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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
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 item: Golpanol ALS wasserfrei
Test item No.: 09/0505-1
Batch identification: 04749356P0
Purity/composition: Mixture, contents of main components: Golpanol ALS 71.3 %, NaCl 26.0 % (for details see analytical data, study code 09L00332)
Physical state, appearance: Solid, white
Storage conditions: Room temperature

Method

Target gene:
His: Salmonella
Trp: E. coli
Species / strain
Species / strain / cell type:
other: Salmonella typhimurium TA 1535, TA 100, TA 1537, TA 98 and E. coli WP2 uvrA
Metabolic activation:
with and without
Metabolic activation system:
S-9 mix
Test concentrations with justification for top dose:
0; 20; 100; 500; 2 500 and 5 000 μg/plate (SPT)
0; 312.5; 625; 1 250; 2 500 and 5 000 μg/plate (PIT)
Vehicle / solvent:
Water
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Positive control substance:
other: see "Details on test system"
Details on test system and experimental conditions:
Positive controls:
With S9 mix:
• 2-aminoanthracene (2-AA)
- 2.5 μg/plate, dissolved in DMSO
- strains: TA 1535, TA 100, TA 1537, TA 98
- 60 μg/plate, dissolved in DMSO
- strain: Escherichia coli WP2 uvrA
Without S9 mix:
• N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)
- 5 μg/plate, dissolved in DMSO
- strains: TA 1535, TA 100
• 4-nitro-o-phenylenediamine (NOPD)
- 10 μg/plate, dissolved in DMSO
- strain: TA 98
• 9-aminoacridine (AAC)
- 100 μg/plate, dissolved in DMSO
- strain: TA 1537
• 4-nitroquinoline-N-oxide (4-NQO)
- 5 μg/plate, dissolved in DMSO
- strain: E. coli WP2 uvrA

Number of Replications: 3 test plates per dose or per control
Evaluation criteria:
Generally, the experiment is considered valid if the following criteria are met:
• The number of revertant colonies in the negative controls was within the range of the historical negative control data for each tester strain
• The sterility controls revealed no indication of bacterial contamination.
• The positive control items both with and without S9 mix induced a distinct increase in the number of revertant colonies within the range of the historical positive control data or above.
• The titer of viable bacteria was ≥ 10e8/mL.
The test item is considered positive in this assay if the following criteria are met:
• A dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontaneous mutation rate in at least one tester strain either without S9 mix or after adding a metabolizing system.
A test item is generally considered non-mutagenic in this test if:
• The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in at least two experiments carried out independently of each other.

Results and discussion

Test results
Species / strain:
other: Salmonella typhimurium TA 1535, TA 100, TA 1537, TA 98 and E. coli WP2 uvrA
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: see "Additional information on results"
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
- Solubility: No test item precipitation was found with and without S9 mix.
- Toxicity: A weak bacteriotoxic effect (slight decrease in the number of his+ revertants) was observed in the standard plate test only with the strain TA 1535 at 5 000 μg/plate. In the preincubation assay no bacteriotoxicity (reduced his- or trp- background growth, decrease in the number of his+ or trp+ revertants, reduction in the titer) was observed up to the highest required concentration.
-Conclusion: The test substance Golpanol ALS wasserfrei is not a mutagenic test item in the bacterial reverse mutation test in the absence and the presence of metabolic activation. See table 1 for further information
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Number of revertants in the control or after treatment with test substance indicated as maximum revertant factor in the a.) Standard Plate Test and b.) Preincubation Test

a.) Standard plate test (20 - 5000 µg/plate)
Strain Metabolic activation system mean revertants in Controls maximum revertant factor dose dependency Assessment
TA 1535 no 18 1.0 no negative
yes 18 1.1 no negative
TA 100 no 113 1.0 no negative
  yes 115 1.2 no negative
TA 1537 no 7 1.1 no negative
yes 9 1.0 no negative
TA 98 no 29 1.0 no negative
  yes 34 0.9 no negative
E. coli WP2 uvrA no 43 1.0 no negative
  yes 48 1.1 no negative

b.) Preincubation test (312.5 - 5000 µg/plate)

Strain Metabolic activation system mean revertants in Controls maximum revertant factor dose dependency Assessment
TA 1535 no 17 1.1 no negative
yes 18 1.1 no negative
TA 100 no 98 1.0 no negative
  yes 110 1.0 no negative
TA 1537 no 9 0.9 no negative
yes 10 1.1 no negative
TA 98 no 28 1.2 no negative
  yes 37 1.0 no negative
E. coli WP2 uvrA no 35 1.1 no negative
  yes 40 1.1 no negative

Applicant's summary and conclusion