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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:
23 Mar - 03 May 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP - Guideline study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report Date:
2006

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
GLP compliance:
yes (incl. certificate)
Remarks:
Groupe Interministeriel des Produit Chimiques, France; Agence Française de Sécurité Sanitaire des Aliments (AFSSA), France
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): pentaisononanoate de dipentaerythritol
- Physical state: colourles viscous liquid
- Analytical purity: > 90%
- Purity test date: 22 March 2006
- Lot/batch No.: 05078603
- Expiration date of the lot/batch: 26 July 2006
- Storage condition of test material: RT

Method

Target gene:
his-operon
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium TA 102
Metabolic activation:
with and without
Metabolic activation system:
cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254
Test concentrations with justification for top dose:
312.5, 625, 1250, 2500 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: ethanol
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: -S9: sodium azide (1 μg/plate, TA1535 and TA100); 9-amino-acridine (50 µg/plate, TA 1537); 2-nitrofluorene (0.5 µg/plate, TA 98); mitomycin C (0.5 µg/plate, TA 102) +S9: 2-amino-anthracene (10 µg/plate, TA 102, 2 μg/plate, all other strains)
Remarks:
the positive controls were dissolved in DMSO, except for mitomycin C which was dissolved in distilled water
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation), experiment 1, with and without S9, experiment 2, without S9; preincubation, experiment 2, with S9
DURATION
- Exposure duration: 48 - 72 h

NUMBER OF REPLICATIONS: 3 replications each in 2 independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: decrease in number of revertant colonies/thinning of bacterial lawn
Evaluation criteria:
Reproducible 2-fold (TA 98, TA 100, TA 102) or 3-fold (TA 1535, TA 1537) increase in number of revertants in comparison to the vehicle control and/or dose-response correlation.

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:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: moderate to marked emulsion at dose levels ≥ 1250 µg/plate

RANGE-FINDING/SCREENING STUDIES: Yes, no cytotoxicity up to the maximum of solubility

COMPARISON WITH HISTORICAL CONTROL DATA: valid
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 ± standard deviation)

Base-pair substitution type

Frameshift type

TA 100

TA1535

TA102

TA98

TA1537

vehicle (ethanol)

134 ± 9

25 ± 5

363 ± 21

27 ± 4

11 ± 5

312.5

98 ± 6

27 ± 4

330 ± 15

38 ± 15

8 ± 3

625

110 ± 20

35 ± 2

438 ± 91

44 ± 11

11 ± 3

1250

103 ± 14

30 ± 6

407 ± 93

34 ± 8

13 ± 6

2500

135 ± 22

31 ± 1

408 ± 22

31 ± 11

13 ± 4

5000

148 ± 63

31 ± 6

324 ± 16

32 ± 14

9 ± 3

Positive controls, –S9

Name

SA

SA

MMC

2NF

9AA

Concentrations

(μg/plate)

1.0

1.0

0.5

0.5

50

Mean No. of colonies/plate

(average of 3 ± SD)

877 ± 38

630 ± 74

2450 ± 103

171 ± 16

966 ± 27

+

vehicle (ethanol)

135 ± 22

30 ± 6

587 ± 10

44 ± 13

16 ± 6

+

312.5

130 ± 39

28 ± 9

533 ± 28

49 ± 4

9 ± 8

+

625

119 ± 6

26 ± 3

565 ± 21

49 ± 5

16 ± 10

+

1250

118 ± 7

28 ± 9

605 ± 38

43 ± 6

20 ± 9

+

2500

111 ± 6

27 ± 6

584 ± 97

43 ± 6

15 ± 6

+

5000

112 ± 5

34 ± 5

571 ± 29

43 ± 10

20 ± 5

Positive controls, +S9

Name

2AA

2AA

2AA

2AA

2AA

Concentrations

(μg/plate)

2.0

2.0

10

2.0

2.0

Mean No. of colonies/plate

(average of 3 ± SD)

1154 ± 180

419 ± 10

4419 ± 435

1208 ± 84

123 ± 16

SA: Sodium azide

9AA : 9-Aminoacridine

2NF : 2-Nitrofluorene

MMC: Mitomycine C

2AA: 2-Aminoanthracene

 

 

Table 2. Test results of experiment 2 (-S9: plate incorporation, +S9: pre-incubation).

With or without S9-Mix

Test substance concentration

(μg/plate)

Mean number of revertant colonies per plate

(average of 3 plates ± Standard deviation)

Base-pair substitution type

Frameshift type

TA 100

TA1535

TA102

TA98

TA1537

vehicle (ethanol)

126 ± 3

12 ± 5

343 ± 75

45 ± 14

6 ± 2

312.5

140 ± 17

15 ± 10

312 ± 40

25 ± 1

10 ± 1

625

183 ± 17

13 ± 6

454 ± 98

33 ± 3

8 ± 4

1250

153 ± 13

10 ± 4

317 ± 41

46 ± 9

5 ± 1

2500

144 ± 26

11 ± 4

359 ± 17

40 ± 13

5 ± 3

5000

175 ± 66

18 ± 44

420 ± 70

56 ± 18

8 ± 3

Positive controls, –S9

Name

SA

SA

MMC

2NF

9AA

Concentrations

(μg/plate)

1.0

1.0

0.5

0.5

50

Mean No. of colonies/plate

(average of 3 ± SD)

527 ± 83

711 ± 104

1258 ± 165

147 ± 22

1442 ± 287

+

vehicle (ethanol)

176 ± 7

9 ± 2

538 ± 13

43 ± 6

9 ± 2

+

312.5

145 ± 28

12 ± 3

728 ± 66

45 ± 3

10 ± 7

+

625

140 ± 21

19 ± 2

491 ± 41

35 ± 8

7 ± 1

+

1250

185 ± 16

11 ± 3

643 ± 128

30 ± 7

8 ± 2

+

2500

166 ± 16

14 ± 3

575 ± 11

35 ± 13

10 ± 4

+

5000

145 ± 22

16 ± 6

622 ± 74

43 ± 2

10 ± 5

Positive controls, +S9

Name

2AA

2AA

2AA

2AA

2AA

Concentrations

(μg/plate)

2.0

2.0

10

2.0

2.0

Mean No. of colonies/plate

(average of 3 ± SD)

705 ± 142

202 ± 25

2678 ± 1940

1259 ± 94

99 ± 27

SA: Sodium azide

9AA : 9-Aminoacridine

2NF : 2-Nitrofluorene

MMC: Mitomycine C

2AA: 2-Aminoanthracene

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative