Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation (Bacterial reverse mutation assay / Ames test): negative with and without activation in Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537Escherichia coliWP2 (OECD TG 471) (Sokolowski, 2008).

Cytogenicity in mammalian cells: negative with and without activation in Chinese hamster V79 cells (OECD TG 473) (Hoffman, 2008).

Mutagenicity in mammalian cells: negative with and without activation in mouse lymphoma L5178Y cells (OECD TG 476) (Trenz, 2012).

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008-01-28 - 2008-02-21
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:
1997
GLP compliance:
yes (incl. certificate)
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:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
Phenobarbital/β-Naphthoflavone induced rat liver S9
Test concentrations with justification for top dose:
33, 100, 333, 1000, 2500 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: acetone
- Justification for choice of solvent: the solvent was chosen because of its solubility properties and its relative non-toxicity to the bacteria.
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
TA1535 and TA100 without metabolic activation 10 µg/plate
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylene-diamine (4-NOPD)
Remarks:
TA98 - 10 µg/plate, TA1537 - 50 µg/plate without metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
WP2 uvrA without metabolic activation 3 µl/plate
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene (2-AA)
Remarks:
TA1535, TA1537, TA98, TA100 - 2.5 µg/plate and WP2 uvrA - 10 µg/plate with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); preincubation

ACTIVATION: S9 mix contained glucose-6-phophate and NADP as co-factors, the S9 supernatant was 10% v/v in the S9 mix of which 0.5 ml were added to 2 ml top agar, giving a final concentration of approximately 2% S9.

DURATION
- Preincubation period: 60 minutes at 37ºC
- Exposure duration: Experiment 1 - 72 hours at 37ºC, experiment II - 72 hours at 37ºC + a further 72 hours at 4ºC


SELECTION AGENT: histidine deficient agar

NUMBER OF REPLICATIONS: triplicate plates, experiment repeated

DETERMINATION OF CYTOTOXICITY
- Method: other: condition of background lawn



Evaluation criteria:
A result is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice (strains TA98, TA100 and WP2 uvrA) or thrice (strains TA1535 and TA1537) the colony count of the corresponding solvent control is observed. A dose dependent increase is considered biologically relevant if the threshold is exceeded at more than one concentration. An increase exceeding the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment.
Statistics:
No statistical analysis was performed.
Key result
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
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
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS

COMPARISON WITH HISTORICAL CONTROL DATA: yes, included in report and all within limits

Results of plate incorporation assay Experiment I; revertants per plate (mean of 3 plates)

[Note, the pre-experiment is reported as Experiment 1, as there were evaluable plates at all concentrations in all strains used.]

Dose level (µg/plate)

Metabolic activation

Mean revertant colony counts

TA 1535

TA 1537

TA 98

TA 100

WP2 uvrA

Solvent control**

-

16

13

22

130

54

Negative control***

-

14

12

24

130

47

3

-

11

14

21

138

50

10

-

11

14

29

130

46

33

-

11

12

21

122

51

100

-

9

14

25

120

45

333

-

16

18

21

136

52

1000

-

15

10

24

138

52

2500

-

15

13

22

137

59

5000

-

13

8

26

158

62

Positive control

-

1935

78

281

2196

1314

Solvent control**

+

15

16

33

172

64

Negative control***

+

18

13

37

154

49

3

+

17

16

33

149

64

10

+

17

15

35

165

57

33

+

17

16

34

177

65

100

+

18

17

38

165

61

333

+

18

18

35

161

64

1000

+

12*

15*

29*

157*

55*

2500

+

11*

13*

25*

156*

55*

5000

+

12*

11*

26*

148*

55*

Positive control

+

234

123

998

1334

231

* non-interfering precipitate

** with acetone

*** Untreated

Results of preincubation assay, Experiment 2; revertants per plate (mean of 3 plates)

 

Dose level µg/plate

Metabolic activation

Mean revertant colony counts

TA 1535

TA 1537

TA 98

TA 100

WP2 uvrA

Solvent control**

-

12

12

30

133

37

Negative control***

-

14

17

30

120

44

33

-

13

13

23

136

42

100

-

14

11

23

135

45

333

-

15

13

31

125

46

1000

-

15

12

26

133

43

2500

-

15

11

21

118

39

5000

-

14

10

31

125

38

Positive control

-

2143

99

335

1941

901

Solvent control**

+

19

15

34

121

60

Negative control***

+

16

18

33

156

49

33

+

18

17

31

142

52

100

+

15

19

33

142

52

333

+

16

19

36

137

60

1000

+

18*

16*

30*

129

60

2500

+

15*

16*

30*

135*

56*

5000

+

17*

14*

30*

154*

49*

Positive control

+

307

195

1206

863

349

 

* non-interfering precipitate

** with acetone

*** Untreated

Conclusions:
1,1,1,3,5,5,5-heptamethyltrisiloxane has been tested for mutagenicity to bacteria in a study which was conducted according to OECD 471 and in compliance with GLP. No evidence of a test-substance related increase in the number of revertants was observed with or without activation in the initial plate incorporation assay or the repeat experiment using the preincubation method. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.
Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008-01-09 - 2008-05-15
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)
Version / remarks:
1997
GLP compliance:
yes (incl. certificate)
Type of assay:
in vitro mammalian chromosome aberration test
Target gene:
- Name of test material (as cited in study report): 1,1,1,3,5,5,5-Heptamethyltrisiloxane
- Substance type: mono-constituent
- Physical state: colourless liquid
- Analytical purity: 98.9% (GLC)
- Molecular weight: 222.5 g/mol
- Lot/batch No.: 14718HC
- Expiration date of the lot/batch: 04 October 2008
- Stability under test conditions: Not indicated by the sponsor
- Storage condition of test material: At room temperature
- Other:
Species / strain / cell type:
Chinese hamster lung fibroblasts (V79)
Details on mammalian cell type (if applicable):
- Type and identity of media: MEM (Minimal Essential Medium) with 10% foetal calf serum (FCS)
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
Metabolic activation:
with and without
Metabolic activation system:
Phenobarbital/β-Naphthoflavone induced rat liver S9
Test concentrations with justification for top dose:
Expt 1: 17.6, 35.2, 70.3 µg/ml (- MA); 8.8, 17.6, 35.2, 70.3 µg/ml (+ MA); Expt 2 17.6, 35.2, 70.3 µg/ml (- MA); 8.8, 17.6, 35.2, 281.3, 562.5, 1125 µg/ml (+MA).
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: acetone
- Justification for choice of solvent/vehicle: solvent chosen for solubility properties and its relative non-toxicity to the cell cultures.
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
ethylmethanesulphonate
Remarks:
without metabolic activation 500µg/ml Experiment II, 900 µg/ml Experiment I
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
cyclophosphamide
Remarks:
with metabolic activation. Experiment I - 1.4 µg/ml, Experiment II - 2 µg/ml
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION

- Exposure duration: without metabolic activation - 4, 18 and 24 hours, with metabolic activation - 18 and 28 hours
- Expression time (cells in growth medium):
- Fixation time (start of exposure up to fixation or harvest of cells):


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

NUMBER OF REPLICATIONS: 2 slides per group, experiment repeated

NUMBER OF CELLS EVALUATED: 100 metaphases per culture

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth; other: cell growth inhibition

OTHER EXAMINATIONS:
- Determination of polyploidy: yes


OTHER: ACTIVATION: Phenobarbital/beta-Naphthoflavone induced rat liver S9, with protein content of 28.3 mg/l (pre-test and expt 1) or 31.0 mg/l (expt 2); S9 mix contained glucose-6-phosphate and NADP as co-factors. Final protein concentration in cultures was 0.75 mg/ml, equivalent to approximately 0.4%.
Evaluation criteria:
The test is considered acceptable if it meets the following criteria: a) The number of structural aberrations found in the solvent controls falls within the range of the laboratory's historical control data: 0.0 - 4.0% aberrant cells, excluding gaps. b) The positive control substances should produce significant increases in the number of cells with structural chromosome aberrations, which are within the range of the laboratory's historical control data.
Statistics:
Fisher's exact test
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 applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: no effect
- Effects of osmolality: no effect
- Water solubility: phase separation occurred in some of the treated cultures
- Precipitation: precipitation in the form of oily drops was observed after 4 hour treatment with 70.3 µg/ml and above in the presence and absence of S9 mix


RANGE-FINDING/SCREENING STUDIES:

COMPARISON WITH HISTORICAL CONTROL DATA: yes and all acceptable

ADDITIONAL INFORMATION ON CYTOTOXICITY:

Summary of results of chromosome aberration study – Experiment 1

 

Exposure period (hrs)

+/- metabolic activation

Prepar-ation interval

Dose level µg/ml

% Polyploidy cells

Cell numbers in % of control

Mitotic indices in % of control

% Aberrant cells

incl gaps

excl gaps

with exchanges

4

-

18

Solvent control

2.0

100.0

100.0

1.5

1.5

1.0

4

-

18

17.6

2.4

96.1

107.2

2.0

1.0

0.0

4

-

18

35.2

2.0

93.4

90.6

2.0

1.0

0.0

4

-

18

70.3P

2.2

92.7

108.6

3.0

1.5

0.0

4

-

18

Positive control

2.4

n.t.

101.1

13.0

12.0

2.5

4

+

18

Solvent control

3.6

100.0

100.0

1.0

1.0

0.0

4

+

18

8.8

2.4

97.7

95.4

2.0

1.5

0.5

4

+

18

17.6

3.1

94.6

81.3

0.5

0.5

0.0

4

+

18

35.2P

2.5

124.7

90.8

2.0

1.5

0.0

4

+

18

Positive control

2.9

n.t.

81.7

12.0

11.0

3.0

 

Summary of results of chromosome aberration study – Experiment II

 

 

Exposure period (hrs)

+/- metabolic activation

Prepar-ation interval

Dose level µg/ml

% Polyploidy cells

Cell numbers in % of control

Mitotic indices in % of control

% Aberrant cells

incl gaps

excl gaps

with exchanges

18

-

18

Solvent control

2.9

100.0

100.0

2.0

1.5

0.5

18

-

18

17.6

2.7

86.1

110.1

1.0

1.0

0.0

18

-

18

35.2

3.0

86.4

109.5

3.5

2.5

0.0

18

-

18

70.3P

2.5

91.9

104.7

2.0

2.0

0.0

18

-

18

Positive control

2.0

n.t.

84.9

18.5

15.0

7.5

28

-

28

Solvent control

2.9

100.0

100.0

1.0

1.0

0.0

28

-

28

35.2

2.5

99.9

107.3

2.5

2.5

0.0

28

-

28

70.3P

3.1

95.4

104.4

1.0

0.5

0.0

28

-

28

Positive control

2.4

n.t.

90.8

32.0

32.0

15.0

4

+

28

Solvent control

2.3

100.0**

100.0

1.5

1.5

0.0

4

+

28

8.8

2.4

79.6**

92.5

3.0

2.0

0.0

4

+

28

17.6

3.2

71.5**

110.9

1.5

1.5

0.0

4

+

28

35.2P

2.4

76.3**

105.2

2.0

0.5

0.0

4

+

28

281.3P

2.5

65.3**

103.7

2.0

1.5

0.0

4

+

28

562.5P

2.4

108.3**

106.7

0.5

0.5

0.0

4

+

28

1125.0P

2.8

93.3**

101.1

0.5

0.5

0.0

4

+

28

Positive control

2.2

n.t.

92.1

14.0

13.0

1.0

 

**  = Cell count on spread slides                      P = Precipitation occurred

n.t. = not tested

Conclusions:
1,1,1,3,5,5,5-Heptamethyltrisiloxane has been tested in an in vitro cytogenicity test to OECD 473, in compliance with GLP. No evidence of a test-substance related increase in chromosome aberrations was observed with or without metabolic activation in the initial or repeat experiment using Chinese Hamster V79 cells. It is concluded that the test substance is negative for cytogenicity under the conditions of the test.
Endpoint:
in vitro gene mutation study in mammalian cells
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2011-08-01 - 2011-10-05
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
according to
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Version / remarks:
1997
GLP compliance:
yes (incl. certificate)
Type of assay:
mammalian cell gene mutation assay
Target gene:
Thymidine kinase
Species / strain / cell type:
mouse lymphoma L5178Y cells
Details on mammalian cell type (if applicable):
- Type and identity of media: RPMI 1640
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically "cleansed" against high spontaneous background: yes
Metabolic activation:
with and without
Metabolic activation system:
Phenobarbital and beta-naphthoflavone induced rat liver S9
Test concentrations with justification for top dose:
With S9 - Exp 1 - 0.1, 0.2, 0.5, 1.25, 2.5, 5.0, 7.5, 10.0 mM Exp 2 - 1, 2, 4, 6, 7, 8, 9, 10 mM
Without S9 Exp 1 - 0.05, 0.1, 0.2, 0.5, 1.25, 2.5, 5.0, 7.5, 9.0, 10.0 mM Exp 2 - 0.1, 0.25, 1.5, 2.5, 3.5, 5.0, 6.0, 7.5, 9.0, 10.0 mM
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: the solvent was compatible with the survival of the cells of the S9 activity.
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Remarks:
200 µg/ml and 300 µg/ml
Positive control substance:
ethylmethanesulphonate
Remarks:
without metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Remarks:
10 µg/ml
Positive control substance:
methylmethanesulfonate
Remarks:
without metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Remarks:
2.5 µg/ml
Positive control substance:
benzo(a)pyrene
Remarks:
with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

ACTIVATION: S9 mix contained glucose-6-phosphate and NADP as co-factors resulting in a final protein concentration of 0.75 mg/ml in the cultures.

DURATION

- Exposure duration: Short term exposure - 4 hrs, long term exposure -24 hrs
- Expression time (cells in growth medium): 2 days at 37ºC
- Selection time (if incubation with a selection agent): at least 6 days
- Fixation time (start of exposure up to fixation or harvest of cells): 14 days

SELECTION AGENT (mutation assays): TFT (Triflourothymidine)

NUMBER OF REPLICATIONS: experiment repeated

DETERMINATION OF CYTOTOXICITY
- Method: other: growth inhibition



Evaluation criteria:
The test item is considered mutagenic if the induced mutant frequency meets or exceeds the Global Evaluation factor (GEF) of 126 per 10x6 cells or a dose-dependent increase in mutant frequency is detected.
Statistics:
The non-parametric Mann-Whitney test is applied to the mutation data to prove the dose groups for any significant difference in mutant frequency compared to the controls.
Key result
Species / strain:
mouse lymphoma L5178Y cells
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
with and without metabolic activation
Cytotoxicity / choice of top concentrations:
other: variable without metabolic activation
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid

   Summary tables of Experiment 1 and 2 without metabolic activation

Treatment (mM)

RTG (%)

MF (mutants/106cells

IMF (mutants/106cells)

Precipitate

 

-S9

-S9

-S9

-S9

Negative control

111.7

74.2

/

-

Negative control

110.7

/

-

Solvent control

100.0

84.2

/

-

Solvent control

/

-

0.05

85.4

82.5

-1.7

-

0.1

67.4

84.4

0.3

-

0.2

10.5

117.3

33.2

-

0.5

9.3

135.8

51.7

-

1.25

100.6

68.4

-15.7

-

2.5

25.0

128.5

44.3

-

5.0

27.0

64.7

-19.5

-

7.5

66.2

93.9

9.8

-

9.0

88.5

98.5

14.3

-

10.0

89.7

97.5

13.4

-

Positive control 1

73.3

530.0

445.9

-

Positive control 2

56.5

537.7

453.6

-

Treatment (mM)

RTG (%)

MF (mutants/106cells

IMF (mutants/106cells)

Precipitate

 

-S9

-S9

-S9

-S9

Negative control

118.7

94.9

/

-

Negative control

120.6

/

-

Solvent control

100.0

67.5

/

-

Solvent control

/

-

0.1

89.7

118.8

51.2

-

0.25

109.5

83.2

15.7

-

1.5

112.8

71.5

4.0

-

2.5

111.2

70.5

2.9

-

3.5

7.5

111.9

44.3

-

5.0

86.2

86.9

19.4

-

6.0

79.1

116.0

48.4

-

7.5

110.2

74.1

6.6

-

9.0

96.9

56.1

-11.5

-

10.0

110.9

49.0

-18.6

-

Positive control 1

41.4

2356.5

2288.9

-

Positive control 2

34.8

1546.6

1479.1

-

Summary tables of Experiment 1 and 2 with metabolic activation

 

Treatment (mM)

RTG (%)

MF (mutants/106cells

IMF (mutants/106cells)

Precipitate

 

+S9

+S9

+S9

+S9

Negative control

106.6

106.8

/

-

Negative control

113.4

/

-

Solvent control

100.0

108.9

/

-

Solvent control

/

-

0.1

96.1

123.8

14.9

-

0.2

100.5

132.6

23.8

-

0.5

102.4

142.9

34.0

-

1.25

111.0

113.0

4.1

-

2.5

107.6

87.9

-21.0

-

5.0

98.6

143.5

34.6

-

7.5

98.0

140.8

31.9

-

10.0

99.4

129.0

20.1

-

Positive control 1

61.4

711.7

602.8

-

Treatment (mM)

RTG (%)

MF (mutants/106cells

IMF (mutants/106cells)

Precipitate

Negative control

100.9

73.5

/

-

Negative control

102.5

/

-

Solvent control

100.0

91.1

/

-

Solvent control

/

-

1

108.1

84.7

-6.4

-

2

111.8

89.4

-1.7

-

4

107.3

81.4

-9.7

-

6

112.7

55.8

-35.3

-

7

104.4

101.9

10.8

-

8

100.7

72.2

-18.9

-

9

103.3

79.5

-11.6

-

10

124.8

83.4

-7.7

-

Positive control 1

81.9

613.1

522.1

-

 

 RTG = Relative total growth

MF = Mutant frequency

IMF = Induced mutant frequency

Conclusions:
1,1,1,3,5,5,5-heptamethyltrisiloxane has been tested for mutagenicity to mammalian cells in a reliable study conducted according to OECD TG 476 and in compliance with GLP. No biologically relevant increase in mutation rate was found in mouse lymphoma L5178Y cells with or without metabolic activation in either the initial or the repeat experiment, up to limit and cytotoxic concentrations. The global evaluation factor was not exceeded at any concentration. In addition, colony sizing showed no clastogenic effects in either experiment. The degree toxicity observed was variable, and not dose related. Appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for mutagenicity to mammalian cells under the conditions of the test.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

1,1,1,3,5,5,5 -Heptamethyltrisiloxane (H-L3) has been tested for mutagenicity to bacteria in a study which was conducted according to OECD 471 and in compliance with GLP (Sokolowski, 2008). No evidence of a test-substance related increase in the number of revertants was observed with or without activation in the initial plate incorporation assay or the repeat experiment using the preincubation method. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.

H-L3 has been tested in an in vitro cytogenicity test to OECD 473, in compliance with GLP (Hoffman, 2008). No evidence of a test-substance related increase in chromosome aberrations was observed with or without metabolic activation in the initial or repeat experiment using Chinese Hamster V79 cells. It is concluded that the test substance is negative for cytogenicity under the conditions of the test.

Information is available for H-L3 from an in vitro study for mutagenicity to mammalian cells conducted according to OECD TG 476 and in compliance with GLP (Trenz, 2012). No evidence for test-substance induced increase in mutant factor was observed when H-L3 was tested in a reliable study conducted according to OECD TG 476 and in compliance with GLP. No biologically relevant increase in mutation rate was found in mouse lymphoma L5178Y cells with or without metabolic activation in either the initial or the repeat experiment, up to limit and cytotoxic concentrations. The global evaluation factor was not exceeded at any concentration. In addition, colony sizing showed no clastogenic effects in either experiment. The degree toxicity observed was variable, and not dose related. Appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for mutagenicity to mammalian cells under the conditions of the test.

In vivo testing is not required as the results of the in vitro assays were all negative.


Justification for classification or non-classification

Based on the available in vitro and in vivo genotoxicity data, H-L3 is not classified for mutagenicity according to Regulation (EC) No 1272/2008.