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EC number: 271-946-1 | CAS number: 68647-35-8 This substance is identified in the Colour Index by Colour Index Constitution Number, C.I. 42535:4.
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- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
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- Toxicological Summary
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- Acute Toxicity
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Endpoint summary
Administrative data
Description of key information
Short term toxicity to fish:
Based on the prediction done using the OECD QSAR toolbox version 3.4 with log kow as the primary descriptor and considering the six closest read across substances, the short term toxicity on fish Cyprinodon variegatus was predicted for Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8). Based on the mortality of fishes, the LC50 value was estimated to be 0.94 mg/l for Cyprinodon variegatus for 96 hrs of duration. Based on this value it can be concluded that the substance Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8) is considered to be toxic to aquatic environment and classified as aquatic acute 1 category as per the criteria mentioned in CLP regulation.
Short term toxicity to aquatic invertebrates:
Based on the prediction done using the OECD QSAR toolbox version 3.4 with log kow as the primary descriptor and considering the six closest read across substances, toxicity on daphnia magna was predicted for Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8). Based on the intoxication of test organism the EC50 value was estimated to be 0.54 mg/l when Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien -1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8) exposed to daphnia magna for 48 hrs. Based on this value it can be concluded that the substance Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8) is considered to be toxic to aquatic environment and classified as aquatic acute 1 category as per the criteria mentioned in CLP regulation.
Short term toxicity to aquatic algae:
Based on the prediction done using the OECD QSAR toolbox version 3.4 with log kow as the primary descriptor and considering the five closest read across substances, toxicity on Pseudokirchneriella subcapitata was predicted for Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8). Based on the growth inhibition of test organism the EC50 value was estimated to be 0.404 mg/l when Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1 -ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8) exposed to pseudokirchneriella subcapitata for 72 hrs. Based on this value it can be concluded that the substance Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8) is considered to be toxic to aquatic environment and classified as aquatic acute 1 category as per the criteria mentioned in CLP regulation.
Toxicity to microorganisms:
Based on the prediction done using the OECD QSAR toolbox version 3.4 with log kow as the primary descriptor and considering the five closest read across substances, toxicity on microorganism Tetrahymena pyriformis predicted for Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8). Based on the growth rate inhibition the IGC50 value was estimated to be 5.85 mg/l when Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8) exposed to Tetrahymena pyriformis for 48hrs.
Additional information
Summarized result of toxicity of Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatesilicates (68647-35-8) on the growth and other activity of fishes, aquatic invertebrates, algae and cyanobacteria and microorganisms by considering the data for target as well as RA chemical which was selected on the basis of structure and functional similarity which are as follows:
Short term toxicity to fish:
Based on the predicted data for the target chemical and experimental data for read across chemicals which was selected on the basis of structure similarity study have been reviewed to determine the toxic nature of Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatesilicates (68647-35-8) on the mortality of fish. The studies are as mentioned below:
In the first predicted weight of evidence report from OECD QSAR toolbox 3.4, 2018 toxicity was measured. Based on the prediction done using the OECD QSAR toolbox version 3.4 with log kow as the primary descriptor and considering the six closest read across substances, the short term toxicity on fish Cyprinodon variegatus was predicted for Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8). Based on the mortality of fishes, the LC50 value was estimated to be 0.94 mg/l for Cyprinodon variegatus for 96 hrs of duration. Based on this value it can be concluded that the substance Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8) is considered to be toxic to aquatic environment and classified as aquatic acute 1 category as per the criteria mentioned in CLP regulation.
Further first predicted study was supported by the second experimental study for read across (83803-79-6) chemical from UERL lab. Fish Acute Toxicity test according to OECD Guideline 203 was conducted for (test item name) [4-[[4-anilino-1-naphthyl][4 -(dimethylamino)phenyl]methylene]cyclohexa-2,5-dien-1-ylidene] dimethylammonium acetate. The test solution was prepared by dissolving 100 mg of the test substance in 100 ml of potable water (passed through reverse osmosis system) with 1 hr. continuous stirring for achieving test concentrations of 1g/L. The nominal concentration selected for the experiment were 0.3125mg/L,0.625mg/L,1.25mg/L,2.5mg/L,5mg/Land test fish were exposed to these concentration for 96 hours. After 96hrs of exposure LC0, LC50 and LC100 was observed. The lethal concentrations LC50 was determine to be >0.3125 mg/L but <0.625 mg/L. LC0 (96 hours) (highest loading at which no mortality was observed) = 0.3125 mg/L. LC50 (96 hours) Experimental = >0.3125 mg/L but <0.625 mg/L. LC100 (96 hours) (lowest loading at which 100% mortality was observed) = 0.625 mg/L. Based on LC50 chemical was toxic and consider as classified in aquatic acute 1 as per the CLP classification criteria.
Similarly for the same read across chemical (83803-79-6) from BASF report Study was conducted to determine the toxic nature of chemical N-(4-{[4-(dimethylamino)phenyl][4-(methylamino)phenyl] methylene}cyclohexa-2,5-dien-1-ylidene)-N-methylmethanaminium acetate on the growth of fish Leuciscus idus. Test was performed according to the OECD guideline. The doses were chosen based on two range finding studies between 0.3 and 1 mg/l) the concentrations, spaced by a factor of about 2.2. Control, 0.01, 0,022, 0.046, 0.1, 0.215, 0464, 1.0, 2.15 mg/L concentration was taken in the study. In the form of an aqueous solution (0.1 g/100 ml) the product was added to the test water, subsequently the fish were placed into the aquaria. Leuciscus idus (golden variety) used as a test organism collected from Fischzucht Paul Eggers, 2354 Hohenwestedt, Germany. Fish does not feed during test. Open glass aquaria having dimeter 30 x 22 x 24 cm, was used having the capacity of 10 liter. 2.8 g fish /L was added. 10 organisms per vessel was used. Chloracetamid added as a reference substance. After the exposure of 96hrs LC50 observed on the basis of Apathy and narcotic-like state was determine at test concentration 2.15 mg/L after 1 hour. In reference substance Chloracetamid the LC50 was 32 mg/L at 48 hr. Based on the mortality of fish the LC50 was 0.158 mg/l and the LC100 was 0.0464 mg/l. Based on the LC50 chemical was consider as toxic and can be consider to be classified as aquatic acute 1 category as per the CLP classification criteria.
Similarly in the fourth weight of evidence study for the read across chemical (6358-36-7) (from UERL 2017) toxicity was conducted on fish to determine the toxic nature of chemical. Fish Acute Toxicity test according to OECD Guideline 203 was conducted for (test item name) 4,4'-carbonimidoylbis [N,Ndiethylaniline] monohydrochloride. The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 1g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with 1 hour continuous stirring. After that analytically detect stock, solubility found 981.56 mg/L and calculations done accordingly. The nominal concentration selected for the experiment were 0.625mg/L,1.25mg/L,2.5mg/L,5mg/L, 10mg/L and test fish were exposed to these concentration for 96 hours. The lethal concentrations LC50 was determine to be 0.625 mg/L. LC50 (96 hours) Experimental = 0.625 mg/L. LC100 (96 hours) (lowest loading at which 100% mortality was observed) = 1.25 mg/L
The fifth study used for the read across chemical (84434-47-9) from UERL lab 2017, fish Acute Toxicity test according to OECD Guideline 203 was conducted for (test item name) [4-[[4 -(dimethylamino) phenyl][4-(methylamino)phenyl]methylene] cyclohexa-2,5-dien- 1-ylidene]dimethylammonium acetate The nominal concentration selected for the experiment were 0.3125mg/L, 0.625mg/L, 1.25mg/L, 2.5mg/L, 5mg/L and test fish were exposed to these concentration for 96 hours. The lethal concentrations LC50 was found to be >0.3125 mg/L but <0.625 mg/L.
LC0 (96 hours) (highest loading at which no mortality was observed) = 0.3125 mg/L
LC50 (96 hours) Experimental = >0.3125 mg/L but <0.625 mg/L
LC100 (96 hours) (lowest loading at which 100% mortality was observed) = 0.625 mg/L
Thus based on the above data sources for target chemical Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatesilicates (68647-35-8) from various experimental studies and predicted studies from QSAR, UERL lab report and BASF data, it was summarized that the chemical was toxic to aquatic organisms fishes and thus can be classified as aquatic acute category 1 as per the CLP criteria.
Short term toxicity to aquatic invertebrates:
Based on the prediction data for the target chemical and experimental data for read across chemical which was selected on the basis of structure similarity study have been reviewed to determine the toxic nature of Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatesilicates (68647-35-8) on the mobility of daphnia magna. The studies are as mentioned below:
In the first predicted weight of evidence report from OECD QSAR toolbox 3.4, 2018 toxicity was measured. Based on the prediction done using the OECD QSAR toolbox version 3.4 with log kow as the primary descriptor and considering the six closest read across substances, toxicity on daphnia magna was predicted for Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8). Based on the intoxication of test organism the EC50 value was estimated to be 0.54 mg/l when Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien -1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8) exposed to daphnia magna for 48 hrs. Based on this value it can be concluded that the substance Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8) is considered to be toxic to aquatic environment and classified as aquatic acute 1 category as per the criteria mentioned in CLP regulation.
Further first predicted study was supported by the second experimental study for read across (83803-79-6) chemical from ABITEC lab. Determination of the inhibition of the mobility of daphnids was carried out with the substance N-(4-{[4-(dimethylamino)phenyl][4-(methylamino)phenyl]methylene}cyclohexa-2,5-dien- 1-ylidene)-N-methylmethanaminium acetate according to OECD Guideline 202. The test substance was tested at the concentrations 0, 0.006, 0.012, 0.025, 0.050, 0.100 mg/l. Effects on immobilisation were observed for 48 hours. The median effective concentration (EC50) for the test substance, N-(4-{[4-(dimethylamino)phenyl][4-(methylamino)phenyl]methylene}cyclohexa-2,5-dien-1-ylidene)-N- methylmethanaminium acetate, in Daphnia magna was determined to be 0.037 mg/L for immobilisation effects. This value indicates that the substance is likely to be hazardous to aquatic invertebrates and can be classified as aquatic acute 1 as per the CLP criteria.
Similarly in the third weight of evidence study for the read across chemical (6358-36-7) (from ABITEC lab report 2017) toxicity was conducted on daphnia to determine the toxic nature of chemical. Determination of the inhibition of the mobility of daphnids was carried out with the substance 4,4'-carbonimidoylbis[N,N-diethylaniline] monohydrochloride according to OECD Guideline 202. The test substance was tested at the concentrations 0, 0.01, 0.02, 0.04, 0.08, 0.16 and 0.32 mg/L. Effects on immobilisation were observed for 48 hours. The median effective concentration (EC50) for the test substance, 4,4'-carbonimidoylbis[N,N-diethylaniline] monohydrochloride, in Daphnia magna was determined to be 0.016 mg/L for immobilisation effects. This value indicates that the substance is likely to be hazardous to aquatic invertebrates and can be classified as Aquatic acute category 1 as per the CLP criteria.
Thus based on the above data sources for target chemical Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatesilicates (68647-35-8) from various experimental studies and predicted studies from QSAR, ABITEC report 2017, it was summarized that the chemical was toxic to aquatic invertebrates and thus can be classified as aquatic acute category 1 as per the CLP criteria.
Short term toxicity to aquatic algae:
Based on the prediction data for the target chemical and experimental data for read across chemical which was selected on the basis of structure similarity study have been reviewed to determine the toxic nature of Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatesilicates (68647-35-8) on the mortality of aquatic algae and cyanobacteria. The studies are as mentioned below:
In the first predicted weight of evidence report from OECD QSAR toolbox 3.4, 2018 toxicity was measured for Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatesilicates (68647-35-8). Based on the prediction done using the OECD QSAR toolbox version 3.4 with log kow as the primary descriptor and considering the five closest read across substances, toxicity on Pseudokirchneriella subcapitata was predicted for Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8). Based on the growth inhibition of test organism the EC50 value was estimated to be 0.404 mg/l when Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1 -ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8) exposed to pseudokirchneriella subcapitata for 72 hrs. Based on this value it can be concluded that the substance Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8) is considered to be toxic to aquatic environment and classified as aquatic acute 1 category as per the criteria mentioned in CLP regulation.
Further first predicted study was supported by the second experimental study for read across (6358-36-7) chemical from ABITEC lab. Freshwater algal growth inhibition test was carried out on Desmodesmus subspicatus with the substance 4,4'-carbonimidoylbis[N,N-diethylaniline] monohydrochloride according to OECD Guideline 201. The stock solution (10 mg/L) was prepared by dissolving yellow powder in OECD growth medium. Test solutions of required concentration were prepared by mixing the stock solution of the test sample with OECD growth medium and inoculum culture and tested at the concentrations 0, 3, 6, 12, 24 and 50 mg/L. Effects on the growth rate of the organism were studied. The median effective concentration (EC50) for the test substance, 4,4'-carbonimidoylbis[N,N-diethylaniline] monohydrochloride, in Desmodesmus subspicatus was determined to be 0.13 mg/L. This value indicates that the substance is likely to be hazardous to aquatic algae and can be classified as Aquatic acute category 1 as per the CLP criteria.
Similarly in the third weight of evidence study for the read across chemical (2465-27-2) (from j-check 2017) toxicity was conducted on algae to determine the toxic nature of chemical. Short term toxicity to green algae study was carried out for 72 hrs. The study was based on the effects of the test compound 4,4'-carbonimidoylbis (N,N-dimethylaniline) hydrochloride (CAS no. 2465 -27 -2) on green algae in a static fresh water system. Based on effect on growth rate of the test organism green algae, the 72 hr EC50 and NOEC value was determined to be 0.34 and 0.026 mg/l, respectively and on the basis of AUG, the 72 hr EC50 and NOEC value was determined to be 0.093 and 0.026 mg/l, respectively. Thus, based on the EC50 value, it can be concluded that the substance 4,4'-carbonimidoylbis(N,N-dimethylaniline) hydrochloride can be considered as toxic to aquatic organisms and thus can be considered to be classified in aquatic acute category 1 as per the CLP classification criteria.
Thus based on the above data sources for target chemical Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatesilicates (68647-35-8) from various experimental studies and predicted studies from QSAR, ABITEC report 2017 and j-check, it was summarized that the chemical was toxic to aquatic algae and cyanobacteria and thus can be classified as aquatic acute category 1 as per the CLP criteria.
Toxicity to microorganisms:
Based on the prediction data for the target chemical and experimental data for read across chemical which was selected on the basis of structure similarity study have been reviewed to determine the toxic nature of Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatesilicates (68647-35-8) on the microorganisms. The studies are as mentioned below:
In the first predicted weight of evidence report from OECD QSAR toolbox 3.4, 2018 toxicity was measured for Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatesilicates (68647-35-8). Based on the prediction done using the OECD QSAR toolbox version 3.4 with log kow as the primary descriptor and considering the five closest read across substances, toxicity on microorganism Tetrahymena pyriformis predicted for Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8). Based on the growth rate inhibition the IGC50 value was estimated to be 5.85 mg/l when Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdate silicates (68647-35-8) exposed to Tetrahymena pyriformis for 48hrs.
Further first predicted study was supported by the second experimental study for read across chemical Basic violet 1 (8004-87-3) from Chemosphere 2003. Determination of toxicity of methyl violet on the growth of 14 gram negative microorganisms. Inhibitory activity of test chemical (methyl violet) was determined by the traditional agar gel method. The conc. of test chemical used for the study was 0.6918 mg/l (1 µM). 14 different gram negative bact. were used for the study was Agrobacterium radiobacter, Agrobacterium tumefaciens, Bradyrhizobium japonicum, Escherichia coli, Erwinia atroseptica, Erwinia herbicola, Erwinia uredovora, Pseudomonas fluorescence, Pseudomonas phaseolicola, Pseudomonas syringae , Rhizobium trifolii, Xanthomonas malvacearum, Xanthomonas phaseoli and X. stewartii. These test organisms were taken from the collection of the Plant Protection Institute, Hungarian Academy of Sciences (Budapest, Hungary). The bacteria were maintained on Nutrient Agar (Oxoid CM3) completed with vitamins pyridoxine. HCl, thiamine. HCl, riboflavine and nicotinamide at 1, 10, 1 and 20 mg/l concentrations. Bacterial suspensions for screening were prepared by washing cells with sterile tap water containing 0.3% peptone from slants of 20 h old cultures grown at (21±1) °C. A layer of 5 mm depth of inoculated medium was dispensed into Petri dishes of 90 mm diameter. Filter paper discs of 5 mm diameter were impregnated with 1 µM solution of test compounds and placed centrally on the surface of the agar plate. Growth inhibition zones were measured after 24 h incubation at 21 ± 1 °C. After incubation colony diameters were determined and growth inhibition was calculated in percentage of dye free colony growth. Each determination was run in quadruplicate. Based on no effect on growth inhibition of gram negative test organisms, the NOEC value of methyl violet was determine to be 0.6918 mg/l for Erwinia herbicola and Pseudomonas syringae. Whereas the growth inhibition was observed on the other organisms so LOEC was 0.6918 mg/l for remaining 12 bacteria.
Similarly in the third weight of evidence study for the same read across chemical (8004-87-3) from Chemosphere 2003 toxicity was measured on the gram positive bacteria. Determination of toxicity of methyl violet on the growth of 8 gram positive microorganisms. Inhibitory activity of test chemical (methyl violet) was determined by the traditional agar gel method. The conc. of test chemical used for the study was 0.6918 mg/l (1 µM). 8 different gram positive bact. were used for the study- Gram – positive bacteria was Corynebacterium fascians, Corynebacterium flaccumfaciens, Corynebacterium michiganense, Corynebacterium oortii, Stapylococcus aureus, Micrococcus luteus, Bacillus subtilis and Bacillus thuringiensis Berliner subp. Kurstaki. These test organisms were taken from the collection of the Plant Protection Institute, Hungarian Academy of Sciences (Budapest, Hungary). The bacteria were maintained on Nutrient Agar (Oxoid CM3) completed with vitamins pyridoxine. HCl, thiamine. HCl, riboflavine and nicotinamide at 1, 10, 1 and 20 mg/l concentrations. Bacterial suspensions for screening were prepared by washing cells with sterile tap water containing 0.3% peptone from slants of 20 h old cultures grown at (21±1) °C. A layer of 5 mm depth of inoculated medium was dispensed into Petri dishes of 90 mm diameter. Filter paper discs of 5 mm diameter were impregnated with 1 µM solution of test compounds and placed centrally on the surface of the agar plate. Growth inhibition zones were measured after 24 h incubation at 21 ± 1 °C. After incubation colony diameters were determined and growth inhibition was calculated in percentage of dye free colony growth. Each determination was run in quadruplicate. Based on growth inhibition of gram positive test organisms, the LOEC value of methyl violet was determine to be 0.6918 mg/l.
Thus based on the toxicity data chemical was consider as toxic to microorganisms.
Based on the overall studies for the target chemical, toxicity on fish, aquatic invertebrates and algae Benzenamine, 4-[(4-aminophenyl)(4-imino-2,5-cyclohexadien-1-ylidene)methyl]-, N-Me derivatives, molybdatesilicates (68647-35-8) chemical was consider as toxic and can be consider to be classified as aquatic acute 1 category as per the CLP classification criteria.
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