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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:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study with acceptable restrictions. No E. coli strain or equivalent S. typhimurium strain was tested; lack of data on test substance.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1988
Report Date:
1988

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
no E. coli strain or equivalent S. typhimurium strain was tested; lack of data on test substance
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): only trade name given
- Physical state: liquid
- Appearance: viscous, amber-coloured liquid
- Analytical purity: no data

Method

Target gene:
his operon
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and TA 1538
Metabolic activation:
with and without
Metabolic activation system:
Cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254.
Test concentrations with justification for top dose:
8, 40, 200, 1000 and 5000 µg/plate with and without metabolic activation
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Tween 80/bidistilled water
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: -S9: sodium azide (2 µg/plate; TA 1535, TA 100); 9-Aminoacridine (80 µg/plate; TA 1537); 4-Nitro-o-phenylenediamine (40 µg/plate; TA 1538, TA 98); +S9: 2-Aminoanthracene (2.5 µg/plate for TA 1535, TA 1537; 5 µg/plate for TA 98, TA 100, TA 1538)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: triplicates each in two independent experiments
Evaluation criteria:
The test material may be considered positive in this test system if the following criteria are met:
For the test item to be considered mutagenic, two-fold (TA 100) or three-fold (all other strains) increases in mean revertant numbers must be observed at more than one dose level. All cultures must demonstrate the characteristic mean number of spontaneous revertants in the vehicle controls.

Results and discussion

Test results
Species / strain:
S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537 and TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
ADDITIONAL INFORMATION ON CYTOTOXICITY: The results show that no cytotoxicity was caused by the test substance.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Test result of experiment 1

Bacterial Reverse Mutation Assay, mean revertants colonies/plate

EXPERIMENT 1

S9-Mix

 

Without

 

Test item (µg/plate)

TA98

TA 100

TA 1535

TA1537

TA 1538

Negative control

18.0 ± 3.5

102.3 ± 15.3

9.5 ± 2.4

5.0 ± 1.2

11.8 ± 3.2

8

13.6 ± 5.0

106.6 ± 13.0

7.3 ± 1.5

6.0 ± 2.0

9.6 ± 0.57

40

16.0 ± 3.0

99.0 ± 2.0

7.6 ± 3.5

7.0 ± 2.0

9.0 ± 1.0

200

21.0 ± 2.0

100.6 ± 8.0

10.3 ± 5.0

7.6 ± 3.5

11.6 ± 0.57

1000

20.6 ± 3.5

92.3 ± 3.2

7.6 ± 1.1

7.0 ± 1.7

8.0 ± 2.0

5000

20.0 ± 5.2

104.0 ± 11.0

8.6 ± 1.5

6.3 ± 2.5

10.6 ± 0.57

Sodium azide

---

492.0 ± 9.8

576.6 ± 31.6

---

---

9-Aminoacridine

---

---

---

372.0 ± 43.3

---

4-Nitro-o-phenylenediamine

807.0 ± 92.6

---

---

---

1970.6 ± 92.0

S9-Mix

 

With

 

Test item (µg/plate)

TA98

TA 100

TA 1535

TA1537

TA 1538

Negative control

25.0 ± 6.2

121.5 ± 11.6

8.5 ± 1.6

7.0 ± 2.0

19.8 ± 4.0

8

22.0 ± 1.7

118.3 ± 13.6

10.0 ± 3.6

6.0 ± 2.6

17.0 ± 4.0

40

19.3 ± 2.5

115.0 ± 3.0

10.3 ± 0.57

5.0 ± 2.6

15.6 ± 3.2

200

30.0 ± 0.0

122.6 ± 3.7

8.0 ± 0.0

5.3 ± 2.3

19.0 ± 8.0

1000

23.3 ± 2.0

117.3 ± 7.0

9.6 ± 2.3

10.0 ± 1.0

10.3 ± 2.0

5000

31.3 ± 8.0

126.3 ± 1.5

11.6 ± 3.7

6.0 ± 1.0

15.6 ± 1.5

2-Aminoanthracene

790.0 ± 2.6

1074.0 ± 39.8

80.3 ± 12.7

57.6 ± 3.2

1097.6 ± 82.0

Table 2: Test result of experiment 2

Bacterial Reverse Mutation Assay, mean revertants colonies/plate

EXPERIMENT 2

S9-Mix

 

Without

 

Test item (µg/plate)

TA98

TA 100

TA 1535

TA1537

TA 1538

Negative control

27.6 ± 2.5

97.1 ± 7.3

8.8 ± 2.6

8.6 ± 1.7

12.5 ± 3.7

8

21.3 ± 4.1

98.6 ± 3.0

6.3 ± 2.0

8.6 ± 0.57

11.0 ± 0.0

40

24.6 ± 12.0

111.0 ± 14.4

7.3 ± 0.57

9.0 ± 2.0

7.3 ± 3.2

200

22.6 ± 0.57

99.6 ± 0.57

8.3 ± 5.7

7.3 ± 5.0

9.3 ± 4.1

1000

26.3 ± 6.8

99.6 ± 15.9

8.6 ± 3.5

6.6 ± 0.57

10.3 ± 2.0

5000

17.3 ± 1.1

90.6 ± 4.0

4.6 ± 1.1

5.6 ± 0.57

10.6 ± 0.57

Sodium azide

---

537.6 ± 25.5

591.3 ± 32.3

---

---

9-Aminoacridine

---

---

---

578.0 ± 61.0

---

4-Nitro-o-phenylenediamine

1242.6 ± 30.2

---

---

---

2475.3 ± 75.0

S9-Mix

 

With

 

Test item (µg/plate)

TA98

TA 100

TA 1535

TA1537

TA 1538

Negative control

30.1 ± 4.1

104.0 ± 8.1

9.1 ± 3.0

6.0 ± 1.8

15.3 ± 1.8

8

32.3 ± 3.5

113.0 ± 1.7

7.6 ± 0.57

6.3 ± 3.0

18.0 ± 2.6

40

31.3 ± 7.5

103.6 ± 4.0

8.6 ± 2.3

11.0 ± 2.6

18.3 ± 4.0

200

30.3 ± 0.57

115.3 ± 10.4

7.6 ± 2.3

6.6 ± 3.2

13.0 ± 3.4

1000

31.0 ± 1.0

101.6 ± 2.5

8.0 ± 1.7

8.6 ± 2.0

15.3 ± 1.1

5000

26.6 ± 1.5

108.0 ± 5.5

6.3 ± 1.1

9.3 ± 3.0

14.0 ± 5.0

2-Aminoanthracene

755.0 ± 205.6

878.0 ± 81.2

101.0 ± 2.0

52.0 ± 14.1

1343.0 ± 80.7

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative