Registration Dossier

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:
2014-12-16~2014-12-19
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
other company data
Title:
Unnamed
Year:
2015
Report Date:
2015

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
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report):Everdirect SH14(DBk112)
- Substance type:Powder
- Composition of test material, percentage of components:88.53%
- Lot/batch No.:4401
- Expiration date of the lot/batch:2016-10-31
- Storage condition of test material:Refrigeration(5 ± 3℃)

Method

Species / strain
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
S9 Mix
Test concentrations with justification for top dose:
5, 2.5, 1.25, 0.625, 0.3125 mg/plate
Controls
Negative controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
2-acetylaminofluorene
4-nitroquinoline-N-oxide
9-aminoacridine
sodium azide
benzo(a)pyrene
mitomycin C
Remarks:
with or without S9 Mix

Results and discussion

Test results
Key result
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity:
not specified
Negative controls valid:
yes
Positive controls valid:
yes

Any other information on results incl. tables

Table 1 : Genotype results for 5 strains of Salmonella typhimurium

Test strains

histidine requirement

uvrB mutation

rfa mutation

ampicillin resistance

TA98

+

+

+

+

TA100

+

+

+

+

TA102

+

Θ

+

+

TA1535

+

+

+

Θ

TA1537

+

+

+

Θ

Table 2 : Mutagenicity results for Salmonella typhimrium TA98

Test groups

Test substance (mg/plate)

Reverse mutagenicity Value (CFU/plate)

Average colony count

1

2

3

Treated with S9 Mix

Negative control

48

48

42

46 ± 3

Positive control

154

161

174

163 ± 10

Test groups

 

5

33

39

28

33 ± 6

2.5

45

31

36

37 ± 7

1.25

48

31

45

41 ± 9

0.625

41

45

36

41 ± 5

0.3125

41

45

33

40 ± 6

Not Treated with S9 Mix

Negative control

42

45

41

43 ± 2

Positive control

230

235

220

228 ± 8

Test groups

 

5

34

48

48

43 ± 8

2.5

47

41

30

39 ± 9

1.25

32

37

41

37 ± 5

0.625

47

59

41

49 ± 9

0.3125

36

32

31

33 ± 3

Table 3 : Mutagenicity results for Salmonella typhimrium TA100

Test groups

Test substance (mg/plate)

Reverse mutagenicity Value (CFU/plate)

Average colony count

1

2

3

Treated with S9 Mix

Negative control

181

189

235

202 ± 29

Positive control

1009

901

1107

1006 ± 103

Test groups

 

5

261

271

280

271 ± 10

2.5

305

247

230

261 ± 39

1.25

293

268

282

281 ± 13

0.625

263

280

263

269 ± 10

0.3125

275

274

261

270 ± 8

Not Treated with S9 Mix

Negative control

218

215

234

222 ± 10

Positive control

429

401

423

418 ± 15

Test groups

 

5

207

232

224

221 ± 13

2.5

252

213

250

238 ± 22

1.25

249

251

222

241 ± 16

0.625

265

252

240

252 ± 13

0.3125

270

303

276

283 ± 18

Table 4 : Mutagenicity results for Salmonella typhimrium TA102

Test groups

Test substance (mg/plate)

Reverse mutagenicity Value (CFU/plate)

Average colony count

1

2

3

Treated with S9 Mix

Negative control

193

172

176

180 ± 11

Positive control

276

245

265

262 ± 16

Test groups

 

5

164

146

157

156 ± 9

2.5

176

163

161

167 ± 8

1.25

148

140

121

136 ± 14

0.625

153

150

147

150 ± 3

0.3125

129

128

141

133 ± 7

Not Treated with S9 Mix

Negative control

109

116

109

111 ± 4

Positive control

356

306

294

319 ± 33

Test groups

 

5

102

126

114

114 ± 12

2.5

130

124

126

127 ± 3

1.25

129

147

129

135 ± 10

0.625

131

106

116

118 ± 13

0.3125

136

146

130

137 ± 8

Table 5 : Mutagenicity results for Salmonella typhimrium TA1535

Test groups

Test substance (mg/plate)

Reverse mutagenicity Value (CFU/plate)

Average colony count

1

2

3

Treated with S9 Mix

Negative control

13

12

13

13 ± 1

Positive control

233

220

214

222 ± 10

Test groups

 

5

10

15

13

13 ± 3

2.5

13

19

16

16 ± 3

1.25

14

10

20

15 ± 5

0.625

18

14

19

17 ± 3

0.3125

20

14

20

18 ± 3

Not Treated with S9 Mix

Negative control

14

18

10

14 ± 4

Positive control

77

80

72

76 ± 4

Test groups

 

5

16

15

12

14 ± 2

2.5

13

18

18

16 ± 3

1.25

17

14

15

15 ± 2

0.625

11

10

8

10 ± 2

0.3125

17

10

10

12 ± 4

Table 6 : Mutagenicity results for Salmonella typhimrium TA1537

Test groups

Test substance (mg/plate)

Reverse mutagenicity Value (CFU/plate)

Average colony count

1

2

3

Treated with S9 Mix

Negative control

8

5

6

6 ± 2

Positive control

376

383

361

373 ± 11

Test groups

 

5

9

5

7

7 ± 2

2.5

11

11

8

10 ± 2

1.25

8

7

7

7 ± 1

0.625

7

8

8

8 ± 1

0.3125

10

8

7

8 ± 2

Not Treated with S9 Mix

Negative control

7

6

6

6 ± 1

Positive control

354

383

371

369 ± 15

Test groups

 

5

9

9

6

8 ± 2

2.5

7

7

10

8 ± 2

1.25

8

7

6

7 ± 1

0.625

6

5

8

6 ± 2

0.3125

6

4

4

5 ± 1

Applicant's summary and conclusion

Conclusions:
No positive criteria was observed when the reverse mutation colonies of all test groups compared with the negative controls, and all test data were within the effective data range. There results indicate that no Salmonella typhimurium reverse mutation occurs.
Executive summary:

According to the OECD Guideline 471 (Bacterial Reverse Mutation Assay), the result of test is "negative response". Therefore, the hazard category in CLP is not classified.