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Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ames assay was performed to determine the mutagenic nature of 2-Furoic acid. The study was perfomed using Salmonella typhimurium strain TA100 and TA98. The test chemical was dissolved in spectral grade DMSO and used at dose levels of 0, 25, 50, 75 or 100µg/plate. The plates were incubated for 40 hrs and then observed for number of revertants/plate.2- Furoic acid did not induce reversion inSalmonella typhumurium TA100 and TA98and hence it is not likely to classify as a gene mutant in vitro.

Link to relevant study records
Reference
Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed publication
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
Ames assay was performed to determine the mutagenic nature of 2-furoic acid
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay
Specific details on test material used for the study:
- Name of test material: 2-furoic acid
- IUPAC name: 2-Furancarboxylic Acid
- Molecular formula : C5H4O3
- Molecular weight: 112.084 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data
Target gene:
Histidine
Species / strain / cell type:
S. typhimurium, other: TA98 and TA100
Details on mammalian cell type (if applicable):
No data
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
not specified
Metabolic activation system:
No data
Test concentrations with justification for top dose:
0, 25, 50, 75 or 100 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Spectral grade DMSO
- Justification for choice of solvent/vehicle: The test chemical dissolved in Spectral grade DMSO
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
spectral grade DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
methylmethanesulfonate
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: No data
- Exposure duration: 40 hrs in dark
- Expression time (cells in growth medium): 40 hrs in dark
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: Duplicates

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other:

OTHER: Compounds showing any ambiguity in the assays were tested twice at concentrations of up to 100 /µg/plate.

The experiments on the effect of pH on the mutagenic activity of the 2-furoate derivatives were performed using a series of base agar plates buffered in the range of pH 6.2-7.2. Agar plates of varying pH were prepared using phosphate buffer.
Rationale for test conditions:
No data
Evaluation criteria:
Chemicals that caused dose-dependent induction of more than twice the number of spontaneous revertants were defined as mutagenic, and mutagenic activity is expressed as the number of revertants per nmol, which was calculated from the linear portion of the dose -response curve.
Statistics:
No data
Species / strain:
S. typhimurium, other: TA98 and TA100
Metabolic activation:
not specified
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No detailed data available
- Effects of osmolality: No data
- Evaporation from medium: No data
- Water solubility: No data
- Precipitation: No data
- Other confounding effects: No data

RANGE-FINDING/SCREENING STUDIES: No data

COMPARISON WITH HISTORICAL CONTROL DATA: No data

ADDITIONAL INFORMATION ON CYTOTOXICITY: No data2- Furoic acid did not induce reversion in Salmonella typhumurium TA100 and TA98 and hence it is not likely to classify as a gene mutant in vitro.
Remarks on result:
other: No mutagenic potential
Conclusions:
2- Furoic acid did not induce reversion in Salmonella typhumurium TA100 and TA98 and hence it is not likely to classify as a gene mutant in vitro.
Executive summary:

Ames assay was performed to determine the mutagenic nature of 2-Furoic acid. The study was perfomed using Salmonella typhimurium strain TA100 and TA98. The test chemical was dissolved in spectral grade DMSO and used at dose levels of 0, 25, 50, 75 or 100µg/plate. The plates were incubated for 40 hrs and then observed for number of revertants/plate.2- Furoic acid did not induce reversion inSalmonella typhumurium TA100 and TA98and hence it is not likely to classify as a gene mutant in vitro.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Gene mutation in vitro:

Data available for the target chemical was reviewed to determine the mutagenic nature of 2 -Furoic acid. The studies are as mentioned below:

Ames assay was performed by Ichikawa et al (Carcinogenesis, 1986) to determine the mutagenic nature of 2-Furoic acid. The study was perfomed using Salmonella typhimurium strain TA100 and TA98. The test chemical was dissolved in spectral grade DMSO and used at dose levels of 0, 25, 50, 75 or 100 µg/plate. The plates were incubated for 40 hrs and then observed for number of revertants/plate.2- Furoic acid did not induce reversion inSalmonella typhumurium TA100 and TA98and hence it is not likely to classify as a gene mutant in vitro.

Soska et al (Mutation Research, 1981) performed Ames assay to determine the mutagenic nature of 2-Furoic acid. The study was performed using the quantitative method using Salmonella typhimurium strain TA100. Spot test was performed as the preliminary study. The reversions were counted, and the counts were plotted against concentrations. From the initial linear part of the dose-response curve, the regression and correlation coefficients of the compound were calculated, and the mutagenic activity was expressed as the number of reversions per nanomole. 2- Furoic acid did not induce reversion inSalmonella typhumurium TA100and hence it is not likely to classify as a gene mutant in vitro.

In the same study performed by Soska et al (Mutation research, 1981), Bacterial repair test was performed to determine the mutagenic nature of 2-furoic acid. The bacteria E. coli B/r strains WP2, WP2 uvrA, WP67, WP100, CM561, CM571 and CM611, at 108per plate, were suspended in M9 medium supplemented with tryptophan. In the middle of the plate, a 5-mm disk of 3 mm Whatman filter paper was placed. 5µl of solution to be tested containing 1000µg was soaked into the paper disk. The concentrations used exhibited a low, or just no, inhibition on the repair-proficient strain WP2. The plates were evaluated after 16 h incubation. The diameters of growth inhibition zones were measured and expressed in mm.2- Furoic acid did not induce mutation inE. coli WP2, WP2 uvrA, WP67, WP100, CM 561, CM 571 and CM 611and hence it is not likely to classify as a gene mutant in vitro.

In vitro unscheduled DNA synthesis was performed by Aaron et al (Mutation Research, 1989) to determine the mutagenic nature of 2-furoic acid. The study was performed using hepatocytes isolated from male Sprague Dawley rats the test chemical was exposed to the rat hepatocytes at dose level of 1, 3, 10, 30, 100, 300 or 1000µg/mL. Rats were housed in polypropylene cages with hardwood-chip bedding and maintained on a 12-h light: 12-h dark cycle. Rats received Purina Rodent Chow No. 5001 (Ralston Purina Co., St. Louis, MO) and water ad libitum. The livers were perfused by inserting a needle in the vena cava and allowing the perfusate to escape from the portal vein. The liver was first perfused with an EGTA wash solution at a flow rate of 40-45 ml/min for about 2-3 min. After perfusion with collagenase the liver was minced, stirred with the collagenase solution and decanted through a sterile gauze pad to remove tissue chunks and debris. The cell suspension was diluted to 50 ml with complete L15 (10% FBS, 50/µg/ml gentamicin, 20 mM Hepes, 10 mM insulin per ml and 0.75 /µg dexamethasone/ml) and allowed to settle on ice 10-12 min. The supernatant was discarded and the cells were resuspended in complete L15. The cell suspension was centrifuged at 50 x g and the cells were resuspended in L15 medium. Viability (trypan blue) and yield (cells/g of liver) were monitored and cell preparation with less than 70% viability was discarded. Typical yield under the conditions described here was 3-5 x 107cells per g liver. Viable cells were plated onto tissue culture coverslips in 6-well plates at 3-5 X 105cells per well and incubated at 73°C for 1-3 h to allow attachment of the cells to the coverslips. Immediately before the assay the test chemical was dissolved in solvent. After cell attachment the hepatocytes were treated with 2-furoic acid. The exposure time was 16-20 h. The final concentration of solvent was maintained at 1% to preclude the possibility of a cytotoxic effect in response to the solvent. Concurrently with the exposure to the test material, the cultures were exposed to 3H-TdR (10µCi/ml). After treatment the cells were washed, swollen, fixed, air-dried and the coverslips were attached to slides. Autoradiography was performed using NTB-2 emulsion. The dipped slides were exposed for 7 days, developed, coded and scored with a semiautomated procedure involving an Artek colony counter connected to an IBM-PC computer. 50 cells were scored per slide. Each nucleus scored was accompanied by scoring of two adjacent cytoplasmic areas. The highest cytoplasmic grain count was subtracted from the nuclear grain count to yield net grains per nucleus (N.G.). 2- Furoic acid did not induce unscheduled DNA synthesis (UDS) in the rat hepatocytes and hence it is not likely to classify as a gene mutant in vitro.

In a study by Kitamura et al (Journal of Pharmacobio-Dyanamics, 1978), Ames assay was performed to determine the mutagenic nature of 2-Furoic acid. The study was perfomed using Salmonella typhimurium strain TA100 by the preincubation assay. The test chemical was dissolved in DMSO and was preincubated for 20 mins with 0.1M sodium phosphate buffer and overnight culture of TA100. Soft molten agar was added and mixed, poured immediately over 1.2% Vogel Bonner minimal medium E agar. The plates were incubated for 2 days at 37˚C and the number of histidine revertant colonies was counted. 2- Furoic acid did not induce reversion inSalmonella typhumurium TA100and hence it is not likely to classify as a gene mutant in vitro.

Based on the data available for the target chemical, 2 -Furoic acid does not exhibit gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant as per the criteria mentioned in CLP regulation.

Justification for classification or non-classification

Based on the data available for the target chemical, 2 -Furoic acid (CAS no 88 -14 -2) does not exhibit gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant as per the criteria mentioned in CLP regulation.