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EC number: 221-359-1 | CAS number: 3077-12-1
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- 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
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Gene mutation in bacterial cells (Ames test): negative in Salmonella typhimurium strains TA100, TA1535, TA102, TA98 and TA1537 with and without metabolic activation
Cytogenicity (chromosome aberration test): negative in cultured peripheral human lymphocytes with and without metabolic activation (based on read-across)
Gene mutation in mammalian cells (mouse lymphoma assay): positive in L5178Y mouse lymphoma cells with and without metabolic activation (based on read-across)
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 26 Oct - 17 Nov 1998
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Remarks:
- (non-standard positive controls were used, which are not recommended in the guideline; single plates were used per concentration instead of triplicate plating)
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- adopted in 1997
- Deviations:
- yes
- Remarks:
- (non-standard positive controls were used, which are not recommended in the guideline; single plates were used per concentration instead of triplicate plating)
- GLP compliance:
- no
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his operon
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
- Metabolic activation:
- with and without
- Metabolic activation system:
- cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254
- Test concentrations with justification for top dose:
- Pre-test for toxicity/Experiment I (plate incorporation): 16, 50, 160, 500, 1600 and 5000 µg/plate with and without metabolic activation
Experiment II (preincubation method): 16, 50, 160, 500, 1600 and 5000 µg/plate with and without metabolic activation - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test substance was soluble in DMSO and formed a clear colorless solution. - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 2-nitrofluorene
- sodium azide
- cumene hydroperoxide
- other: 2-aminoanthracene: +S9, 3 µg/plate for all strains; nitrofurantoin: - S9, 0.2 µg/plate for TA100; 4-nitro-1,2-phenylene diamine: -S9, 0.5 and 10 µg/plate for TA1537 and TA98, respectively
- Remarks:
- Each batch of S9 mix was checked for its metabolizing capacity by using reference mutagenes (not further specified); appropriate activity was demonstrated.
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation, Experiment I); preincubation (Experiment II)
DURATION
- Preincubation period: 20 min (Experiment II)
- Exposure duration: 48 h (Experiment I and II)
NUMBER OF REPLICATIONS: Single plates per concentration each in two independent experiments
DETERMINATION OF CYTOTOXICITY
- Method: background growth, marked and dose-dependent reduction in mutant count, titre count - Evaluation criteria:
- A reproducible and dose-related increase in mutant counts of at least one strain is considered to be a positive result. For TA1535, TA100 and TA98 this increase should be about twice that of negative controls, whereas for TA1537, at least a threefold increase should be reached. For TA102 an increase about 150 mutants should be reached. Otherwise, the result is evaluated as negative. However, these guidelines may be overruled by good scientific judgement. In case of questionable results, investigations should continue, possibly with modifications, until a final evaluation is possible.
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at 5000 µg/plate with and without metabolic activation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at 5000 µg/plate with and without metabolic activation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at 5000 µg/plate with and without metabolic activation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at 5000 µg/plate with and without metabolic activation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 102
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at 160 and 500 µg/plate with and without metabolic activation, respectively
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation of the test substance was not observed.
RANGE-FINDING/SCREENING STUDIES: Experiment I (plate incorporation) served as pre-test for toxicity.
HISTORICAL CONTROL DATA
Only single plates were counted in the present study for each control and concentration level. Moreover only mean values without standard deviation were given in the historical control data, therefore it cannot be confirmed if the positive or negative control values are within the range of the historical control data. However, the positive control values from the study and from the historical control data increased mutant counts well over those of the negative controls. - Conclusions:
- Based on the results of the conducted study, the test substance was not considered to reveal mutagenic properties in Salmonella typhimurium strains TA1535, TA1537, TA89, TA100 and TA102 with and without metabolic activation, respectively.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- refer to analogue justification provided in IUCLID section 13
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Species / strain:
- lymphocytes: Cultured peripheral human lymphocytes
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- A chromosome aberration study with the source substance Reaction mass of 2,2'-[(4-methylphenyl)imino]bisethanol and Ethanol 2-[[2-(2-hydroxyethoxy)ethyl](4-methylphenyl)amino]- (EC 911-490-9) was performed according to OECD 473 guideline and GLP principles, in cultured peripheral human lymphocytes in two independent experiments. It is concluded that the source substance is not clastogenic in human lymphocytes. Applying the read-across approach, similar results are expected for the target substance N,N-Dihydroxyethyl-p-toluidine (CAS 3077-12-1).
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- refer to analogue justification provided in IUCLID section 13
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- without
- Genotoxicity:
- positive
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at and above 3330 μg/mL (3 h treatment); at and above 250 μg/mL (24 h treatment)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at and above 1650 μg/mL (3 h treatment)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not specified
- Positive controls validity:
- valid
- Conclusions:
- The mutagenic activity of the source substance Reaction mass of 2,2'-[(4-methylphenyl)imino]bisethanol and Ethanol 2-[[2-(2-hydroxyethoxy)ethyl](4-methylphenyl)amino]- (EC 911-490-9) was evaluated in an in vitro mammalian cell gene mutation test with L5178Y mouse lymphoma cells, performed according to OECD 476 and following GLP principles.The test substance induced mutations in the absence of metabolic activation, but not with metabolic activation, which was confirmed in an independent repeat experiment.
It is concluded that the test substance is positive in the in vitro mammalian cell gene mutation test. Applying the read-across approach, similar results are expected for the target substance N,N-Dihydroxyethyl-p-toluidine (CAS 3077-12-1).
Referenceopen allclose all
Table 1 Results of Experiment I (plate incorporation)
With or without S9-Mix | Test substance concentration (μg/plate) | Revertant colonies per plate | ||||
Base-pair substitution type | Frameshift type | |||||
TA100 | TA1535 | TA102 | TA98 | TA1537 | ||
– | DMSO | 64 | 11 | 115 | 9 | 9 |
– | 16 | 63 | 10 | 139 | 9 | 8 |
– | 50 | 75 | 11 | 131 | 16 | 14 |
– | 160 | 71 | 9 | 162 | 11 | 6 |
– | 500 | 68 | 9 | 109 | 9 | 8 |
– | 1600 | 77 | 12 | 93B | 11 | 5 |
5000 | 77 | 14 | 60B | 8 | 4 | |
Positive controls, –S9 | Name | NF | SA | Cumene | 4-NPDA | 4-NPDA |
Concentrations (μg/plate) | 0.2 | 10 | 50 | 0.5 | 10 | |
Revertant colonies per plate | 293 | 714 | 277 | 148 | 129 | |
+ | DMSO | 141 | 9 | 203 | 31 | 12 |
+ | 16 | 96 | C | 195 | 14 | 16 |
+ | 50 | 104 | 7 | 198 | 27 | 17 |
+ | 160 | 104 | 10 | 151 | 35 | 9 |
+ | 500 | 119 | 6 | 161B | 28 | 8 |
+ | 1600 | 145 | 11 | 117B | 31 | 5 |
+ | 5000 | 145 | 5 | 51B | 18 | 2 |
Positive controls, –S9 | Name | 2-AA | 2-AA | 2-AA | 2-AA | 2-AA |
Concentrations (μg/plate) | 3 | 3 | 3 | 3 | 3 | |
Revertant colonies per plate | 1658 | 379 | 492 | 1214 | 382 |
NF = Nitrofurantoin
SA = Sodium azide
Cumene = Cumene hydroperoxide
4-NPDA = 4-Nitro-1,2-phenylene diamine
2AA = 2-Aminoanthracene
C = contaminated
B = reduced background lawn
Table 2: Results of Experiment II (preincubation method)
With or without S9-Mix | Test substance concentration (μg/plate) | Revertant colonies per plate | ||||
Base-pair substitution type | Frameshift type | |||||
TA100 | TA1535 | TA102 | TA98 | TA1537 | ||
– | DMSO | 73 | 14 | 178 | 16 | 7 |
– | 16 | 96 | 14 | 216 | 22 | 7 |
– | 50 | 100 | 8 | 211 | 18 | 11 |
– | 160 | 72 | 6 | 215 | 16 | 7 |
– | 500 | 114 | 8 | 202 | 19 | 9 |
– | 1600 | 106 | 9 | 179 | 18 | 4 |
5000 | 83B | 5B | 75B | 5B | 2B | |
Positive controls, –S9 | Name | NF | SA | Cumene | 4-NPDA | 4-NPDA |
Concentrations (μg/plate) | 0.2 | 10 | 50 | 0.5 | 10 | |
Revertant colonies per plate | 347 | 650 | 523 | 152 | 146 | |
+ | DMSO | 125 | 9 | 239 | 27 | 9 |
+ | 16 | 94 | 12 | 266 | 22 | 13 |
+ | 50 | 101 | 12 | 237 | 19 | 9 |
+ | 160 | 112 | 10 | 266 | 22 | 11 |
+ | 500 | 133 | 8 | 252 | 16 | 11 |
+ | 1600 | 155 | 13 | 168 | 9 | 8 |
+ | 5000 | 151B | 10B | 79B | 17B | 3B |
Positive controls, –S9 | Name | 2-AA | 2-AA | 2-AA | 2-AA | 2-AA |
Concentrations (μg/plate) | 3 | 3 | 3 | 3 | 3 | |
Revertant colonies per plate | 1798 | 198 | 439 | 1587 | 184 |
NF = Nitrofurantoin
SA = Sodium azide
Cumene = Cumene hydroperoxide
4-NPDA = 4-Nitro-1,2-phenylene diamine
2AA = 2-Aminoanthracene
C = contaminated
B = reduced background lawn
Table 3: Historical control values (January - December 1996): plate incorporation
With or without S9-Mix | Compound | TA100 | TA1535 | TA102 | TA98 | TA1537 |
– | DMSO | 86 | 9 | 292 | 20 | 8 |
– | SA | -- | 739 | -- | -- | -- |
– | NF | 253 | -- | -- | -- | -- |
– | 4-NPDA | -- | -- | -- | 155 | 139 |
– | Cumene | -- | -- | 624 | -- | -- |
+ | DMSO | 103 | 11 | 340 | 30 | 10 |
+ | 2-AA | 1540 | 204 | 829 | 1461 | 281 |
NF = Nitrofurantoin
SA = Sodium azide
Cumene = Cumene hydroperoxide
4-NPDA = 4-Nitro-1,2-phenylene diamine
2AA = 2-Aminoanthracene
Table 4: Historical control values (January - December 1996): preincubation method
With or without S9-Mix | Compound | TA100 | TA1535 | TA102 | TA98 | TA1537 |
– | DMSO | 80 | 9 | 271 | 22 | 8 |
– | SA | -- | 747 | -- | -- | -- |
– | NF | 278 | -- | -- | -- | -- |
– | 4-NPDA | -- | -- | -- | 183 | 127 |
– | Cumene | -- | -- | 602 | -- | -- |
+ | DMSO | 94 | 11 | 337 | 28 | 8 |
+ | 2-AA | 1318 | 172 | 601 | 1239 | 164 |
NF = Nitrofurantoin
SA = Sodium azide
Cumene = Cumene hydroperoxide
4-NPDA = 4-Nitro-1,2-phenylene diamine
2AA = 2-Aminoanthracene
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (positive)
Genetic toxicity in vivo
Description of key information
Gene mutation in vivo (Comet assay): negative in liver, duodenum and glandular stomach tissues of male rats (based on read-across)
Link to relevant study records
- Endpoint:
- in vivo mammalian cell study: DNA damage and/or repair
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Remarks:
- Summary of available data used for the endpoint assessment of the target substance
- Adequacy of study:
- key study
- Justification for type of information:
- refer to analogue justification provided in IUCLID section 13
- Reason / purpose for cross-reference:
- read-across source
- Reason / purpose for cross-reference:
- read-across source
- Key result
- Sex:
- male
- Genotoxicity:
- negative
- Toxicity:
- yes
- Vehicle controls validity:
- valid
- Negative controls validity:
- not examined
- Positive controls validity:
- valid
- Remarks on result:
- other: Source: EC 911-490-9
- Conclusions:
- It is concluded that the comet assay in liver, duodenum and glandular stomach was valid and the source substance Reaction mass of 2,2'-[(4-methylphenyl)imino]bisethanol and Ethanol 2-[[2-(2-hydroxyethoxy)ethyl](4-methylphenyl)amino]- (EC 911-490-9) does not provoke DNA damage in the Comet assay up to and including a dose of 750 mg/kg bw (the maximum tolerated dose in accordance with current regulatory guidelines). Applying the read-across approach, similar results are expected for the target substance N,N-Dihydroxyethyl-p-toluidine (CAS 3077-12-1).
Reference
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Gene mutation in bacterial cells: Ames test
Reliable data are available regarding genetic toxicity in bacterial cells for N,N-Dihydroxyethyl-p-toluidine (CAS 3077-12-1).
An Ames test was performed similar to OECD 471 with Salmonella typhimurium strains TA100, TA1535, TA102, TA98 and TA1537, with and without metabolic activation (Bayer AG, 1998). No test substance precipitation was observed when tested up to 5000 μg/plate. The pre-test for cytotoxicity served as Experiment 1. Cytotoxicity was observed at 5000 µg/plate with and without metabolic activation for S. typhimurium strains TA100, TA1535, TA98 and TA1537, respectively. In S. typhimurium TA102 the test substance showed cytotoxic properties at 160 and 500 µg/plate with and without metabolic activation, respectively. The positive controls showed the expected increase in mutant colonies when compared to the corresponding negative control. Evidence of mutagenic activity of N,N-Dihydroxyethyl-p-toluidine (CAS 3077-12-1) was not seen, since no biologically relevant increase in the mutant count in comparison with the negative controls were observed. Therefore, based on these data it is concluded that N,N-Dihydroxyethyl-p-toluidine (CAS 3077-12-1) is not mutagenic in bacterial cells under the conditions of this test.
No data is available regarding gene mutation and cytogenicity in mammalian cells and genetic toxicity in vivo for N,N-Dihydroxyethyl-p-toluidine (CAS 3077-12-1). Therefore, read-across from an appropriate structural analogue substance is conducted in accordance with Regulation (EC) No 1907/2006, Annex XI, 1.5. in order to fulfil the standard data requirements defined in Regulation (EC) No 1907/2006, Annex VIII, 8.4. Structural similarities and similarities in properties and/or activities of the source and target substance are the basis of read-across. A detailed justification for the analogue read-across approach is provided in the technical dossier (see IUCLID Section 13). Information from the analogue substance Reaction mass of 2,2'-[(4-methylphenyl)imino]bisethanol and 2-[[2-(2-hydroxyethoxy)ethyl](4-methylphenyl)amino]-ethanol (EC 911-490-9) will be taken into account to fulfil the standard data requirements defined in Regulation (EC) No 1907/2006, Annex VIII, 8.4.
Cytogenicity in mammalian cells: In vitro chromosome aberration test
A chromosome aberration study with the source substance Reaction mass of 2,2'-[(4-methylphenyl)imino]bisethanol and 2-[[2-(2-hydroxyethoxy)ethyl](4-methylphenyl)amino]-ethanol (EC 911-490-9) was performed according to OECD 473 and GLP principles, in cultured peripheral human lymphocytes in two independent experiments (WIL Research, 2013d). No precipitation of the substance was observed, but the test substance was tested up to and beyond cytotoxic concentrations. Both in the absence and presence of S9-mix, the source substance did not induce a statistically significant or biologically relevant increase in the number of cells with chromosome aberrations. It can be concluded that Reaction mass of 2,2'-[(4-methylphenyl)imino]bisethanol and 2-[[2-(2-hydroxyethoxy)ethyl](4-methylphenyl)amino]-ethanol (EC 911-490-9) does not disturb mitotic processes and cell cycle progression and does not induce numerical chromosome aberrations under the experimental conditions described in this report.
In vitro gene mutation test in mammalian cells: mouse lymphoma assay:
An in vitro mammalian cell gene mutation test with L5178Y mouse lymphoma cells, performed according to OECD 476 and following GLP principles is available. The source substance Reaction mass of 2,2'-[(4-methylphenyl)imino]bisethanol and 2-[[2-(2-hydroxyethoxy)ethyl](4-methylphenyl)amino]-ethanol (EC 911-490-9) was tested up to cytotoxic concentrations (WIL Research, 2013e). The positive control data and the solvent control data validated the study. In the presence of S9-mix, the source substance did not induce a significant increase in the mutation frequency in two experiments. In the absence of S9-mix, the test substance induced a 5.3-fold increase in the mutation frequency at the prolonged treatment period of 24 hours. This was verified in a further experiment, in which the substance induced an up to 3.3-fold increase in the mutation frequency. The mutation frequency of both the small and large colonies was significantly increased. Based on these data, it is concluded that the test substance is positive in the in vitro mammalian cell gene mutation test.
Alkaline in vivo Comet assay in liver, stomach and duodenum of rats:
The alkaline in vivo Comet assay with the source substance Reaction mass of 2,2'-[(4-methylphenyl)imino]bisethanol and 2-[[2-(2-hydroxyethoxy)ethyl](4-methylphenyl)amino]-ethanol (EC 911-490-9) was performed according to OECD 489 and GLP principles (Charles River, 2017a). One experiment was performed for liver and two experiments were performed for glandular stomach and duodenum since the first experiment did not pass the acceptance criteria for the control group. Groups of 5 male rats were dosed once daily via oral gavage with vehicle or with 187.5, 375 and 750 mg of the test substance per kg body weight for three consecutive days. A positive control group was included. The animals that were dosed with 750 mg/kg bw/day showed clinical signs including lethargy, tremors, hunched posture and ataxia. Approximately 3-4 hours after the second dose of ethylmethanesulphonate (EMS) and third dose of the vehicle or the test substance, liver, glandular stomach, and duodenum tissue were collected, and single cell suspensions were made followed by Comet slide preparation. No biologically relevant increase in the mean Tail Intensity (%) was observed in liver cells. No statistically significant increase in the mean Tail Intensity (%) was observed in glandular stomach and duodenum cells. As the acceptance criteria for the control group were not passed for glandular stomach and duodenum, no conclusion could be drawn about the potential DNA damaging properties of the test substance in these tissues. Therefore the in vivo alkaline Comet assay was repeated for the organs duodenum and glandular stomach, because the Tail Intensity of the negative control did not meet the acceptance criteria.
It is concluded that the Comet assay in liver is valid and the source substance Reaction mass of 2,2'-[(4-methylphenyl)imino]bisethanol and 2-[[2-(2-hydroxyethoxy)ethyl](4-methylphenyl)amino]-ethanol (EC 911-490-9) does not provoke DNA damage in the Comet assay up to and including a dose of 750 mg/kg bw (the maximum tolerated dose in accordance with current regulatory guidelines).
The alkaline in vivo Comet assay with the source substance Reaction mass of 2,2'-[(4-methylphenyl)imino]bisethanol and 2-[[2-(2-hydroxyethoxy)ethyl](4-methylphenyl)amino]-ethanol (EC 911-490-9), performed according to OECD 489 and GLP principles, was repeated for the organs duodenum and glandular stomach, because the Tail Intensity (%) of the vehicle control group determined in the initial experiment (Charles River, 2017a) was considered to be invalid. Groups of 5 male rats were dosed once daily via oral gavage with vehicle or with 187.5, 375 and 750 mg of the test substance per kg body weight for three consecutive days (Charles River, 2018). A positive control group and a vehicle control were included. All animals of the low (187.5 mg/kg bw) and mid dose group (375 mg/kg bw) were lethargic. After approximately 21 hour these animals showed no abnormalities. Within one hour after the second and third treatment the animals showed no treatment related effects with exception of 1/5 animal in the mid dose group and 2/5 animals in the low dose group, which were lethargic after the second and third dosing. In addition, 1/5 animals of the low dose group showed ataxia after the third dose. 1/5 animals of the high dose group were lethargic and showed ataxia after the first dosing. After approximately 21 hour the animal showed no abnormalities. The same animal was lethargic and showed ataxia after the second and third dosing as well. The effects in another animal of the high dose group were the same with the exception that the animal showed only lethargy after the first dosing. The remaining animals treated with 750 mg/kg bw showed mortality (3/5), one after the first dosing and two after the second dosing. No statistically significant increase in the mean Tail Intensity (%) was observed in stomach cells and in duodenum cells of test item-treated male animals at any of the dose levels tested compared to the vehicle treated animals. The mean Tail Intensity (%) of vehicle treated male rats was 0.91 ± 0.63 % in stomach cells and 2.47 ± 0.65 % in duodenum cells. The mean vehicle control Tail Intensity was within the 95% control limits of the distribution of the historical negative control database and the mean Tail Intensity (%) of the vehicle control was below 20% for both organs. The positive control EMS revealed an increase of the Tail Intensity, 52-fold induction in glandular stomach cells and 18-fold induction in duodenum cells, respectively. Thus all negative and positive control values in glandular stomach and duodenum met the acceptance criteria.
In conclusion, the test is valid and Reaction mass of 2,2'-[(4-methylphenyl)imino]bisethanol and 2-[[2-(2-hydroxyethoxy)ethyl](4-methylphenyl)amino]-ethanol (EC 911-490-9) does not cause DNA damage in the Comet assay in glandular stomach and duodenum cells when sampled approximately 3-4 hours post dosing, of male rats that were dosed via oral gavage for three consecutive days up to a dose of 750 mg/kg bw (the maximum tolerated dose in accordance with current regulatory guidelines) under the experimental conditions described in this report.
Justification for classification or non-classification
Reliable data with the registration substance and from a structural analogue substance regarding in vitro and in vivo genetic toxicity indicate that the registration substance does not meet the classification criteria according to Regulation (EC) 1272/2008, and are therefore conclusive but not sufficient for classification.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.