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Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Based on the results of these experiments and on standard evaluation criteria, it is concluded that FAT 40554 did not induce genetic toxicity.

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1991
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Justification for type of information:
None
Reference:
Composition 1
Qualifier:
according to
Guideline:
other: EEC B.14 (Ames test)
Principles of method if other than guideline:
None
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Test material information:
Composition 1
Specific details on test material used for the study:
None
Target gene:
None
Species / strain:
other: TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell lines (if applicable):
E. coli WP2uvrA
Additional strain characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
Rat-liver post mitochondrial supernatant (S9 fraction)
Test concentrations with justification for top dose:
0.05, 0.5, 5, 50, 500, 5000 μg/plate for pretest0.015, 0.05, 0.15, 0.5, 1.5, 5 μg/plate for TA100 without S9 activation on main test I & II0.5, 1.5, 5, 15, 50, 150 μg/plate for TA100 with S9 activation on main test I & II156, 313 ,625, 1250, 2500, 5000 μg/plate for TA 1535, TA 1537, TA 98 and WP2uvrA on main test I & II
Vehicle:
Solvent:DMSO
Negative controls:
yes
Remarks:
DMSO
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
benzo(a)pyrene
Details on test system and conditions:
None
Rationale for test conditions:
None
Evaluation criteria:
Does not use statistical approach for data analysis. A number of revertant colonies treated with test sample solution shows twice or more as many as a number of revertant colonies treated with solvent. And it increase concentration-dependent.Th result is judged as "Positive" for mutagenicity
Statistics:
Does not use statistical approach for data analysis.
Key result
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Genotoxicity:
positive
Cytotoxicity:
no
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Additional information on results:
None

None

Conclusions:
FAT 40554 is considered to be mutagenic in this Salmonella typhimurium reverse mutation assay.
Executive summary:

With or without S9 mix, the number of revertant colonies treated with test sample solution for TA100 shows more than twice as many as the one treated with solvent. And it increased concentration-dependent.

Pretest organized with 0.05, 0.5, 5, 50, 500, 5000 μg/plate.

With or without S9 mix, fatal effect was not observed. Without S9 mix, precipitation was observed on the concentration of 50μg/plate and more. With S9 mix, precipitation was observed on the concentration of 500μg/plate and more. With or without S9 mix, the number of revertant colonies treated with test sample solution for TA100 shows more than twice as many as the one treated with solvent.

Based on pretest result, TA100 without S9mix takes 0.015, 0.05, 0.15, 0.5, 1.5, 5 μg/plate and TA100 with S9mix takes 0.5, 1.5, 5, 15, 50, 150 μg/plate for main test I and II. (Table 2 and 4) Other species with or without S9 mix takes 156, 313 ,625, 1250, 2500, 5000 μg/plate. (Table 3 and 5) Based on main test result, with or without S9 mix, the number of revertant colonies treated with test sample solution for TA100 shows more than twice as many as the one treated with solvent. And it increased concentration-dependent. And without S9 mix, the number of revertant colonies treated with test sample solution for TA98 shows almost twice as many as the one treated with solvent. Based on above result, the test substance is judged as "Positive" for mutagenicity.

In conclusion, FAT 40554 is considered to be mutagenic in this Salmonella typhimurium reverse mutation assay.

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1991
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Justification for type of information:
None
Reference:
Composition 1
Qualifier:
according to
Guideline:
EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
Principles of method if other than guideline:
None
GLP compliance:
not specified
Type of assay:
in vitro mammalian chromosome aberration test
Test material information:
Composition 1
Specific details on test material used for the study:
None
Target gene:
Not applicable
Species / strain:
Chinese hamster lung fibroblasts (V79)
Additional strain characteristics:
not specified
Remarks:
Not reported within the SNIF file
Metabolic activation:
with and without
Metabolic activation system:
Rat-liver post mitochondrial supernatant (S9 fraction)
Test concentrations with justification for top dose:
'0, 20, 39, 78, 156, 313, 625, 1250, 2500, 5000 μg/ml
Vehicle:
- Solvent used: CMC -Na (carboxymethyl cellulose)-1%
Negative controls:
yes
Remarks:
CMC -Na (carboxymethyl cellulose)-1%
Solvent controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
cyclophosphamide
mitomycin C
Remarks:
with and without metabolic activation
Details on test system and conditions:
Exposure period (with metabolic activation): 6 hours
Rationale for test conditions:
None
Evaluation criteria:
In each concentration, frequency of number of cells which has structural abnormality and polyploid excluding number of GAP is used to judge.In case the frequency is below 5%, the result is judged as "Negative".In case the frequency is 5% to 10%, exclusive 10%giyousei , the result is judged as "Pseudo-positive".In case the frequency is 10% or more, the result is judged as "Positive".In case frequency of cells which has structural abnormality is clearly increased compare to solvent control. And the function has repeatability or dose-response, it is judged the test substance has clastogenicity.
Statistics:
None
Key result
Species / strain:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no
Vehicle controls valid:
yes
Negative controls valid:
yes
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.
Additional information on results:
None

Cell reproductive rate.
Direct methods
(w/o metabolic activity)
Conc
(mg/ml)
  Reproductive rate
(%)
24hr
treatment
48hr
treatment
0 (solvent) 100.0 100.0
0.020 * 96.7 90.5
0.039 * 97.4 94.5
0.078 * 93.2 94.1
0.156 * 92.6 89.0
0.313 * 84.4 96.1
0.625 * 77.4 85.9
1.250 * 71.0 67.8
2.500 * 72.4 65.1
5.000 * 66.3 60.8
Direct methods
(w/o metabolic activity)
Conc
(μg/ml)
  Reproductive rate
(%)
6hr
treatment
+S9
6hr
treatment
-S9
0 (solvent) 100.0 100.0
20 * 96.6 95.7
39 * 94.0 97.9
78 * 91.7 96.5
156 * 90.0 96.8
313 * 97.2 96.4
625 * 90.8 92.5
1250 * 91.6 93.1
2500 * 89.2 90.6
5000 * 88.6 88.2
* test substnce is a suspension in medium
Conclusions:
In conclusion, it can be stated that in the study described and under the experimental conditions reported, the test article FAT 40554 did not induce structural chromosome aberrations in V79 cells (Chinese hamster cell line).
Executive summary:

An in vitro screening assay was performed to assess the potential of FAT 40554 to induce structural chromosome aberrations.

The test substance does not induce frequency of cells which has structural abnormality and polyploid cells, even if metabolic activation method or direct methods .

Based on inhibition test result of cell proliferation, test substance has weak inhibition for Direct methods (without metavolic activation) 1250, 2500, 5000 μg/ml is selected for chromosomal aberration test with and without metabolic activation. In all concentration, test substance in medium is suspended.

In conclusion, it can be stated that in the study described and under the experimental conditions reported, the test article FAT 40554 did not induce structural chromosome aberrations in V79 cells (Chinese hamster cell line).

Endpoint:
in vitro gene mutation study in mammalian cells
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1992
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Justification for type of information:
None
Reference:
Composition 1
Qualifier:
according to
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
yes
Remarks:
The incubation period of all cultures for cytotoxicity was5 days instead of 6 days.
Principles of method if other than guideline:
None
GLP compliance:
not specified
Type of assay:
mammalian cell gene mutation assay
Test material information:
Composition 1
Specific details on test material used for the study:
None
Target gene:
V79 Chinese hamster cells were obtained. Rat liver S9 was prepared.
Species / strain:
Chinese hamster lung fibroblasts (V79)
Details on mammalian cell lines (if applicable):
Not reported within the SNIF file
Additional strain characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
Liver S9 mix of induced rats
Test concentrations with justification for top dose:
Concentration range in the main test (with metabolic activation): 625 ... 5000 µg/mlConcentration range in the main test (without metabolic activation): 625 ... 5000 µg/ml
Vehicle:
- Vehicle(s)/solvent(s) used: CMC (carboxymethyl cellulose)
Details on test system and conditions:
Exposure period (with metabolic activation): 5 hExposure period (without metabolic activation): 5 hcell cutlure: 7 d
Rationale for test conditions:
None
Evaluation criteria:
The chemical was judged to be mutagenic when all the following requirements are fulfilled:1. The mutation frequencies of the chemical-treated cultures are higher than threefold of that of the corresponding vehicle control.2. The mutation frequencies of the chemical-treated culture are higher than the highest historical background data.3. The mutation frequency inctreases dose-dependently. to examine the dose-relationship, linear regression analysis is performed.4. Above 3 responses are reproduced in the two independent assays.
Statistics:
The plating efficiency and the mutation frequency were calculated.
Key result
Species / strain:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
(> 5000 µg/ml)
Vehicle controls valid:
not specified
Negative controls valid:
not specified
Positive controls valid:
yes
Remarks on result:
other:
Remarks:
Migrated from field 'Test system'.
Additional information on results:
None

None

Conclusions:
Negative with and without metabolic activation.
Executive summary:

The test substance FAT 40554 was examined for its mutagenecity activity by assayinh the induction of 6- thioguanine resistant mutants of cultured V79 Chinese hamster cells in the presence and absence of metabolic activation (S9 mix) according to OECD Guideline 476 (In vitro Mammalian Cell Gene Mutation Test).

The cells were exposed to the test compound for 5 hr at doses of 625, 1250, 2500 and 5000 µg/ml both with and without metabolic activation. The test compound did not induce any increase in the mutation frequency as compared with those of the vehicle controls either in the presence or absence of metabolic activation. On the other hand, positive control chemicals, ethyl methanesulfonate (EMS, 200 µg/ml, without metabolic activation) and 9, 10 -dimethyl-1,2 -benzanthracene (DMBA, 5 µg/ml, with metabolic activation), induced marked increase in the mutation frequency.

Based on the above findings, it is concluded that the substance is not mutagenic under the conditions tested.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

Based on the results of micronucleus assay and on standard evaluation criteria, it is concluded that FAT 40554 did not induce genetic toxicity.

Link to relevant study records
Reference
Endpoint:
in vivo mammalian germ cell study: gene mutation
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
None
Reference:
Composition 1
Qualifier:
according to
Guideline:
other: Japanese Industrial Safety and Health Law guideline
Principles of method if other than guideline:
None
GLP compliance:
yes
Type of assay:
micronucleus assay
Test material information:
Composition 1
Specific details on test material used for the study:
None
Species:
mouse
Strain:
not specified
Details on species / strain selection:
None
Sex:
male
Details on test animals and environmental conditions:
Specific Pathogen Free(SPF) male mice of BDF1(C57BL/6xDBA/2) strain Pilot study Main Study a) Receipt of animals Feb. 15, 1990 March 15, 1990 (age) (8 weeks old) (8 weeks old)b) Number ordered Male: 50 Male: 45c) Date of administration Feb. 26, 1990 March 26, 1990 (age) (9 weeks old) (9 weeks old)d) Body weight at 25.7-27.9g 24.3-28.7g administration period
Route of administration:
intraperitoneal
Vehicle:
Corn oil
Details on exposure:
None
Duration of treatment / exposure:
None
Frequency of treatment:
None
Post exposure period:
None
No. of animals per sex per dose:
Female: 1250 mg/kg; No. of animals: ; Sacrifice times: hoursFemale: 2500 mg/kg; No. of animals: ; Sacrifice times: hoursFemale: 5000 mg/kg; No. of animals: ; Sacrifice times: hours
Positive control(s):
Mitomycin C with saline
Tissues and cell types examined:
None
Details of tissue and slide preparation:
None
Evaluation criteria:
None
Statistics:
None
Key result
Sex:
female
Toxicity:
yes
Remarks:
Doses producing toxicity:>= 2500 mg/kg
Additional information on results:
Observations:The test substance induced significant cytotoxicity in mouse bone marrow cells at 5000 mg/kg (p<0.005). However, the incidence of MNPCE at any of the dose levels was comparable to that in the control group and there was no statistically significant increase.Mitomycin C-treated (2mg/kg) mice, the positive control, caused a statistically significant increase in the frequency of MNPCE. clinical symptoms: no lethal effects.

None

Conclusions:
Negative is the in vivo chromosome aberration test.
Executive summary:

The in-vivo mouse micronucleus test was conducted using BDF (C57BL/6xDBA/2) hybrid to assess the potential of D-523 to induce micronucleated erythrocyte. Dose levels of 1250, 2500 and 5000 mg/kg were selected on the basis of a erythrocytes (MNPCE) and polychromatic erythrocytes (PCE) were counted 24 hours after a single intraperitoneal injection of FAT 40554.

The test substance, FAT 40554, induced significant cytotoxicity in mouse bone marrow cells at 5000 mg/kg (p<0.05). However, the incidence of MNPCE at any of the dose levels and comparable to that in the solvent control group and there was no statisticakky significant increase.

Additional information

In-vitro:

An in vitro screening assay was performed to assess the potential of FAT 40554 to induce structural chromosome aberrations.

The test substance does not induce frequency of cells which has structural abnormality and polyploid cells, even if metabolic activation method or direct methods .

Based on inhibition test result of cell proliferation, test substance has weak inhibition for Direct methods (without metabolic activation) 1250, 2500, 5000 μg/ml is selected for chromosomal aberration test with and without metabolic activation. In all concentration, test substance in medium is suspended.

In conclusion, it can be stated that in the study described and under the experimental conditions reported, the test article FAT 40554 did not induce structural chromosome aberrations in V79 cells (Chinese hamster cell line).

An in vitro screening assay was performed to assess the potential of FAT 40554 using Ames test. With or without S9 mix, the number of revertant colonies treated with test sample solution for TA100 shows more than twice as many as the one treated with solvent. And it increased concentration-dependent. Pretest organized with 0.05, 0.5, 5, 50, 500, 5000 μg/plate.

With or without S9 mix, fatal effect was not observed. Without S9 mix, precipitation was observed on the concentration of 50μg/plate and more. With S9 mix, precipitation was observed on the concentration of 500μg/plate and more. With or without S9 mix, the number of revertant colonies treated with test sample solution for TA100 shows more than twice as many as the one treated with solvent.

Based on pretest result, TA100 without S9mix takes 0.015, 0.05, 0.15, 0.5, 1.5, 5 μg/plate and TA100 with S9mix takes 0.5, 1.5, 5, 15, 50, 150 μg/plate for main test I and II. (Table 2 and 4) Other species with or without S9 mix takes 156, 313 ,625, 1250, 2500, 5000 μg/plate. (Table 3 and 5) Based on main test result, with or without S9 mix, the number of revertant colonies treated with test sample solution for TA100 shows more than twice as many as the one treated with solvent. And it increased concentration-dependent. And without S9 mix, the number of revertant colonies treated with test sample solution for TA98 shows almost twice as many as the one treated with solvent. Based on above result, the test substance is judged as "Positive" for mutagenicity. In conclusion, FAT 40554 is considered to be mutagenic in this Salmonella typhimurium reverse mutation assay.

The test substance FAT 40554 was examined for its mutagenecity activity by assaying the induction of 6- thioguanine resistant mutants of cultured V79 Chinese hamster cells in the presence and absence of metabolic activation (S9 mix) according to OECD Guideline 476 (In vitro Mammalian Cell Gene Mutation Test).

The cells were exposed to the test compound for 5 hr at doses of 625, 1250, 2500 and 5000µg/ml both with and without metabolic activation. The test compound did not induce any increase in the mutation frequency as compared with those of the vehicle controls either in the presence or absence of metabolic activation. On the other hand, positive control chemicals, ethyl methanesulfonate (EMS, 200µg/ml, without metabolic activation) and 9, 10 -dimethyl-1,2 -benzanthracene (DMBA, 5µg/ml, with metabolic activation), induced marked increase in the mutation frequency. Based on the above findings, it is concluded that the substance is not mutagenic under the conditions tested.

In-vivo:

The in vivo mouse micronucleus test was conducted using BDF (C57BL/6xDBA/2) hybrid to assess the potential of D-523 to induce micronucleated erythrocyte. Dose levels of 1250, 2500 and 5000 mg/kg were selected on the basis of a erythrocytes (MNPCE) and polychromatic erythrocytes (PCE) were counted 24 hours after a single intraperitoneal injection of FAT 40554.

The test substance, FAT 40554, induced significant cytotoxicity in mouse bone marrow cells at 5000 mg/kg (p<0.05). However, the incidence of MNPCE at any of the dose levels and comparable to that in the solvent control group and there was no statisticakky significant increase.

Based on the available in vitro (positive - Ames test, negative - in vitro mammalian cell gene mutation test and negative - in vitro mammalian chromosome aberration test)

and in vivo (negative - chromosome aberration test in mouse) studies, FAT 40554 can be deemed as non genotoxic.

Justification for classification or non-classification

Based on the above stated genotoxic assessment, FAT 40554 does not need to be classified according to CLP (EC No 1272/2008) Regulation Of The European Parliament And Of The Council as implementation of UN-GHS in the EU.