1-Dodecanamine, reaction products with 1,1'-methylenebis[4-isocyanatobenzene] and 1-octanamine

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

In vitro genetic toxicity studies have been conducted on four substances in the MDI category. Bacterial reverse mutation assays (Ames tests) have been conducted on 3,3'-dioctadecyl-1,1'-methylenebis(4,1-phenylene)diurea (EC 406-690-3), 3,3'-dicyclohexyl-1,1'-methylenebis(4,1-phenylene)diurea (EC 406-370-3), N,N''-(methylenedi-4,1-phenylene)bis[N'-octyl]urea (EC 445-760-8), and a mixture of: 3,3'-dicyclohexyl- 1,1'-methylenebis(4,1-phenylene)diurea; 3-cyclohexyl-1-(4-(4-(3-octadecylureido)benzyl)phenyl)urea; 3,3'-dioctadecyl-1,1'-methylenebis(4,1-phenylene)diurea (EC 406-530-2),. The results from all tests concluded that the substances were not mutagenic in the presence or absence of metabolic activation. Additionally, an in vitro mammalian chromosome aberration test was also performed on N,N''-(methylenedi-4,1-phenylene)bis [N'-octyl]urea (EC 445-760-8). The study concluded that no chromosomal abnormalities were observed in either the presence or absence of metabolic activation during the two replicate tests, and therefore the conclusion is that this substance is not a clastogen.

In addition, three in vivo micronucleus clastogenicity studies were undertaken on 3,3'-dioctadecyl-1,1'- methylenebis(4,1-phenylene)diurea (EC 406-690-3), 3,3'-dicyclohexyl-1,1'-methylenebis(4,1-phenylene)diurea (EC 406-370-3), and a mixture of: 3,3'-dicyclohexyl-1,1'-methylenebis(4,1-phenylene)diurea; 3-cyclohexyl- 1-(4-(4-(3-octadecylureido)benzyl)phenyl)urea; 3,3'-dioctadecyl-1,1'-methylenebis(4,1-phenylene)diurea (EC 406-530-2). No adverse effects were observed and no chromosomal abnormalities detected based on either the P/N ratio or micronucleus formation; though in the preliminary tests some minor toxic effects were observed at 5000 mg/kg in the studies conducted on 3,3'-dicyclohexyl-1,1'-methylenebis(4,1-phenylene)diurea (EC 406-370-3), and a mixture of: 3,3'-dicyclohexyl- 1,1'-methylenebis(4,1-phenylene)diurea; 3-cyclohexyl- 1-(4-(4-(3-octadecylureido)benzyl)phenyl)urea; 3,3'-dioctadecyl-1,1'-methylenebis(4,1-phenylene)diurea (EC 406-530-2). Hence it was confirmed that noin vivoclastogenicity was evident as a result of treatment with the test substances.

  

Under the conditions of the tests, the four substances did not demonstrate a mutagenic or clastogenic response, and thus the other MDI category members are also considered to be non-mutagenic.

 

 

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Remarks:

This study is included in a NONS registration and therefore has been evaluated by a relevant competent authority and is considered to be reliable.

Qualifier:

according to guideline

Guideline:

other: Annex V (Ames)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Remarks:

and TA 1538

Metabolic activation:

with and without

Metabolic activation system:

Aroclor induced rat liver S9

Test concentrations with justification for top dose:

Concentration range in the main test (with metabolic activation): 10 ... 5000 µg/plate
Concentration range in the main test (without metabolic activation): 10 ... 5000 µg/plate

Vehicle / solvent:

Solvent: Dimethylsulfoxide

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

other: 4-nitro-ortho-ohenylenediamine and 2-aminoanthracene

Details on test system and experimental conditions:

Concentration of the test substance resulting in precipitation: 333 µg/plate

Species / strain:

other: Strains as specified above, preliminary test

Metabolic activation:

with

Genotoxicity:

not specified

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

>5000 µg/plate

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

primary culture, other: Strains as specified above, preliminary test

Metabolic activation:

without

Genotoxicity:

not specified

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

> 5000 µg/plate

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

other: Strains as specified above, main test

Metabolic activation:

with

Genotoxicity:

not specified

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

> 5000 µg/plate

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

other: Strains as specified above, main test

Metabolic activation:

without

Genotoxicity:

not specified

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

>5000 µg/plate

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

Observations: The plates incubated with the test substance showed normal background growth at concentrations up to 5000 µg/plate. No increases in the numbers of revertant colonies were seen at any of the concentrations of the test substance with or without metabolic activation.

Positive control substances sodium azide, 4-nitro-ortho-ohenylenediamine) and 2-aminoanthracene led to increases in induce revertant colonies in the appropriate bacterial strains.

Conclusions:

The Ames test was negative with metabolic activation and without metabolic activation.

Executive summary:

The test item was investigated for in vitro genetic toxicity at the concentration in an Ames test with a concentration range of 10 to 5000 µg/plate, with and without metabolic activation, following a standard guideline. The study included TA98, TA100, TA1535, TA1537 and TA1538 strains. Positive control substances were also included. The Ames test was negative with metabolic activation and negative without metabolic activation as no increases in the numbers of revertant colonies were seen at any of the concentrations of the test substance, with or without metabolic activation.

The study is a GLP compliant, guideline study and is acceptable with restrictions for assessment of this endpoint.

Reference 2

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Remarks:

This study is included in a NONS registration and therefore has been evaluated by a relevant competent authority and is considered to be reliable.

Qualifier:

according to guideline

Guideline:

other: Annex V (Ames)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

Aroclor induced rat liver S9

Test concentrations with justification for top dose:

Concentration range in the main test (with metabolic activation): 10 ... 5000 µg/plate
Concentration range in the main test (without metabolic activation): 10 ... 5000 µg/plate

Vehicle / solvent:

Solvent: Dimethylsulfoxide

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

not specified

Details on test system and experimental conditions:

Concentration of the test substance resulting in precipitation: 333 µg/plate

Species / strain:

other: Strains specified above

Remarks:

Preliminary test

Metabolic activation:

with

Genotoxicity:

not specified

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

>5000 ug/plate

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Species / strain:

other: Strains specified above

Remarks:

Preliminary test

Metabolic activation:

without

Genotoxicity:

not specified

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

> 5000 ug/plate

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Species / strain:

other: Strains specified above

Remarks:

Main test

Metabolic activation:

with

Genotoxicity:

not specified

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

> 5000 ug/plate

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Species / strain:

other: Strains specified above

Remarks:

Main test

Metabolic activation:

without

Genotoxicity:

not specified

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

>5000 ug/plate

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

Observations: No increases in the number of revertant colonies were seen in any of the bacterial strains treated with the test substance, either in the presence or absence of S9.
Positive control substance led to significant increases in the number of revertant colonies compared with controls.

Conclusions:

The Ames test was negative with metabolic activation and negative without metabolic activation.

Executive summary:

The test item was investigated in an in vitro genetic toxicity study (Ames test) at a concentration range of 10 to 5000 µg/plate, along with a positive control and solvent control. The strains TA98, TA100, TA1535 and TA1537 were included in the test. The test was conducted with and without metabolic activation. The Ames test was negative with metabolic activation and negative without metabolic activation.

The study is a GLP compliant, guideline study and is acceptable with restrictions for assessment of this endpoint.

Reference 3

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Remarks:

This study is included in a NONS registration and therefore has been evaluated by a relevant competent authority and is considered to be reliable.

Qualifier:

according to guideline

Guideline:

other: Annex V

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain / cell type:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

Aroclor induced rat liver S9

Test concentrations with justification for top dose:

Concentration range in the main test (with metabolic activation): 10 ... 5000 µg/plate
Concentration range in the main test (without metabolic activation): 10 ... 5000 µg/plate
Concentration of the test substance resulting in precipitation: 5000 µg/plate

Vehicle / solvent:

Solvent: DMSO

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Key result

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Toxic at 5000 µg/plate, with and without metabolic activation (in one experiment only)

Additional information on results:

RANGE-FINDING/SCREENING STUDIES: No increase in revertants was observed up to > 5000 µg/plate in the preliminary test. No toxicity observed up to 5000 µg/plate for TA98 and TA100, with and without metabolic activation.

Conclusions:

The Ames test concluded that the test item was negative with and without metabolic activation.

Executive summary:

The test item was investigated for in vitro genetic toxicity in Salmonella typhimurium bacteria strains TA98, TA100, TA1535, TA1537 and TA1538 following EU Annex V guidelines. Bacteria were exposed to the test item in the concentration range of 10 to 5000 µg/plate, with and without metabolic activation. The test item was observed to precipitate at the highest concentration of 5000 µg/plate. The test item was cytotoxic at 5000 µg/plate for TA1538, however this was observed in one experiment only. The test concluded that the test item was negative with and without metabolic activation.

The study is a GLP compliant, guideline experimental study and is acceptable with restrictions for assessment of this endpoint.

Reference 4

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Remarks:

This study is included in a NONS registration and therefore has been evaluated by a relevant competent authority and is considered to be reliable.

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Remarks:

and TA 102

Metabolic activation:

with and without

Metabolic activation system:

S9 mix prepared from the microsomal fraction (S9 mix fractions) of rat liver induced by Aroclor 1254.

Test concentrations with justification for top dose:

Concentration range in the main test (with metabolic activation): 3.13 ... 50 µg/plate
Concentration range in the main test (without metabolic activation): 3.13 ... 50 µg/plate

Vehicle / solvent:

Solvent: Ethanol

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Details on test system and experimental conditions:

The product is insoluble in vehicles usually used in this test. A suspension of homogeneous appearance could not be obtained, therefore an extract produced in ethanol has been tested. The doses are therefore expressed in µL of ethanol extract per plate.
The highest dose tested is 50 µL per plate which corresponds to the highest volume used for this vehicle in this type of test in the laboratory

Species / strain:

other: main test

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Species / strain:

other: Main test

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Conclusions:

The Ames test concluded that the test item was negative with metabolic activation and negative without metabolic activation.

Executive summary:

The test item was investigated for in vitro genetic toxicity at the concentration range of 3.13 to 50 µg/plate in an Ames test. The study was conducted with and without metabolic activation following the OECD 471 and EU Method B13/14 guidelines. The Ames test concluded that the test item was negative with metabolic activation and negative without metabolic activation.

The study is a GLP compliant, guideline experimental study and is acceptable with restrictions for assessment of this endpoint.

Reference 5

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Remarks:

This study is included in a NONS registration and therefore has been evaluated by a relevant competent authority and is considered to be reliable.

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Qualifier:

according to guideline

Guideline:

EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)

GLP compliance:

yes

Type of assay:

in vitro mammalian cell micronucleus test

Species / strain / cell type:

human lymphoblastoid cells (TK6)

Metabolic activation:

with and without

Metabolic activation system:

S9 mix prepared from the microsomal fraction (S9 mix fractions) of rat liver induced by Aroclor 1254.

Test concentrations with justification for top dose:

Concentration range in the main test (with metabolic activation): 12 ... 15 µg/mL
Concentration range in the main test (without metabolic activation): 12 ... 15 µg/mL

Vehicle / solvent:

Solvent: Ethanol

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Details on test system and experimental conditions:

Exposure period (with metabolic activation): 3 hours
Exposure period (without metabolic activation): 44 hours
Expression time:
First test: 3 hours exposure with and without S9 mix.
Second test: exposure of 20 and 44 hours without S9 mix.
Exposure of 3 hours with S9 mix.
Selection time:
First test: collection 20 hours after the start of treatment (about 1 and a half normal cell cycle).
Second test: collection 20 and 44 hours after the start of treatment (about 1 and a half normal cell cycle, and 24 hours later).

Species / strain:

other: main test

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Species / strain:

other: Main test

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

No dose-dependent anomaly observed during the two trials

Conclusions:

The study concluded that the test was negative with metabolic activation and negative without metabolic activation.

Executive summary:

The test item was investigated for in vitro genetic toxicity at a concentration range of 12 to 15 µg/mL. The test was conducted with and without metabolic activation following the OECD 473 and EU Method B10 guidelines. The test item is insoluble in the vehicles usually used in this test and no suspension of homogeneous appearance could be obtained, therefore an extract of the product in ethanol was tested. The doses are therefore expressed in µL of ethanol extract per tube (containing 5.5 mL of final treatment volume). The highest dose tested is 15 µL per tube which corresponds to the largest volume used for this vehicle in this type of test in the laboratory.

No dose-dependent anomaly was observed during these two trials. The study concluded that the test item was negative with metabolic activation and negative without metabolic activation.

The study is a GLP compliant, guideline experimental study and is acceptable with restrictions for assessment of this endpoint.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

In vivo genetic toxicity studies were conducted on three MDI category substances, 3,3'-dioctadecyl-1,1'- methylenebis(4,1-phenylene)diurea (EC 406-690-3), 3,3'-dicyclohexyl-1,1'-methylenebis(4,1-phenylene)diurea (EC 406-370-3), and a mixture of: 3,3'-dicyclohexyl-1,1'-methylenebis(4,1-phenylene)diurea; 3-cyclohexyl- 1-(4-(4-(3-octadecylureido)benzyl)phenyl)urea; 3,3'-dioctadecyl-1,1'-methylenebis(4,1-phenylene)diurea (EC 406-530-2), as discussed in the key information above.

Link to relevant study records

Reference

Reference 1

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Remarks:

This study is included in a NONS registration and therefore has been evaluated by a relevant competent authority and is considered to be reliable.

Qualifier:

according to guideline

Guideline:

other: Annex V (Micronucleus)

GLP compliance:

yes

Type of assay:

other: micronucleus assay

Species:

mouse

Strain:

NMRI

Sex:

male/female

Route of administration:

oral: unspecified

Vehicle:

Corn oil

Details on exposure:

The dose of the test substance was judged to be the maximum attainable.

Duration of treatment / exposure:

Male: 5000 mg/kg; No. of animals: 5; Sacrifice time: 24 hours
Male: 5000 mg/kg; No. of animals: 5; Sacrifice time: 48 hours
Male: 5000 mg/kg; No. of animals: 5; Sacrifice time: 72 hours
Female: 5000 mg/kg; No. of animals: 5; Sacrifice times: 24 hours
Female: 5000 mg/kg; No. of animals: 5; Sacrifice times: 48 hours
Female: 5000 mg/kg; No. of animals: 5; Sacrifice times: 72 hours

Frequency of treatment:

Not specified

Post exposure period:

Not specified

Dose / conc.:

5 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

15 male and 15 females

Control animals:

not specified

Positive control(s):

Cyclophosphamide

Tissues and cell types examined:

Not specified

Evaluation criteria:

Micronucleus formation

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

not specified

Negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

No increase in micronucleus formation was observed. The positive control substance cyclophosphamide led to an increase in the incidence of micronuclei.
Toxicity: yes. Doses producing toxicity: Mitotic index: 5000 mg/kg; In a preliminary experiment 2 males and 2 females were treated with 5000 mg/kg of the test substance and exhibited a reduction in spontaneous activity.

Conclusions:

The in vivo genetic toxicity of the test item was assessed in a micronucleus assay. The result was negative for the test item.

Executive summary:

The in vivo genetic toxicity of the test item was assessed in a micronucleus assay. The result was negative for the test item.

In vivo genetic toxicity was assessed in a micronucleus assay following a standard guideline. 15 male and female mice were exposed at a concentration of 5000 mg/kg, with 5 of each sex being sacrificed after 24, 48 and 72 hours. The study was negative, as no increase in micronucleus formation was observed.

In a preliminary experiment, 2 males and 2 females were treated with 5000 mg/kg of the test substance and exhibited a reduction in spontaneous activity.

The study is a GLP compliant, guideline experimental study and is acceptable with restrictions for assessment.

Reference 2

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Remarks:

This study is included in a NONS registration and therefore has been evaluated by a relevant competent authority and is considered to be reliable.

Qualifier:

according to guideline

Guideline:

other: Annex V (B12, Micronucleus)

GLP compliance:

yes

Type of assay:

other: micronucleus assay

Species:

mouse

Strain:

NMRI

Sex:

male/female

Route of administration:

oral: unspecified

Vehicle:

Corn oil

Duration of treatment / exposure:

Male: 5000 mg/kg; No. of animals: 5; Sacrifice time: 24 hours
Male: 5000 mg/kg; No. of animals: 5; Sacrifice time: 48 hours
Male: 5000 mg/kg; No. of animals: 5; Sacrifice time: 72 hours
Female: 5000 mg/kg; No. of animals: 5; Sacrifice times: 24 hours
Female: 5000 mg/kg; No. of animals: 5; Sacrifice times: 48 hours
Female: 5000 mg/kg; No. of animals: 5; Sacrifice times: 72 hours

Frequency of treatment:

Not specified

Post exposure period:

Not specified

Dose / conc.:

5 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

15 male and 15 females

Control animals:

not specified

Positive control(s):

Not specified

Tissues and cell types examined:

Not specified

Evaluation criteria:

P/N Ratio

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

not specified

Negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

Doses producing toxicity: No change in the P/N ratio.
In a preliminary toxicity study in which 2 males and 2 females were given 5000 mg/kg, the only sign of toxicity observed was apathy and a decrease in spontaneous activity for up to 24 hours post-treatment

Conclusions:

An in vivo micronucleus assay with the test item was negative, as no change in the P/N ratio was observed.

Executive summary:

The in vivo genetic toxicity of the test item was assessed using NMIR rats (male and female) in a micronucleus assay following a standard guideline. 15 male and female mice were exposed at a concentration of 5000 mg/kg, with 5 of each sex sacrificed after 24, 48 and 72 hours. The P/N ratio was investigated. The study was negative. as no change in the P/N ratio was observed. In a preliminary toxicity study in which 2 males and 2 females were given 5000 mg/kg, the only sign of toxicity observed was apathy and a decrease in spontaneous activity for up to 24 hours post-treatment.

The study is a GLP compliant, guideline experimental study and is acceptable with restrictions for assessment of this endpoint.

Reference 3

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Remarks:

This study is included in a NONS registration and therefore has been evaluated by a relevant competent authority and is considered to be reliable.

Qualifier:

according to guideline

Guideline:

other: Annex V

GLP compliance:

yes

Type of assay:

mammalian bone marrow chromosome aberration test

Species:

mouse

Strain:

NMRI

Sex:

male/female

Route of administration:

other: oral (not specified)

Vehicle:

- Vehicle(s)/solvent(s) used: corn oil

Duration of treatment / exposure:

24, 48 and 72 hours

Dose / conc.:

5 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

5 per sex per dose

Control animals:

not specified

Positive control(s):

Not specified

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Remarks:

Based on P/N ratio

Additional information on results:

Other toxic signs: Dosing at 5000 mg/kg in a preliminary test for toxicity using 20 mL/kg b.w. resulted in a reduction in activity in all animals up to 6 hours post-dosing but these effects are not clearly substance related. No signs of toxicity were stated for the main study.

Conclusions:

The test item was concluded to be negative for mammalian chromosome aberration. No signs of toxicity or cytotoxicity (based on P/N ratio) were observed.

Executive summary:

The test item was investigated for in vivo chromosome aberration in male and female NMRI mice following EU Annex V guidelines (micronucleus test). Test animals were administered 5000 mg/kg test item by oral gavage and sacrificed at 24, 48 and 72 hours. The test item was concluded to be negative for mammalian chromosome aberration. No signs of toxicity or cytotoxicity (based on P/N ratio) were observed.

The study is a GLP compliant, guideline experimental study and is acceptable with restrictions for assessment of this endpoint.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Justification for classification or non-classification

In vitro genetic toxicity studies were conducted on four substances, and in vivo micronucleus assays were conducted on three substances. No adverse effects were observed and it is concluded that the substances tested and the other substances in the MDI category are not mutagenic or clastogenic and are not classified for this endpoint.