Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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:
10 Sep - 05 Oct 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report Date:
1992

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
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): LUCULLUS 235/02
- Physical state / appearance: brown pasty mass
- Analytical purity: 100%
- Lot/batch No.: Lot 235/04
- Storage condition of test material: at +4°C

The test substance is a proteinic basis, whose intended use being for the preparation of reaction flavours.

Method

Target gene:
his operon
Species / strain
Species / strain / cell type:
other: TA 1535, TA 1537, TA 102, TA 98, TA 100
Metabolic activation:
with and without
Metabolic activation system:
S9 mix, S9 fraction comes from liver homogenates from rats induced with Aroclor 1254 (500 mg/kg)
Test concentrations with justification for top dose:
312.5, 625, 1250, 2500 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: distilled water
- Concentration of the test substance in solvent: 50 mg/mL
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: without S9 mix: sodium azide (TA 1535, TA 100), 9-Aminoacridine (TA 1537), 2-Nitrofluorene (TA 98), mitomycin C (TA 102); with S8 mix: 2-Anthramine (TA 1535, TA 1537, TA 98, TA 100), Danthron (TA 102)
Remarks:
with and without S9 mix
Details on test system and experimental conditions:
METHOD OF APPLICATION: direct in agar (plate incorporation) both tests without S9 mix, first test with S9 mix
preincubation method: second test with S9 mix described by Maron and Ames, 1983

DURATION
- Preincubation period: 60 minutes at 37°C
- Exposure duration: 48 to 72 hours

NUMBER OF REPLICATIONS: 2 per dose

DETERMINATION OF TOXICITY
- Method: relative total growth, preliminary test

Evaluation criteria:
For determining a positive response:
- a reproducible and significant dose relationship using a linear regression analysis, considered as significant if p <= 0.05 (for n = 18 values, the correlation coefficient must be r >= 0.47)
and/or
- a reproducible and significant increase (i.e. a doubling in the number of revertants for at least one of the tested strains when compared to that of the negative and/or solvent controls) for at least one of the tested concentrations.

A test substance is considered as non-mutagenic in this test system if the above two criteria are not fully met.
Statistics:
Biological and statistical significances were considered during the evaluation.

Results and discussion

Test results
Species / strain:
other: TA 1535, TA 1537, TA 102, 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:
valid
Positive controls validity:
valid
Additional information on results:

RANGE-FINDING/SCREENING STUDIES: preliminary toxicity test (10, 100, 1000, 2500 and 5000 µg/plate)

COMPARISON WITH HISTORICAL CONTROL DATA: yes, in the range

ADDITIONAL INFORMATION ON TOXICITY: The test substance was not toxic at the concentrations used.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

The number of revertants obtained in the presence of LUCULLUS 235/02 - Batch No 235/04, with and without S9 mix, for the 5 strains was equivalent to that of the negative (untreated) and/or solvent controls.

Table 1: Lucullus 235/02 Batch No. 235/04: Results of the first and second mutagenicity test with and without metabolic activation

1st experiment

number of revertants: mean value of negative control

number of revertants: mean value of positive control

max. number of revertants and concentration max. number appeared at [µg/plate]

without S9-mix

 

 

TA 1535

9 ± 2

 576 ± 20

12 ± 2 [625]

TA 1537

7 ± 1

 224 ± 33

10 ± 1 [2500]

TA 102

315 ± 18

1668 ± 88

 358 ± 49 [625]

TA 98

18 ± 1

159 ± 7

25 ± 4 [0]

TA 100

115 ± 3

458 ± 13

111 ± 10 [5000]

with S9-mix

 

 

 

TA 1535

12 ± 2

411 ± 57

13 ± 2 [625]

TA 1537

8 ± 2

456 ± 22

10 ± 1 [1250]

TA 102

369 ± 27

931 ± 69

400 ± 12 [5000]

TA 98

25 ± 8

1927 ± 59

21 ± 2 [625]

TA 100

125 ± 2

2104 ± 99

126 ± 12 [1250]

2nd experiment

number of revertants: mean value of negative control

number of revertants: mean value of positive control

 

max. number of revertants and concentration max. number appeared at [µg/plate]

without S9-mix

 

 

TA 1535

9 ± 2

359 ± 17

11 ± 3 [312.5, 625, 5000]

TA 1537

9 ± 2

186 ± 12

13 ± 5 [2500, 5000]

TA 102

226 ± 9

1490 ± 157

295 ± 49 [1250]

TA 98

25 ± 3

170 ± 13

31 ± 4 [0]

TA 100

105 ± 9

603 ± 66

119 ± 18 [0]

with S9-mix

 

 

 

TA 1535

12 ± 2

118 ± 9

14 ± 5 [5000]

TA 1537

7 ± 1

155 ± 19

9 ± 5 [2500]

TA 102

259 ± 3

933 ± 55

395 ± 8 [5000]

TA 98

19 ± 1

969 ± 62

20 ± 3 [5000]

TA 100

94 ± 7

1392 ± 76

111± 17 [1250]

Under the experimental conditions the test substance did not show mutagenic activity in the Ames test.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative