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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:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2003
Report Date:
2003

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: aqueous formulation
Details on test material:
- Name of test material (as cited in study report): Genagen LAB, CAS 66455-29-6, Hoe S 3838
- Substance type: surfactant
- Physical state: colourless, liquid
- Analytical purity: not specified
- Composition of test material, percentage of components: Alkyldimethylbetaine (ca. 30 % in water)
- Lot/batch No.: DEGE 069380
- Expiration date of the lot/batch: January 2004
- Stability under test conditions: stability in water not specified
- Storage condition of test material: room temperature, protected from moisture and light
- Other: density 1.05 g/cm³ (20°C, DIN 51757), pH ca. 7 (10 g/L, 20°C), provided by sponsor

Method

Target gene:
His-operon
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA97a, TA98, TA100, TA1535, TA102
Metabolic activation:
with and without
Metabolic activation system:
Phenobarbital and β-Naphtoflavone induced rat liver S9
Test concentrations with justification for top dose:
1. Study: 0.016, 0.05, 0.16, 0.50, 1.60 mg/plate (TA97a, TA100 ± S9; TA1535 - S9); 0.05, 0.16, 0.50, 1.60, 5.00 mg/plate (TA98, TA102 ± S9; TA1535 + S9);
2. Study: 0.005, 0.016, 0.05, 0.16, 0.50 mg/plate (TA97a, TA102 ± S9; TA100, TA1535 - S9); 0.016, 0.05, 0.16, 0.50, 1.60 mg/plate (TA98, TA102 ± S9; TA100, TA1535 + S9)
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
aqua bidest
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: ICR 191, 0.5 µg/plate
Remarks:
TA97a without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
aqua bidest
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4-Nitro-o-phenylene-diamine, 0.5 µg/plate
Remarks:
TA98 without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
aqua bidest
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: Nitrofurantoine, 0.2 µg/plate
Remarks:
TA100 without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
aqua bidest
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
TA1535 without S9

Migrated to IUCLID6: 0.25 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
aqua bidest
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Aminoanthracene, 2 µg/plate
Remarks:
TA97a, TA98, TA100, TA1535 with S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
aqua bidest
True negative controls:
no
Positive controls:
yes
Positive control substance:
cumene hydroperoxide
Remarks:
TA102 without S9

Migrated to IUCLID6: 100 µg/plate
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
aqua bidest
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: Danthron, 30 µg/plate
Remarks:
TA102 with S9
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 hours


SELECTION AGENT (mutation assays): histidine prototrophy


NUMBER OF REPLICATIONS: triplicates


DETERMINATION OF CYTOTOXICITY
- Method: other: reduction of background lawn
Evaluation criteria:
The test item was interpreted as mutagenic if a concentration effect relationship occurred and the induction rate was equal to or greater than 2.
Spontaneous revertants/plate had to be within historical ranges, and the induction rates of the positive controls had to be equal to or greater than 2 to consider the test valid.
Statistics:
Arithmetic mean values and standard deviations were calculated from colonies per plate of three replicates. For evaluation of the results the induction rate of the mean values was calculated by division of the number of revertant colonies of the test item by the number of revertant colonies of the corresponding control.

Results and discussion

Test results
Species / strain:
S. typhimurium, other: TA97a, TA98, TA100, TA1535, TA102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
Lowest cytotoxic concentration without and with S9 (mg/plate): TA97a (0.5 and 1.6, resp.); TA98 (1.6 and 5.0, resp.); TA100 (1.6 without S9 only); TA102 (5.0 and 5.0, resp.); TA1535 (1.6 and 5.0, resp.)
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES:
Test concentrations were selected based on the results of a preliminary test (non-GLP)

COMPARISON WITH HISTORICAL CONTROL DATA:
The spontaneous revertants/plate (negative controls) had to be within the following ranges:
TA97a ± S9: 150 - 450;
TA98 ± S9: 15 - 50;
TA100 ± S9: 60 - 200;
TA102 ± S9: 300 - 600;
TA1535 ± S9: 5 - 30

Any other information on results incl. tables

Study 1:

Maximum number of revertants (mean ± SD):

Strain

-S9

+S9

Control

Test group (mg/plate)

Induction rate

Control

Test group (mg/plate)

Induction rate

TA97a

251 ± 10.5

288 ± 1.1 (0.16)

1.1

282 ± 15.3

252 ± 14.5 (0.16)

0.9

TA98

35 ± 7.1

33 ± 7.5 (0.05)

1.0

38 ± 6.1

39 ± 12.0 (0.16)

1.0

TA100

93 ± 8.2

104 ± 12.2 (0.016)

1.1

73 ± 8.2

82 ± 9.5 (0.5)

1.1

TA102

429 ± 8.3

444 ± 36.7 (0.5)

1.0

519 ± 16.3

519 ± 11.5 (0.16)

1.0

TA1535

18 ± 4.0

28 ± 3.1 (0.05)

1.5

7 ± 2.5

14 ± 3.6 (0.05)

1.9

 

Study 2:

Maximum number of revertants (mean ± SD):

Strain

-S9

+S9

Control

Test group (mg/plate)

Induction rate

Control

Test group (mg/plate)

Induction rate

TA97a

281 ± 26.1

239 ± 12.2 (0.05)

0.8

298 ± 41.6

273 ± 21.1 (0.05)

0.9

TA98

22 ± 10.4

41 ± 5.5 (1.6)

1.4

33 ± 2.6

35 ± 1.5 (0.5)

1.1

TA100

127 ± 14.2

111 ± 13.1 (0.016)

0.9

91 ± 8.0

91 ± 7.1 (0.5)

1.0

TA102

349 ± 20.4

355 ± 41.7 (0.05)

1.0

451 ± 34.3

462 ± 19.1 (0.5)

1.0

TA1535

29 ± 2.5

32 ± 1.7 (0.005)

1.1

11 ± 2.1

10 ± 3.1 (0.016)

0.9

 

Conclusion:

The test substance was not mutagenic to bacteria under the conditions chosen.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative