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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
Study period:
1990-08-28 to 1991-01-25
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well conducted GLP study but only 4 strains were tested.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1991
Report Date:
1991

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
4 strains tested
Principles of method if other than guideline:
Not applicable
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): 2-pentenenitrile, (Z)
- Physical state: colorless liquid
- Stability under test conditions: assumed to be stable, no evidence of instability was observed

Method

Target gene:
Histidine revertant
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA 97, TA 98, TA 100, TA 1535
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S-9 mix from male Crl:CD rat injected i.p. with Aroclor 1254
Test concentrations with justification for top dose:
0, 500, 1000, 2000, 3000, 4000 and 5000 µg/plate.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO (Baker Lot B51634)
- Justification for choice of solvent/vehicle: based on information supplied with the test material
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene (Sigma Lot 33F-0816); 2-nitrofluorene (Aldrich Lot JPO3222JJ); sodium azide (Sigma Lot 26F-0434); ICR-191 Acridine (Raylo Chemicals Lot 178)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Incubation period: 48 hours at 37 °C

NUMBER OF REPLICATIONS: 3

Evaluation criteria:
A test sample is classified as positive when the average number of revertants in any strain at any test sample concentration studied is at least two
times greater than the average number of revertants in the solvent control and there is a positive dose-response relationship in that same strain.
Statistics:
Doses with and without activation were ranked and results from a strain were analyzed individually by were multiple linear regression. Comparisons
were made between each dose/concentration and the solvent control (0 rank) using the mean square error estimate. All comparisons were at the 95 %level of confidence.

Results and discussion

Test results
Species / strain:
S. typhimurium, other: TA 1535, TA 97, TA 98, 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
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
No information
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 7.6.1/1: Results of mutagenicity assay with Cis-2-pentenenitrile and tester strain TA 1535

Concentration [µg/plate]

No. of revertants per plate**

No. of revertants per plate**

Cytotoxicity

- MA

+ MA

 

0*

14 ± 3

13 ± 5

No

500

12 ± 4

11 ± 2

No

1000

12 ± 4

11 ± 3

No

2000

13 ± 1

10 ± 5

No

3000

11 ± 1

10 ± 4

No

4000

13 ± 5

14 ± 4

No

5000

13 ± 2

16 ± 5

No

Positive control

239 ± 15

261 ± 9

 

*solvent control with DMSO

** mean of 3 plates

 

Table 7.6.1/2: Results of mutagenicity assay with Cis-2-pentenenitrile and tester strain TA 97

Concentration [µg/plate]

No. of revertants per plate**

No. of revertants per plate**

Cytotoxicity

- MA

+ MA

 

0*

102 ± 5

114 ± 18

No

500

105 ± 4

125 ± 8

No

1000

101 ± 9

126 ± 33

No

2000

98 ± 19

134 ± 14

No

3000

101 ± 7

130 ± 16

No

4000

103 ± 7

122 ± 25

No

5000

98 ± 10

108 ± 23

No

Positive control

1265 ± 34

1482 ± 72

 

*solvent control with DMSO

** mean of 3 plates

 

Table 7.6.1/3: Results of mutagenicity assay with Cis-2-pentenenitrile and tester strain TA 98

Concentration [µg/plate]

No. of revertants per plate**

No. of revertants per plate**

Cytotoxicity

- MA

+ MA

 

0*

13 ± 4

19 ± 2

No

500

12 ± 3

28 ± 4

No

1000

13 ± 6

28 ± 6

No

2000

12 ± 2

21 ± 1

No

3000

14 ± 2

27 ± 9

No

4000

14 ± 5

25 ± 2

No

5000

14 ± 4

21 ± 5

No

Positive control

1590 ± 80

2096 ± 72

 

*solvent control with DMSO

** mean of 3 plates

 

Table 7.6.1/4: Results of mutagenicity assay with Cis-2-pentenenitrile and tester strain TA 100

Concentration [µg/plate]

No. of revertants per plate**

No. of revertants per plate**

Cytotoxicity

- MA

+ MA

 

0*

100 ± 2

131 ± 4

No

500

96 ± 15

117 ± 13

No

1000

101 ± 4

110 ± 15

No

2000

104 ± 9

122 ± 9

No

3000

100 ± 11

115 ± 10

No

4000

94 ± 9

108 ± 7

No

5000

98 ± 8

114 ± 14

No

Positive control

413 ± 9

1871 ± 75

 

*solvent control with DMSO

** mean of 3 plates

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

There was no evidence of induced mutant colonies over background in this study
Executive summary:

In a reverse gene mutation assay in bacteria performed according to the OECD test guideline No. 471 and in compliance with GLP, strains TA 1535, TA 97, TA 98 and TA 100 of S. typhimurium were exposed to Cis-2-pentenenitrile (99.97 % pure) in DMSO at concentrations of 0, 500, 1000, 2000, 3000, 4000 and 5000 µg/plate in the presence and absence of mammalian metabolic activation (S-9 mix from male Crl:CD rat injected i.p. with Aroclor 1254).Cis-2-pentenenitrile was tested up to limit concentration. The positive controls induced the appropriate responses in the corresponding strains.  There was no evidence of induced mutant colonies over background.

 

This study is classified as acceptable and satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenesis (bacterial reverse gene mutation) data.