Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ames test (OECD 471): negative in S. typhimurium TA 1535, TA 1537, TA 98 and TA 100 and E. coli WP2 uvrA with and without metabolic activation

RA from source substance Ditridecyl adipate (CAS 16958 -92 -2) and Hexanedioic acid, di-C16-18 (even numbered)-alkyl esters (CAS 92969-90-9)

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP - Guideline study
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
The Departement Of Health Of The Government Of The United Kingdom, UK
Type of assay:
bacterial reverse mutation assay
Target gene:
his operon and trp operon
Species / strain / cell type:
other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats induced with Phenobarbitone/β-Naphthoflavone at 80/100 mg/kg/day, orally, for 3 days prior to preparation on day 4
Test concentrations with justification for top dose:
Preliminary toxicity test (in TA 100 and WP2 uvr A): 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500 and 5000 μg/plate with and without metabolic activation
Main Assay (Experiment I and II): 50, 150, 500, 1500 and 5000 μg/plate with and without metabolic activation

Vehicle / solvent:
- Vehicle(s)/solvent(s) used: acetone
- Justification for choice of solvent/vehicle: The test item was insoluble in sterile distilled water and dimethyl sulphoxide at 50 mg/mL, but was fully soluble in dimethyl formamide at the same concentration and in acetone at 100 mg/mL and tetrahydrofuran at 200 mg/mL. Acetone was selected as the vehicle.
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Remarks:
-S9: ENNG (2-5 µg/mL for WP2 uvr A, TA 100, TA 1535); 9-AA (80 μg/plate for TA 1537); 4NQO (0.2 μg/plate for TA98); +S9: 2-AA (1-10 µg/plate for TA 100, TA 1535, TA 1537 and WP2 uvr A); BP (5 μg/plate for TA 98)
Positive control substance:
4-nitroquinoline-N-oxide
9-aminoacridine
N-ethyl-N-nitro-N-nitrosoguanidine
benzo(a)pyrene
other: 2-aminoanthracene
Remarks:
ENNG: N-ethyl-N'-nitro-N-nitrosoguanidine; 9-AA:9-Aminoacridine; 4-NQO: 4-nitroquinoline-1-oxide; 2-AA: 2-aminoanthracene; BP: benzo(a)pyrene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium (Preliminary toxicity test and Experiment I); preincubation (Experiment II)

DURATION
- Preincubation period: 20 min (Experiment II)
- Exposure duration: 48 h (all experiments)

NUMBER OF REPLICATIONS: triplicates in two independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: determination of the number of revertant colonies and inspection of the bacterial background lawn
Evaluation criteria:
There were several criteria for determining a positive result. Any, one, or all of the following were used to determine the overall result of the study:
- a dose-related increase in mutant frequency over the dose range tested.
- a reproducible increase at one or more concentrations.
- biological relevance against historical control ranges.
- statistical analysis of data as determined by UKEMS.
- fold increase greater than two times the concurrent solvent control for any tester strain (especially if accompanied by an out-of-historical range response).
A test item was considered non-mutagenic (negative) in the test system if the above criteria were not met.
Statistics:
Mean values and standard deviations of the mean number of revertants were calculated. Statistical analysis of data was performed according to guidelines from UKEMS.
Key result
Species / strain:
other: S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Remarks:
/ precipitation was observed starting at 1500 μg/plate, with and without metabolic activation
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: A test item precipitate (globular in appearance) was noted by eye at and above 1500 μg/plate, this observation did not prevent the scoring of revertant colonies.

RANGE-FINDING/SCREENING STUDIES: In the Preliminary toxicity test, the test item was non-toxic to strains TA 100 and WP2 uvrA up to the maximum concentration of 5000 µg/plate.

COMPARISON WITH HISTORICAL CONTROL DATA: The mean number of revertants of vehicle and positive controls was within the normal range of the respective historical control data.

Table 1. Test results of experiment I (plate incorporation)

Bacterial Reverse Mutation Assay, mean revertant colonies/plate (mutation factor) (n=3 ± SD)

EXPERIMENT I (plate incorporation)

S9-Mix

Without

 

Concentration (per plate)

TA 100

TA1535

WP2 uvrA

TA98

TA 1537

SC

108 ± 14

14 ± 2

19 ± 2

21 ± 4

10 ± 1

Test material

 

 

 

 

 

50 µg

91 ± 12

13 ± 2

18 ± 6

19 ± 2

7 ± 4

150 µg

95 ± 21

15 ± 2

22 ± 2

18 ± 5

8 ± 4

500 µg

91 ± 7

14 ± 4

21 ± 5

16 ± 1

8 ± 2

1500 µg

94 ± 6

13 ± 1

18 ± 10

22 ± 8

6 ± 2

5000 µg

93 ± 3

10 ± 2

20 ± 2

16 ± 3

8 ± 3

PC

 

 

 

 

 

ENNG

614 ± 28

442 ± 54

937 ± 42

-

-

4-NQO

-

-

-

239 ± 30

-

9-AA

-

-

-

-

912 ± 96

S9-Mix

 

With

Concentration (per plate)

TA 100

TA 1535

WP2 uvr A

TA98

TA 1537

NC

91 ± 2

11 ± 3

26 ± 8

27 ± 3

11 ± 0

Test material

 

 

 

 

 

50 µg

101 ± 12

11 ± 2

25 ± 4

24 ± 4

9 ± 3

150 µg

96 ± 11

11 ± 1

28 ± 6

31 ± 2

10 ± 1

500 µg

104 ± 11

10 ± 1

29 ± 4

23 ± 8

10 ± 4

1500 µg

89 ± 3

11 ± 1

22 ± 7

23 ± 10

12 ± 5

5000 µg

99 ± 10

12 ± 1

24 ± 8

24 ± 6

10 ± 2

PC

 

 

 

 

 

2-AA

1238 ± 270

231 ± 11

286 ± 14

-

237 ± 12

BP

-

-

-

199 ± 11

-

SC = Solvent control; PC = Positive control substances; SD = standard deviation;

ENNG: N-ethyl-N'-nitro-N-nitrosoguanidine; 9-AA: 9-Aminoacridine; 4-NQO: 4-nitroquinoline-1-oxide; 2-AA: 2-aminoanthracene; BP: benzo(a)pyrene

Table 2. Test results of experiment II (pre-incubation)

Bacterial Reverse Mutation Assay, mean revertant colonies/plate (mutation factor) (n=3 ± SD)

EXPERIMENT II (pre-incubation)

S9-Mix

Without

 

Concentration (per plate)

TA 100

TA1535

WP2 uvrA

TA98

TA 1537

SC

79 ± 6

15 ± 5

26 ± 3

15 ± 3

10 ± 4

Test material

 

 

 

 

 

50 µg

72 ± 2

12 ± 2

24 ± 5

26 ± 12

9 ± 1

150 µg

82 ± 13

12 ± 2

27 ± 5

16 ± 3

11 ± 3

500 µg

66 ± 1

8 ± 2

18 ± 1

22 ± 5

11 ± 1

1500 µg

74 ± 3

13 ± 4

27 ± 5

16 ± 1

5 ± 1

5000 µg

79 ± 1

11 ± 4

23 ± 6

21 ± 7

11 ± 4

PC

 

 

 

 

 

ENNG

728 ± 131

892 ± 86

946 ± 35

-

-

4-NQO

-

-

-

118 ± 8

-

9-AA

-

-

-

-

534 ± 206

S9-Mix

 

With

Concentration (per plate)

TA 100

TA1535

WP2 uvrA

TA98

TA 1537

NC

75 ± 5

14 ± 6

34 ± 9

24 ± 4

9 ± 2

Test material

 

 

 

 

 

50 µg

75 ± 4

12 ± 3

25 ± 0

21 ± 1

7 ± 3

150 µg

83 ± 22

10 ± 2

22 ± 9

26 ± 2

12 ± 4

500 µg

70 ± 18

10 ± 7

29 ± 4

21 ± 2

8 ± 2

1500 µg

76 ± 12

11 ± 6

24 ± 2

21 ± 3

11 ± 2

5000 µg

76 ± 22

10 ± 2

29 ± 5

26 ± 3

10 ± 3

PC

 

 

 

 

 

2-AA

1506 ± 102

201 ± 30

246 ± 22

-

316 ± 32

BP

-

-

-

130 ± 7

-

SC = Solvent control; PC = Positive control substances; SD = standard deviation;

ENNG: N-ethyl-N'-nitro-N-nitrosoguanidine; 9-AA: 9-Aminoacridine; 4-NQO: 4-nitroquinoline-1-oxide; 2-AA: 2-aminoanthracene; BP: benzo(a)pyrene

Conclusions:
Based on the results of the conducted study the test substance did not exhibit mutagenic properties in bacterial cells.
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
28 Aug - 26 Sep 2016
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:
adopted July 1997
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Health Care Inspectorate of the Ministry of Health, Welfare and Sport, Utrecht, The Netherlands
Type of assay:
bacterial reverse mutation assay
Target gene:
his operon; trp operon
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:
Cofactor supplemented post-mitochondrial factor (S9 mix), prepared from the livers of male rats treated with Aroclor.
Test concentrations with justification for top dose:
Dose range finding test / Experiment I (TA 100 and WP2 uvr A): 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 µg/plate with and without metabolic activation
Experiment I (TA 1537, TA 1535 and TA 98): 52, 164, 512, 1600 and 5000 µg/plate with and without metabolic activation
Experiment II (all strains): 17, 52, 164, 512 and 1600 µg/plate with and without metabolic activation
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: A solubility test was performed. The test item was observed to be insoluble in water and dimethyl sulfoxide. In ethanol, the test item was fully soluble at 50 mg/mL. Based on these solubility findings, ethanol was selected as vehicle.
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
2-nitrofluorene
sodium azide
methylmethanesulfonate
other: -S9: acridine mutagen ICR-191 (2.5 µg/plate) for TA 1537 (Exp. I); +S9: 2-amino-anthracene (1, 2.5 or 5 µg/plate) for all strains
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation, Experiment I) and preincubation (Experiment II)

DURATION
- Preincubation period: 30 min (Experiment II)
- Exposure duration: 48 ± 4 h (Experiment I and II)

NUMBER OF REPLICATIONS: triplicates each in 2 independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: reduction in the mean number of revertants, reduction in the background lawn and the increase in the size of the microcolonies
Evaluation criteria:
A test item is considered negative (not mutagenic) in the test if:
a) The total number of revertants in tester strain TA 100 or WP2 uvr A is not greater than twice the concurrent control, and the total number of revertants in tester strains TA 1535, TA 1537 or TA 98 is not greater than three times the concurrent control.
b) The negative response should be reproducible in at least one follow up experiment.

A test item is considered positive (mutagenic) in the test if:
a) The total number of revertants in tester strain TA 100 or WP2 uvr A is greater than twice the concurrent control, or the total number of revertants in tester strains TA 1535, TA 1537 or TA 98 is greater than three times the concurrent control at one or more concentrations with or without metabolic activation and/or a concentration-related increase over the range tested.
b) In case a repeat experiment is performed when a positive response is observed in one of the tester strains, the positive response should be reproducible in at least one follow-up experiment.
Statistics:
Mean values and standard deviations were calculated.
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Remarks:
precipitation was observed at 1600 and 5000 µg/plate, with and without metabolic activation
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Remarks:
precipitation was observed at 1600 and 5000 µg/plate, with and without metabolic activation
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Remarks:
precipitation was observed at 1600 and 5000 µg/plate, with and without metabolic activation
Vehicle controls validity:
valid
Untreated 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:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Remarks:
precipitation was observed at 1600 and 5000 µg/plate, with and without metabolic activation
Vehicle controls validity:
valid
Untreated 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:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Remarks:
precipitation was observed at 1600 and 5000 µg/plate, with and without metabolic activation
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: In the dose range finding study precipitation of the test material on the plates was observed at the start and at the end of the incubation period at concentrations of 1600 and 5000 µg/plate. In Experiment I the test substance precipitated on the plates at the start of the incubation period at 5000 µg/plate and at 1600 and 5000 µg/plate at the end of the incubation period. Precipitation at 5000 µg/plate was observed as oily droplets at the start and the end of the incubation period. In Experiment II the test substance precipitated at the start of the incubation period at concentrations of 512 and 1600 µg/plate in TA 98 without metabolic activation only. At the end of the incubation period precipitation of the test substance was observed at 1600 µg/plate in all tester strains with and without metabolic activation. Slight precipitate did not influence automated counting of the plate.

RANGE-FINDING/SCREENING STUDIES: The test substance was initially tested in TA100 and WP2 uvr A as a dose range finding test with concentrations of 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 μg/plate in the absence and presence of S9-mix. No reduction of the bacterial background lawn and no biologically significant decrease in the number of revertants were observed in the dose range finding test. The results of the range-finding test were included in Experiment I.

HISTORICAL CONTROL DATA (ranges)
- Positive historical control data:
TA 1535 (-S9: 78-1381; +S9: 78-1058)
TA 1537 (-S9: 55-1565; +S9: 55-1112)
TA 98 (-S9: 410-2057; + S9: 263-1907)
TA 100 (-S9: 549-1848; +S9: 620-2651)
WP2 uvr A (-S9: 127-1951; +S9: 85-1390)
The positive controls were within the range of the historical control data.

- Negative (solvent/vehicle) historical control data:
TA 1535 (-S9: 4-36; +S9: 3-34)
TA 1537 (-S9: 3-25; +S9: 3-28)
TA 98 (-S9: 9-50; + S9: 9-57)
TA 100 (-S9: 63-153; +S9: 60-456)
WP2 uvr A (-S9: 12-68; +S9: 12-70)
The solvent control was within the range of the historical control data.

Tabe 1: Summary of the dose range finding test / Experiment I

  TA 100   WP2 uvr A  
  -S9 mix + S9 mix -S9 mix + S9 mix
Ethanol 86 ± 6 93 ± 5 22 ± 3 21 ± 5
Test substance
[µg/plate]
       
1.7 72 ± 18 88 ± 23 24 ± 5 16 ± 2
5.4 68 ± 10 71 ± 3 32 ± 4 16 ± 0
17 87 ± 16 72 ± 6 22 ± 5 19 ± 4
52 65 ± 8 85 ± 15 25 ± 2 22 ± 2
164 73 ± 10 79 ± 6 21 ± 5 13 ± 3
512 73 ± 20 73 ± 5 20 ± 2 19 ± 7
1600 68 ± 8 SP 71 ± 3 SP 28 ± 5 SP 18 ± 3SP
5000 82 ± 19 SP 87 ± 20 SP 21 ± 2 SP 21 ± 4SP
Positive controls methylmethane sulfonate 2-aminoanthracene 4-nitroquinoline N-oxide 2-aminoanthracene
  1102 ± 106 749 ± 105 433 ± 60 1230 ± 94

SP: Slight precipitation

Table 2: Summary of Experiment I

  TA 1535   TA 1537   TA 98  
  -S9 mix + S9 mix -S9 mix + S9 mix -S9 mix + S9 mix
Solvent control 6 ± 2 10 ± 5 5 ± 2 11 ± 4 15 ± 4 15 ± 4
Test substance
[µg/plate]
           
52 8 ± 5 7 ± 3 7 ± 3 12 ± 2 10 ± 6 15 ± 4
164 9 ± 2 10 ± 6 4 ± 4 10 ± 4 9 ± 6 17 ± 3
512 6 ± 3 10 ± 2 7 ± 3 5 ± 2 14 ± 3 18 ± 2
1600 4 ± 3 SP 7 ± 2 SP 4 ± 1 SP 6 ± 1 SP 16 ± 5 SP 20 ± 5 SP
5000  11 ± 3 SP 8 ± 3 SP 11 ± 8 SP 6 ± 4 SP 13 ± 3 SP 15 ± 9 SP
Positive control sodium azide 2-aminoanthracene ICR-191 2-aminoanthracene 2-nitrofluorene 2-aminoanthracene
  753 ± 67 170 ± 20 932 ± 72 230 ± 54 1283 ± 54 982 ± 46

SP: Slight precipitation

Table 3: Summary of Experiment II

  TA 1535   TA 1537   TA 98   TA 100   WP2 uvr A  
  -S9 mix + S9 mix -S9 mix + S9 mix -S9 mix + S9 mix -S9 mix + S9 mix -S9 mix + S9 mix
Solvent control 9 ± 5 8 ± 2 20 ± 21 23 ± 29 19 ± 5 27 ± 6 126 ± 4 114 ± 20 28 ± 9 29 ± 9
Test substance
[µg/plate]
                   
17 13 ± 7 6 ± 2 5 ± 2 10 ± 2 21 ± 2 25 ± 8 110 ± 15 112 ± 8 37 ± 11 23 ± 4
52 8 ± 1 13 ± 1 14 ± 11 5 ± 2 19 ± 7 43 ± 35 124 ± 23 102 ± 3 32 ± 6 25 ± 9
164 5 ± 3 9 ± 6 5 ± 2 9 ± 8 21 ± 1 20 ± 0 118 ± 6 106 ± 18 28 ± 10 25 ± 9
512 7 ± 4 12 ± 2 5 ± 2 9 ±6 17 ± 3 39 ± 32 98 ± 11 106 ± 8 35 ± 11 33 ± 9
1600 8 ± 4 SP 11 ± 4 SP 5 ± 2 SP 14 ± 11 SP 19 ± 5 SP 28 ± 5 SP 9 ± 8 SP 112 ± 6 SP 26 ± 2 SP 33 ± 7 SP
Positive control sodium azide 2-aminoanthracene ICR-191 2-aminoanthracene 2-nitrofluorene 2-aminoanthracene methylmethane sulfonate 2-aminoanthracene 4-nitroquinoline N-oxide 2-aminoanthracene
  875 ± 31 161 ± 8 100 ± 15 221 ± 47 1312 ± 52 671 ± 56 735 ± 35 1390 ± 112 225 ± 159 520 ± 26

SP: Slight precipitation

Conclusions:
Based on the results of the conducted study the test substance did not exhibit mutagenic properties in bacterial cells.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for read-across

There are no substance-specific data available in regard to genetic toxicity in bacteria for Fatty acids, C18 -unsaturated, 1 -6 Hexanediol Diester.

To fulfil the standard data requirements defined in Regulation (EC) No 1907/2006, Annex VII, 8.4, read-across from appropriate substances is conducted in accordance with Regulation (EC) No 1907/2006, Annex XI, 1.5.

According to Article 13 (1) of Regulation (EC) No 1907/2006, "information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set out in Annex XI are met”. In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests, which includes the use of information from structurally related substances (grouping or read-across) “to avoid the need to test every substance for every endpoint”.

 

For each specific endpoint the source substance(s) structurally closest to the target substance is/are chosen for read across, with due regard to the requirements of adequacy and reliability of the available data. Structural similarities and similarities in properties and/or activities of the source and target substances are the basis of read-across. A detailed justification for the analogue read- across approach is provided in the technical dossier (see IUCLID Section 13).

 

As no reliable measured/experimental data are available on genetic toxicity in bacterial cells for Fatty acids, C18 -unsaturated, 1 -6 Hexanediol Diester

, read-across to reliable data on the analogue substances Ditridecyl adipate (CAS 16958-92-2) and Hexanedioic acid, di-C16-18 (even numbered)-alkyl esters (CAS 92969-90-9) was conducted.

 

CAS 16958-92-2

The in vitro genetic toxicity of the test substance was assessed in a bacterial reverse mutation assay (Ames test) according to OECD 471 and in compliance with GLP (Thompson, 2013). The mutagenic potential of the test substance was assessed in S. typhimurium tester strains including TA 98, 100, 1535 and 1537 and in E. coli WP2 uvr A bacterial cells at concentrations up to 5000 µg/plate in 2 independent experiments, including plate incorporation and preincubation method. Precipitates were observed starting at a concentration of 1500 µg/plate (all tester strains). The test substance did not exhibit mutagenic properties in the absence or presence of metabolic activation. Cytotoxicity was not observed. The vehicle and positive controls were valid and lay within the range of historical control data, proving validity of the assay. Based on the results of the conducted study, the test substance is not considered to exhibit mutagenic properties in bacterial cells.

CAS 92969-90-9

The in vitro genetic toxicity of the test substance was assessed in a bacterial reverse mutation assay (Ames test) according to OECD 471 and in compliance with GLP ( Verspeek-Rip, 2017). The mutagenic potential of the test substance was assessed in S. typhimurium tester strains including TA 98, 100, 1535 and 1537 and in E. coli WP2 uvr A bacterial cells at concentrations up to 5000 µg/plate in the plate incorporation test and up to 1600 µg/plate in the preincubation test. Precipitates were observed in both tests starting at a concentration of 1600 µg/plate (all tester strains). The test substance did not exhibit mutagenic properties in the absence or presence of metabolic activation. Cytotoxicity was not observed. The vehicle and positive controls were valid and lay within the range of historical control data, proving validity of the assay. Based on the results of the conducted study, the test substance is not considered to exhibit mutagenic properties in bacterial cells.

 

Conclusion on genetic toxicity

The available measured data for the read-across analogue substances Ditridecyl adipate (CAS 16958-92-2) and Hexanedioic acid, di-C16-18 (even numbered)-alkyl esters (CAS 92969-90-9) reveal no genetic toxicity. Thus, no potential for genetic toxicity is expected for Fatty acids, C18 -unsaturated, 1 -6 Hexanediol Diester.

Justification for classification or non-classification

Based on the analogue read-across approach and on the available data, there is no indication that the substance induces genetic toxicity. Nevertheless, no final decision on classification for genetic toxicity according to Regulation (EC) 1272/2008 can be made, as no information on chromosomal aberration and mutagenicity in mammalian cells/in vivo is available.