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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
10 - 29 Sep 1983
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
no information on historical data, acceptance and evaluation criteria; only 1 assay performed, tested up to 8000 µg/plate

Data source

Reference
Reference Type:
other: Summary of translation of japanese report
Title:
Unnamed
Year:
1983

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
adopted in 1997
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent

Method

Target gene:
his operon, trp operon
Species / strainopen allclose all
Species / strain:
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 male rats, induced with PCB (KC-500)
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Metabolic activation system:
cofactor-supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of male rats, induced with PCB (KC-500)
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
cofactor-supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of male rats, induced with PCB (KC-500)
Test concentrations with justification for top dose:
Following concentrations were used in main experiment (preincubation):

all strains: 1, 5, 10, 50, 100, 500, 1000, 5000, 8000 μg/plate (with and without metabolic activation)
Vehicle:
- Vehicle(s)/solvent(s) used: DMSO
Controls
Negative controls:
no
Solvent controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
9-aminoacridine
benzo(a)pyrene
other: AF-2; ethyl-N-nitro-N'- nitrosoguanidine (ENNG), 2-aminoanthracene (2AA)
Details on test system and conditions:
METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period:
20 min
- Exposure duration:
48 h

NUMBER OF REPLICATIONS:
duplicate

DETERMINATION OF CYTOTOXICITY
- Method: inspection of bacterial background lawn

Results and discussion

Test results
Species / strain:
other: S. typhimurium TA 98, TA 100, TA 1535, TA 1537, TA 1538, E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
no, but tested up to precipitating concentrations
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation:
Precipitation of the test substance was observed at a concentration of 1000 µg/plate.

Any other information on results incl. tables

Table 1: Test results (experiment 1, preincubation)

With or without S9 Mix

Test substance concentration (μg/plate)

Number of revertant colonies per plate
(single and average of 2 plates)

Frameshift type

Base-pair substitution type

TA 98

TA 1537

TA 1538

TA 100

TA 1535

WP2 uvr A (pKM 101)

-

Solvent control (DMSO)

25

26

12

16

30

32

139

142

34

37

34

35

27

20

34

145

40

35

1

24

27

19

21

30

30

107

127

33

34

25

25

29

22

30

147

34

24

5

39

31

12

12

23

31

144

147

37

37

26

29

23

12

39

150

37

32

10

27

27

15

15

30

26

137

145

39

38

24

31

27

15

22

153

36

38

50

24

27

13

14

24

24

115

136

38

31

28

27

30

14

24

157

23

26

100

21

24

12

14

30

30

113

129

18

22

25

27

27

15

29

145

26

28

500

24

26

10

12

30

31

131

137

35

33

22

27

28

14

31

142

31

31

1000 P

25

29

13

16

22

29

126

141

31

35

28

33

32

19

36

155

39

38

5000 P

33

29

18

21

29

27

153

155

33

40

35

34

24

24

25

157

47

33

8000 P

29

29

-

-

148

149

-

-

29

150

Positive controls (µg/plate)

AF-2 (0.05)

9AA (80)

4NQO (0.25)

AF-2 (0.01)

ENNG (5)

AF-2 (0.05)

No. of colonies/plate

263

255

957

843

186

187

471

435

1916

2048

987

1027

246

729

188

399

2180

1066

+

Solvent control (DMSO)

34

32

19

19

31

32

180

169

18

14

28

26

30

19

32

157

9

24

1

40

44

12

12

35

36

163

158

12

14

26

29

48

12

36

152

15

31

5

42

43

9

8

36

37

136

137

19

18

24

23

43

7

38

138

16

21

10

33

35

23

17

27

27

137

142

11

14

20

24

36

11

27

147

16

27

50

41

43

13

21

24

33

150

149

14

17

23

28

45

28

41

148

20

32

100

34

34

19

21

33

35

140

151

19

19

35

34

34

23

36

162

19

32

500

35

38

10

12

43

46

135

136

16

13

33

29

41

14

48

137

9

25

1000 P

44

45

16

17

39

41

156

160

15

17

41

40

45

17

42

163

18

39

5000 P

24

41

15

17

38

43

149

152

11

13

33

33

40

18

47

155

15

33

8000 P

45

46

-

-

150

151

-

-

47

151

Positive controls (µg/plate)

B(a)P (5)

2AA(5)

2AA(5)

No. of colonies/plate

853

801

289

320

270

276

1051

1021

764

780

256

245

749

350

281

990

796

234

2AA = 2-aminoanthracene

4NQO = 4-nitroquinoline-N-oxide

9AA = 9-aminoacridine

B(a)P = benzo(a)pyrene

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

P = precipitate

Applicant's summary and conclusion

Conclusions:
Based on the results of the conducted study the test substance did not exhibit mutagenic properties in bacterial cells.