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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 from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Study period:
09 Jan - 07 Feb 2002
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP -guideline study with acceptable restrictions (no analytical purity reported)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report date:
2002

Materials and methods

Test guidelineopen allclose all
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
no analytical purity reported
Qualifier:
equivalent or similar to guideline
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
adopted in 2000
Deviations:
yes
Remarks:
no analytical purity reported
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Fatty acids, C16-18 and C18-unsatd., branched and linear, esters with trimethylolpropane
Cas Number:
403507-18-6
Molecular formula:
not applicable, substance is a UVCB
IUPAC Name:
Fatty acids, C16-18 and C18-unsatd., branched and linear, esters with trimethylolpropane
Details on test material:
- Name of test material (as cited in study report): only trade name given
- Physical state: dark amber, slightly viscous liquid
- Analytical purity: no data
- Stability under test conditions: room temperature, in the dark

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 phonobarbitone/beta-naphthoflavone
Test concentrations with justification for top dose:
First and second experiment: 50, 150, 500, 1500 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: acetone
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: -S9: N-ethyl-N´-nitro-N-nitrosoguanidine (ENNG, 3 or 5 µg/plate) for TA 100 and TA 1535; 9-Aminoacridine (9AA, 80 µg/plate) for TA 1537; mitomycin C (MMC, 0.5 µg/plate) for TA 102; 4-Nitroquinoline-1-oxide (4NQO, 0.2 µg/plate) for TA98
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: +S9: 2-Aminoanthracene (2AA, 1 or 2 µg/plate) for TA 100, TA 1535 and TA 1537; Benzo(a)pyrene (BP, 5 µg/plate) for TA 98; 1,8-Dihydroxyanthraquinone (DAN, 10 µg/plate) for TA 102
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3 replications each in 2 independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: Inspection of the bacterial background lawn

Evaluation criteria:
The test material may be considered positive mutagenic in this test system if the following criteria are met: the test item should induce a reproducible, dose-related and statistically significant increase in the revertant count in at least one strain of bacteria.
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:
no cytotoxicity, but tested up to precipitating 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, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
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

0

80 ± 5.7

24 ± 6.1

294 ± 18.1

24 ± 4.6

18 ± 2.5

50

81 ± 6.1

22 ± 5.9

297 ± 19.3

23 ± 1.0

14 ± 2.1

150

86 ± 3.1

23 ± 3.2

310 ± 18.2

21 ± 4.6

15 ± 4.7

500

82 ± 11.0

23 ± 2.1

330 ± 31.5

22 ± 4.2

19 ± 4.4

1500

89 ± 1.5

23 ± 4.0

311 ± 14.0

29 ± 5.7

16 ± 3.2

5000

85 ± 5.9 P

30 ± 2.6 P

321 ± 34.6 P

28 ± 4.2 P

16 ± 4.6 P

Positive controls, –S9

Name

ENNG

ENNG

MMC

4NQO

9AA

Concentrations

(μg/plate)

3

5

0.5

0.2

80

Mean No. of colonies/plate

(average of 3 ± SD)

439 ± 33.5

464 ± 29.4

740 ± 113.4

96 ± 5.9

1510 ± 150.1

+

0

92 ± 10.4

25 ± 4.7

339 ± 24.3

40 ± 3.2

23 ± 0.6

+

50

109 ± 11.5

23 ± 7.2

375 ± 14.6

39 ± 9.2

18 ± 2.0

+

150

99 ± 18.0

19 ± 1.5

359 ± 22.4

45 ± 1.2

24 ± 3.1

+

500

94 ± 17.7

15 ± 2.6

358 ± 23.7

45 ± 4.6

26 ± 1.5

+

1500

102 ± 9.6

23 ± 5.6

358 ± 32.1

35 ± 2.0

21 ± 5.2

+

5000

102 ± 6.8

22 ± 4.7

369 ± 72.2

39 ± 4.6

25 ± 2.1

Positive controls, +S9

Name

2AA

2AA

DAN

BP

2AA

Concentrations

(μg/plate)

1

2

10

5

2

Mean No. of colonies/plate

(average of 3 ± SD)

2112 ± 97.7

269 ± 37.8

706 ± 32.6

232 ± 19.2

217 ± 12.4

 

Table 2. Test results of experiment 2 (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

0

82 ± 8.1

21 ± 4.0

249 ± 20.7

18 ± 0.6

10 ± 2.6

50

70 ± 13.1

15 ± 2.6

269 ± 9.8

24 ± 1.5

12 ± 5.5

150

79 ± 15.5

18 ± 1.5

279 ± 34.8

24 ± 4.5

6 ± 2.3

500

81 ± 7.8

19 ± 6.4

268 ± 2.3

22 ± 6.1

7 ± 1.0

1500

88 ± 8.0

22 ± 6.1

275 ± 8.3

27 ± 7.0

13 ± 3.5

5000

75 ± 12.1 P

21 ± 3.6 P

290 ± 50.2 P

24 ± 2.9 P

10 ± 6.4 P

Positive controls, –S9

Name

ENNG

ENNG

MMC

4NQO

9AA

Concentrations

(μg/plate)

3

5

0.5

0.2

80

Mean No. of colonies/plate

(average of 3 ± SD)

679 ± 29.4

933 ± 340.6

1167 ± 40.7

188 ± 11.7

2357 ± 179.5

+

0

87 ± 21.1

10 ± 4.4

330 ± 21.2

40 ± 2.1

19 ± 3.8

+

50

86 ± 7.2

13 ± 4.0

369 ± 18.9

33 ± 4.6

17 ± 2.6

+

150

105 ± 10.4

11 ± 4.4

349 ± 27.5

42 ± 1.0

14 ± 7.1

+

500

80 ± 8.0

15 ± 6.1

364 ± 8.3

35 ± 2.5

18 ± 3.6

+

1500

85 ± 6.7

12 ± 3.0

369 ± 34.9

38 ± 2.0

18 ± 1.5

+

5000

88 ± 12.9 P

11 ± 2.5 P

365 ± 15.0 P

36 ± 8.2 P

19 ± 4.4 P

Positive controls, +S9

Name

2AA

2AA

DAN

BP

2AA

Concentrations

(μg/plate)

1

2

10

5

2

Mean No. of colonies/plate

(average of 3 ± SD)

3159 ± 145.2

313 ± 14.5

1021 ± 104.5

312 ± 32.4

222 ± 10.2

 

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

MMC = mitomycin C

4NQO = 4-nitroquinoline-N-oxide

9AA = 9-aminoacridine

2AA = 2 -aminoanthracene

BP = benzo(a)pyrene

DAN = 1,8 -dihydroxanthraquinone

P = Precipitate

 

 

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative