Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to a test protocol that is comparable to the appropriate OECD test guideline. There is no data on GLP compliance. Details of positive control substances are not included in the publication Supporting material.

Data source

Reference
Reference Type:
publication
Title:
A compilation of two decades of mutagenicity test results with the Ames Salmonella typhimurium and L5178Y mouse lymphoma cell mutation assays.
Author:
Seifried, H.E. et al
Year:
2006
Bibliographic source:
Chem. Res. Toxicol 19, 627-644

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
full details of positive controls are not given in the publication; information on condition of background lawn not included. Highest concentration outside normal dose range.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
not specified
Details on test material:
- Name of test material (as cited in study report): 2-methyltetrahydrofuran, CAS 96-47-8

Method

Target gene:
Histidine operon
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
Aroclor induced rat liver S9
Test concentrations with justification for top dose:
10-10000 μg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
not specified
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 hours

SELECTION AGENT (mutation assays): selective top agar

NUMBER OF REPLICATIONS: triplicate plates


DETERMINATION OF CYTOTOXICITY
- Method: no data

ACTUIVATION: S9 mix included NADP and glucose-6-phosphate, and 10% S9. 0.5 ml of S9 mix was added to a final volume 2.6 ml, giving a final concentration of approximately 2% S9.
Evaluation criteria:
A dose-dependent doubling (TA 98, TA 100, TA 1535) or 3-fold increase (other strains) in mean number of revertants per plate was considered positive.
Statistics:
No information

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Remarks:
tested to 10000 mg/ml; doses based on preliminary toxicity test with TA 100, and no reduction in number of revertants.
Vehicle controls validity:
not specified
Untreated negative controls validity:
not applicable
Positive controls validity:
not specified
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

 Salmonella Test Data revertants per plate (mean of three plates ± S.D.)

Concentration (µg/plate) TA98 TA100 TA102 TA1535 TA1537
no S9 rat S9 Ham'r S9 no S9 rat S9 Ham'r S9 no S9 rat S9 Ham'r S9 no S9 rat S9 Ham'r S9 no S9 rat S9 Ham'r S9
DMSO 19 ± 6 21 ± 4 19 ± 4 120 ± 9 113 ± 16 97 ± 2 223 ± 41 351 ± 8 354 ± 38 15 ± 3 14 ± 6 17 ± 7 11 ± 3 15 ± 4 6 ± 1
100 13 ± 1 26 ± 6 23 ± 5 114 ± 12 104 ± 15 99 ± 12 249 ± 53 348 ± 44 402 ± 11 19 ± 4 15 ± 8 13 ± 5 9 ± 2 10 ± 3 8 ± 3
333 12 ± 4 20 ± 6 27 ± 4 118 ± 20 115 ± 26 125 ± 13 291 ± 62 376 ± 49 373 ± 63 20 ± 6 20 ± 4 25 ± 3 10 ± 4 15 ± 4 8 ± 3
1000 14 ± 2 26 ± 6 17 ± 5 114 ± 6 107 ± 9 101 ± 11 295 ± 18 395 ± 58 369 ± 29 16 ± 5 21 ± 6 15 ± 4 6 ± 4 11 ± 3 6 ± 2
3333 12 ± 2 26 ± 4 22 ± 2 116 ± 5 122 ± 5 94 ± 9 296 ± 67 370 ± 41 317 ± 24 19 ± 2 18 ± 1 17 ± 4 4 ± 3 14 ± 3 6 ± 3
10000 13 ± 3 25 ± 1 23 ± 7 117 ± 6 119 ± 14 90 ± 7 345 ± 2 422 ± 24 312 ± 17 17 ± 2 19 ± 2 12 ± 3 7 ± 2 12 ± 6 9 ± 3
Positive control 429 ± 54 265 ± 53 1018 ± 169 421 ± 16 367 ± 18 846 ± 64 1402 ± 102 1419 ± 239 1398 ± 61 228 ± 35 104 ± 38 245 ± 164 977 ± 366 87 ± 19 311 ± 95

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with and without metabolic activation

Tetrahydro-2-methylfuran has been tested for mutagenicity to bacteria using a method that is comparable to OECD TG 471. The result is reported as negative with and without metabolic activation in Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102. Solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.
Executive summary:

Reliable data are available for tetrahydro-2-methylfurna from studies on in vitro mutagenicity to bacterial and mammalian cells. Information for cytogenicity is available from two studies, one in vitro and the other in vivo, where the reliability is not assignable as there are insufficient details in the publication.

Tetrahydro-2-methylfuran has been tested for mutagenicity to bacteria using a method that is comparable to OECD TG 471 (Seifried, H.E. et. al. (2006)). The result is reported as negative with and without metabolic activation in Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and TA 102. Solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test. This result is supported by a study in which tetrahydro-2-methylfuran was tested for mutagenicity to bacteria using Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2 uvrA (Antonucci, V. et. al. (2011)). No substantial increases in revertant colony counts were obtained in the presence or absence of metabolic activation. The information on method and results included in the publication is limited, so the reliability is 4. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.

An investigation into the mutagenicity of tetrahydro-2-methylfuran has been reported (Seifried, H.E. et. al. (2006)). The method used is similar to OECD TG 476, and the study is considered to be reliability 2. The authors of the publication report that the test substance was negative for mutagenicity to L5178Y cells without metabolic activation, but that the result with metabolic activation was inconclusive; however, the data presented do not include clear evidence of a substance-related, dose-dependent increase in mutant frequency, so it is the conclusion of the reviewer is that the test substance is negative for mutagenicity to mammalian cells under the conditions of the test.

Tetrahydro-2-methylfuran has been tested for cytogenicity in peripheral human lymphocytes (Antonucci, V. et. al. (2011)). No statistically significant increase in the proportion of cells with chromosome damage was observed. The information on method and results included in the publication is limited, so the reliability is 4. It is concluded that the test substance is negative for clastogenicity (induction of chromosome aberrations) under the conditions of the test.

Tetrahydro-2-methylfuran has been tested for induction of micronuclei in vivo (Antonucci, V. et. al. (2011)). The information on method and results included in the publication is limited, so the reliability is 4. No induction of micronuclei was observed, and no effect on the proportion of polychromatic erythrocytes was noted in male or female mice after three months treatment by oral gavage. It is concluded that the test substance is negative for the induction of micronuclei under the conditions of the test.