Registration Dossier
Registration Dossier
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 202-625-6 | CAS number: 97-99-4
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Gene mutation (Bacterial reverse mutation assay / Ames test): negative
with and without metabolic activation in Salmonella typhimurium strains
TA98, TA100, TA1535 and TA1537 Escherichia coli WP2 (equivalent to OECD
TG 471) (Mitsubishi, 2004a).
Cytogenicity in mammalian cells: negative with and without metabolic
activation in cultured human lymphocytes (OECD TG 473) (Mitsubishi,
2004b).
Mutagenicity to mammalian cells: negative with and without metabolic
activation in mouse lymphoma L5178Y cells (OECD TG 476) (Flanders, L,
2012)
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- not reported
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
- Remarks:
- The Japanese relevant authority has stated clearly in secondary summaries that this study was conducted in compliance with GLP. However, some pages, which may contain the confirmation of GLP compliance, are missing from the published study report. Overall, the studies are the part of safety evaluation by the national authority of Japan (from Japan Existing Chemical Data Base (JECDB)). The data are collected under the control of the authority and approved by the GLP authority, which are under national control. Therefore we consider that the studies are in compliance with GLP although GLP or QA statement is not included in the available study reports.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- yes
- Remarks:
- 2AA- only control with metabolic activation
- GLP compliance:
- yes
- Remarks:
- evidence of GLP is not included in the report
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- S. typhimurium TA100 and TA1535; his G (base-pair substitution)
S. typhimurium TA98; his D (frameshift)
S. typhimurium TA 1537; his C (frameshift)
E. coli WP2uvrA/pK101; trp E (base-pair substitute) - Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- Phenobarbital and tetrahydrofurfuryl alcohol induced rat liver S9
- Test concentrations with justification for top dose:
- Tester strains; TA 100, TA1535, WP2uvrA/pK101, TA 98 and TA 1537: 5000, 2500, 1250, 625 and 313 µg/plate (with or without S9 mix)
Positive substance;
with S9 mix: TA 100 (2-aminoanthracene (2-AA); 1 µg/plate), TA1535 (2-AA; 2 µg/plate), WP2uvrA/pK101 (2-AA; 2 µg/plate), TA 98 (2-AA; 0.5 µg/plate) and TA 1537 (2-AA; 2 µg/plate)
without S9 mix: TA 100 (furfurylamide (AF-2); 0.01 µg/plate), TA1535 (sodium azide (NaN3); 0.5 µg/plate), WP2uvrA/pK101 (ENNG; 2 µg/plate), TA 98 (AF-2; 0.5 µg/plate) and TA 1537 (9-AA; 2 µg/plate) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Water for injection (Otsuka Pharmaceutical Factory, Inc., Lot No.; K0C78)
- Justification for choice of solvent/vehicle: The test substance was soluble in water for injection (DW) at the concentration of 50 mg/ml in the preliminary test for selection of solvent. No exothermic, bubbling and discoloration was observed when DW was added. Based on these results, DW was used for solvent. - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- distilled water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- furylfuramide
- Remarks:
- AF-2 (Lot No.; CAP0185, content; 98.9%, Wako Pure Chemical Industries, Ltd.), Solvent: DMSO
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- distilled water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- NaN3 (Lot No.; KWE6685, content; 96.5%, Wako Pure Chemical Industries, Ltd.), Solvent: DW
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- distilled water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- N-ethyl-N-nitro-N-nitrosoguanidine
- Remarks:
- ENNG (Lot No. 56F-3651, content; 99.0%, Sigma Chemical Company), Solvent: DMSO
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DW
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- 9-AA (Lot No., 80F-0186, content; >99%, Sigma Chemical Company), Solvent: DMSO
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- sterile distilled water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene (2-AA)
- Remarks:
- 2-AA (Lot No. TWH2355, content; 98.0%, Wako Pure Chemical Industries, Ltd.), Solvent: DMSO
- Details on test system and experimental conditions:
- 1. Tester strain;
Five tester strains, Salmonella typhimurium strains TA100, TA1535, TA98 and TA1537 which were obtained from the University of California on May 27, 1983 and Escherichia coli strain WP2uvrA/pKM101 which was obtained from Japan Bioassay Research Center were used. These strains are widely used for bacterial reverse mutation test, and are recommended by OECD Guidelines and the guideline in Japanese Chemical Substances Control Law.
2. Bacterial suspension;
Each frozen tester strains were thawed at use, and aliquot of 20 µl was taken and inoculated to 10 µl of liquid complete medium. They were incubation under shaking culture for 8 hours at 37°C (90 agitation/min.). L-shaped tube (capacity; 22 ml) was used for culturing vessel. After incubation period was finished, turbidity of bacterial suspension was measured with turbidity meter. Viable bacteria count was calculated by conversion from turbidity. The appropriate bacterial concentration (10 E+09/ml or more) was confirmed in the suspension and then used for the test (Note 2).
3. Reverse mutation test;
The test was conducted by pre-incubation method with and without S9 mix. For each dosage, 0.1 ml of test substance solution or negative (solvent) control material was added into sterilized tube, and then 0.5 ml of 0.1 mol/l sodium-phosphate buffer (pH 7.4) (without S9 mix) or 0.5 ml of S9 mix (with S9 mix) was added. Then 0.1 ml of bacterial suspension was added and was shaking-incubated at 37°C for 20 minutes. After pre-incubation, 2 ml of top agar was added to the mixture and was multilayered onto minimum glucose agar plate medium. After solidification of multilayered top agar, it was incubated at 37°C for 48 hours. Growth of bacterial flora was observed with stereoscopic microscope to investigate degree of bacterial growth inhibition by test material and then the presence of precipitation was investigated by visual inspection. Number of revertant colonies on the plate was counted with automatic colony counter (CA-11, System Science Co., Ltd.; correction method: area and miscount correction). Tests for the positive control materials were conducted similarly. - Evaluation criteria:
- If more than 2-fold increase is noted in the number of revertant colonies (mean value) compared with negative (solvent) control with our without S9 mix in any tester strain and also if reproducibility is noted in such increase, test substance is judged to have mutagenicity (positive). For other case, it is judged as negative.
- Statistics:
- No statistical method was used for judgment of test results.
- Key result
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- The numbers of revertant colonies were below 2 fold of negative (solvent) control value at the dosages of 5000, 1250, 313, 78.1, 19.5, 4.88 and 1.22 µg/plate for all tester strains with or without S9 mix as the result of the preliminary test. No growth inhibition and no precipitation were observed for any tester strain with or without S9 mix.
Based on above results, main tests were conducted at 5 dosages of 5000, 2500, 1250, 625 and 313 µg/plate for all tester strains.
As the result of 2 main tests, the numbers of revertant colonies for all tester strains were below 2 folds of negative (solvent) control value with or without S9 mix. No growth inhibition and no precipitation were observed for any tester strain with or without S9 mix.
From sterility tests for the test substance solution at the highest concentration and S9 mix, no contamination by bacteria or mould which affected the results of the tests was observed.
Based on the results of preliminary test, main tests were conducted with the highest concentration of 5000 µg/plate.
Number of revertant colonies was below 2 folds of negative (solvent) control value for all tester strains with
or without S9 mix.
It was confirmed that the test was appropriately conducted because negative (solvent) control value and positive
control value in this test are within the normal range which calculated from historical background data, and because
positive control material showed positive result with significant increase of over 2 fold in the number of revertant
colonies which was induced by the positive control material in each tester strains with or without S9 mix compared
with the number of negative (solvent) control in each tester strains.
Based on above results, it was concluded that tetrahydrofurfuryl alcohol has no mutagenicity (negative) for
bacterial reverse mutation test. - Conclusions:
- Tetrahydrofurfuryl alcohol has been tested according to a Japanese national method that is equivalent to OECD TG 471. No evidence of test-substance induced increases in the number of revertants was obtained using S. typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2, with and without metabolic activation, in either the initial or the repeat experiment, both of which used the pre-incubation method. Appropriate 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.
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2004
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
- Remarks:
- The Japanese relevant authority has stated clearly in secondary summaries that this study was conducted in compliance with GLP. However, some pages, which may contain the confirmation of GLP compliance, are missing from the published study report. Overall, the studies are the part of safety evaluation by the national authority of Japan (from Japan Existing Chemical Data Base (JECDB)). The data are collected under the control of the authority and approved by the GLP authority, which are under national control. Therefore we consider that the studies are in compliance with GLP although GLP or QA statement is not included in the available study reports.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Qualifier:
- according to guideline
- Guideline:
- JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
- Deviations:
- no
- GLP compliance:
- yes
- Remarks:
- Evidence of GLP is not included in the study report.
- Type of assay:
- in vitro mammalian chromosome aberration test
- Species / strain / cell type:
- other: CHL/IU cell line derived from lung of female Chinese hamster
- Details on mammalian cell type (if applicable):
- Cell line was purchased from Dainippon Pharma Co. Ltd., on July 17, 2001 (number of passage at purchasing: 14), and DMSO (lot number: 210G1441, Kanto Chemical Co., Inc.,) was added to the cell suspension to obtain the final concentration of 10 v/v%, and the solution was dispensed to 1 ml and stored frozen in liquid nitrogen (number of passage at freezing: 17). The frozen lot of cells was supplied for characteristic inspection. It was confirmed that they had chromosome number of 25, cell cycle of 12.5 hours and to negative in mycoplasma contamination. The frozen lot of cells was thawed, incubated and used for the test within 4 weeks after thawing (number of passage: 18 - 24). Incubation of cells was conducted on plastic plate (6 cm or 10 cm of diameter, Becton Dickinson and Company) in CO2 incubator (set to 5% of CO2 and 37°C of temperature, with humidification, Type 7300 or 6301C, NAPCO).
- Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-mix
- Test concentrations with justification for top dose:
- Test substance;
Short-term treatment (6-hrs treatment - 18-hrs recovery incubation); 257.5, 515 and 1030 µg/ml
24-hours continuous treatment; 257.5, 515 and 1030 µg/ml
Positive control;
Short-term treatment; without S9-mix: MMC; 0.1 µg/ml, with S9-mix: BP; 20 µg/ml
24-hours continuous treatment; MMC; 0.05 µg/ml - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Physiological saline (PS) (Otsuka Pharmaceutical Factory, Inc., Lot No.; K2A79)
- Justification for choice of solvent/vehicle: The test substance was soluble in PS at the concentration of 50 mg/ml in the test for selection of solvent. No exothermic, bubbling and discoloration was observed when PS was added. Based on these results, PS was used for solvent. - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- mitomycin C
- Remarks:
- without S9-mix: abbreviated as MMC, Lot No. 342AJH, content; 103%, Kyowa Hakko Kogyo Co.
- Untreated negative controls:
- yes
- Remarks:
- sterility control
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- benzo(a)pyrene
- Remarks:
- with S9-mix: abbreviated as BP, Lot No. GG01, content; 95.6%, Tokyo Chemical Industry Co., Ltd.
- Details on test system and experimental conditions:
- (1) Cell line and selection rationale;
The CHL/IU cell line derived from lung of female Chinese hamster was used. The cell is widely used in chromosome aberration test for noted advantages, which include that the number of chromosomes is only 25, and that chromosomes are relatively large, contributing to easier specimen observation.
(2) Culture medium;
Approximately 8.3 g of Eagle MEM Medium “Nissui” (1) (lot number: 460201, Nihon Pharmaceutical Co., Ltd.,) was dissolved in 880 ml of purified water, autoclaved (121°C, 20 minutes) and 8.8 ml of 2.92 w/v% L-glutamine aqueous solution and 11.2 ml of 10 w/v% sodium bicarbonate which were sterilized separately were added, respectively in order to prepare a MEM solution. Then, 100 ml of inactivated (heated at 56°C for 30 minutes) bovine serum (Lot No. 296130, GIBCO BRL) was added to 900 ml of MEM solution. The obtained solution was used as MEM medium.
(3) Manufacture of S9-mix;
(a) S9;
S9 was purchased from Kikkoman Corporation on March 5, 2002. S9 was derived from liver of 7-week old male SD strain rat (body weight: 214 to 239 g) which was enzyme-induced with phenobarbital (single intraperitoneal administration at 30 mg/kg on Day 1, daily single intraperitoneal administration at 60 mg/kg for 3 days after Day 2) and 5,6-benzoflavone (single intraperitoneal administration at 80 mg/kg on Day 3). S9 of lot number RAA-459 (final protein concentration: 1.26 mg/ml, manufactured on February 22, 2002) was used for the test. Purchased S9 was stored frozen in Deep Freezer with controlled temperature of below -80°C (measured temperature: -84 to -82°C) until use.
(b) S9-mix;
Composition of S9-mix per 1 ml was shown in Table 3 and prepared when needed. S9-mix was stirred in ice until use.
(4) Preparation of test substance solution and positive control substance solution;
(a) Test substance solution;
Designated quantity of the test substance was weighed and dissolved in added PS by shaking stirring. This solution was used as the highest concentration solution.
The highest concentration solution was serially diluted with PS to prepare 10-fold concentration of the test substance solutions for each dose level. Test substance solution was prepared at use, and weighing, dilution, dispense and storage after preparation of test substance was done under yellow light. The storage period before treatment was 10 to 20 minutes in cell growth inhibition test, 5 to 15 minutes in chromosome aberration test (short term treatment) and 5 minutes in chromosome aberration test (continuous treatment).
(b) Positive control substance solution;
MMC in 2 mg-volume vial was dissolved in 5 ml of water for injection (Lot No.: K2A84, Otsuka Pharmaceutical Factory, Inc.,) at use. The solution was serially diluted with PS (Lot No.: K2A79, Otsuka Pharmaceutical Factory, Inc.,) to prepare solution at 10-fold concentration of treatment dose in each treatment condition (short term treatment: 1 µg/ml, continuous treatment: 0.5 µg/ml).
For BP, solution was prepared at 200-fold concentration of treatment dose by dissolving in DMSO (Lot No.: 207G1673, Kanto Chemical Co., Inc.,) at 4 mg/ml and was stored frozen until use.
(5) Cell growth inhibition test;
(a) Dose of test substance;
Prior to cell growth inhibition test, preliminary test was conducted by 24-hour treatment with and without S9 mix. Preliminary test (dose level: 10.3, 103, 1030 µg/ml [equivalent to 0.1, 1 and 10 mmol]) was conducted using 1 plate per each dose level. After the completion of the treatment, plates were observed by phase contrast inverted microscope and relative cell density was determined with the cell density in the negative control as 100%. As a result, the cell density in each dose level was showed 100%. According to above result, dose levels in cell growth inhibition test were set as follows:
-S9 mix: 64.4, 128.8, 257.5, 515, 1030 µg/ml
+S9 mix: 64.4, 128.8, 257.5, 515, 1030 µg/ml
24-hour treatment: 64.4, 128.8, 257.5, 515, 1030 µg/ml
(b) Treatment of cells;
Each 5 ml of cell suspension prepared at 4x10E3/ml was inoculated onto 6 cm plate contained MEM medium and incubated for 3 days.
After removal of MEM medium, solution for cell treatment of the following composition (Table 4) was added to 2 plates per 1 dose level, and cells were treated for 24 hours (continuous treatment) or 6 hours (short term treatment). In short term treatment, cell surface was washed with MEM after 6-hour treatment, 5 ml of fresh MEM medium was added and incubation was continued for further 18 hours. The solvent of the test substance was used as negative control material and similarly treated.
(c) Measurement of cell growth rate;
Surface of cell was washed with Dulbecco's phosphate buffered saline (referred as PBS(-) hereinafter, Nissui Pharmaceutical Co., Ltd.,) which was free of Ca2+ and Mg2+. Then cells were fixed with methanol for 10 minutes, stained with 3 v/v% Giemsa solution for 10 minutes and washed gently with water and dried. The cell growth rate was measured using mono layer culture cell density meter (Monocellater, Olympus Optical Co., Ltd.,) for each stained plate. As a result, 50% inhibitory concentration (IC50) of the test substance could not be calculated because cell growth inhibition greater than 50% was not observed at any treatment conditions.
(6) Chromosome aberration test;
Initially, short term treatment only was conducted for chromosome aberration test. The result of short term treatment was negative and continuous treatment was conducted consequently.
(a) Dose level for test substance and positive control substance;
Based on result of cell growth inhibition test, following dose levels were set in chromosome aberration test. No precipitation or deposition was observed at test substance treatment at any treatment condition and any test substance treatment group.
-S9 mix: 257.5, 515 and 1030 µg/ml
+S9 mix: 257.5, 515 and 1030 µg/ml
24-hour treatment: 257.5, 515 and 1030 µg/ml
For dose levels of positive control materials, MMC was set at 0.1 µg/ml in -S9 mix and BP was set at 0.05 µg/ml in 24-hour treatment. 20 µg/ml of BP was used in +S9 mix. These dose levels are known to induce clastogenicity.
(b)Treatment of cells;
Cells were treated similarly as in cell growth inhibition test.
For positive control, cells were treated similarly with cell treatment solution with the following composition (Table 5).
For negative control group and test substance treatment group, 4 plates were used per each treatment condition (2 plates for specimen preparation and 2 plates for measurement of cell growth rate). For positive control group, 2 plates were used per each treatment condition because the measurement of cell growth inhibition test was not conducted.
(c) Preparation of specimen;
Colcemid was added on each plate for specimen preparation at 2 hours prior to specimen preparation to make the final dose of 0.1 µg/ml and metaphase cells were accumulated. After treatment was completed, surface of cells was washed with PBS(-) and cells were separated by treatment of 0.25 w/v% trypsin. The solution was taken into centrifuge tube and cells were collected by centrifugation (1000 rpm, 5 minutes; following procedure was done under the same condition). After removal of supernatant, 4 ml of 0.075 mol/L potassium chloride solution was taken into each centrifuge tube and hypotonically treated (37°C, 15 minutes). 0.5 ml of cooled fixative (mixture of methanol and acetic acid [3:1, v/v]; following procedure was done under the same condition) was added and mixed, and then centrifuged to remove supernatant. 4 ml of fixative was added and the same procedure was repeated for 2 times. Then, cells were suspended in appropriate volume of fixative and the solution was dropped onto 2 positions of slide glass which was placed on wetted towel and the slide glass was dried. Specimen was stained with 3 v/v% Giemsa stain for 20 minutes, washed with water, dried and enclosed to obtain specimen of chromosome. 2 specimens were prepared per plate. - Evaluation criteria:
- 1. Structural and numerical aberration;
Observation was performed by blind test on 100 specimens per plate, i.e., 200 metaphases per 1 dose level. Cells with well-spread chromosome were observed in the metaphase observation.
Structural aberration was observed based on the following criteria:
Cells with centromere of 25 ± 2 or over 35 were excluded.
Chromatid break
Chromatid exchange
Chromosome break
Chromosome exchange (dicentric chromosome, circular chromosome and etc)
Fragmentation
Gap was defined as a condition that width of achromatic region of chromatid is smaller than width of chromatid. Gap was recorded separately from other aberrations and not included in structural aberration.
Numerical aberration was defined as cells with centromeres over 35 and polyploid cells including endoreduplicated cells. Cells with numerical aberrations were counted.
2. Acceptance criteria for the test results:
Cells with 1 or more structural aberration were counted as structural chromosome aberration. Specimens with less than 50 observable metaphases per plate were excluded from counting.
3. Criteria for judgment of clastogenicity in each treatment condition are as follows:
- Negative (-): frequencies of both structural and numerical aberration are less than 5%
- Pseudo-Positive [equivocal] (+/-): frequencies of structural or numerical aberration or both are more than 5% and less than 10%.
- Positive (+):frequencies of structural or numerical aberration or both are more than 10%. - Statistics:
- Statistical method was not used for evaluation of the results.
- Key result
- Species / strain:
- other: CHL/IU cell line derived from lung of female Chinese hamster
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Chromosome aberration test using cultured mammalian cells was conducted to investigate clastogenicity of tetrahydrofurfuryl alcohol.
Frequencies of cells with structural and numerical aberration were less than 5% in both short term treatment and continuous treatment.
Frequencies of cells with structural and numerical aberration were less than 5% in all negative control groups and frequency of cells with structural aberration was more than 10% in all positive control groups. Therefore, it was confirmed that this study was valid technically.
Based on above, it is concluded that tetrahydrofurfuryl alcohol does not induce chromosomal aberration for CHL/IU cells under the conditions of this study. - Conclusions:
- Tetrahydrofurfuryl alcohol has been tested according to a Japanese national protocol that is equivalent to OECD 473. No substance-induced increase in the number of cells with aberrations was observed in CHL/IU cells when tested with and without metabolic activation. Appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for the induction of chromosome aberrations under the conditions of the test.
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5300 - In vitro Mammalian Cell Gene Mutation Test
- GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- thymidine kinase
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: RPMI 1640 Glutamax-1 and HEPES buffer (20 mM) supplemented with Penicillin (100 units/ml), Streptomycin (100 μg/ml), Sodium pyruvate (1 mM), Amphotericin B (2.5 μg/ml) and 10% donor horse serum (giving R10 media) at 37 °C with 5% CO 2 in air.
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: no information
- Periodically "cleansed" against high spontaneous background: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- phenobarbital/beta-naphthoflavone induced rat liver S9
- Test concentrations with justification for top dose:
- 0, 63.75, 127.5, 255, 510, 765, 1020 μg/ml +/- MA, both experiments
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: R0 medium
- Justification for choice of solvent/vehicle: following solubility checks - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- without metabolic activation
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- Remarks:
- with metabolic activation
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Preincubation period: none
- Exposure duration: 4 hours (-MA, expt 1, +MA expts 1 and 2); 24 hours (-MA, expt 2)
- Expression time (cells in growth medium): 2 days
- Selection time (if incubation with a selection agent): 10-14 days
SELECTION AGENT (mutation assays): 4 μg/ml 5-trifluorothymidine (TFT)
NUMBER OF REPLICATIONS: duplicate cultures, experiment repeated. 96-well microtitre plates used for selection/viability assessment
DETERMINATION OF CYTOTOXICITY
- Method: relative total growth
OTHER EXAMINATIONS:
- Other: numbers of small and large colonies were recorded. - Evaluation criteria:
- A result is considered positive if the mutant frequency exceeds the solvent mutant frequency by the Global Evaluation Factor (GEF) 126 x 10E-06, and demonstrates a positive linear trend and is reproducible.
- Statistics:
- UKEMS statistical package
- Key result
- Species / strain:
- mouse lymphoma L5178Y cells
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: none (see Table 1 below)
- Effects of osmolality: none (see Table 1 below)
RANGE-FINDING/SCREENING STUDIES: no toxicity observed up to the 10 mM limit dose of 1020 μg/ml.
COMPARISON WITH HISTORICAL CONTROL DATA: results were within the range of historical control
ADDITIONAL INFORMATION ON CYTOTOXICITY: see Table 2 below) - Conclusions:
- Tetrahydrofurfuryl alcohol has been tested for mutagenicity to mouse lymphoma L5178Y cells in a study conducted according to OECD 476 and in compliance with GLP. No increase mutant frequency or in the ratio of small to large colonies was observed in the presence or absence of metabolic activation in either the first experiment (4 hour exposure with and without metabolic activation) or the independent repeat experiment (24 hour exposure without metabolic activation, 4 h exposure with metabolic activation). No cytotoxicity was observed up to the limit concentration of 1020 μg/ml (10 mM). Appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for mutagenicity to mammalian cells under the conditions of the test.
Referenceopen allclose all
Pre-experiment for toxicity
Whether or not there is metabolic activation |
Dosage level of the test substance (µg/plate) |
Number of reverse mutations (number of colonies/plate) |
|||||
Base pair substitution type |
Frame shift type |
||||||
TA100 |
TA1535 |
WP2uvrA/pKM101 |
TA98 |
TA1537 |
|||
－S9 mix |
Negative control |
114 |
9 |
81 |
16 |
17 |
|
1.22 |
130 |
10 |
87 |
23 |
23 |
||
4.88 |
109 |
8 |
84 |
24 |
24 |
||
19.5 |
123 |
13 |
67 |
26 |
29 |
||
78.1 |
116 |
14 |
71 |
24 |
24 |
||
313 |
126 |
10 |
83 |
18 |
22 |
||
1250 |
118 |
8 |
65 |
27 |
24 |
||
5000 |
105 |
13 |
73 |
26 |
25 |
||
+S9 mix |
Negative control |
113 |
8 |
78 |
32 |
24 |
|
1.22 |
117 |
10 |
86 |
26 |
31 |
||
4.88 |
126 |
14 |
114 |
35 |
20 |
||
19.5 |
120 |
11 |
112 |
25 |
24 |
||
78.1 |
128 |
8 |
89 |
34 |
30 |
||
313 |
123 |
9 |
125 |
39 |
22 |
||
1250 |
114 |
8 |
104 |
32 |
27 |
||
5000 |
108 |
11 |
88 |
33 |
25 |
||
Positive control |
Positive control groups for which S9 mix is not necessary |
Name |
AF-2 |
NaN3 |
ENNG |
AF-2 |
9-AA |
Dosage (µg/plate) |
0.01 |
0.5 |
2 |
0.1 |
80 |
||
(Number of colonies/plate) |
551 |
467 |
3659 |
697 |
263 |
||
Positive control groups for whichS9mix is necessary |
Name |
2-AA |
2-AA |
2-AA |
2-AA |
2-AA |
|
Dosage (µg/plate) |
1 |
2 |
2 |
0.5 |
2 |
||
(Number of colonies/plate) |
1298 |
182 |
625 |
400 |
184 |
||
Results of main experiment 1, preincubation; revertants per plate (mean and s.d.)
Whether or not there is metabolic activation |
Dosage level of test substance (µg/plate) |
Number of reverse mutations (number of colonies/plate) |
||||||||||
Base pair substitution type |
Frame shift type |
|||||||||||
TA100 |
TA1535 |
WP2uvrA/ pKM101 |
TA98 |
TA1537 |
||||||||
－S9 mix |
Negative control |
103 123 113 |
(113) (± 10) |
14 11 11 |
(12) (± 2) |
82 75 87 |
(81) (± 6) |
22 20 22 |
(21) (± 1) |
17 16 17 |
(17) (± 1) |
|
313 |
97 103 122 |
(107) (± 13) |
11 8 10 |
(10) (± 2) |
93 73 92 |
(86) (± 11) |
17 23 21 |
(20) (± 3) |
16 21 15 |
(17) (± 3) |
||
625 |
112 122 108 |
(114) (± 7) |
10 8 12 |
(10) (± 2) |
76 76 98 |
(83) (± 13) |
21 20 19 |
(20) (± 1) |
15 18 14 |
(16) (± 2) |
||
1250 |
110 134 96 |
(113) (± 19) |
14 11 10 |
(12) (± 2) |
63 77 95 |
(78) (± 16) |
31 18 25 |
(25) (± 7) |
18 20 15 |
(18) (± 3) |
||
2500 |
116 109 111 |
(112) (± 4) |
11 9 9 |
(10) (± 1) |
93 81 92 |
(89) (± 7) |
20 17 24 |
(20) (± 4) |
17 11 17 |
(15) (± 3) |
||
5000 |
118 119 102 |
(113) (± 10) |
10 16 11 |
(12) (± 3) |
87 70 91 |
(83) (± 11) |
17 21 17 |
(18) (± 2) |
17 23 15 |
(18) (± 4) |
||
+S9 mix |
Negative control |
128 127 102 |
(119) (± 15) |
13 8 15 |
(12) (± 4) |
99 106 99 |
(101) (± 4) |
33 24 23 |
(27) (± 6) |
25 24 22 |
(24) (± 2) |
|
313 |
114 120 119 |
(118) (± 3) |
9 11 11 |
(10) (± 1) |
87 111 92 |
(97) (± 13) |
40 23 29 |
(31) (± 9) |
26 27 21 |
(25) (± 3) |
||
625 |
137 98 125 |
(120) (± 20 |
13 10 12 |
(12) (± 2) |
92 102 77 |
(90) (± 13) |
33 30 23 |
(29) (± 5) |
27 23 22 |
(24) (± 3) |
||
1250 |
108 126 106 |
(113) (± 11) |
12 11 17 |
(13) (± 3) |
104 111 112 |
(109) (± 4) |
30 31 37 |
(33) (± 4) |
22 27 24 |
(24) (± 3) |
||
2500 |
113 120 123 |
(119) (± 5) |
10 16 15 |
(14) (± 3) |
109 95 112 |
(105) (± 9) |
29 29 33 |
(30) (± 2) |
26 24 18 |
(23) (± 4) |
||
5000 |
101 120 113 |
(111) (± 10) |
10 18 13 |
(14) (± 4) |
86 113 116 |
(105) (± 17) |
23 33 23 |
(26) (± 6) |
25 30 29 |
(28) (± 3) |
||
Positive control |
Positive control groups for which S9 mix is not necessary |
Name |
AF-2 |
NaN3 |
ENNG |
AF-2 |
9-AA |
|||||
Dosage (µg/plate) |
0.01 |
0.5 |
2 |
0.1 |
80 |
|||||||
(Number of colonies/plate) |
564 555 533 |
(551) (± 16)
|
520 476 468 |
(488) (± 28) |
4412 4259 3992 |
(4221) (± 213) |
632 644 625 |
(634) (± 10) |
292 237 346 |
(292) (± 55) |
||
Positive control groups for whichS9mix is necessary |
Name |
2-AA |
2-AA |
2-AA |
2-AA |
2-AA |
||||||
Dosage (µg/plate) |
1 |
2 |
2 |
0.5 |
2 |
|||||||
(Number of colonies/plate) |
1381 1446 1443 |
(1423) (± 37) |
169 149 171 |
(163) (± 12) |
935 955 925 |
(938) (± 15) |
436 437 451 |
(441) (± 8) |
171 167 175 |
(171) (± 4) |
||
AF-2:2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide
NaN3: Sodium azide
ENNG:N-ethyl-N’-nitro-N-nitrosoguanidine,
9-AA:9-Aminoacridine hydrochloride,
2-AA:2-Aminoanthracene
Results of main experiment 2, preincubation; revertants per plate (mean and s.d.)
Whether or not there is metabolic activation |
Dosage level of test substance (µg/plate) |
Number of reverse mutations (number of colonies/plate) |
||||||||||
Base pair substitution type |
Frame shift type |
|||||||||||
TA100 |
TA1535 |
WP2uvrA/ pKM101 |
TA98 |
TA1537 |
||||||||
－S9 mix |
Negative control |
121 118 102 |
(114) (± 10) |
16 15 14 |
(15) (± 1) |
82 85 88 |
(85) (± 3) |
19 22 22 |
(21) (± 2) |
18 14 11 |
(14) (± 4) |
|
313 |
127 111 108 |
(115) (± 10) |
14 11 12 |
(12) (± 2) |
96 94 86 |
(92) (± 5) |
24 24 26 |
(25) (± 1) |
17 16 15 |
(16) (± 1) |
||
625 |
95 106 107 |
(103) (± 7) |
9 10 10 |
(10) (± 1) |
95 86 89 |
(90) (± 5) |
16 24 25 |
(22) (± 5) |
16 16 15 |
(16) (± 1) |
||
1250 |
110 112 117 |
(113) (± 4) |
10 11 12 |
(11) (± 1) |
76 90 88 |
(85) (± 8) |
17 25 21 |
(21) (± 4) |
14 18 17 |
(16) (± 2) |
||
2500 |
101 105 105 |
(104) (± 2) |
13 9 15 |
(12) (± 3) |
86 90 91 |
(89) (± 3) |
16 17 20 |
(18) (± 2) |
15 13 17 |
(15) (± 2) |
||
5000 |
121 98 130 |
(116) (± 17) |
8 11 14 |
(11) (± 3) |
93 87 81 |
(87) (± 6) |
26 18 23 |
(22) (± 4) |
12 10 12 |
(11) (± 1) |
||
+S9 mix |
Negative control |
110 127 108 |
(115) (± 10) |
11 11 16 |
(13) (± 3) |
93 95 87 |
(92) (± 4) |
32 29 35 |
(32) (± 3) |
23 23 15 |
(20) (± 5) |
|
313 |
123 114 126 |
(121) (± 6) |
12 13 13 |
(13) (± 1) |
88 111 91 |
(97) (± 13) |
24 31 27 |
(27) (± 4) |
20 17 22 |
(20) (± 3) |
||
625 |
124 119 110 |
(118) (± 7) |
14 16 14 |
(15) (± 1) |
90 87 103 |
(93) (± 9) |
39 21 24 |
(28) (± 10) |
18 22 29 |
(23) (± 6) |
||
1250 |
121 115 112 |
(116) (± 5) |
10 13 10 |
(11) (± 2) |
87 95 102 |
(95) (± 8) |
25 28 33 |
(29) (± 4) |
22 25 22 |
(23) (± 2) |
||
2500 |
108 99 116 |
(108) (± 9) |
14 10 13 |
(12) (± 2) |
90 96 108 |
(98) (± 9) |
24 27 30 |
(27) (± 3) |
22 16 20 |
(19) (± 3) |
||
5000 |
108 114 102 |
(108) (± 6) |
9 13 10 |
(11) (± 2) |
101 87 108 |
(99) (± 11) |
25 25 28 |
(26) (± 2) |
22 19 19 |
(20) (± 2) |
||
Positive control |
Positive control groups for which S9 mix is not necessary |
Name |
AF-2 |
NaN3 |
ENNG |
AF-2 |
9-AA |
|||||
Dosage (µg/plate) |
0.01 |
0.5 |
2 |
0.1 |
80 |
|||||||
(Number of colonies/plate) |
563 520 482 |
(552) (± 41)
|
369 387 440 |
(399) (± 37) |
4358 4439 4275 |
(4357) (± 82) |
632 618 590 |
(613) (± 21) |
242 209 209 |
(220) (± 19) |
||
Positive control groups for whichS9mix is necessary |
Name |
2-AA |
2-AA |
2-AA |
2-AA |
2-AA |
||||||
Dosage (µg/plate) |
1 |
2 |
2 |
0.5 |
2 |
|||||||
(Number of colonies/plate) |
1398 1398 1301 |
(1366) (± 56) |
150 147 138 |
(145) (± 6) |
1347 1373 1072 |
(1264) (± 167) |
432 428 423 |
(428) (± 5) |
200 182 194 |
(192) (± 9) |
||
Table 1.; Result of chromosome aberration test (short-term treatment)
Treatment-recovery period (h) |
S9mix |
Dose of test substance (µg/ml) |
Number of cells showing structural chromosome aberration (incidence, %) |
Number of gap appearances |
Cell proliferation rate (%) |
Number of cells showing numerical chromosome aberration (incidence, %) |
|||||||||
Number of cells observed |
Chromatid break |
Chromatid exchange |
Chromosome break |
Chromosome exchange |
Fragment |
Total number of aberrations (%) |
Number of cells observed |
Polyploid |
endoreduplication |
Total number of aberrant cells (%) |
|||||
6-18 |
- |
Negative control (NSS) |
100 100 200 |
0 1 1 (0.5) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 1 1 (0.5) |
0 0 0 |
101 99 100 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
6-18 |
- |
257.5 |
100 100 200 |
1 1 2 (1.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 1 1 (0.5) |
1 0 1 (0.5) |
2 2 4(2.0) |
0 0 0 |
107 103 105 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
6-18 |
- |
515 |
100 100 200 |
1 0 1(0.5) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
1 0 1 (0.5) |
0 0 0 |
105 103 104 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
6-18 |
- |
1030 |
100 100 200 |
0 1 1 (0.5) |
1 0 1 (0.5) |
1 0 1 (0.5) |
0 0 0 (0.0) |
0 0 0 (0.0) |
1 1 2 (1.0) |
0 0 0 |
105 107 106 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
6-18 |
- |
Positive control (MMC 0.1) |
100 100 200 |
15 19 34 (17.0) |
23 28 51 (25.5) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
36 46 82 (41.0) |
0 0 0 |
|
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
6-18 |
+ |
Negative control (NSS) |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
1 0 1 (0.5) |
0 0 0 (0.0) |
0 0 0 (0.0) |
1 0 1 (0.5) |
0 1 1 |
111 89 100 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
6-18 |
+ |
257.5 |
100 100 200 |
0 0 0 (0.0) |
1 0 1 (0.5) |
1 0 1 (0. 5) |
0 0 0 (0.0) |
0 0 0 (0.0) |
2 0 2 (1.0) |
0 0 0 |
111 105 108 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
6-18 |
+ |
515 |
100 100 200 |
0 0 0 (0.0) |
1 0 1 (0.5) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
1 0 1 (0.5) |
0 0 0 |
106 108 107 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
6-18 |
+ |
1030 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 |
109 114 112 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
6-18 |
+ |
Positive control (BP20) |
100 100 200 |
15 9 24 (12.0) |
78 79 157 (78.5) |
1 1 2 (1.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
81 82 163 (81.5) |
0 1 1 |
|
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
NSS: Normal saline solution
DMSO: Dimethyl sulfoxide, MMC: mitomycin C, BP: benzo(a)pyrene
Table 2.; Result of chromosome aberration test (continuous treatment)
Treatment-recovery period (h) |
Dose of test substance (µg/ml) |
Number of cells showing structural chromosome aberration (incidence, %) |
Number of gap appearances |
Cell proliferation rate (%) |
Number of cells showing numerical chromosome aberration (incidence, %) |
|||||||||
Number of cells observed |
Chromatid break |
Chromatid exchange |
Chromosome break |
Chromosome exchange |
Fragment |
Total number of aberrations (%) |
Number of cells observed |
Polyploid |
endoreduplication |
Total number of aberrant cells (%) |
||||
24-0 |
Negative control (NSS) |
100 100 200 |
0 1 1 (0.5) |
0 0 0 (0.0) |
0 1 1 (0.5) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 2 2 (1.0) |
1 0 1 |
106 94 100 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
24-0 |
257.5 |
100 100 200 |
0 0 0 (0.0) |
1 0 1 (0.5) |
1 0 1 (0.5) |
1 0 1 (0.5) |
0 0 0 (0.0) |
3 0 3 (1.5) |
0 0 0 |
104 102 103 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
24-0 |
515 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 |
108 96 102 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
24-0 |
1030 |
100 100 200 |
0 0 0 (0.0) |
2 0 2 (1.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
2 0 2 (1.0) |
0 0 0 |
101 101 101 |
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
24-0 |
Positive control (MMC 0.05) |
100 100 200 |
14 7 21 (10.5) |
18 15 33 (16.5) |
0 1 1 (0.5) |
0 0 0 (0.0) |
0 0 0 (0.0) |
30 23 53 (26.5) |
0 0 0 |
|
100 100 200 |
0 0 0 (0.0) |
0 0 0 (0.0) |
0 0 0 (0.0) |
NSS: Normal saline solution
MMC: Mitomycin C
Table 1 pH and osmolality readings from the solubility test
μg/ml |
0 |
3.99 |
7.97 |
15.94 |
31.88 |
63.75 |
127.5 |
255 |
510 |
1020 |
pH |
7.31 |
7.31 |
7.31 |
7.33 |
7.33 |
7.33 |
7.33 |
7.33 |
7.33 |
7.34 |
mOsm |
297 |
296 |
300 |
298 |
297 |
298 |
299 |
299 |
304 |
308 |
Table 2 Preliminary Toxicity Test
Dose (μg/ml) |
% RSG (-S9) 4-Hour Exposure |
% RSG (+S9) 4-Hour Exposure |
% RSG (-S9) 24-Hour Exposure |
0 |
100 |
100 |
100 |
3.99 |
89 |
104 |
95 |
7.97 |
87 |
103 |
94 |
15.94 |
96 |
100 |
79 |
31.88 |
88 |
100 |
93 |
63.75 |
85 |
100 |
90 |
127.5 |
94 |
109 |
72 |
255 |
85 |
110 |
86 |
510 |
87 |
109 |
80 |
1020 |
87 |
108 |
81 |
%RSG= Relative Suspension Growth
RTG = Relative Total Growth
Table 3 Results experiment 1
Treatment (μg/ml) |
4 Hours -MA |
Treatment (μg/ml) |
4 Hours +MA |
||||
%RSG |
RTG |
MF* |
%RSG |
RTG |
MF* |
||
0 |
100 |
1.00 |
145.64 |
0 |
100 |
1.00 |
177.18 |
63.75 |
98 |
1.03 |
185.92 |
63.75 |
109 |
1.09 |
206.57 |
127.5 |
98 |
1.00 |
197.08 |
127.5 |
101 |
1.06 |
205.50 |
255 |
101 |
1.05 |
180.91 |
255 |
108 |
1.10 |
190.59 |
510 |
99 |
1.10 |
154.04 |
510 |
110 |
1.02 |
200.68 |
765 |
106 |
1.07 |
181.93 |
765 |
106 |
1.02 |
191.57 |
1020 |
102 |
1.06 |
175.32 |
1020 |
105 |
1.08 |
177.59 |
Linear trend |
NS |
Linear trend |
NS |
||||
EMS 400 |
58 |
0.35 |
1334.46 |
CP 2 |
58 |
0.33 |
1254.84 |
%RSG= Relative Suspension Growth
RTG = Relative Total Growth
*Positive wells per tray, 96 wells plated
NS = not significant
Table 4 Results experiment 2
Treatment (μg/ml) |
24 Hours -MA |
Treatment (μg/ml) |
4 Hours +MA |
||||
%RSG |
RTG |
MF* |
%RSG |
RTG |
MF* |
||
0 |
100 |
1.00 |
141.18 |
0 |
100 |
1.00 |
131.90 |
63.75 |
95 |
1.02 |
147.90 |
63.75 |
93 |
1.02 |
152.72 |
127.5 |
112 |
1.15 |
92.03 |
127.5 |
98 |
1.05 |
121.28 |
255 |
106 |
1.07 |
125.96 |
255 |
96 |
0.98 |
141.72 |
510 |
97 |
0.87 |
154.35 |
510 |
99 |
1.13 |
142.60 |
765 |
107 |
1.10 |
136.45 |
765 |
95 |
0.98 |
140.60 |
1020 |
105 |
0.92 |
148.69 |
1020 |
102 |
1.13 |
124.91 |
Linear trend |
NS |
Linear trend |
NS |
||||
EMS 150 |
35 |
0.35 |
971.89 |
CP 2 |
75 |
0.53 |
1179.29 |
%RSG= Relative Suspension Growth
RTG = Relative Total Growth
*Positive wells per tray, 96 wells plated
NS = not significant
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Data are available from reliable studies on mutagenicity to bacterial cells and mammalian cells, and cytogenicity in mammalian cells.
Tetrahydrofurfuryl alcohol has been tested according to a Japanese national method that is equivalent to OECD TG 471 (Mitsubishi, 2004a). No evidence of test-substance induced increases in number of revertants was obtained using S. typhimurium strains TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2, with and without metabolic activation, in either the initial or the repeat experiment, both of which used the pre-incubation method. Appropriate 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.
Tetrahydrofurfuryl alcohol has been tested according to a Japanese national protocol that is equivalent to OECD 473 (Mitsubishi, 2004b). No substance-induced increase in the number of cells with aberrations was observed in CHL/IU cells when tested with and without metabolic activation. Appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for the induction of chromosome aberrations under the conditions of the test.
Tetrahydrofurfuryl alcohol has been tested for mutagenicity to mouse lymphoma L5178Y cells in a study conducted according to OECD 476 and in compliance with GLP (Flanders, L, 2012). No increase mutant frequency or in the ratio of small to large colonies was observed in the presence or absence of metabolic activation in either the first experiment (4 hour exposure with and without metabolic activation) or the independent repeat experiment (24 hour exposure without metabolic activation, 4 h exposure with metabolic activation). No cytotoxicity was observed up to the limit concentration of 1020 μg/ml (10 mM). Appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for mutagenicity to mammalian cells under the conditions of the test.
Proposal of in vivo testing is not required as the results of all the in vitro studies were negative.
Justification for classification or non-classification
Based on the available in vitro genotoxicity data, tetrahydrofurfuryl alcohol is not classified for mutagenicity according to Regulation (EC) No 1272/2008.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
