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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:
weight of evidence
Study period:
1977
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Meets general accepted scientific standards

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1977

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
: strain TA 102 is missing; detection of cross-linking and oxidizing substances not possible; no justification for maximal test concentration
Principles of method if other than guideline:
according the method of Ames et al. 1975
GLP compliance:
not specified
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): Trigonox 36-CD-75 (bis(3,5,5-trimethylhexanoyl) peroxide)
- Substance type: organic peroxide
- Physical state: clear colourless liquid
- Analytical purity: 75% solution in aromatic free mineral spirit
- Storage condition of test material: -20°C

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
n.a.
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
spot test: 0 and 3 mg/ 30µl DMSO
plate incorporation assay: 0, 0.2, 2, 20, 500 µg/0.1 ml DMSO/plate
Vehicle / solvent:
DMSO
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
Migrated to IUCLID6: N-methyl-N-nitro-N-nitrosoguanidine, 4-aminobiphenyl, 9-aminoacridine
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); spotting onto the plate surface
DURATION
- Exposure duration: 2 days

SELECTION AGENT (mutation assays): histidine

All determinations were made in duplicate and appropriate controls were included in each assay.
Statistics:
no data

Results and discussion

Test results
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:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
not specified
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Results of the spot test

Trigonox 36-CD-75 spotted/plate (mg/30 µl DMSO)

S9 (µl/plate)

His+revertants in each of two plates with

TA 1535

TA 1537

TA 98

TA 100

0

50

0;1

4;4

24;24

15;15

3

50

4;6

4;9

25;35

15;22

0

0

0;2

1;0

8;12

9;11

3

0

2;8

0;2

4;4

18;19

 

 

Table 2: Results of the plate incorporation assay

Trigonox 36-CD-75 (µg/plate)

S9 (µl/plate)

His+revertants in each of two plates with

TA 1535

TA 1537

TA 98

TA 100

0

50

3;7

5;5

18;20

35;36

0.2

50

6;7

3;6

15;22

21;27

2

50

7;11

4;7

10;16

23;28

20

50

9;11

5;7

12;16

24;33

500

50

6;6

4;7

20;26

27;30

0

0

3;6

7;7

11;16

27;27

0.2

0

5;7

6;8

8;13

24;26

2

0

8;10

4;9

7;10

27;28

20

0

6;8

2;4

8;11

19;31

500

0

4;6

3;5

9;11

22;25

 

 

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Under conditions of this study the test item did not reveal mutagenic activity up to non-inhibitory levels in the Salmonella/microsome mutagenicity test either with or without metabolic activation.
Executive summary:

In a reverse gene mutation assay in bacteria, strains TA1535, TA1537, TA100 and TA98 of S. typhimurium were exposed to bis (3,5,5-trimethylhexanoyl) peroxide (75% solution in aromatic free mineral spirit) at concentrations of 0, 0.2, 2, 20 and 500 µg/plate in the presence and absence of mammalian metabolic activation (S9 mix) in a plate incorporation. In addition a spot test was conducted with 0 and 3 mg/30 µl DMSO spotted per plate in the absence and presence of S9-mix.

Up to the highest concentration tested (500 µg/plate and 3 mg/spot) no cytotoxic effects were observed. There was no evidence of induced mutant colonies over background. This study is considered as acceptable.