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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:
16 Jun 2017 - 10 Jul 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Test material form:
liquid
Details on test material:
Appearance: Clear colourless liquid
Purity/Composition: UVCB
Test item storage: At room temperature protected from light
Stable under storage conditions until: 31 March 2018 (expiry date)

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
Metabolic activation:
with and without
Metabolic activation system:
Rat liver microsomal enzymes (S9 homogenate), prepared from male Sprague Dawley rats that had been injected intraperitoneally with Aroclor 1254 (500 mg/kg body weight).
Test concentrations with justification for top dose:
Dose-range Finding Test
Selection of an adequate range of doses was based on a dose-range finding test with the strains TA100 and the WP2uvrA, both with and without S9-mix. Eight concentrations, 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 µg/plate were tested in triplicate.

The highest concentration of the test item used in the subsequent mutation assays was 5000 μg/plate. At least five different doses (increasing with approximately half-log steps) of the test item were tested in triplicate in each strain in the absence and presence of S9-mix.
Vehicle / solvent:
The vehicle of the test item was Milli-Q water (Millipore Corp., Bedford, MA., USA).
Controlsopen allclose all
Untreated negative controls:
yes
Remarks:
Milli-Q water
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
2-nitrofluorene
sodium azide
methylmethanesulfonate
other: ICR-191 (Sigma), 2-aminoanthracene (2AA)
Details on test system and experimental conditions:
Salmonella typhimurium bacteria and Escherichia coli bacteria
Rationale for test conditions:
Recommended test system in international guidelines (e.g. OECD, EC).

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Untreated negative controls validity:
valid
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 nor precipitates, but tested up to recommended limit concentrations
Untreated negative controls validity:
valid
Positive controls validity:
valid

Any other information on results incl. tables

Dose-Range Finding Test: Mutagenic Response of MEA Polyborate 1:1 in the Salmonella typhimurium Reverse Mutation Assay and in the Escherichia coli Reverse Mutation Assay

Direct plate assay


Dose

(µg/plate)


Mean number of revertant colonies/3 replicate plates (
±S.D.) with one Salmonella typhimurium and one Escherichia colistrain.

 


TA100


WP2uvrA



 

Without S9-mix

Positive control

722

±

32

 

1272

±

76

Solvent control

101

±

7

 

19

±

6

1.7

99

±

7

 

19

±

1

5.4

108

±

5

 

18

±

3

17

101

±

25

 

25

±

6

52

117

±

9

 

23

±

0

164

111

±

29

 

21

±

5

512

100

±

16

 

22

±

8

1600

110

±

6

 

20

±

5

5000

128

±

11

n NP

19

±

6

n NP

With S9-mix

Positive control

475

±

106

 

420

±

13

Solvent control

92

±

6

 

29

±

10

1.7

107

±

15

 

27

±

4

5.4

95

±

12

 

25

±

4

17

110

±

12

 

28

±

8

52

111

±

16

 

29

±

11

164

112

±

14

 

24

±

5

512

107

±

9

 

27

±

6

1600

101

±

11

 

27

±

2

5000

126

±

17

n NP

38

±

4

n NP

NP

No precipitate

n

Normal bacterial background lawn

Experiment 1: Mutagenic Response of MEA Polyborate 1:1 in the Salmonella typhimurium Reverse Mutation Assay

Direct plate assay

Dose

(µg/plate)


Mean number of revertant colonies/3 replicate plates (
±S.D.) with
different strains of Salmonella typhimurium.

 


TA1535


TA1537

 


TA98

 

 Without S9-mix

Positive control

790

±

47

 

985

±

99

 

1193

±

86

 

Solvent control

11

±

2

 

6

±

2

 

16

±

3

 

52

16

±

3

 

6

±

2

 

11

±

6

 

164

11

±

4

 

4

±

1

 

15

±

5

 

512

11

±

7

 

6

±

2

 

25

±

5

 

1600

11

±

4

 

6

±

2

 

16

±

9

 

5000

6

±

4

n NP

4

±

1

n NP

16

±

2

n NP

 

 

 

 

 

 

 

 

 

 

 

 

 

 

With S9-mix

Positive control

328

±

21

 

546

±

104

 

1022

±

61

 

Milli-Q water

10

±

3

 

8

±

4

 

24

±

11

 

52

13

±

2

 

7

±

2

 

29

±

5

 

164

10

±

2

 

6

±

3

 

25

±

9

 

512

12

±

1

 

7

±

3

 

30

±

6

 

1600

13

±

3

 

5

±

0

 

24

±

3

 

5000

7

±

2

n NP

8

±

4

n NP

21

±

2

n NP

 

 

 

 

 

 

 

 

 

 

 

 

 

 

NP

No precipitate

n

Normal bacterial background lawn

Experiment 2: Mutagenic Response of MEA Polyborate 1:1 in the Salmonella typhimurium Reverse Mutation Assay and in the Escherichia coli Reverse Mutation Assay


Pre-incubation assay


Dose

(µg/plate)


Mean number of revertant colonies/3 replicate plates (
±S.D.) with
different strains ofSalmonella typhimuriumand oneEscherichia colistrain.

 

TA1535


TA1537

 


TA98


TA100


WP2uvrA

Without S9-mix

Positive control

886

±

21

 

1183

±

75

 

1211

±

169

 

708

±

171

 

202

±

35

 

 

Solvent control

7

±

2

 

9

±

1

 

20

±

5

 

118

±

4

 

36

±

9

 

 

52

13

±

1

 

8

±

2

 

34

±

21

 

114

±

13

 

25

±

10

 

 

164

12

±

2

 

8

±

4

 

19

±

7

 

104

±

51

 

29

±

4

 

 

512

15

±

5

 

8

±

3

 

18

±

4

 

112

±

20

 

31

±

4

i

 

1600

9

±

6

 

11

±

6

 

12

±

4

 

149

±

8

 

31

±

13

 

 

5000

9

±

3

n SP

5

±

2

n SP

15

±

5

n SP

179

±

17

n SP

38

±

17

n SP

 

 

With S9-mix

Positive control

217

±

3

 

177

±

16

 

495

±

86

 

1008

±

343

 

541

±

13

 

Milli-Q water

19

±

5

 

4

±

0

 

26

±

2

 

118

±

21

 

42

±

9

 

52

8

±

1

 

8

±

2

 

28

±

13

 

116

±

10

 

41

±

6

 

164

11

±

4

 

6

±

5

 

28

±

2

 

135

±

27

 

61

±

7

 

512

10

±

2

 

8

±

4

 

25

±

7

 

145

±

27

 

54

±

9

 

1600

13

±

1

 

10

±

8

 

23

±

9

 

148

±

11

 

48

±

9

 

5000

7

±

2

n SP

5

±

3

n SP

21

±

4

n SP

189

±

18

n SP

52

±

5

n SP

 

SP

Slight Precipitate

i

Plate infected

n

Normal bacterial background lawn

 

Applicant's summary and conclusion

Conclusions:
Based on the results of a GLP OECD 471 study it is concluded that MEA Polyborate 1:1 is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.