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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:
21 March to 04 May, 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report date:
2018

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
1997
GLP compliance:
yes
Remarks:
US FDA (21 CFR Parts 58, 210, 211, and 820) Regulations
Type of assay:
bacterial reverse mutation assay

Test material

1
Chemical structure
Reference substance name:
Octan-1-ol, reaction products with diphosphorus pentaoxide, potassium salts
EC Number:
807-789-8
Cas Number:
111062-42-1
Molecular formula:
C8-H18-O.K.O5-P2
IUPAC Name:
Octan-1-ol, reaction products with diphosphorus pentaoxide, potassium salts
Specific details on test material used for the study:
Test Article: Agent 447C; 1-octanol, reaction products with phosphorus oxide (P2Os), potassium salts (CAS 111062-42-1 ; EC 807-789-8)

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium, other: TA97A, TA98, TA100, TA1535
Species / strain / cell type:
E. coli WP2
Metabolic activation:
with and without
Metabolic activation system:
S-9, rat liver
Test concentrations with justification for top dose:
5 μL, 1.6 μL, 0.5 μL, 0.16 μL, 0.05 μL/plate

The concentrations tested were based on the OECD 471 highest recommended concentration for non-cytotoxic compounds.
Vehicle / solvent:
The test article was diluted in sterile water
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
sterile water
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
sodium azide
methylmethanesulfonate
other: 4-nitro-0-phenylenediamine (tested without S-9 metabolic activation only), 2-aminofluorene (tested with S-9 metabolic activation only), 2-aminoanthracene (tested with S-9 metabolic activation only)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)
- Cell density at seeding: the titers of each strain culture had concentrations of approximately 10E8 CFU/mL or higher

DURATION
- Exposure duration: The plates were incubated for growth at 37 ± 2°C for 48-72 hours.

NUMBER OF REPLICATIONS: 3 replicates for each test article or control were prepared.
Evaluation criteria:
Criteria for a Mutagen:
1) A reversion rate greater than 200% of the solvent control in strains TA97a and TA 100. A reversion rate greater than 300% of the solvent control in strains TA98,TA1535, and WP2.
2) Demonstration of a clear dose related response when dilutions are tested.

Criteria for a Non-Mutagen:
1) A reversion rate less than or equal to 200% of the solvent control in strains TA97a and TA 100. A reversion rate less than or equal to 300% of the solvent control in strains TA98, TA 1535, and WP2.
2) No dose related response when dilutions are tested.
Statistics:
The results were calculated using a validated computer program. Manual calculations may have differed slightly due to rounding. All results greater than 300 colony forming units (CFU) were considered estimates.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium, other: TA97a
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
At the highest test article concentration (5 µL/plate) without metabolic activation
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
E. coli WP2
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid

Any other information on results incl. tables

The criteria for acceptance of the test and criteria for determination of a mutagen were:

1) Tested strains for phenotype verification and achieved the appropriate responses.

2) All chemical controls included in the test gave the appropriate responses.

               a) for TA97a: % of control results >200 qualify as a positive

               b) for TA98: % of control results >300 qualify as a positive

               c) for TA100: % of control results >200 qualify as a positive

               d) for TA1535: % of control results >300 qualify as a positive

               e) for WP2: % of control results >300 qualify as a positive

3) The reversion rates for each tester strain were within the historical ranges (from 2016)

Applicant's summary and conclusion

Conclusions:
The test article concentrations of 5 μL, 1.6 μL, 0.5 μL, 0.16 μL, 0.05 μL/plate did not produce a two-fold or three-fold increase in the number of revertants nor produce a clear dose related response in any of the 5 tester strains (S. typhimurium tester strains TA97a, TA98, TA100, and TA1535, and E. coli test strain WP2). The spot tests showed no zone of increased reversion or of toxicity. In summary, the test article concentrations tested against the five strains did not meet the criteria for a potential mutagen.

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