210 mL) until the eluent reached within 1 cm of the top of theplate. The plate was removed from the chamber and air dried for30 min. The bands labeled fractions F1–F6 were individually removedfrom theplate using a razor blade. The individual fractionswere extractedusing methanol (20 mL), filtered through Celite, and the Celite/silica gelcake was washed with methanol until no color remained. The solventwas evaporated under vacuum, and each fraction was again dilutedwithmethanol (1 mL),filtered through a syringefilter to remove residualsilica gel, and the solvent was evaporated. Thus were obtained F1(1.9 mg), F2 (1.1 mg), F3 (1.8 mg), F4 (23.3 mg), F5 (1.7 mg), and F6(0.9 mg). Thin layer chromatography plates used for Rf determinationscontained silica gel G, 250 μm thick. The Rf values for the six fractionswere 0.84, 0.75, 0.68, 0.33, 0.15, and 0.03, respectively. The six compo-nents were identified using proton NMR and high resolution mass spec-trometric analysis.
The dispersing agent used in this study was obtained by separationfrom the colored components. A 10 mg sample of the commercial dyein 200 mL ethyl acetate (99.5%, F. Maia, Brazil) was sonicated in anultrasound bath (135 W, 40 kHz, Unique 1400, Brazil) for seven
10-min cycles at 25 °C. Optical density was monitored using a UV–visspectrophotometer (GBC, Cintra 6), at λmax 483 nm, and 10 mmquartzcuvettes. The dispersing agentwas collected by filtration and air drying,and corresponded to 20% of the commercial dye mass.
2.2. Chlorination of commercial dye
A 50 mg/L solution of Disperse Red 1 in ultrapure water was treatedwith chlorine gas generated from the careful addition of 100 mL of 12 MHCl to 500 mg of potassium permanganate (KMnO4). The gas formedwas conducted through a tube and bubbled into a flask containing500 mL of dye solution. Color removal was monitored in a UV–vis spec-trophotometer, with a 200 to 800 nmscan every 2 min. The chlorinationprocess was terminated when an absorbance in the visible region wasnot detected (~30 min). Residual free chlorine was measured at theend of the process using the N,N′-diethyl-para-phenylenediamine(DPD) method (APHA, 1995).
2.2.1. Identification of the chlorination productsThe chlorination products were separated and identified using a
liquid chromatography–electrospray ionization-mass spectrometry(LC–ESI-MS/MS Qtrap). Full scan (EMS— enhancedmass spectrometry)using enhanced resolution (ER)modewas employed for the determina-tion of isotopic ratios and enhanced product ion (EPI) scanning used toidentify the main product.
Samples of product mixtures were dissolved in 50:50 methanol:water containing 0.1% formic acid before each injection and separatedon an Agilent Technologies Zorbax C-18 column (5 mm, 150×4.6 mm)using an Agilent 1200 auto sampler and Agilent1200 HPLC pump.
HPLC analysis employed formic acid (0.1%) in water (solvent A)and methanol (solvent B) under a linear gradient of 10–80% B for0–8 min, 80% B for the next 8–15 min, 80–100% B for the next15–18 min, and 100% B for the final 18–25 min. The flow rate was500 μL/min.
Mass spectrometric analysis of dyeswas carried out using an ion-trapmass spectrometer (Applied Biosystems) equipped with a turbo ionspray connected to the HPLC system. The vaporizer was set at 500 °Cand a voltage of 4.5 kV for ion spray generation in the positive ionizationmode. The desolvation potential was set at 50 V and ultra-pure N2 gaswas used as the collision gas. The experiments were performed withER at a scan rate of 250 s−1 for 50 ms. For EPI analysis, the scanningspeedwas 4000 s−1, using a collision energy of 30 Vwith a 10 V spreadand 20 ms for capturing ions. Experiments were performed in the50–600 Da range, with an 8 V potential for entry and Q0 trappingenabled.
2.3. Ecotoxicity testing
The commercial dye, its individual components, and the chlorina-tion products were evaluated for acute toxicity to D. similis, accordingto OECD 202 guidelines (OECD, 2004). Stock solutions of the commer-cial dye were prepared in deionized water and the dye components inwater containing 1% methanol to facilitate dissolution. Negative con-trols were tested accordingly. Twenty neonates (b24 h old) from 2 to3 week-old mothers were placed in each concentration of a test solu-tion. Experiments, including negative controls, were performed in fourreplicates at 21±0.3 °C in the dark. After 48 h, the number of immobileorganisms was recorded. The results were statistically analyzed usingthe trimmed Spearman–Karber method for estimating the median im-mobilization concentration — EC50 (Hamilton et al., 1977).
Because of the high sensitivity of Hydra to chlorinated compounds(Ake et al., 2003; Mayura et al., 1991) acute tests using Hydra attenuata(Trottier et al., 1997) were included, to evaluate the commercial dyeand its chlorination products. The test was conducted in 12-wellmicroplates,with 3 replicatewells for each concentration, 5 mLof samplein each well, and 3 animals for each replica. Organisms were exposed for
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96 h and lethal effects were recorded. The results were statistically ana-lyzed using the trimmed Spearman–Karber method for estimating themedian effective concentration — EC50 (Hamilton et al., 1977). Testswith the chlorinated solution were initiated only when free chlorinewas not detected.
2.4. Mutagenicity testing
The commercial dye, its chlorination products, and the six DisperseRed 1 components were evaluated in the Salmonella/microsome assayusing the ISO 16240:2005 procedure (ISO, 2005) TA98 strain (hisD3052,rfa, Δbio, ΔuvrB, and pKM101) and YG1041 (a derivative of the TA98that overproduces nitroreductase and O-acetyltransferase). The testswere performed in the presence and absence of a 4% (v/v) lyophilizedAroclor-1254-induced rat liver S9 fraction (Moltox Inc., Boone, NC) withcofactors, using dimethylsulfoxide as a solvent and negative control. Foreach plate, 0.5 mL of sodium phosphate buffer or S9 mix and 1.0 mL ofsample solution were added. After 66 h of incubation at 37 °C, the colo-nies were manually counted under a stereoscope. Toxicity was also eval-uated by visual observation under a stereoscope. For TA98 testing, thepositive controls were 0.5 μg/plate of 4-nitroquinoline-oxide (4NQO)(Sigma-Aldrich) and 2.5 μg/plate of 2-aminoanthracene (2AA) (Sigma-Aldrich), both dissolved in dimethylsulfoxide. For YG1041, the positivecontrols were 10 μg/plate of 4-nitro-ortho-phenylenediamine (4NOP)(Sigma-Aldrich) and 0.0625 μg/plate of 2-aminoanthracene (2AA), bothdissolved in dimethylsulfoxide. The negative control for all tests was ster-ile ultrapure Milli-Q water. Five different doses ranging from 1 to 50 μg/plate of samples were tested, along with negative and positive controls.Doses were tested in duplicates. Results were statistically analyzedusing the Bernstein model (Bernstein et al., 1982). Samples were consid-ered positive when significant ANOVA (pb0.05) and positive dose re-sponse was obtained. The results were expressed as the number ofrevertants/μg of each sample tested. In the case of the chlorination prod-ucts, the results were expressed per μg equivalent of the chlorinated dye.Before each test, samples were checked for contamination using nutrientagar plates. Also, the residual free chlorine level was measured and theanalysis was initiated only when no chlorine was detected.
3. Results
3.1. Analysis of commercial dye
High resolution mass spectra and molecular structures of the sixcolored components isolated from commercial Disperse Red 1 arepresented in Fig. 1. Each mass spectrum contained an M+1 peak thatcorresponded to the exact mass of F1–F6. The main fraction (F4)corresponded to the Colour Index structure for Disperse Red 1. Theother colored components were subsidiary dyes arising from themethodof synthesis of the target dye and its precursors.
3.2. Chlorination of commercial dye
Fig. 2 shows the results from the chlorination of commercial DisperseRed 1 over a 30-minute period. Reduction in intensity of the absorptionpeak at λmax=428 nm was consistent with destruction of the azo chro-mophore. LC–MS (positive ESI) was used to identify the main productsproduced by this process. Retention time (tR) and molecular mass(MM) values observed for the product ions are summarized in Table 1.The protonated ion peak for nitro-substituted compounds (Product Aand commercial dye fraction F4) was intense. Both substances producedsignals resulting from the elimination of the NO2 radical, which is consis-tent with previous reports (Dron et al., 2008; Holcapek et al., 2010;Levsen et al., 2007; Tai et al., 2006; Thevis et al., 2008). The completespectra are provided in Fig. 3. Product A arose from cleavage of theC\N bond on the right side of the azo group, producing Product B andthe corresponding diazo compound (O2N–C6H4–N2
+). This transient
species was attacked by a chloride ion, liberating molecular nitrogen(N2) and forming Product A. Chlorinated aromatic amines (Products Cand D) were formed upon further chlorination of electron-rich positionsortho to the tertiary amino group of Product B. Bearing in mind that thepresence of chlorine atoms leads to isotopic peaks for 35Cl and 37Cl havinga 3:1 ratio, ER (enhanced resolution) experiments were used to deter-mine isotopic contributions (Levsen et al., 2007; Tai et al., 2006;Holcapek et al., 2010). The results were consistent with the structuresof the four chlorinated products.
Peakswere also observed in the total ion current (TIC) chromatogramat retention times of 7.46 and 11.31 min (data not shown). Results of MSanalysis indicated that the associated compounds were mono- or di-chlorinated aromatic amines or nitrobenzenes but the precise molecularstructure of these minor components was unclear.
3.3. Ecotoxicity testing
3.3.1. Commercial dye and individual componentsThe commercial dye was tested for acute toxicity using D. similis
and H. attenuata. The EC50 was 0.13 mg/L for D. similis and 1.9 mg/Lfor H. attenuata. The dispersing agent was tested only using D. similisand the EC50 was 21 mg/L, which corresponded to the equivalentquantity present in 100 mg/L of the commercial dye. Therefore, it isevident that the surfactant did not contribute to the toxicity of thecommercial product, because it was at least one hundred times lesstoxic than the dye itself.
Due to limited amounts of the individual dye components, com-pounds F1–F6 were tested only with D. similis. It was found that onlyF4 could account for the toxicity of the commercial product (Fig. 4).This explainswhy the ecotoxicity of the commercial dye in theD. similisacute test was similar to the pure Disperse Red 1 (Ferraz et al., 2011).
3.3.2. Chlorine treated commercial dyeThe EC50 of the chlorine treated commercial dye was 4.3 mg/L using
D. similis and 0.7 mg/L using H. attenuata. Because insufficient amountsof F1–F6were available to chlorinate them separately, to make possiblecomparisons the toxicity of chlorination products A, B, C andD (Table 1)were estimated using the ECOSAR (Ecological Structure Activity Rela-tionships) program (US EPA, 2012). The predicted values for acute tox-icity to daphnids were 29.1, 46.5, 16.2 and 5.5 mg/L for A, B, C and D,respectively. The estimated EC50 of Product D was similar to the EC50 ofthe chlorine treated commercial dye obtained in theD. similis acute toxic-ity test. Although Product A was present at the highest concentration thetoxicity of themixture usingDaphnia could also be attributed to the othercompounds, especially product D, which gave the lowest predicted EC50.
3.4. Mutagenicity testing
3.4.1. Mutagenicity of individual dye componentsThe dye components exhibited positive mutagenic activity to the
YG1041 strain in the absence and presence of metabolic activation(Table 2). Component F1 showed the highest mutagenicity but be-cause it corresponded only to 5% of the mass it accounted for 26.5%(with S9) and 24.6% (without S9) of the mutagenicity of the commer-cial dye (Table 2). The main component (F4) accounted for the major-ity of the mutagenic activity of the mixture (64.2% with S9 and 49.7%without S9). The other components contributed only 9.3% (with S9)and 25.7% (without S9). With S9, components F1–F6 accounted forthe mutagenicity of the commercial dye. Without S9, the six compo-nents accounted for 67.5% of the mutagenicity. Therefore, mutagenicnon-azo dye compounds that were not isolated could be present inthe commercial dye.
3.4.2. Mutagenicity of the commercial dye and its chlorination productsCommercial Disperse Red 1 was non-mutagenic in the absence of
metabolic activation (S9) in TA98 and gave positive responses in
Fig. 1. High resolution mass spectra and structures characterizing commercial Disperse Red 1 components (F1– .
Fig. 2. Absorption spectral changes during the chlorination of commercial Disperse Red 1.
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TA98 with S9 and in YG1041 with and without S9 (Fig. 5). The chlori-nation products were mutagenic in both strains and the potencieswere higher than the commercial dye. Similar results were obtainedwhen pure Disperse Red 1 was chlorinated (Oliveira et al., 2010).
Product A (1-chloro-4-nitrobenzene) was the main identified chlo-rination product and seemed to be responsible for the mutagenicity ofthe chlorine-treated dye mixture. While this compound was previouslyreported to be mutagenic in TA98 (IUCLID, 2000), higher mutagenicityin YG1041 than in TA98 and the decreased response when S9 wasadded (Fig. 5) are known behaviors of nitro-compounds (Umbuzeiro
Table 1Summary of mass spectral data for Disperse Red 1 (F4) and its chlorination products.
SubstanceMM (Da)
tR(min)
Ions observed in first-order MS(m/z — structure)
Produ(m/z —
Initial dyea
MM=314.3313.06 315.14 — [M+H]+ 299 —
285 —
269 —
224 —
Product Ab
MM=157.5514.01 158.00 — [M+H]+ 142 —
128 —
112 —
Product Bb
MM=199.6710.40 200.00 — [M+H]+ 155 —
182 —
112 —
Product Cb
MM=234.1216.25 234.04 — [M+H]+ 216 —
217 —
199 —
189 —
155 —
146 —
189 —
88 —
Product Db
MM=268.5612.70 268.00 — [M+H]+ 223 —
250 —
180 —
88 —
MM = molecular mass; Da = Daltons; tR = retention time.a The sample gave the same fragmentation profile and retention time as commercial Disb Degradation products formed from the chlorination process.
et al., 2011). Since chlorinated aromatics were found in the effluentsof an azo dye manufacturer, as a result of chlorine treatments (Sarasaet al., 1998), it seems that 1-chloro-4-nitrobenzene is an importantaquatic contaminant.
Dispersing agents can also be transformed into mutagenic com-pounds during chlorination treatments, but in that case a good strain/condition for evaluation would be TA100 without S9 (Umbuzeiro et al.,2011). It was assumed that lignin sulfonate was the dispersing agentin Disperse Red 1, due to its dominance in disperse dye formulations(Hamprecht and Hunger, 2003); therefore chlorinated compounds de-rived from lignin (Rapson et al., 1980;Moller et al., 1986) could also con-tribute to the mutagenicity of the product mixture. An assay usingTA100 in the presence and absence of S9 was performed and the resultswere compared to those obtained using the commercial dye. An increasein mutagenicity without S9 was observed after chlorination (data notshown) which would suggest that mutagenic compounds were formedfrom chlorination of the dispersing agent (Umbuzeiro et al., 2011).Morestudies are being conducted to establish the specific contribution ofchlorinated compounds arising from the presence of lignin sulfonate tothe mutagenicity of the commercial dye.
4. Conclusions
Commercial Disperse Red 1 is composed of 60% dye component(N-ethyl-N-(2-hydroxyethyl)-4-(4-nitrophenylazo)), 20% subsidiarycolorants, and20%dispersing agent as a dyeing auxiliary. All six colorantsin the commercial dye are hydrophobic monoazo compounds that differin the number and types of alkyl groups attached to the amino N-atom.
The ecotoxicity of the commercial dye seems to arise mainly fromthe principal component (F4), the substance typically associated with
Fig. 3. Mass spectra (pos. ESI) of products A, B, C, and D from the chlorination of commercial Disperse Red 1.
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Disperse Red 1. The dispersing agent and the subsidiary colorants donot seem to contribute to the ecotoxicity of the commercial dye, atleast towards D. similis. Although the main component was not theonly mutagenic fraction, it contributed at least 49% of the totalmutagenicity.
The chlorine-treated commercial dye, although less toxic than theuntreated commercial product toD. similis, wasmore toxic toH. attenuataandmoremutagenic in the Salmonella/microsome assay. In this aspect of
the study, 1-chloro-4-nitrobenzene (A), 2-((4-chlorophenyl)(ethyl)amino)ethanol (B), 2-((2,4-dichlorophenyl)(ethyl)amino)ethanol (C),and 2-(ethyl(2,4,6-trichlorophenyl)amino)-ethanol (D) were detectedin the chlorine treated commercial dye mixture. Based on the ECOSARanalysis of each product, compound D seems to be the main contributorto the ecotoxicity of the mixture, along with one or more unidentifiedproducts. Further studies are required to determine which compoundsare responsible for the toxicity detected with the Hydra.
0 200 400 600 800 1000
0
5
10
15
20
Num
ber
of im
mob
ile o
rgan
ism
s
Concentration (µg/L)
F1 F2 F3 F4 F5 F6
Dye
Fig. 4. Comparison of the acute toxicity of commercial Disperse Red 1 components toDaphnia similis.
Table 2Contribution of components F1–F6 to the mutagenic activity obtained using YG1041strain in the presence and absence of metabolic activation.
308 F.I. Vacchi et al. / Science of the Total Environment 442 (2013) 302–309
Bearing in mind that the chlorination of commercial Disperse Red1 can generate para-chloronitrobenzene and other as yet unidentifiedcompounds that can be more harmful to the environment than thecommercial product, it is evident that disinfection or decolorationusing chlorine should be avoided in effluents containing azo dyessuch as Disperse Red 1.
TA98 -S9 TA98 +S9 YG1041 -S9 YG1041 +S9
0
50
100
150
200
250
300
350
400
100
25
400
0
40
0 418
Rev
./µg
Commercial Dye Chlorinated Dye
Fig. 5.Mutagenicity of the commercial dye and its chlorination products in the Salmonellastrains TA98 and YG1041, in the absence and presence of metabolic activation.
Acknowledgments
The authors thank FAPESP (2008/10449-7 and 2009/12739-5) forfinancial support and Jaqueline G. Honório for help with Ames testing.
References
Ake CL, Wiles MC, Huebner HJ, Mcdonald TJ, Cosgriff D, Richardson MB, et al. Porousorganoclay composite for the sorption of polycyclic aromatic hydrocarbons andpentachlorophenol from groundwater. Chemosphere 2003;51:835–44.
American Public Health Association (APHA). Standardmethods for the examination of waterand wastewater. 19th edition. Washington, D.C.: American Public Health Association,American Water Works Association, and Water Pollution Control Federation; 1995
Bae JS, Freeman HS. Aquatic toxicity evaluation of new direct dyes to the Daphniamagna. Dyes Pigment 2007;73(1):81–5.
Bafana A, Devi SS, Chakrabarti T. Azo dyes: past, present and the future. Environ Rev2011;19:350–70.
Bernstein L, Kaldor J,McCann J, PikeMC. Anempirical approach to the statistical analysis ofmutagenesis data from the Salmonella test. Mutat Res 1982;97:267–81.
Colour Index International. Society of Dyers and Colourists and American Association ofTextile Chemists and ColoristsOnline edition. ; 2011.
Dron J, Abidi E, El Haddad I, Marchand N, Wortham H. Precursor ion scanning-massspectrometry for the determination of nitro functional groups in atmospheric par-ticulate organic matter. Anal Chim Acta 2008;618(2):184–95.
Ferraz ERA, Umbuzeiro GA, De-Oliveira G, Caloto-Oliveira A, Chequer FM, Zanoni MV,Dorta D, Oliveira DP. Differential toxicity of Disperse Red 1 and Disperse Red 13in the Ames test, HepG2 cytotoxicity assay, and Daphnia acute toxicity test. EnvironToxicol 2011;26(5):489–97.
Gottlieb A, Shaw C, Smith A, Wheatley A, Forsythe S. The toxicity of textile reactive azodyes after hydrolysis and decolourisation. J Biotechnol 2003;101(1):49–56.
Hamilton MA, Russo RC, Thurfton RB. Trimmed Spearman–Karber method for estimatingmedian lethal concentration in toxicity bioassays. Environ Sci Technol 1977;11(7):714–9.
Hamprecht R, Hunger K. Dye classes for practical applications. In: Hunger K, editor. Indus-trial dyes: chemistry, properties, applications. Wiley-VCH; 2003. p. 146.
Holcapek M, Jirasko R, Lisa M. Basic rules for the interpretation of atmospheric pressureionization mass spectra of small molecules. J Chromatogr A 2010;1217(25):3908–21.
International Organization for Standardization. ISO 16240. Water quality — determina-tion of the genotoxicity of water and waste water — Salmonella/microsome test(Ames test); 2005 [20 pp.].
IUCLID. International Uniform Chemical Information Database. European Commission;2000 [101 pp.].
Kummrow F, Umbuzeiro GA. 2-Fenilbenzotriazóis (PBTA): uma nova classe decontaminantes ambientais. Quim Nova 2008;31(2):401–6.
Laitinen HA, Boyer KW. Simultaneous determination of bromine and chlorine withmethyl orange. Anal Chem 1972;44(6):920–6.
Levsen K, Schiebel HM, Terlouw JK, Jobst KJ, Elend M, Preib A, et al. Even-electron ions:a systematic study of the neutral species lost in the dissociation of quasi-molecularions. Mass Spectrom 2007;42(8):1024–44.
Mayura K, Smith EE, Clement BA, Phillips TD. Evaluation of the developmental toxicityof chlorinated phenols utilizing Hydra attenuata and postimplantation rat embryosin culture. Toxicol Appl Pharmacol 1991;108(2):253–66.
Moller M, Carlberg GE, Soteland N. Mutagenic properties of spent bleaching liquorsfrom sulphite pulps and a comparison with kraft pulp bleaching liquors. MutatRes 1986;172:89–96.
Neal RA. Evaluation of mixtures. In: Vouk VB, Butler GC, Upton AC, Parke DV, Asher SC,editors. Methods for assessing the effects of mixtures of chemicals. SCOPE. JohnWiley & Sons; 1987. p. 543–53.
Novotný C, Dias N, Kapanen A, Malachová K, Vándrovcová M, Itävaara M, et al. Compar-ative use of bacterial, algal and protozoan tests to study toxicity of azo- and anthra-quinone dyes. Chemosphere 2006;63(9):1436–42.
Nukaya H, Yamashita J, Tsuji K, Terao Y, Ohe T, Sawanishi H, et al. Isolation and chemical–structural determination of a novel aromatic amine mutagen in water from theNishitakase River in Kyoto. Chem Res Toxicol 1997;10:1061–6.
OECD. Daphnia sp. acute immobilisation test. Organisation for Economic Co-operationand Development — Guideline for testing of chemicals, no. 202; 2004 [12 pp.].
Oliveira DP, Carneiro PA, Rech CM, Zanoni MVB, Claxton LD, Umbuzeiro GA. Mutageniccompounds generated from the chlorination of disperse azo-dyes and their presencein drinking water. Environ Sci Technol 2006;40:6682–9.
Oliveira GAR, Ferraz ERA, Chequer FMD, Grando MD, Angeli JPF, Tsuboy MS, et al. Chlo-rination treatment of aqueous samples reduces, but does not eliminate, the muta-genic effect of the azo dyes Disperse Red 1, Disperse Red 13 and Disperse Orange 1.Mutat Res 2010;703(2):200–8.
Osugi ME, Rajeshwar K, Ferraz ERA, Oliveira DP, Araújo AR, Zanoni MVB. Comparison ofoxidation efficiency of disperse dyes by chemical and photoelectrocatalytic chlori-nation and removal of mutagenic activity. Electrochim Acta 2009;54:2086–93.
Rapson WH, Nazar MA, Butsky VV. Mutagenicity produced by aqueous chlorination oforganic compounds. Bull Environ Contam Toxicol 1980;24(1):590–6.
Reife A, Freeman HS. Environmental chemistry of dyes and pigments. N.Y.: John Wileyand Sons; 1995 [352 pp.].
Sarasa J, RocheMP, OrmadMP, Gimeno E, Puig A, Ovelleiro JL. Treatment of a wastewaterresulting from dyes manufacturing with ozone and chemical coagulation. Water Res1998;32:2721–7.
309F.I. Vacchi et al. / Science of the Total Environment 442 (2013) 302–309
Sponza DT. Toxicity studies in a chemical dye production industry in Turkey. J HazardMater 2006;138(3):438–47.
Stolz A. Basic and applied aspects in the microbial degradation of azo dyes. ApplMicrobiol Biotechnol 2001;56:69–80.
Tai Y, Pei S, Wan J, Cao X, Pan Y. Fragmentation study of protonated chalcones by atmo-spheric pressure chemical ionization and tandem mass spectrometry. RapidCommun Mass Spectrom 2006;20(6):994-1000.
Thevis M, Kohler M, Thomas A, Schlorer N, Schanzer W. Doping control analysis of tri-cyclic tetrahydroquinoline-derived selective androgen receptor modulators usingliquid chromatography/electrospray ionization tandem mass spectrometry. RapidCommun Mass Spectrom 2008;22(16):2471–8.
Trottier S, Blaise C, Kusui T, Johnson EM. Acute toxicity assessment of aqueous samples usingamicroplate-basedHydra attenuata assay. Environ ToxicolWater Qual 1997;12:265–71.
Umbuzeiro GA, Roubicek DA, Rech CM, Sato MIZ, Claxton LD. Investigating the sourcesof the mutagenic activity found in a river using the Salmonella assay and differentwater extraction procedures. Chemosphere 2004;54(11):1589–97.
Umbuzeiro GA, Freeman HS, Warren SH, Oliveira DP, Terao Y, Watanabe T, et al. Thecontribution of azo dyes to the mutagenic activity of Cristais River. Chemosphere2005;60(1):55–64.
Umbuzeiro GA,MachalaM,Weiss J. Diagnostic tools for effect-directed analysis ofmutagens,AhR agonists and endocrine disruptors. In: Brack,Werner. (Org.). Effect-directed analysisof complex environmental contamination. 1st ed. Nova York: Springer, v. 15; 2011, p.70–100.
US EPA. Estimation Programs Interface Suite™ forMicrosoft®Windows, v 4.10.Washington,DC, USA: United States Environmental Protection Agency; 2012.
Venkataraman K. The chemistry of synthetic dyes, v7. New York: Academic Press; 1978[372 pp.].
Wang Y, Chen J, Ge L, Wang D, Cai X, Huang L, Hao C. Experimental and theoreticalstudies on the photoinduced acute toxicity of a series of anthraquinone derivativestowards the water flea (Daphnia magna). Dyes Pigment 2009;83(3):276–80.
