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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:
23 April 1991 to 28 May 1991
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
other: Ames et al. (1975)
Deviations:
no
Principles of method if other than guideline:
not applicable
GLP compliance:
yes (incl. certificate)
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): Diacid 1550
- Physical state: Liquid
- Lot/batch No.: Confidential

Method

Target gene:
His-gene (HisG 46, HisC 3076, HisD 3052)
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
not applicable
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
S. typhimurium TA 1538
Details on mammalian cell type (if applicable):
not applicable
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
mammalian liver S9
Test concentrations with justification for top dose:
2.5 and 8 µL of a 10 mg/L solution
2.5, 8, and 25 µL of a 100 mg/L
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Dimethylsulfoxide
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
other: 2-anthramine, N-methyl-N-nitro-N-nitrosoguanidine
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: not applicable
- Exposure duration: 48 ±4 hours

SELECTION AGENT (mutation assays): Agar containing Histidine

NUMBER OF REPLICATIONS: experiments performed in triplicate

NUMBER OF CELLS EVALUATED: no data available

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth (number of revertant colonies per plate) in TA100
Evaluation criteria:
The average number of revertant colonies (± the standard deviation) per plate is tabulated. A positive reponse is indicated if a sample substance produces a consistent bacterial response which is two times that of the solvent or spontaneous reversions for TA 98 and TA 100. For the strains TA 15335, TA 1537 and TA 1538, a sample producing a consistent bacterial reponse three times that of the solvent or spontaneous control is indicated as positive.
Statistics:
No information available

Results and discussion

Test resultsopen allclose all
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 nor precipitates, but tested up to recommended limit concentrations
Remarks:
100 mg/mL induced cytotoxicity in TA 100
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not determined
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: sample was not soluble above 100 mg/mL in dimethylsulfoxide

RANGE-FINDING/SCREENING STUDIES: Sample was diluted in dimethylsulfoxide and tested for cytoxicity in TA 100; 100 mg/mL induced cytotoxicity
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Average test results per strain ± standard deviation with activation:

Tester Strain

Solvent control

Positive control

10 mg/mL

100 mg/mL

2.5 µL

8 µL

2.5 µL

8 µl

25 µl

TA 98

28±2

3842±935

28±5

23±6

28±6

30±8

28±6

TA 100

88±3

2894±176

82±12

75±15

56±9

61±16

53±13

TA 1535

13±1

589±200

14±2

8±3

8±2

11±6

11±3

TA 1537

15±2

135±32

14±4

19±0

31±5

14±6

2±1

TA 1538

28±3

461±254

24±3

16±5

24±3

20±5

41±25

 

Average test results per strain ± standard deviation without activation:

Tester Strain

Solvent control

Positive control

10 mg/mL

100 mg/mL

2.5 µL

8 µL

2.5 µL

8 µl

25 µl

TA 98

14±5

1443±93

10±3

16±3

11±4

11±4

11±5

TA 100

43±2

2764±79

33±7

29±4

32±10

32±10

27±6

TA 1535

32±4

1386±33

26±6

20±3

4±3

5±2

8±5

TA 1537

38±16

1973±606

42±7

19±6

10±3

4±2

3±2

TA 1538

28±20

1024±213

6±1

4±2

4±2

2±2

2±1

 

Applicant's summary and conclusion

Conclusions:
In this Ames test, Diacid 1550 did not show any mutagenic activity in the Salmonella Typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538 with and without activitation under the conditions of this test.
Executive summary:

In this study the ability of Diacid 1550 to induce mutations in the Salmonella Typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538 was investigated. The study procedures used in this study were based on the Ames Test as described by Ames et al. (1975).

The samples were diluted in dimethylsulfoxide and were incubated on agar plates for 48 ±4 hours at 37 °C (plate incorporation assay). After incubation, the agar plates were examined for the average number of revertant colonies. Prior to the Ames test, a cytoxicity test was performed in TA100 restulting in a cytoxic dose of 100 mg/mL.

Diacid 1550 dit not show any mutagenic activity up to limit concentrations in the Salmonella Typhimurium strains TA98, TA100, TA1535, TA1537 and TA1538 with and without activitation under the conditions of this test.