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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:
experimental study
Adequacy of study:
key study
Study period:
20 Oct 1999 - 11 Nov 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP - Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1999
Report Date:
1999

Materials and methods

Test guidelineopen allclose all
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
Qualifier:
according to
Guideline:
EPA OPPTS 870.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
UK GLP Monitoring Authority, Department of Health
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Neopentylglycol dioctanoate- Physical state: liquid, colourless- Lot/batch No.: 0709N- Storage condition of test material: RT, protected from light- Analytical purity: 98%

Method

Target gene:
his operon
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of male rats treated with phenobarbitone/b-naphthoflavone
Test concentrations with justification for top dose:
Pretest: 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate without metabolic activationMain test: 50, 150, 500, 1500 and 5000 µg/plate with and without metabolic activation
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: acetone
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: N-ethyl-N-nitro-N-nitrosoguanidine (ENNG; 3 µg/plate for TA 100 and 5 µg/plate for TA 1535); 9-aminoacridine (9AA; 80 µg/plate for TA 1537); mitomycin C (MMC; 0.5 µg/plate for TA 102); 4-nitroquinoline-N-oxide ( 4NQO; 0.2 µg/plate for TA 98)
Remarks:
without metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: aminoanthracene (2AA; 1 µg/plate for TA100 and 2 µg/plate for TA 1535 and TA 1537); benzo(a)pyrene (BP; 5 µg/plate for TA 98); dihydroxyanthraquinone (DANTHRON; 10 µg/plate for TA 102)
Remarks:
with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)DURATION- Exposure duration: appr. 48 hNUMBER OF REPLICATIONS: triplicates in two independet experiments DETERMINATION OF CYTOTOXICITY- Method: relative total growth
Evaluation criteria:
The test material may be considered to be positive in this test system if the following criteria are met:The test material should have induced a reproducible, dose-related and statistically significant increase in the revertant count in at least one strain of bacteria.
Statistics:
Dunnett´s method of linear regression

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Remarks:
at and above 1500 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS- Precipitation: Greasy, oily precipitate at and above 1500 µg/plate. This did not prevent the scoring of revertant coloniesRANGE-FINDING/SCREENING STUDIES: Yes- Test test material was non-toxic to the strain TA100. The test material formulation and the S9-mix used were both shown to be sterile.COMPARISON WITH HISTORICAL CONTROL DATA: Yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Mutagenicity on bacteria - Experiment I

 

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

Frameshift type

TA 100

TA1535

TA102

TA98

TA1537

 

Vehicle

99

14

303

28

15

-

50

99

14

300

31

11

-

150

110

22

318

27

18

-

500

114

21

311

29

14

-

1500

101

22

307

33

17

-

5000

111

18

300

26

14

Positive

controls

- S9

Name

ENNG

ENNG

MMC

4NQO

9AA

Concentrations

(μg/plate)

3.0

5.0

0.5

0.2

80

Number of colonies/plate

492

428

979

143

455

+

Vehicle

100

13

315

37

18

+

8

89

12

329

28

16

+

40

97

13

338

31

16

+

200

96

11

336

29

17

+

1000

109

14

318

26

15

+

5000

98

13

320

35

13

Positive

controls

+ S9

Name

2AA

2AA

DAN

BP

2AA

Concentrations

(μg/plate)

1.0

2.0

10

5

2

Number of colonies/plate

753

228

939

161

556

 

ENNG = N-ethyl-N+-nitro-nitrosoguanidine

4NQO= 4-nitroquinoline-1-oxide

9AA = 9-Aminoacridine

2AA = 2-Aminoanthracene

BP = Benzo(a)pyrene

DAN = 1,8-Dihydroxyanthraquinone

 

 

 

Table 2: Mutagenicity on bacteria - Experiment II

 

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

Frameshift type

TA 100

TA1535

TA102

TA98

TA1537

 

Vehicle

114

24

283

36

9

-

50

100

22

279

34

8

-

150

122

23

302

35

13

-

500

120

28

306

32

14

-

1500

117

26

291

29

12

-

5000

121

28

278

28

10

Positive

controls

- S9

Name

ENNG

ENNG

MMC

4NQO

9AA

Concentrations

(μg/plate)

3.0

5.0

0.5

0.2

80

Number of colonies/plate

465

370

885

173

1004

+

Vehicle

107

20

295

32

14

+

8

125

22

300

28

16

+

40

115

26

294

35

18

+

200

112

18

297

24

15

+

1000

116

22

321

29

20

+

5000

106

20

313

28

14

Positive

controls

+ S9

Name

2AA

2AA

DAN

BP

2AA

Concentrations

(μg/plate)

1.0

2.0

10

5

2

Number of colonies/plate

1791

156

649

201

469

ENNG = N-ethyl-N+-nitro-nitrosoguanidine

4NQO= 4-nitroquinoline-1-oxide

9AA = 9-Aminoacridine

2AA = 2-Aminoanthracene

BP = Benzo(a)pyrene

DAN = 1,8-Dihydroxyanthraquinone

Under the tested experimental conditions the test substance did not induce gene mutations in S. typhimurium strains up to the maximum dose. Therefore it is not considered to be mutagenic in this bacterial mutagenicity test in vitro.

Applicant's summary and conclusion

Conclusions:
it was concluded that 3-[(2-ethylhexanoyl)oxy]-2,2-dimethylpropyl 2-ethylheptanoate CAS: 28510-23-8 is not considered to be mutagenic based on the negative results obtained from the Ames test.
Executive summary:

A study was conducted to assess the mutagenic potential of 3-[(2-ethylhexanoyl)oxy]-2,2-dimethylpropyl 2-ethylheptanoate CAS: 28510-23-8. The test was done in an accredited GLP laboratory, in accordance with internationally recognised guidelines:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

EPA OPPTS 870.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)

The bacterial reverse mutation assay was done so using numerous concentrations of the test substance, on S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102. Under the conditions of the study the 3-[(2-ethylhexanoyl)oxy]-2,2-dimethylpropyl 2-ethylheptanoate CAS: 28510-23-8 did not elicit a positive response, and as such it not considered to be mutagenic.