Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

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:
12 February 2014 - 7 March 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Test method according to OECD 471. GLP study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report date:
2014

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
3-[(1S)-1-(methylamino)ethyl]phenol
EC Number:
805-518-8
Cas Number:
894079-42-6
Molecular formula:
C9H13NO
IUPAC Name:
3-[(1S)-1-(methylamino)ethyl]phenol
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material: (S)-3-(1-Methylamino-ethyl)-phenol
- Physical state: White or slightly coloured powder
- Analytical purity: 98.5%
- Lot/batch No.: I13213A
- Storage condition: Controlled room temperature

Method

Target gene:
Histidine-requiring gene in Salmonella typhimurium and Tryptophan-requiring gene in the strain of Escherichia coli.
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Metabolic activation system:
S9 mix from rat liver
Test concentrations with justification for top dose:
5000, 1581, 500, 158.1, 50, 15.81 and 5 μg/plate.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The solubility of the test item was examined using Distilled water, Dimethyl sulfoxide (DMSO) and N,N-dimethylformamide (DMF). The test item was insoluble in Distilled water and partial dissolution was observed in DMF. However the formulations at 100 mg/mL concentration using DMSO or DMF as vehicle were suitable for the test. Therefore, DMSO was selected as vehicle of the study.
Controlsopen allclose all
Negative solvent / vehicle controls:
yes
Remarks:
DMSO and distilled water
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
TA100, TA1535, -S9 (2 µg/plate)
Positive controls:
yes
Positive control substance:
other: 4-nitro-1,2-phenylene-diamine
Remarks:
TA98, -S9 (4 µg/plate)
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
TA1537, -S9 (50 µg/plate)
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
WP2 uvrA, -S9 (2 µg/plate)
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
all Salmonella strains, +S9 (2 µg/plate) and WP2 uvrA, +S9 (50 µg/plate)
Details on test system and experimental conditions:
METHOD OF APPLICATION:
Plate incorporation (initial mutation test):
Molten top agar was prepared and kept at 45°C. 2 mL of top agar was aliquoted into individual test tubes (3 tubes per control or concentration level). The equivalent number of minimal glucose agar plates was properly labelled. The test item (or controls) and other components (top agar, overnight culture of test strain, phosphate buffer (pH 7.4) or S9 mix) were prepared freshly and added to the overlay (45°C). This solution was mixed and poured on the surface of minimal agar plates. After preparation, the plates were incubated at 37°C for 48 hours.

Pre-incubation (confirmatory mutation test):
Before the overlaying, the test item formulation (or vehicle/solvent or reference control), the bacterial culture and the S9 mix or phosphate buffer was added into appropriate tubes to provide direct contact between bacteria and the test item (in its vehicle/solvent). The tubes (3 tubes per control or concentration level) were gently mixed and incubated for 20 min at 37ºC in a shaking incubator. After the incubation period, 2 mL of molten top agar was added to the tubes; the content was mixed up and poured onto minimal glucose agar plates as described for the standard plate incorporation method. After preparation, the plates were incubated at 37°C for 48 hours.

NUMBER OF REPLICATIONS: 3 replicates per dose and controls.

EVALUATION OF EXPERIMENTAL DATA:
The colony numbers were determined by manual counting.
Visual examination of the plates was performed, precipitation or signs of growth inhibition (if any) were recorded.
The mean number of revertants per plate, the standard deviation and the mutation factor values were calculated.
Evaluation criteria:
Criteria for a Positive Response:
A test item was considered mutagenic if:
- a dose–related increase in the number of revertants occurred and/or;
- a reproducible biologically relevant positive response for at least one of the dose groups occurred in at least one strain with or without metabolic activation.
An increase was considered biologically relevant if:
- the number of reversions is more than two times higher than the reversion rate of the negative (solvent) control in Salmonella typhimurium TA98, TA100 and Escherichia coli WP2 uvrA bacterial strains;
- the number of reversions is more than three times higher than the reversion rate of the negative (solvent) control in Salmonella typhimurium TA1535 and TA1537 bacterial strains.
According to the guidelines, statistical method may be used as an aid in evaluating the test results. However, statistical significance should not be the only determining factor for a positive response.
Criteria for a Negative Response:
A test article was considered non-mutagenic if it produced neither a dose-related increase in the number of revertants nor a reproducible biologically relevant positive response at any of the dose groups, with or without metabolic activation.
Statistics:
The mean number of revertants per plate, the standard deviation and the mutation factor values were calculated for each concentration level of the test item and for the controls using Microsoft ExcelTM software.

Results and discussion

Test results
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:
cytotoxicity
Remarks:
(confirmatory test, all strains at 5000 µg/plate, +/- S9)
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES:
The observed numbers of revertant colonies were in the normal range and within the historical control range in all cases.
No precipitate, inhibitory or cytotoxic effect of the test item was detected in the Preliminary Range Finding Test.

ADDITIONAL INFORMATION ON CYTOTOXICITY: Inhibitory, cytotoxic effect of the test item (reduced / slightly reduced background lawn development and / or reduced number of revertant colonies, in some cases pinpoint colonies were also detected) were observed in the Confirmatory Mutation Test in all tester strains at 5000 μg/plate concentration with and without metabolic activation.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

In the Initial Mutation Test and Confirmatory Mutation Test, none of the observed revertant colony numbers were above the respective biological threshold value. There were no dose-related trends and no indication of any treatment effect. In all test item treated groups, the numbers of revertant colonies did not exceed the biological relevance when compared to the solvent control and were within the normal biological variability of the test system.

Initial Mutation Test:

Concentrations
(µg/plate)

Mean
values of revertants / Mutation factor (MF)

Salmonella typhimurium tester strains

Escherichia coli

TA98

TA100

TA1535

TA1537

WP2uvrA

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

5000

Mean

18.3

24.3

95.7

121.7

10.7

12.7

8.0

9.3

35.0

37.0

MF

1.08

1.00

1.13

1.09

1.10

1.46

1.04

0.85

1.50

1.13

1581

Mean

20.0

27.3

81.0

115.7

7.3

9.0

7.3

10.3

25.0

26.3

MF

1.18

1.12

0.96

1.04

0.76

1.04

0.96

0.94

1.07

0.81

500

Mean

22.0

23.7

95.7

97.7

9.3

13.0

8.7

6.7

26.7

34.0

MF

1.29

0.97

1.13

0.88

0.97

1.50

1.13

0.61

1.14

1.04

158.1

Mean

22.0

24.7

84.0

115.3

10.3

12.3

7.0

8.3

30.3

26.7

MF

1.29

1.01

0.99

1.04

1.07

1.42

0.91

0.76

1.30

0.82

50

Mean

15.7

25.7

97.3

99.7

11.3

12.3

8.0

9.3

25.3

33.0

MF

0.92

1.05

1.15

0.90

1.17

1.42

1.04

0.85

1.09

1.01

15.81

Mean

16.0

24.0

87.7

114.0

11.3

12.0

9.3

8.3

25.7

37.3

MF

0.94

0.99

1.04

1.02

1.17

1.38

1.22

0.76

1.10

1.14

5

Mean

21.3

21.7

101.0

115.3

8.0

12.3

5.3

9.0

23.0

33.0

MF

1.25

0.89

1.19

1.04

0.83

1.42

0.70

0.82

0.99

1.01

Confirmatory Mutation Test:

Concentrations
(µg/plate)

Mean
values of revertants / Mutation factor (MF)

Salmonella typhimurium tester strains

Escherichia coli

TA98

TA100

TA1535

TA1537

WP2uvrA

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

5000

Mean

10.7

29.7

47.0

86.7

5.7

13.0

5.0

7.3

27.0

38.0

MF

0.43

0.93

0.54

0.80

0.61

1.03

0.79

1.05

0.92

0.91

1581

Mean

25.0

36.3

92.3

109.3

10.0

9.7

6.0

8.3

37.7

38.3

MF

1.01

1.14

1.05

1.01

1.07

0.76

0.95

1.19

1.28

0.92

500

Mean

21.7

26.3

86.7

106.7

13.7

14.0

4.3

7.3

28.0

37.3

MF

0.88

0.82

0.99

0.98

1.46

1.11

0.68

1.05

0.95

0.90

158.1

Mean

32.0

36.7

86.3

117.3

11.7

11.3

6.0

6.0

28.0

33.0

MF

1.30

1.15

0.98

1.08

1.25

0.89

0.95

0.86

0.95

0.79

50

Mean

21.3

28.0

89.7

129.3

7.3

9.7

3.7

9.0

36.0

34.3

MF

0.86

0.88

1.02

1.19

0.79

0.76

0.58

1.29

1.23

0.82

15.81

Mean

24.0

30.0

97.3

109.3

6.3

10.3

5.7

8.0

25.0

35.0

MF

0.97

0.94

1.11

1.01

0.68

0.82

0.89

1.14

0.85

0.84

5

Mean

18.0

33.7

83.0

102.7

6.3

10.7

4.7

9.3

27.7

33.0

MF

0.73

1.05

0.95

0.95

0.68

0.84

0.74

1.33

0.94

0.79

Untreated, negative (vehicle/solvent) and positive controls were run concurrently. The mean values of revertant colony numbers of untreated, negative (vehicle/solvent) and positive control plates were within the historical control range. At least five analyzable concentrations were presented in all strains of the main tests.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative (with and without metabolic activation)

The test item had no mutagenic activity in the applied bacterium tester strains under the test conditions used in this study.
Executive summary:

The test item was tested for potential mutagenic activity using the Bacterial Reverse Mutation Assay according to OECD Guideline 471 (GLP study). The experiments were carried out using Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 and Escherichia coli WP2 uvrA in the presence and absence of metabolic activation (S9 fraction prepared from the livers of rats). Based on the results of the Solubility Test, the test item was dissolved in DMSO. Concentrations of 5000; 2500; 1000; 316; 100; 31.6 and 10 μg/plate were examined in the Range Finding Test. Based on the results of the Range Finding Test, the test item concentrations in the main tests were 5000, 1581, 500, 158.1, 50, 15.81 and 5 μg test item/plate. In the Initial Mutation Test and Confirmatory Mutation Test, none of the observed revertant colony numbers were above the respective biological threshold value. There were no dose-related trends and no indication of any treatment effect. In all test item treated groups, the numbers of revertant colonies did not exceed the biological relevance when compared to the solvent control and were within the normal biological variability of the test system. Inhibitory, cytotoxic effect of the test item were observed in the Confirmatory Mutation Test in all tester strains at 5000 μg/plate with and without metabolic activation. The tests were considered to be valid. In conclusion, the test item had no mutagenic activity in the bacterium tester strains under the test conditions used in this study.