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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:
disregarded due to major methodological deficiencies
Study period:
No data available
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Compared to OECD test guidelines, this study has significant shortcomings, possibly affecting the outcome of the study, and therefore it is not considered reliable.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1977
Report date:
1977

Materials and methods

Test guideline
Qualifier:
no guideline followed
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
(1992)
Deviations:
yes
Remarks:
The highest recommended concentration of 5000 µg/plate was not tested; The positive controls in the absence of S9 were not acceptable; Only one plate per concentration was tested and no independent repeat was performed; recommended 5th strain not included
GLP compliance:
no
Remarks:
performed before GLP came into force.
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Tetrasodium hexacyanoferrate
EC Number:
237-081-9
EC Name:
Tetrasodium hexacyanoferrate
Cas Number:
13601-19-9
Molecular formula:
C6FeN6.4Na
IUPAC Name:
tetrasodium hexacyanoferrate
Details on test material:
- Name of test material (as cited in study report): Manox Sodium Ferrocyanide
- The substance was received from Manox Ltd. in January 1977
- Physical state: Pale yellow crystals soluble in water
- Structural formula: Na4Fe(CN)6

Method

Target gene:
Histidine gene
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium TA 1538
Metabolic activation:
with and without
Metabolic activation system:
Rat liver S9-mix induced by Aroclor 1254
Test concentrations with justification for top dose:
4, 20, 100, 500 and 2500 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Phosphate buffer, pH 7.0
- Justification for choice of solvent/vehicle: The substance is soluble in water
Controls
Untreated negative controls:
yes
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene for all tester strains
Remarks:
with S9
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 hour

NUMBER OF REPLICATIONS: No replicates, one plate per concentration

DETERMINATION OF CYTOTOXICITY
- Method: The reduction of the bacterial background lawn, the increase in the size of the microcolonies and the reduction of the revertant colonies.

OTHER EXAMINATIONS:
- The presence of precipitation of the test compound on the plates was determined.
Evaluation criteria:
At least a doubling of the mean negative control values was looked for at some concentration of the test substance if a mutagenic effect was to be suspected. Known mutagens normally produce at least a four-fold increase in one of the strains at optimal concentrations.
Statistics:
Not performed.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
other: Yes, except for TA100 and TA1537 at which no doubling compared to the mean negative control values was observed.
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
not valid
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Positive controls validity:
not valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: The substance was completely dissolved on the plates at all concentrations.

Applicant's summary and conclusion