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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 based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
May and June 1997
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP-Guideline study with acceptable restrictions (limited data on test substance available)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report Date:
1997

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
limited data on test substance
Qualifier:
according to
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
Deviations:
yes
Remarks:
limited data on test substance
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
yes
Remarks:
limited data on test substance
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Molecular formula (if other than submission substance): C9H11O6-R3 (approximately C60H116O6), R (cannot be quantified)
- Physical state: yellow liquid
- Analytical purity: not indicated, treated as 100% pure
- Lot/batch No.: N558876
- Storage condition of test material: at 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:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
co-factor supplemented post-mitochondrial fraction (S9 mix), prepared from rats treated with Aroclor
Test concentrations with justification for top dose:
3, 10, 33, 100, 333, 1000, 3330 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
TA 1535 (-S9 mix), 1µg/plate

Migrated to IUCLID6: 1 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
TA 1537 (-S9 mix), 60 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: daunomycine
Remarks:
TA98 (-S9 mix), 4 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Remarks:
TA100 (-S9 mix), 650 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4-nitroquinoline
Remarks:
WP2uvrA (-S9 mix) , 10 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene
Remarks:
for all strains (+S9 mix); 1 µg/ plate (TA 1535, TA98, TA 100), 2.5 µg/plate (TA1537), 5 µg/plate (WP2 uvrA)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 h

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
Remarks:
at 3330 µg/plate and above
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Remarks:
at 3330 µg/plate and above
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
ADDITIONAL INFORMATION ON CYTOTOXICITY: No cytotoxicity was observed up to the maximum dose tested.
COMPARISON WITH HISTORICAL CONTROL DATA: The number of revertants for TA1537 and WP2 uvr A (+S9) were just outside the historical control range in the second experiment which was not considered to affect the validity of the study.
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 experiment 1:

With or without S9-Mix

Test substance concentration

(μg/plate)

Mean number of revertant colonies per plate

(average of 3 plates)

Base-pair substitution type

Frameshift type

TA 1535

TA1537

TA98

TA100

WP2 uvrA

-

Vehicle1

11

5

17

87

15

-

3

 

 

 

84

17

-

10

 

 

 

89

20

-

33

11

3

13

97

19

-

100

10

3

15

94

18

-

333

14

4

12

83

17

-

1000

13

4

17

103

19

-

33302

11

5

18

104

15

-

50002

 

 

 

98

16

Positive

controls

- S9

Name

SA

9AA

DM

MMS

4NQO

Concentrations

(μg/plate)

1

60

4

650

10

Number of colonies/plate

332

590

603

844

1829

+

Vehicle1

13

5

13

90

18

+

3

 

 

 

102

22

+

10

 

 

 

98

19

+

33

13

6

18

95

16

+

100

6

6

20

75

16

+

333

12

5

19

96

24

+

1000

16

7

14

102

17

+

33302

12

5

21

105

20

+

50002

 

 

 

91

17

Positive

controls

+ S9

Name

2AA

2AA

2AA

2AA

2AA

Concentrations

(μg/plate)

1

2.5

1

1

5

Number of colonies/plate

1594

227

1472

1273

734

1) 0.1 mL DMSO

2) precipitate visible

SA: sodium azide

4NQO: 4-nitroquinoline

9AA : 9-aminoacridine

DM : daunomycine

MMS: methylmethanesulfonate

2-AA: 2-aminoanthracene

 

 

Table 2: Results of experiment 2:

With or without S9-Mix

Test substance concentration

(μg/plate)

Mean number of revertant colonies per plate

(average of 3 plates)

Base-pair substitution type

Frameshift type

TA 1535

TA1537

TA98

TA100

WP2uvrA

-

Vehicle1

7

5

17

74

22

-

33

7

2

15

76

19

-

100

6

5

11

72

22

-

333

7

6

17

76

20

-

1000

4

4

16

73

23

-

33302

7

6

17

79

13

Positive

controls

- S9

Name

SA

9AA

DM

MMS

4NQO

Concentrations

(μg/plate)

1

60

4

650

10

Number of colonies/plate

230

266

788

793

1040

+

Vehicle1

6

2

18

77

25

+

33

7

3

14

72

19

+

100

8

4

16

82

18

+

333

9

5

15

82

16

+

1000

6

5

16

75

18

+

33302

9

3

11

73

19

Positive

controls

+ S9

Name

2AA

2AA

2AA

2AA

2AA

Concentrations

(μg/plate)

1

2.5

1

1

5

Number of colonies/plate

224

466

1165

1874

303

1) 0.1 mL DMSO

2) precipitate visible

SA: sodium azide

4NQO: 4-nitroquinoline

9AA : 9-aminoacridine

DM : daunomycine

MMS: methylmethanesulfonate

2-AA: 2-aminoanthracene

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative