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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1984-08-13 to 1985-01-07
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study followed standard methodology, but not all details of methodology included in report.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1985
Report Date:
1985

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
other: as described in Ames, B.N. et al, Mutation Research 31: pp. 347-364, 1975
GLP compliance:
yes (incl. certificate)
Remarks:
GLP Regulations stated in Federal Register, Vol. 43, No. 247, 12/22/1978.
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): AN 701
- Physical state: a yellow-brown liquid

Method

Target gene:
Histidine locus in selected strains
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:
Aroclor 1254 induced rat liver S-9 fraction
Test concentrations with justification for top dose:
2.5 x 10-2, 5 x 10-3, 2.5 x 10-3, 5 x 10-4, 2.5 x 10-4 mg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: acetone
- Justification for choice of solvent/vehicle: solubility of test article was best in this solvent
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
not specified
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)
NUMBER OF REPLICATIONS: three

Evaluation criteria:
Positive response would be an increase in mutant colonies greater than three times the rate seen in the solvent control for that test strain. In the case of TA 98, the solvent control test with activation showed a rate below the spontaneous rate and the historical control. For that test, the spontaneous rate was used for comparison rather than the solvent control results.

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:
other: The highest dose rate (2.5 x 10-2 mg/plate) showed cytotoxicity for TA 1535, TA 1537, and TA 100 with and without metabolic activation.
Vehicle controls validity:
other: Solvent control data acceptable, except for TA 98 with activation. Mutation rate was below spontaneous and historical control for that test. Spontaneous rate used for comparison in that test.
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

Mutant colony data for Salmonella typhimurium Ames test for AN 701

Strain

Activation

Solvent Control

Positive Control

2.5 x 10-2 mg/plate

5 x 10-3 mg/plate

2.5 x 10 -3 mg/plate

5 x 10-4 mg/plate

2.5 x 10 -4 mg/plate

TA1535

-

+

6

4

35*

28*

6

4

7

5

4

5

6

8

8

8

TA1537

-

+

6

9

116*

718*

6

10

7

10

10

10

9

8

8

7

TA98

-

+

9

15**

275*

628

12

9

11

9

21

17

24

19

17

12

TA100

-

+

89

98

3175*

TNTC*

110

111

110

104

116

111

107

113

95

100

                 

* > 3 times solvent control

** Spontaneous rate

TNTC too numerous to count

Applicant's summary and conclusion

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

Test article did not induce a dose related increase in mutant colonies in any Salmonella typhimurium strains tested (TA 1535, TA1537, TA98, TA100) with or without metabolic activation. Therefore, AN 701 was not genetically active in the Salmonella/Microsomal Assay.
Executive summary:

Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 were tested in a plate incorporation Ames test in the presence and absence of metabolic activation (S-9 fraction from livers of rats induced with Arocolor 1254) with test article dissolved in acetone. Dose levels were 2.5 x 10-2, 5 x 10-3, 2.5 x 10-3, 5 x 10-4, 2.5 x 10-4 mg AN 701/plate. An 701 was soluble at all dose levels tested. Each dose was treated in triplicate. An untreated control, solvent control, and positive control (appropriate to the strain and activation state) were tested concurrently. The highest concentration of AN 701 demonstrated toxicity in strains TA 1535, TA 1537, and TA 100 with and without metabolic activation. AN 701 did not induce a dose related increase in mutant colonies in any strains with or without activation.