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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP - Guideline study. According to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1996
Reference Type:
secondary source
Title:
Dibutyl adipate CAS N°: 105-99-7
Author:
OECD
Year:
1996
Bibliographic source:
SIDS Initial Assessment Report for SIAM 4; Tokyo, Japan, 20-22 May 1996

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
(adopted in 1983)
Deviations:
no
Remarks:
the current guildeline - adopted in 1997 - requires other positive controls +S9 beside 2-aminoanthracene
Qualifier:
according to guideline
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
Version / remarks:
(adopted in 1983)
Deviations:
no
Qualifier:
according to guideline
Guideline:
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Dibutyl adipate
EC Number:
203-350-4
EC Name:
Dibutyl adipate
Cas Number:
105-99-7
IUPAC Name:
dibutyl adipate
Details on test material:
- Name of test material (as cited in study report): Dibutyl adipate
- Analytical purity: 99.8%
- Lot/batch No.: N-41001
- Storage condition of test material: At room temperature

Method

Target gene:
his operon and trp operon
Species / strainopen allclose all
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 fraction (S9 mix), prepared from the livers of rats treated with phenobarbital and 5,6-Benzoflavone.
Test concentrations with justification for top dose:
312.5, 625, 1250, 2500 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: -S9: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (0.01 µg/plate) for TA100, WP2uvrA, (0.1 µg/plate) for TA98; sodium azide (0.5 µg/plate) for TA1535; 9-aminoacridine (80 µg/plate) for TA1537; +S9: 2-aminoanthracene (up to 10 µg/plate) all strains
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: triplicates each in two independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: Inspection of the bacterial background lawn,
In a preliminary experiment employing all five applied tester strains triplicates of controls and single plates with test substance (50, 150, 500, 1500 or 5000 µg/plate) were incubated to determine cytotoxicity in the presence or absence of S9.
Evaluation criteria:
If the number of colonies with revertants was as double as that of control group and dose-related response was observed, the interpretation of the result was positive.
Statistics:
Mean values and standard deviation were calculated.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
TA 100 at 5000 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING: In a preliminary experiment employing all five applied tester strains triplicates of controls and single plates with test substance (50, 150, 500, 1500 or 5000 µg/plate) were incubated to determine cytotoxicity in the presence or absence of S9. No cytotoxicity was observed in the pretest.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Test results of experiment 1 (plate incorporation)

With or without S9-Mix

Test substance concentration (µg/plate)

Mean number of revertant colonies per plate (average of 3 plates)

 

 

Base-pair substitution type

Cross-linking type

Frameshift type

 

 

TA 100

TA 1535

WP2

TA 98

TA 1537

0

135

13

22

19

7

312.5

114

10

20

21

7

650

118

12

18

22

6

1250

105

11

20

18

4

2500

104

9

18

21

4

5000

90*

8

22

17

4

Positive controls,

-S9

Name

AF2

SA

AF2

AF2

9AA

 

Concentrations (μg/plate)

0.01

0.5

0.01

0.1

80

 

Mean No. of colonies/plate (average of 3)

523

349

156

629

1114

+

0

129

12

24

29

9

+

312.5

124

10

29

34

13

+

650

138

15

29

38

12

+

1250

138

13

28

32

13

+

2500

128

18

29

31

11

+

5000

142

17

29

25

12

Positive controls,

+S9

Name

2AA

2AA

2AA

2AA

2AA

 

Concentrations (μg/plate)

1

2

10

0.5

2

 

Mean No. of colonies/plate (average of 3)

1015

205

1510

290

207

 

Table 2: Test results of experiment 2 (plate incorporation)

With or without S9-Mix

Test substance concentration

Mean number of revertant colonies per plate (average of 3 plates)

 

 

Base-pair substitution type

Cross-linking type

Frameshift type

 

 

TA 100

TA 1535

WP2

TA 98

TA 1537

0

119

14

28

26

9

312.5

113

8

20

26

6

650

127

9

29

22

4

1250

121

9

19

28

6

2500

114

9

21

24

6

5000

90*

8

20

24

5

Positive controls, –S9

Name

AF2

SA

AF2

AF2

9AA

 

Concentrations (μg/plate)

0.01

0.5

0.01

0.1

80

 

Mean No. of colonies/plate (average of 3)

612

450

143

870

1882

+

0

125

13

21

34

15

+

312.5

134

9

22

33

13

+

650

145

14

24

40

14

+

1250

138

10

28

39

12

+

2500

149

12

24

36

13

+

5000

131

16

28

34

14

Positive controls, + S9

Name

2AA

2AA

2AA

2AA

2AA

 

Concentrations (μg/plate)

1

2

10

0.5

2

 

Mean No. of colonies/plate (average of 3)

1106

310

1512

310

300

AF2 = 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide

9AA = 9-aminoacridine

2AA = 2-aminoanthracene

SA = sodium azide

* = Inhibition was observed against growth of the bacteria.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative