Registration Dossier

Diss Factsheets

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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study, GLP study (only TA 98, TA 100, TA 1535 and TA 1537 tested)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report date:
1992

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
(adopted 1983)
Deviations:
no
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
(adopted 1997)
Deviations:
yes
Remarks:
(only 4 strains; only 2-aminoanthracene as positive control with metabolic activation)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Suberonitrile
EC Number:
211-089-2
EC Name:
Suberonitrile
Cas Number:
629-40-3
Molecular formula:
C8H12N2
IUPAC Name:
suberonitrile
Details on test material:
- Name of test substance (as cited in study report): Korksaeuredinitril
- Analytical purity: 99.18%
- Substance No.: 91/319
- Batch No.: KW 7376 107-5 P 5
- Physical state: yellowish liquid
- Storage: room temperature

Method

Target gene:
His-operon
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
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:
20, 100, 500, 2500, 5000 µg/plate
Vehicle / solvent:
DMSO
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: (+ S9): 2-aminoanthracene (2AA; all strains); (- S9): N-methyl-N'-nitro-N-nitrosoguanidine (MNNG; TA 100 and TA 1535), 4-nitro-o-phenylendiamine (NOPD; TA 98), 9-aminoacridine chloride monohydrate (AAC; TA 1537)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation): experiment 1

DURATION
- Exposure duration: 48 h (37 °C in the dark)

METHOD OF APPLICATION: preincubation: experiment 2

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h (37 °C in the dark)

NUMBER OF REPLICATIONS: 3 plates per dose per experiment

DETERMINATION OF CYTOTOXICITY
- Method: reduction in the number of revertant colonies; clearing or diminution of the background lawn; degree of survival of treated cultures
Evaluation criteria:
In general, a substance has to fulfill the follwoing requirements to be characterized as positvie in the Ames test:
- doubling of the spontaneous mutation rate
- dose-response relationship
- reproducibility of the results.

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:
cytotoxicity
Remarks:
(TA 1537 and TA 100 (5000 µg/plate, (-S9); TA 100 (2500 µg/plate, (+S9))
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: no precipitation occurred up to a concentration of 5000 µg/plate.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Standard plate test

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

TA98

TA1537

DMSO

133 ± 5

16 ± 6

35 ± 4

12 ± 2

20

125 ± 17

16 ± 1

28 ± 2

9 ± 3

100

123 ± 15

13 ± 3

20 ± 3

8 ± 1

500

139 ± 15

11 ± 2

23 ± 5

7 ± 3

2500

102 ± 1

14 ± 2

29 ± 8

9 ± 6

5000

83 ± 6

12 ± 2

29 ± 2

5 ± 3

Positive controls, –S9

Name

MNNG

MNNG

NOPD

AAC

Concentrations

(μg/plate)

5

5

10

100

Mean No. of colonies/plate

(average of 3 ± SD)

1187 ± 49

1220 ± 69

890 ± 200

683 ± 137

+

DMSO

142 ± 6

17 ± 3

38 ± 5

15 ± 3

+

20

126 ± 15

18 ± 2

48 ± 4

12 ± 3

+

100

130 ± 12

15 ± 5

42 ± 8

6 ± 1

+

500

95 ± 18

18 ± 4

37 ± 9

9 ± 2

+

2500

77 ± 7

15 ± 4

31 ± 2

12 ± 6

+

5000

96 ± 16

16 ± 3

27 ± 7

9 ± 2

Positive controls, +S9

Name

2AA

2AA

2AA

2AA

Concentrations

(μg/plate)

2.5

2.5

2.5

2.5

Mean No. of colonies/plate

(average of 3 ± SD)

1393 ± 29

185 ± 69

680 ± 34

106 ± 8

MNNG = N-methyl-N'-nitro-N-nitrosoguanidine

NOPD = 4-nitro-o-phenylenediamine

AAC = 9-aminoacridine

2AA = 2-aminoanthracene

 

Table 2: Preincubation test

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

TA98

TA1537

DMSO

140 ± 9

22 ± 2

27 ± 2

10 ± 4

20

139 ± 12

24 ± 5

28 ± 5

12 ± 1

100

151 ± 5

22 ± 3

28 ± 6

11 ± 1

500

129 ± 16

23 ± 4

27 ± 3

8 ± 1

2500

131 ± 15

20 ± 4

27 ± 7

11 ± 3

5000

122 ± 6

21 ± 6

28 ± 4

9 ± 1

Positive controls, –S9

Name

MNNG

MNNG

NOPD

AAC

Concentrations

(μg/plate)

5

5

10

100

Mean No. of colonies/plate

(average of 3 ± SD)

915 ± 75

601 ± 67

1247 ± 6

982 ± 227

+

DMSO

130 ± 7

20 ± 3

40 ± 6

13 ± 3

+

20

131 ± 20

23 ± 5

38 ± 4

13 ± 5

+

100

143 ± 20

20 ± 4

35 ± 3

12 ± 3

+

500

120 ± 11

22 ± 4

42 ± 2

13 ± 1

+

2500

103 ± 2

21 ± 3

44 ± 3

12 ± 2

+

5000

120 ± 18

21 ± 1

34 ± 4

9 ± 2

Positive controls, +S9

Name

2AA

2AA

2AA

2AA

Concentrations

(μg/plate)

2.5

2.5

2.5

2.5

Mean No. of colonies/plate

(average of 3 ± SD)

877 ± 36

163 ± 20

536 ± 8

149 ± 13

MNNG = N-methyl-N'-nitro-N-nitrosoguanidine

NOPD = 4-nitro-o-phenylenediamine

AAC = 9-aminoacridine

2AA = 2-aminoanthracene

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Categories Display