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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
Study period:
04 July 2002 - 21 February 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to OECD, EU, US FDA and Japanese (EPA, MHLW and METI) test guidelines, and in compliance with GLP.

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)
Version / remarks:
(Commission Directive 2000/32/EC; L136, 57-64, Annex 4D)
Deviations:
no
Qualifier:
according to
Guideline:
other: US FDA Administration Center for Food Safey & Applied Nutrition Office of Premarket Approval, Redbook 2000, Toxicological Principles for the Safety of Food Ingredients, IV.C.1.a Bacterial Reverse Mutation Test, July 07, 2000.
Deviations:
no
Qualifier:
according to
Guideline:
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Version / remarks:
"Kanpoan 287 - EPA", "Eisei 127 - MHLW", and "Heisei 09/10/31 Kikyoku 2 - METI"
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: liquid
Details on test material:
- Name of test material (as cited in study report): MTF
- Substance type: organic
- Physical state: liquid
- Analytical purity: 99.1%
- Lot/batch No.: 010920
- Stability under test conditions: > 3 days in water
- Storage condition of test material: Approx. 4°C. The container has to be tightly sealed with nitrogen. Each time, the test item is used, the container has to be refilled with nitrogen.

Method

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 102
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
rat-liver post mitochondrial supernatant (S9 fraction)
Test concentrations with justification for top dose:
Range finding test:
strains S. typhimurium TA 100 and E. coli WP2 uvrA (with and without methabolic activation): 20.6, 61.8, 185.2, 555.6, 1666.7 and 5000 µg/plate


without metabolic activation:
strains TA 100, TA 102: 1.0, 3.3, 10, 33 and 100 µg/plate
strain TA 1537: 10, 33, 100, 333 and 1000 µg/plate
strains TA 1535, TA 98, WP2 uvrA: 33, 100, 333, 1000 and 2500 µg/plate

with metabolic activation (pre-incubation assay):
strains TA 100, TA 1537, TA 102: 10, 33, 100, 333 and 1000 µg/plate
strains TA 1535, TA 98, WP2 uvrA: 33, 100, 333, 1000 and 2500 µg/plate

Due to a wide range of toxicity in the pre-incubation assay with microsomal activation, additional concentrations were tested with metabolic activation:
strains TA 100, TA 1537: 0.3, 1.0, 3.3, 10 and 33 µg/plate
strains T A 1535, T A 98, T A 102: 3.3, 10, 33, 100 and 333 µg/plate.
Controlsopen allclose all
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
Without metabolic activation
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
Without metabolic activation
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
Without metabolic activation
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
Remarks:
Without metabolic activation
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
mitomycin C
Remarks:
Without metabolic activation
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
cyclophosphamide
Remarks:
With metabolic activation
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
other: 2-AA
Remarks:
With metabolic activation

Results and discussion

Test resultsopen allclose all
Key result
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Positive controls validity:
valid
Key result
Species / strain:
S. typhimurium TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Positive controls validity:
valid
Key result
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Positive controls validity:
valid

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

During the described mutagenicity test and under the experimental conditions reported, the test item did not induced gene mutations by base-pair changes or frameshifts in the genome of the strains used.


Therefore, MTF is considered to be non-mutagenic in this Salmonella typhimurium and Escherichia coli reverse mutation assay.