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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: Guideline study
Cross-reference
Reason / purpose:
reference to same study

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1999
Reference Type:
secondary source
Title:
Phthalimide - CAS No: 85-41-6
Author:
OECD SIDS
Year:
2005
Bibliographic source:
SIDS Initial Assessment Report for 20th SIAM, UNEP Publications

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
GLP compliance:
yes
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): Phthalimide
- Analytical purity: 99.9%

Method

Target gene:
his operone
Species / strain
Species / strain / cell type:
other: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537; E. coli WP2 uvrA
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 from male rat liver, induced with phenobarbital and 5,6-benzoflavone
Test concentrations with justification for top dose:
0, 313, 625, 1250, 2500, 5000 µg/plate
Vehicle / solvent:
- Vehicle/solvent used: DMSO
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: without S9-mix: 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (TA98, TA100), sodium azide (TA1535), 9-aminoacridine (TA1537), N-ethyl-N'-nitro-N-nitrosoguanidine (WP2); with S9-mix; 2-aminoanthracene (all five strains)
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation
NUMBER OF REPLICATIONS: 2


Results and discussion

Test results
Species / strain:
other: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, and E. coli WP2 uvrA
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Remarks:
No cytotoxicity was observed up to 5000 µg/plate, the highest concentration tested
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
Phthalimide did not induce mutations in the S. typhimurium and E. coli strains. No toxicity was observed up to a concentration of 5000 µg/plate, with or without metabolic activation. The positive controls had a marked mutagenic effect, as was seen by a relevant increase in mutant colonies compared to the corresponding negative controls
Remarks on result:
other: other: Salmonella typhimurium TA 98, TA 100, TA 1535, TA 1537, and E. coli WP2 uvrA
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Results of reverse mutation test of phthalimide on bacteria (I)

 

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 100

TA1535

WP2uvrA

TA98

TA1537

-

0

162 ± 12

8 ± 2

28 ± 2

16 ± 6

8 ± 3

-

313

158 ± 7

8 ± 3

28 ± 8

21 ± 4

7 ± 2

-

625

161 ± 24

10 ± 3

33 ± 3

18 ± 3

7 ± 2

-

1250

159 ± 7

11 ± 4

29 ± 3

19 ± 3

9 ± 1

-

2500

168 ± 12

11 ± 1

32 ± 3

15 ± 3

5 ± 1

-

5000

143 ± 9

11 ± 1

27 ± 3

15 ± 2

6 ± 2

Positive

controls

- S9

Name

AF2

NaN3

ENNG

AF2

9AA

Concentrations

(µg/plate)

0.01

0.5

2

0.1

80

Number of revertants

531 ± 26

517 ± 22

392 ± 21

372 ± 32

197 ± 20

+

0

187 ± 11

15 ± 4

33 ± 6

27 ± 10

12 ± 1

+

313

188 ± 13

12 ± 3

35 ± 2

30 ± 4

7 ± 2

+

625

164 ± 30

16 ± 1

33 ± 1

30 ± 1

9 ± 2

+

1250

170 ± 20

11 ± 3

33 ± 4

20 ± 1

7 ± 2

+

2500

164 ± 18

9 ± 2

31 ± 8

27 ± 4

8 ± 1

+

5000

154 ± 9

16 ± 1

33 ± 7

27 ± 10

8 ± 2

Positive

controls

+ S9

Name

2AA

2AA

2AA

2AA

2AA

Concentrations

(µg/plate)

1

2

10

0.5

2

Number of revertants

1127±50

390±31

1555±119

299±47

178±36

 

AF2 = 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide

NaN3=Sodium azide

9AA = 9-aminoacridine

2AA = 2-Aminoanthracene

ENNG: N-ethyl-N´-nitro-N-nitrosoguanidine

Table 2: Results of reverse mutation test of phthalimide on bacteria (II)

 

With or with out 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 100

TA1535

WP2uvrA

TA98

TA1537

-

0

132 ± 10

11 ± 2

23 ±7

19 ± 6

8 ± 2

-

313

120 ± 8

11 ± 2

29 ±7

17 ± 2

7 ± 3

-

625

119 ± 6

9 ± 1

27 ±1

18 ± 3

6 ± 3

-

1250

130 ± 7

9 ± 1

20 ±1

19 ± 2

5 ± 2

-

2500

122 ±12

9 ± 1

19 ±2

23 ± 6

8 ± 2

-

5000

105 ± 5

10±2

22 ±5

15 ± 3

8 ± 3

Positive

controls

- S9

Name

AF2

NaN3

ENNG

AF2

9AA

Concentrations

(µg/plate)

0.01

0.5

2

0.1

80

Number of revertants

462 ± 13

525 ± 15

630 ± 8

355 ± 20

246 ± 34

+

0

144 ± 13

13 ± 6

27 ± 2

32 ± 3

13 ± 5

+

313

151 ± 15

19 ± 4

28 ± 2

24 ± 4

9 ± 2

+

625

154 ± 5

10 ± 2

27 ± 5

35 ± 7

13 ± 3

+

1250

142 ± 17

16 ± 5

30 ± 7

30 ± 6

11 ± 1

+

2500

139 ± 12

14 ± 3

26 ± 6

28 ± 5

8 ± 3

+

5000

138 ± 12

9 ± 1

24 ± 2

28 ± 7

9 ± 1

Positive

controls

+ S9

Name

2AA

2AA

2AA

2AA

2AA

Concentrations

(µg/plate)

1

2

10

0.5

2

Number of revertants

1033 ± 77

297 ± 32

1246 ± 70

341 ± 4

145 ±16

 

AF2 = 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide

NaN3=Sodium azide

9AA = 9-aminoacridine

2AA = 2-Aminoanthracene

ENNG: N-ethyl-N´-nitro-N-nitrosoguanidine

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation
Executive summary:

MHW Japan (1999):

Salmonella typhimurium strains TA100, TA1535, TA98, TA1537, and E.coli WP2 uvrA were treated with the test material diluted in DMSO using the Ames plate incorporation method according to OECD Guideline No. 471 at five dose levels, in duplicate, both with and without the addition of a rat liver homogenate metabolising system. The dose range was from 313 to 5000 µg/plate for both of two consecutive experiments. No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test material, either with or without metabolic activation.