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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:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study, no data on GLP, adequate for assessment

Data source

Reference
Reference Type:
publication
Title:
Mutagenicity Test Data of Existing Chemical Substances, January 1996
Author:
JETOC
Year:
1996
Bibliographic source:
Japan Chemical Industry Ecology-Toxicology & Information Center, Japan

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
This test has been held according to the Notification stipulating the standard provided by the Minister of Labour based on the Industrial Safety and Health Law Article 57-2 Paragraph 1 (Notification No.77 of the Ministry of Labour, dated September 1, 1988) and related Notice (No.603, dated September 16, 1988) .
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): 2-Aminoethanol
- Supplier: Tokyo Kasei Kogyo Co. Ltd.
- Analytical purity: extra pure reagent
- Lot. No. AQ01

Method

Target gene:
his
Species / strainopen allclose all
Species / strain / cell type:
other: TA98, TA 100, TA 102, TA 104, TA 1535, TA 1537 and TA1538
Species / strain / cell type:
other: WP2uvrA and WP2uvrA/pKM101
Metabolic activation:
with and without
Metabolic activation system:
rat liver S9 (Sodium phenobarbital and 5,6-benzofravone-induced)
Test concentrations with justification for top dose:
50 - 5000 µg/plate
Vehicle / solvent:
water
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: see below
Details on test system and experimental conditions:
The preincubation procedure was performed as described previously (Matsushima et al., 1980). The test compound dissolved in 0.05 or 0.1 ml of solvent was supplemented with 0.5 ml of S9 mix (metabolic activation method) or 0.1M phosphate buffer pH 7.4 (direct method) and 0.1 ml of tester strains which had been cultured in nutrient broth. The mixture was incubated for 20 min at 37°C, then rapidly mixed with 2 ml of molten top agar containing 0.05 µmol/ml of L-histidine and biotin for the Salmonella test. In the E. coli test 0.05 µmol/ml of L-tryptophan was used instead of L-histidine and biotin. Then the top agar mixture was rapidly poured onto a 30 ml of Vogel -Bonner minimal agar plate. All plates were incubated for 48 hours at 37°C and the numbers of revertant colonies were scored.

Evaluation criteria:
Two-hold rule criteria was used for data evaluation (Ames et al., 1975). The chemicals are considered to be mutagenic when a dose-related increase in revertant colony count is observed and the number of revertant colonies per plate with the test substance is more than twice that of the negative control (solvent control) and when a reproducibility of test result is observed. Mutagenic potency was calculated by following equation and maximum value of mutagenic potency was expressed as a specific acitivity on the data sheet: mutagenic potency (induced revertants / mg test substance) _(number of induced revertants on the dose X - number of revertant on the solvent control) = mg of test chemical on the dose X .

Results and discussion

Test resultsopen allclose all
Species / strain:
other: TA98, TA 100, TA 102, TA 104, TA 1535, TA 1537 and TA1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
>=2000 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
other: WP2uvrA and WP2uvrA/pKM101
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
>=2000 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

control values (mean ± SD):

Tester strain

Solvent control

positive control

- S9

+ S9

- S9

+ S9

TA100

138 ± 27

139 ± 29

728 ± 196

1011 ± 210

TA1535

14 ± 5

13 ± 4

300 ± 81

255 ± 70

TA98

20 ± 8

26 ± 7

413 ± 82

404 ± 118

TA1538 (2NF)

16 ± 3

23 ± 5

376 ± 79

556 ± 216

TA1538 (4NQO)

16 ± 3

23 ± 5

343 ± 34

556 ± 216

TA1537

8 ± 2

11 ± 4

497 ± 254

196 ± 70

TA102

260 ± 49

317 ± 52

758 ± 175

1676 ± 562

TA104

269 ± 40

332 ± 48

1973 ± 755

1196 ± 252

WP2uvrA

29 ± 11

34 ± 11

273 ± 126

879 ± 177

WP2uvrA/pKM101

141 ± 43

198 ± 49

2080 ± 884

928 ± 250

 

results with 2 -aminoethanol

conc (µg/plate)

Number of revertants/plete (mean of 2 Experiments)

 

Base-substitution

Frame-shift

Strain

TA100

TA1535

WP2uvrA

TA98

TA1537

TA1539

Metabolic acitvation

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

pos control

942

462

258

172

274

1071

293

301

307

255

296

227

neg control

205

197

17

11

23

22

23

31

11

9

9

16

50

203

215

26

12

29

26

19

38

9

22

11

13

100

198

208

30

10

15

28

21

32

9

13

9

15

200

199

218

17

12

23

23

16

32

10

14

9

20

500

205

220

22

12

19

21

19

30

6

20

7

15

1000

188

191

15

19

26

26

22

28

6

17

8

8

2000

72

148

12

15

15

29

14

21

4

8

6

8

5000

0

0

0

0

0

0

0

0

0

0

0

0

Judgement

-

-

-

-

-

-

-

-

-

-

-

-

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative