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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:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
06 Feb 1996 - 16 Feb 1996
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions, only four strains tested.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report Date:
1996

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
adopted May 23, 1983
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
adopted December, 1992
Deviations:
no
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): Prisorine 3505
- Physical state: liquid
- Analytical purity: not indicated by sponsor; treated as 100 % pure
- Lot/batch No.: N541439
- Stability under test conditions: stable
- Storage condition of test material: at room temperature in the dark
- Other: pale yellow

Method

Target gene:
his operon
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
rat liver microsomal enzymes from adult male Wistar rats (S9-mix) obtained from Aroclor 1254-induced animals.
Test concentrations with justification for top dose:
3, 10, 33, 100, 333, 1000, 3330, 5000 μg/plate (range finder)
3, 10, 33, 100, 333, 1000, 3330 μg/plate (main study)
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
Saline, DMSO
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: -S9-mix: sodium azide(1 µg/plate), 9-aminoacridine (60 µg/plate), daunomycine (4 µg/plate), methylmethanesulfonate (650 µg/plate); +S9-mix: 2-aminoanthracene (0.5 and 5 µg/plate)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in top agar (plate incorporation)

DURATION
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: triplicate in two experiments

Evaluation criteria:
A test substance is considered negative (not mutagenic) in the test if:
a) The total number of revertants in any tester strain at any concentration is not greater than two times the solvent control value, with or without metabolic activation.
b) The neagtive response should be reproducible in at least one independently repeated experiment.

A test substance is considered positive (mutagenic) in the test if:
a) It reduces at least 2-fold, dose related increase in the number of revertants with respect tot he number induced by the solvent control in any of the tester strains, either with or without metabolic activation. However, any mean plate count of less than 20 is considered to be not significant.
b) The positive response should be reproducible in at least one independently repeated experiment.

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Prisorine 3505 precipitated in the top agar at 1000 and 3330 µg/plate. Precipitation of Prisorine 3505 on the plated was observed at the start and at the end of the incubation period at the concentration of 3330 µg/plate in all tester strains.


RANGE-FINDING/SCREENING STUDIES:
Prisorine 3505 was tested in strain TA 100 with concentrations of 3, 10, 33, 100, 333, 1000, 3330 and 5000 µg/plate in the absence and presence of S9-mix.
Prisorine 3505 precipitated in the top agar at 1000 µg/plate and upwards. Precipitation of Prisorine 3505 was observed at the start and at the end of the incubation period at 3330 and 5000 µg/plate in tester strain TA 100.
No reduction of the bacterial lawn was observed.
In the absence of S9-mix a decrease in the number of revertants, less than 50% compared to the mean number of revertants of the solvent control, was observed at 33 µg/plate and upwards.
In the presence of S9-mix a decrease in the number of revertants was observed at 5000 µg/plate.
Based on these data, Prisorine 3505 was tested in the mutation assay up to and including a concentration of 3330 µg/plate in the absence and presence of S9-mix.


ADDITIONAL INFORMATION ON CYTOTOXICITY:
In the absence of S9-mix a decrease in the number of revertants, less than 50% compared to the mean number of revertants of the solvent control, was observed at 33 µg/plate and upwards in tester strain TA100, and at 1000 µg/plate and upwards in tester strain TA98.
In the presence of S9-mix a decrease in the number of revertants was observed at 333 µg/plate and upwards in tester strain TA100, and at 1000 µg/plate and upwards in tester strain TA 98.
All other concentraitons showed no decrease in the amount of revertants. The tester strains 1535 and TA 1537 showed no decrease in the amount of revertants.
The bacterial background lawn was not reduced at all concentrations tested.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Mutagenic response of Prisorine 3505 in the Salmonella thyphimurium reverse mutation assay

Experiment 1

Dose (µg/plate)

Mean number of revertants/plate ± SD (n=3)

 

TA1535

TA 1537

TA 98

TA 100

 

Without S9-mix

3

 

 

 

81 ± 3

10

 

 

 

87 ± 10

33

11 ± 4

8 ± 1

19 ± 2

65 ± 3

100

11 ± 3

8 ± 2

24 ± 2

64 ± 6

333

9 ± 3

7 ± 1

26 ± 242

60 ± 11

1000

8 ± 3

4 ± 2

11 ± 3

53 ± 11

3330 SP

8 ± 2

6 ± 2

14 ± 4

43 ± 5

5000 SP

 

 

 

49 ± 7

Solvent control1

8 ± 1

4 ± 1

24 ± 3

93 ± 5

Positive control

149 ± 13

149 ± 25

301 ± 50

908 ± 24

 

Dose (µg/plate)

Mean number of revertants/plate ± SD (n=3)

 

TA1535

TA 1537

TA 98

TA 100

 

With S9-mix

3

 

 

 

86 ± 8

10

 

 

 

77 ± 4

33

10 ± 2

5 ± 1

27 ± 0

88 ± 6

100

7 ± 1

7 ± 1

29 ± 6

76 ± 6

333

9 ± 3

4 ± 2

26 ± 3

67 ± 2

1000

8 ± 3

5 ± 2

24 ± 5

70 ± 5

3330 SP

6 ± 2

5 ± 2

24 ± 3

70 ± 5

5000 SP

 

 

  

61 ± 15

Solvent control1

11 ± 2

6 ± 1

27 ± 5

88 ± 5

Positive control

119 ± 21

236 ± 36

635 ± 21

997 ± 20

 

Experiment 2

Dose (µg/plate)

Mean number of revertants/plate ± SD (n=3)

 

TA1535

TA 1537

TA 98

TA 100

 

Without S9-mix

33

12 ± 3

8 ± 7

20 ± 3

57 ± 1

100

10 ± 6

7 ± 3

18 ± 4

51 ± 10

333

7 ± 3

8 ± 5

19 ± 3

47 ± 7

1000

8 ± 6

5 ± 2

16 ± 2

38 ± 12

3330 SP

11 ±3

5 ± 1

21 ± 92

33 ± 5

Solvent control1

12 ± 5

6 ± 2

23 ± 2

70 ± 7

Positive control

192 ±29

217 ± 23

455 ± 84

1030 ± 58

 

Dose (µg/plate)

Mean number of revertants/plate ± SD (n=3)

 

TA1535

TA 1537

TA 98

TA 100

 

With S9-mix

33

8 ± 1

7 ± 2

23 ± 7

68 ± 4

100

11 ± 5

8 ± 4

22 ± 7

69 ± 6

333

12 ± 3

5 ± 1

22 ± 7

58 ± 11

1000

8 ± 5

6 ± 3

13 ± 1

49 ± 3

3330 SP

8 ± 5

4 ± 0

16 ± 3

41 ± 2

Solvent control1

11 ± 4

7 ± 82

32 ± 17

77 ± 7

Positive control

354 ± 46

797 ± 24

711 ± 47

1151 ± 80

 

10.1 mL DMSO

2Plate infected

SP: Test substance precipitated slightly on the plates

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Based on the results of this study it is concluded that Prisorine 3505 is not mutagenic in the Salmonella typhimurium reverse mutation assay.