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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:
03 - 26 July 2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and guideline compliance, no relevant deviations

Data source

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

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)
Version / remarks:
adopted 2000
Deviations:
no
GLP compliance:
yes (incl. certificate)
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): PET-3-MP
- Physical state: liquid
- Analytical purity: 97.3%
- Purity test date: 2002-06-24
- Lot/batch No.: 2518
- Expiration date of the lot/batch: no data
- Stability under test conditions: guaranteed by the sponsor
- Storage condition of test material: approximately 4°C in the dark, under nitrogen

Method

Target gene:
his operon
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
S. typhimurium TA 102
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
rat liver S9, induced with phenobarbitone/ß-naphthoflavone (80/100 mg per kg per day)
Test concentrations with justification for top dose:
50 to 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: established vehicle for substances with poor water solubility
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
N-ethyl-N-nitro-N-nitrosoguanidine
Remarks:
without activation

Migrated to IUCLID6: 3 µg/plate for TA100 and 5 µg/plate for TA1535
Positive control substance:
9-aminoacridine
Remarks:
without activation

Migrated to IUCLID6: 80 µg/plate for TA1537
Positive control substance:
mitomycin C
Remarks:
without activation

Migrated to IUCLID6: 0.5 µg/plate for TA102
Positive control substance:
4-nitroquinoline-N-oxide
Remarks:
without activation

Migrated to IUCLID6: 0.2 µg/plate for TA98
Positive control substance:
other: 2-aminoanthracene: 1 µg/plate for TA100, 2 µg/plate for TA1535 and TA1537
Remarks:
with activation
Positive control substance:
benzo(a)pyrene
Remarks:
Migrated to IUCLID6: 5 µg/plate for TA98
Positive control substance:
other: 1,8-Dihydroxyanthraquinone, 10 µg/plate for TA102
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)


DURATION
- Preincubation period: n.a.
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3 per concentration per experiment
Evaluation criteria:
The test material may be considered positive in this test system if the following criteria are met:
The test material should have induced a reproducible, dose-related and statistically (Dunnett's method of linear regression) significant increase in the revertant count in at least one strain of bacteria.
Statistics:
Dunnett's method of linear regression

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
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: at 5000 µg/plate
- Other confounding effects:


RANGE-FINDING/SCREENING STUDIES:
The test material was non-toxic to the strain of Salmonella used (TA100).

COMPARISON WITH HISTORICAL CONTROL DATA:
pos. and neg. controls within historical range
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Experiment 1 - Without Metabolic Activation

S9 Mix

Test substance concentration (µg/ plate)

Number of revertants (mean number of colonies per plate)

Base-pair substitution type

Frameshift type

TA100

TA1535

TA102

TA98

TA1537

0

82

25

383

18

12

50

77

22

352

15

10

150

75

22

359

17

7

500

80

21

365

16

9

1500

71

21

346

18

9

5000

74 P

33 P

325 P

18 P

7 P

Pos controls
‑S9

Name

ENNG

ENNG

MMC

4NQO

9AA

Conc. (µg/plate)

3

5

0.5

0.2

80

No. of revertants per plate

270

444

1186

171

1287

ENNG N-ethyl-N'-nitro-N-nitrosoguanidine

4NQO 4-Nitroquinoline-1-oxide

9AA    9-Aminoacridine

MMC  Mitomycin C

P        Precipitate

 

 

Table 2: Experiment 1 - With Metabolic Activation

S9 Mix

Test substance concentration (µg/ plate)

Number of revertants (mean number of colonies per plate)

Base-pair substitution type

Frameshift type

TA100

TA1535

TA102

TA98

TA1537

+

0

87

13

375

46

19

+

50

84

9

365

37

13

+

150

80

10

356

35

18

+

500

77

13

345

30

14

+

1500

62

14

319

24

13

+

5000

51 P

16 P

233 P

32 P

8 P

Pos controls +S9

Name

2AA

2AA

DAN

BP

2AA

Conc. (µg/plate)

1

2

10

5

2

No. of revertants per plate

1853

351

784

225

329

2AA    2-Aminoanthracene

BP     Benzo(a)pyrene

DAN   1,8-Dihydroxyanthraquinone

P        Precipitate

 

 

Table 3: Experiment 2 - Without Metabolic Activation

S9 Mix

Test substance concentration (µg/ plate)

Number of revertants (mean number of colonies per plate)

Base-pair substitution type

Frameshift type

TA100

TA1535

TA102

TA98

TA1537

0

76

22

353

15

10

50

81

23

368

14

7

150

69

29

373

14

8

500

78

30

351

18

9

1500

90

30

343

23

8

5000

94 P

36 P

280 P

24 P

4 P

Pos controls
‑S9

Name

ENNG

ENNG

MMC

4NQO

9AA

Conc. (µg/plate)

3

5

0.5

0.2

80

Avg. no. of revertants per plate

473

449

998

110

847

ENNG N-ethyl-N'-nitro-N-nitrosoguanidine

4NQO 4-Nitroquinoline-1-oxide

9AA    9-Aminoacridine

MMC  Mitomycin C

P        Precipitate

 

 

Table 4: Experiment 2 - With Metabolic Activation

S9 Mix

Test substance concentration (µg/ plate)

Number of revertants (mean number of colonies per plate)

Base-pair substitution type

Frameshift type

TA100

TA1535

TA102

TA98

TA1537

+

0

87

15

376

30

12

+

50

75

15

395

23

15

+

150

76

17

384

22

11

+

500

69

17

382

22

9

+

1500

60

16

338

19

6

+

5000

54 P

12 P

157 P

21 P

9 P

Pos controls +S9

Name

2AA

2AA

DAN

BP

2AA

Conc. (µg/plate)

1

2

10

5

2

No. of revertants per plate

2950

168

748

152

264

2AA    2-Aminoanthracene

BP     Benzo(a)pyrene

DAN   1,8-Dihydroxyanthraquinone

P        Precipitate

 

 

 

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

PETMP is negative in the Ames test, with and without metabolic activation