Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

OECD 471 (RA): not mutagenic in bacteria

OECD 473: not clastogenic in mammalian cells

OECD 476: not mutagenic in mammalian cells

Link to relevant study records

Referenceopen allclose all

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:
25 October 2017 until 16 January 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
in vitro mammalian chromosome aberration test
Species / strain / cell type:
lymphocytes: human
Details on mammalian cell type (if applicable):
- Type and identity of media: Dulbeccos's modified Eagle's medium/Ham's F12 medium
- Properly maintained: yes
Metabolic activation:
with and without
Metabolic activation system:
rat liver S9
Test concentrations with justification for top dose:
With metabolic activation:
Experiment I: 4.4, 7.7, 13.5, 23.7, 41.5, 72.6, 127, 222, 667, 2000 µg/mL

Without metabolic activation:
Experiment I: 1.4, 2.5, 4.4, 7.7, 13.5, 23.7, 41.5, 72.6, 127, 222, 667, 2000 µg/mL

Experiment IIA: 1.4, 2.4, 4.2, 7.3, 12.8, 22.4, 39.2, 68.6, 120 µg/mL
Experiment IIB: 16.2, 32.3, 64.6, 74.3, 85.5, 98.3, 113, 130 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: solubility and relatively low cytotoxicity in accordance to the OECD Guideline 473
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
ethylmethanesulphonate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
cyclophosphamide
Details on test system and experimental conditions:
Three independent experiments were performed. In Experiment I the exposure period was 4 hours with and without metabolic activation. In Experiment IIA and IIB the exposure period was 4 hours with S9 mix and 22 hours without S9 mix. The chromosomes were prepared 22 hours after start of treatment with the test item. Evaluation of two cultures per dose group.

METHOD OF APPLICATION: in culture medium

DURATION
- Exposure duration: 4 hours (+/- S9 mix) and 22 hours (- S9 mix)
- Fixation time (start of exposure up to fixation or harvest of cells): 22 hours


SPINDLE INHIBITOR (cytogenetic assays): Colcemid
STAIN (for cytogenetic assays): Giemsa


NUMBER OF REPLICATIONS: about 1.5


NUMBER OF CELLS EVALUATED: 150 per culture, except for the positive control in Experiment IIB, where only 50 metaphases were evaluated


DETERMINATION OF CYTOTOXICITY
- Method: mitotic index
Evaluation criteria:
Evaluation of the cultures was performed (according to standard protocol of the "Arbeitsgruppe der Industrie, Cytogenetik") using NIKON microscopes with 100x oil immersion objectives. Breaks, fragments, deletions, exchanges, and chromosome disintegrations were recorded as structural chromosome aberrations. Gaps were recorded as well but not included in the calculation of the aberration rates. 150 well spread metaphases per culture were scored for cytogenetic damage on coded slides.
Only metaphases with characteristic chromosome numbers of 46 ± 1 were included in the analysis. To describe a cytotoxic effect the mitotic index (% cells in mitosis) was determined.
In addition, the number of polyploid cells in 500 metaphases per culture was determined (% polyploid metaphases; in the case of this aneuploid cell line polyploid means a near tetraploid karyotype). Additionally the number of endomitotic cells scored at the evaluation of polyploid cells was noticed and reported (% endomitotic metaphases).
Statistics:
The statistical significance is confirmed by the Fisher’s exact test (modified) (p < 0.05) using a validated test script of “R”, a language and environment for statistical computing and graphics.
Key result
Species / strain:
lymphocytes: human
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
The test item, dissolved in DMSO, was assessed for its potential to induce chromosomal aberrations in human lymphocytes in vitro in the absence and presence of metabolic activation by S9 mix.
Three independent experiments were performed. In Experiment I, the exposure period was 4 hours with and without S9 mix. In Experiment IIA and IIB, the exposure period was 22 hours without S9 mix. The chromosomes were prepared 22 hours after start of treatment with the test item.
In each experimental group two parallel cultures were analyzed. 150 metaphases per culture were evaluated for structural chromosomal aberrations, except for the positive control in Experiment IIB, where only 50 metaphases were evaluated due to strong clastogenic effects. 1000 cells were counted per culture for determination of the mitotic index.
The highest treatment concentration in this study, 2000 µg/mL was chosen with respect to the OECD Guideline for in vitro mammalian cytogenetic tests.
In Experiment I, precipitation of the test item in the culture medium was observed at 13.5 µg/mL and above in the absence of S9 mix and at 41.5 µg/mL and above in the presence of S9 mix. In addition, precipitation occurred in Experiment IIA, in the absence of S9 mix, at 120 µg/mL and in Experiment IIB in the absence of S9 mix, at 64.6 µg/mL and above.
No relevant influence on osmolarity or pH was observed.
In the absence and presence of S9 mix, no cytotoxicity was observed up to the highest evaluated concentrations, which showed precipitation.
Either with or without metabolic activation neither a statistically significant nor a biologically relevant increase in the number of cells carrying structural chromosomal aberrations was observed after treatment with the test item.
No evidence of an increase in polyploid metaphases was noticed after treatment with the test item as compared to the control cultures.
Either EMS (770 or 660 µg/mL) or CPA (5.0 µg/mL) were used as positive controls and showed distinct increases in cells with structural chromosome aberrations.

Summary of results of the chromosomal aberration study

Exp.

Preparation
interval

Test item
concentration
(µg/mL)

Mitotic indices
(%
of control)

Aberrant cells(%)
incl. gaps*

Aberrant cells(%)
excl. gaps*

Aberrant cells(%)
carrying exchanges

Exposure period 4 hrs without S9 mix

I

22 hrs

Solvent control1

100.0

2.3

2.0

0.0

 

 

Positive control2

66.5

9.3

9.3S

2.0

 

 

4.4

99.2

1.7

1.7

0.0

 

 

7.7

103.1

1.0

0.7

0.0

 

 

13.5P

97.1

1.7

1.7

0.0

Exposure period 22 hrs without S9 mix

II

22 hrs

Solvent control1

100.0

1.7

1.7

0.0

 

 

Positive control3#

39.6

40.0

40.0S

14.0

 

 

16.2

105.7

1.0

1.0

0.0

 

 

32.3

104.0

0.7

0.7

0.0

 

 

64.6P

61.6

1.0

1.0

0.0

Exposure period 4 hrs with S9 mix

I

22 hrs

Solvent control1

100.0

1.3

1.0

0.0

 

 

Positive control4

32.0

11.0

11.0S

0.7

 

 

13.5

97.0

1.7

1.7

0.0

 

 

23.7

89.3

0.7

0.7

0.0

 

 

41.5P

98.1

2.3

2.3

0.0

*      Including cells carrying exchanges

#       Evaluation of 50 metaphases per culture

P      Precipitation occurred at the end of treatment

S       Aberration frequency statistically significant higher than corresponding control values

1       DMSO      0.5 % (v/v)
2         EMS          770 µg/mL
3
         EMS          660 µg/mL
4
       CPA           5.0 µg/mL

Conclusions:
In conclusion, it can be stated that under the experimental conditions reported, the test item did not induce structural chromosomal aberrations in human lymphocytes in vitro.
Therefore, the test is considered to be non-clastogenic in this chromosome aberration test, when tested up to precipitating concentrations.
Executive summary:

The test item, dissolved in DMSO, was assessed for its potential to induce structural chromosomal aberrations in human lymphocytesin vitro in three independent experiments. The following study design was performed:

 

Without S9 mix

With S9 mix

 

Exp. I

Exp. IIA & IIB

Exp. I

Exposure period

4 hrs

22 hrs

4 hrs

Recovery

18 hrs

-

18 hrs

Preparation interval

22 hrs

22 hrs

22 hrs

In each experimental group two parallel cultures were analyzed. Per culture 150 metaphases were evaluated for structural chromosomal aberrations, except for the positive control in Experiment IIB, where only 50 metaphases were evaluated.

The highest applied concentration in this study (2000 µg/mL of the test item) was chosen with respect to the current OECD Guideline 473.

Dose selection of the cytogenetic experiment was performed considering the toxicity data and the occurrence of test item precipitation in accordance with OECD Guideline 473.

In the absence and presence of S9 mix, no cytotoxicity was observed up to the highest evaluated concentrations, which showed precipitation.

Either with or without metabolic activation neither a statistically significant nor a biologically relevant increase in the number of cells carrying structural chromosomal aberrations was observed after treatment with the test item.

No evidence of an increase in polyploid metaphases was noticed after treatment with the test item as compared to the control cultures.

Appropriate mutagens were used as positive controls. They induced statistically significant increases in cells with structural chromosome aberrations.

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:
05 October 2017 until 02 November 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
mammalian cell gene mutation assay
Target gene:
HPRT
Species / strain / cell type:
Chinese hamster lung fibroblasts (V79)
Details on mammalian cell type (if applicable):
- Type and identity of media: MEM
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
- Periodically "cleansed" against high spontaneous background: yes
Metabolic activation:
with and without
Metabolic activation system:
Phenobarbital/Beta-Naphtoflavone induced Rat liver S9
Test concentrations with justification for top dose:
4 hours treatment without S9 mix: 1.95; 3.9; 7.8; 15.6; 31.3(P) µg/mL
4 hours treatment with S9 mix: 1.95; 3.9; 7.8; 15.6; 31.3; 62.5(P) µg/mL

P = Precipitation visible to the unaided eye at the end of treatment

Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: solubility properties
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
ethylmethanesulphonate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
7,12-dimethylbenzanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium


DURATION
- Exposure duration: Experiment I: 4 hours with and without metabolic activation
- Expression time (cells in growth medium): 72 hours
- Selection time (if incubation with a selection agent): 10 days

SELECTION AGENT (mutation assays): 6-Thioguanine


NUMBER OF REPLICATIONS: 2


NUMBER OF CELLS EVALUATED: >1,5x10exp. 6


DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency

Evaluation criteria:
A test item is classified as positive if it induces a concentration-related increase of the mutant frequency exceeding the historical solvent control range.
A test item producing no concentration-related increase of the mutant frequency above the historical solvent control range is considered to be non-mutagenic in this system.
A mutagenic response is described as follows:
The test item is classified as mutagenic if it induces with at least one of the concentrations in both parallel cultures a mutation frequency that exceeds the historical negative and solvent control data range (95% confidence interval limits).
The increase should be significant and dose dependent as indicated by statistical analysis (linear regression, least squares).
Statistics:
Statistical Analysis:
A linear regression (least squares, calculated using a validated excel spreadsheet) was performed to assess a possible dose dependent increase of mutant frequencies. The numbers of mutant colonies generated with the test item were compared to the solvent control groups. A trend is judged as significant whenever the p-value (probability value) is below 0.05.
A t-Test was performed using a validated test script of “R”, a language and environment for statistical computing and graphics, to evaluate an isolated increase of the mutation frequency at a test point exceeding the 95% confidence interval. Again a t-test is judged as significant if the p-value (probability value) is below 0.05.
However, both, biological and statistical significance were considered together.
Key result
Species / strain:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: Not effected (pH 7.35 in the solvent control versus pH 7.38 at 2000 µg/mL)
- Effects of osmolality: No relevant increase (404 mOsm in the solvent control versus 381 mOsm at 2000 µg/mL)
- Evaporation from medium: Not examined
- Precipitation: determined at 31.3 µg/mL without metabolic activation and at 62.5 µg/mL with metabolic activation
- Other confounding effects: None

RANGE-FINDING/SCREENING STUDIES:
The pre-experiment was performed in the presence and absence of metabolic activation. Test item concentrations between 15.6 µg/mL and 2000 µg/mL were used. The highest concentration was chosen with respect to the current OECD Guideline 476.
In the pre-experiment relevant toxic effects were observed after 4 hours treatment at 62.5 µg/mL and above without metabolic activation.
The test medium was checked for precipitation or phase separation at the end of each treatment period (4 hours) before the test item was removed. Precipitation occurred at 62.5 µg/mL and above with and without metabolic activation.
There was no relevant shift of pH and osmolarity of the medium even at the maximum concentration of the test item.
The concentrations used in the main experiment were selected based on precipitation observed in the pre-experiment. The individual concentrations were spaced by a factor of 2.
To overcome problems with possible deviations in toxicity the main experiment was started with more than four concentrations.

COMPARISON WITH HISTORICAL CONTROL DATA: Complies

ADDITIONAL INFORMATION ON CYTOTOXICITY:
No cytotoxic effects indicated by an adjusted cloning efficiency I below 50% was observed neither in absence nor presence of metabolic activation.
Summary Table
Culture I
        relative relative rel. adjusted mutant 95%
  conc. P S9 cloning cell cloning colonies/ confidence
  µg/mL mix efficiency I density efficiency I 106cells interval
        % % %    
Solvent control with DMSO - 100.0 100.0 100.0 9.4 1.7 - 30.2
Positive control (EMS) 300 - 76.2 79.3 60.4 252.7 1.7 - 30.2
Test item 1.95 - 86.4 106.4 91.9 24.3 1.7 - 30.2
Test item 3.9 - 94.7 83.8 79.3 21.0 1.7 - 30.2
Test item 7.8 - 102.4 73.4 75.2 18.5 1.7 - 30.2
Test item 15.6 - 91.1 66.5 60.5 9.8 1.7 - 30.2
Test item 31.3 P - 80.2 84.6 67.8 18.3 1.7 - 30.2
Solvent control with DMSO + 100.0 100.0 100.0 16.5 2.0 - 29.4
Positive control (DMBA) 2.3 + 98.8 79.5 78.5 99.6 2.0 - 29.4
Test item 1.95 + 99.5 72.8 72.4 27.8 2.0 - 29.4
Test item 3.9 + 102.3 76.6 78.4 13.6 2.0 - 29.4
Test item 7.8 + 96.3 71.8 69.2 23.8 2.0 - 29.4
Test item 15.6 + 102.2 66.2 67.7 14.2 2.0 - 29.4
Test item 31.3 + 99.0 68.1 67.4 22.3 2.0 - 29.4
Test item 62.5 P + 97.6 69.2 67.5 17.5 2.0 - 29.4
Culture II
        relative relative rel. adjusted mutant 95%
  conc. P S9 cloning cell cloning colonies/ confidence
  µg/mL mix efficiency I density efficiency I 106cells interval
        % % %    
Solvent control with DMSO - 100.0 100.0 100.0 23.1 1.7 - 30.2
Positive control (EMS) 300 - 89.0 86.4 76.9 292.7 1.7 - 30.2
Test item 1.95 - 102.3 95.5 97.7 21.3 1.7 - 30.2
Test item 3.9 - 97.9 99.0 96.9 16.2 1.7 - 30.2
Test item 7.8 - 101.0 93.0 94.0 35.1 1.7 - 30.2
Test item 15.6 - 95.4 96.0 91.6 12.1 1.7 - 30.2
Test item 31.3 P - 102.3 104.4 106.8 12.1 1.7 - 30.2
Test item 62.5 P - # # # # #
Test item 125 P - # # # # #
Test item 250 P - # # # # #
Solvent control with DMSO + 100.0 100.0 100.0 5.5 2.0 - 29.4
Positive control (DMBA) 2.3 + 75.6 118.6 89.7 74.3 2.0 - 29.4
Test item 1.95 + 70.3 137.2 96.5 10.4 2.0 - 29.4
Test item 3.9 + 51.3 125.1 64.2 22.2 2.0 - 29.4
Test item 7.8 + 45.3 139.3 63.1 9.0 2.0 - 29.4
Test item 15.6 + 71.9 134.5 96.6 31.0 2.0 - 29.4
Test item 31.3 + 54.5 131.5 71.6 23.2 2.0 - 29.4
Test item 62.5 P + 58.9 118.8 70.0 14.7 2.0 - 29.4
Test item 125 P + # # # # #
Test item 250 P + # # # # #
P = Precipitation visible at the end of treatment
Conclusions:
In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells.
Therefore, the test item is considered to be non-mutagenic in this HPRT assay.
Executive summary:

The study was performed to investigate the potential of test item to induce gene mutations at the HPRT locus in V79 cells of the Chinese hamster. The treatment period was 4 hours with and without metabolic activation. The maximum test item concentration of the pre-experiment (2000 µg/mL) was chosen with the respect to the current OECD guideline. The highest concentration in the main experiment was limited by precipitation observed in the pre-experiment. No relevant and reproducible increase in mutant colony numbers/106cells was observed in the main experiment up to the maximum concentration.

No cytotoxic effects indicated by an adjusted cloning efficiency I below 50% was observed neither in absence nor presence of metabolic activation.

Appropriate reference mutagens, used as positive controls, induced a distinct increase in mutant colonies and thus, showed the sensitivity of the test system and the activity of the metabolic activation system.

Conclusion

In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells.

Therefore, the test item is considered to be non-mutagenic in this HPRT assay.

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2006-11-01 to 2006-11-10
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
07-1997
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
Official Journal of the European Communities L136, 8. June 2000
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay
Target gene:
HIS operon (S. thyphimurium)
TRP operon (E. coli)
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
liver S9 mix from Aroclor 1254-pretreated rats with standard co-factors
Test concentrations with justification for top dose:
The test material concentrations used were selected according to the EC and OECD guidelines for this test system and the requirements of the Labor Ministry of Japan:
1st series: 5, 15.8, 50, 158, 500, 1580 and 5000 µg/plate
2nd series: 50, 88.9, 158, 281 and 500 μg/plate
In the two series with S9 mix, 10 or 30 % S9 in the S9 mix were used in the 1st and 2nd series, respectively.
Vehicle / solvent:
DMSO
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
9-aminoacridine
sodium azide
cumene hydroperoxide
other: daunomycin
Remarks:
without S9 mix
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
benzo(a)pyrene
other: 2-aminoanthracene
Remarks:
with S9 mix
Details on test system and experimental conditions:
The assessment of test material-induced effects is dependent on the number of spontaneous revertants of each bacterial strain (solvent controls) and the increase in the number of revertants at the test material concentration which shows the highest number of colonies. The following criteria, based upon the historical controls of the laboratory and statistical considerations, are established:

Mean Number of Colonies Maximal Mean Number of Colonies over
(Solvent Control) the Actual Solvent Control (Test Material)
----------------------------------------------------------------------------------------------
<=10 <=9 >=30
<=30 <=19 >=40
<=80 <=29 >=80
<=200 <=49 >=120
<=500 <=79 >=200
Assessment: No increase Clear increase
----------------------------------------------------------------------------------------------

All further results, ranging between "no" and "clear", are assessed as "weak in-creases".

Interpretations:

A test material is defined as non-mutagenic in this assay if:
- "no" or "weak increases" occur in the first and second series of the main experiment. ("Weak increases" randomly occur due to experimental variation.)

A test material is defined as mutagenic in this assay if:
- a dose-related (over at least two test material concentrations) increase in the number of revertants is induced, the maximal effect is a "clear increase", and the effects are reproduced at similar concentration levels in the same test system;
- "clear increases" occur at least at one test material concentration, higher concentrations show strong precipitation or cytotoxicity, and the effects are reproduced at the same concentration level in the same test system.

In all further cases, a third test series with the bacterial strain in question should be performed. If the criteria for a positive test result are not fulfilled in at least two out of the three series, the test material is defined as being non-mutagenic in this test system.
Evaluation criteria:
For details see results.
Statistics:
n.a.
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
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
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
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
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
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Conclusions:
With and without addition of S9 mix as the external metabolizing system, the test material was not mutagenic under the experimental conditions described.
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
For this endpoint information from structural similar compounds is available. The studies for these similar compounds were performed according to GLP and the methods applied are fully compliant with OECD TG 471. See chapter 13 report for a more detailed justification.
Reason / purpose:
read-across source
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
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
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
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
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
True negative controls validity:
not examined
Positive controls validity:
valid
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

Based on the provided information there is no need for classification according to the EU Regulation (EC) No 1272/2008 on Classification,Labelling and Packaging of Substances and Mixtures.