Registration Dossier
Registration Dossier
Diss Factsheets
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: 224-658-5 | CAS number: 4439-24-1
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data

Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
A reverse mutation experiment was undertaken in line with the Maron and Ames (1983) pre-incubation technique to determine the mutagenic potential of 2-iso-butoxyethanol in Salmonella typhimurium TA100, TA1535, TA98, and TA1537 and Escherichia coli WP2 uvrA. No significant increase in revertant colony frequency was observed with and without metabolic activation (S9 mix) in any bacterial strain, at any concentration. Under the experimental conditions, 2-iso-butoxyethanol was established not to be a mutagen. This conclusion has been supported by an in vitro mammalian chromosome aberration experiment with hamster lung (CHI/IU) cells, in which no significant inhibition of cell proliferation, nor significant structural aberration or polyploidy, was observed in chromosomal DNA with and without metabolic activation following 6- and 24-hour exposure to 2-iso-butoxyethanol. Classification is not required (CLP Regulation (EC) No. 1272/2008).
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:
- March 15, 2013
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
- Qualifier:
- according to guideline
- Guideline:
- other: Japanese: Methods of Testing New Chemical Substances
- Version / remarks:
- 2011
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: Standard Concerning Testing Laboratories Implementing Tests for New Chemical Substances
- Version / remarks:
- 2011
- Deviations:
- no
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Details on mammalian cell type (if applicable):
- Provided by Japan Bioassay Research Centre on August 7, 1997
- Additional strain / cell type characteristics:
- other: Histidine-dependent
- Species / strain / cell type:
- E. coli WP2 uvr A
- Details on mammalian cell type (if applicable):
- Provided by Japan Bioassay Research Centre on April 9, 1997
- Additional strain / cell type characteristics:
- other: Tryptophan-dependent
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- Dose finding test: 1.50, 5.00, 15.0, 50.0, 150, 500, 1500 and 5000 μl/plate
Main test I and II: 313, 625, 1250, 2500 and 5000 μl/plate - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Distilled water (Hikari Pharmaceutical Co., Ltd; Lot No. C23VS1)
- Justification for choice of solvent/vehicle: Able to dissolve 2-iso-butoxyethanol for the purpose of injection - Untreated negative controls:
- yes
- Remarks:
- Distilled water
- Negative solvent / vehicle controls:
- no
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- sodium azide
- benzo(a)pyrene
- furylfuramide
- other: 2-Aminoanthracene (Lot No. EPM0250; Purity: 96.3%)
- Details on test system and experimental conditions:
- - Method: Preincubation method by Matsuyama et al. (1980), based on Maron and Ames (1983)
- Composition of S9 mix (in 1 ml): S9 from 7-week old Sprague-Dawley male rat liver: 10 vol % (0.1 ml); 0.2 mole/l Na-Phosphoric acid buffer: 100 μmole/ml (0.5 ml); Coenzyme solution 0.38 ml; KCl: 33 μmole/ml; Glucose-6-phosphate: 5 μmole/ml; Nicotinamide-adenine dinucleotide (reduced form, disodium salt): 4 μmole/ml; Nicotinamide-adenine dinucleotide phosphate (reduced form, tetrasodium salt): 4 μmole/ml; 0.4 mole/l MgCl solution: 8 μmole/ml (0.02 ml)
EXPERIMENTAL PROCEDURE:
- Protocol: In the experimental condition without metabolic activation (S9 mix absence), 0.1 ml test substance solution, 0.5 ml 0.1 mole/l Na-phosphoric acid buffer (pH7.4) and 0.1 ml bacteria were gently mixed. In the experimental condition with metabolic activation (S9 mix presence), 0.1 ml test substance solution and 0.5 ml S9 mix were gently mixed. 20 minutes after incubation at 37 °C, 2 ml of top agar was added, mixed, and poured onto the surface of minimum glucose agar.
0.1 ml distilled water for injection and positive control solution were used as the negative and positive control, respectively. Cultivation spanned 48 hours at 37 °C. The number of mutant colonies was counted by visual observation and a Colony Analyser (CA-11 System Science). Presence of sediment was checked by visual observation. Growth inhibition and the status of bacterial flora on the surface of agar was determined by visual observation and a stereo microscope. The mean mutant colony value for the negative and positive control plates was calculated.
- Contamination check: 1 ml test substance solution and 0.5ml 0.1 mole/l Na-phosphoric acid buffer (pH7.4) (or 1 ml test substance solution and S9 mix 0.5 ml) were gently mixed and incubated at 37 °C for 20 minutes. The solution was then gently mixed with 2 ml top agar for S. typhimurium and poured onto the surface of a minimum glucose agar. Bacterial contamination in agar was checked 48 hours after cultivation at 37 °C. No bacterial contamination was detected in the test substance solution or S9 mix - Evaluation criteria:
- 2-Iso-butoxyethanol was considered to have mutagenicity when the average number of mutant colonies in the plates containing the test item was more than twice that of the negative control and where reproducibility or dose-dependency was detected.
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Untreated negative controls validity:
- valid
- 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
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- 2-Iso-butoxyethanol did not inhibit growth or induce mutagenicity in Salmonella typhimurium TA 98, TA 100, TA 1535, and TA 1537, and Escherichia coli WP2 uvrA in the dose finding test, nor in the two main tests, with or without metabolic activation.
- Conclusions:
- Following a reverse mutation experiment in Salmonella typhimurium and Escherichia coli, it has been concluded that 2-iso-butoxyethanol does not possess mutagenic potential. Based on the results obtained under the conditions of this study, classification in line with CLP Regulation (EC) No. 1272/2008 is not required.
- Executive summary:
A reverse mutation study was performed to test the effect of 2-iso-butoxyethanol in Salmonella typhimurium TA100, TA1535, TA98, and TA1537 and Escherichia coli WP2 uvrA. The test procedure was according to Good Laboratory Practise (GLP) and the Japanese Methods of Testing New Chemical Substances (2011) and Japanese Standard Concerning Testing Laboratories Implementing Tests for New Chemical Substances (2011) without deviation. This methodology is considered to be similar / equivalent to OECD Guideline 471 (Bacterial Reverse Mutation Assay). The purpose of the experiment was to determine the mutagenic potential of 2-iso-butoxyethanol with and without metabolic activation (S9 mix) using the preincubation technique described by Maron and Ames (1983).
A dose finding test was performed at 1.50, 5.00, 15.0, 50.0, 150, 500, 1500 and 5000 μl/plate of the registered substance. No inhibition of bacterial growth was observed. Two subsequent tests were performed as part of the main experiment using 313, 625, 1250, 2500, and 5000 μl/plate of 2-iso-butoxyethanol. 0.1 ml distilled water (solvent) was used as a negative control and 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide, sodium azide, 9-aminoacridine, benzo[a]pyrene, and 2-aminoanthracene were selected as positive controls. Cultivation was permitted for 48 hours at 37°C. Where the sum of mutated colonies exposed to 2-iso-butoxyethanol was more than twice that of the negative control and where reproducibility or dose-dependency was detected, the registered substance could be categorised as a mutagen.
Reverse mutation inS. typhimuriumandE. colistrains revealed consistently negative results whereby colony numbers at all concentrations were less than double those of the negative control with and without metabolic activation. No bacterial contamination was detected in the solution containing 2-iso-butoxyethanol, nor the S9 mix, and mutagenicity arising from exposure to the positive controls was detected in all bacteria. In addition, the value of the positive and negative controls was within the range of background data (average ±3 x SD). It can be concluded, therefore, that 2-iso-butoxyethanol does not have the potential for genetic toxicity under these experimental conditions.
- Endpoint:
- in vitro cytogenicity / micronucleus study
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- March 27, 2013
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
- Qualifier:
- according to guideline
- Guideline:
- other: Japanese: Methods of Testing New Chemical Substances
- Version / remarks:
- 2011
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: Standard Concerning Testing Laboratories Implementing Tests for New Chemical Substances
- Deviations:
- no
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- GLP compliance:
- yes
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- Chromosome
- Species / strain / cell type:
- other: Hamster lung (CHI/IU) cells
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix from the liver of 7-week old male Sprague-Dawley rats
- Test concentrations with justification for top dose:
- Dose finding concentrations: 0.0188, 0.0375, 0.0750, 0.150, 0.300, 0.600, 1.200 mg/ml.
Although cell proliferation decreased to 79% in the 24h continous test, cell proliferation inhibition did not exceed 50% in all conditions with 1.2 mg/ml. Therefore the following test concentrations were used for the abberration test: 0.15, 0.30, 0.60, 1.20 mg/ml - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Distilled water (Hikari Pharmaceutical Co., Ltd; Lot No. C23VS1)
- Justification for choice of solvent/vehicle: Able to dissolve 2-iso-butoxyethanol for the purpose of injection - Untreated negative controls:
- yes
- Remarks:
- Distilled water
- Negative solvent / vehicle controls:
- no
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- mitomycin C
- Details on test system and experimental conditions:
- Procedure: The test consisted of a 6-hour experiment with and without metabolic activation (S9 mix) and a 24-hour continuous experiment. Four concentration groups from 0.15, 0.30, 0.60, and 1.2mg/ml (common ratio: 2) were analysed at all treatment conditions. Four dishes were prepared per each concentration group, with two dishes for chromosome sampling and the rest for cell proliferation rate.
- Positive control: Mitomycin as a positive control for a 6-hour treatment without S9 mix and for a 24-hour continuous treatment at 15 µl/dish (final concentration:0.1 µl/ml) and 12.5 µl/dish (final concentration:0.05 µl/ml), respectively. Cyclophosphamide was used as a positive control for the 6-hour treatment with the presence of S9 mix at 30 µl/dish (final concentration: 10 µl/ml). It is known that these concentrations of mitomycin and cyclophosphamide induce structural abnormality of chromosome. Only two dishes were prepared for chromosome sampling and the culture solution was replaced by 10%CS/MEN or S9.
- Negative control: Distilled water for injection was used as a negative control and added at the level of 10 vol% per dish.
Other
- 10%CS/MEN: Culture solution containing calf serum (Lot No. 522123, Biological Industries; 990250, GIBCO) at the level of 10 vol% of Eagle’s Minimal Essential Medium (EMEM, Nissui Pharmaceutical Co. Ltd.).
- S9 mix: S9 (Lot No. RAA-653 and RAA-655, Kikkoman Corp.) was prepared from the liver of 7-week old male Sprague-Dawley rats into which phenobarbital and 5,6-benzoflavone were injected. The final S9 mixture contained 5 vol% S9, 0.83mmol/L glucose-6-phosphate, 0.67mmole/L β-NADP+, 0.83mmol/L MgCl2, 5.5 mmol/L KCl, and 0.67mmol/L HEPES.- Evaluation criteria:
- Type and number of structural abnormalities were examined in the 200 metaphase cells (chromosome number: 23 - 27) per group (100 cell/dish, 25 cell/observer). Polyploid cell numbers were counted in the 800 metaphase cells (chromosome number: 38 or more than 38) per group (400 cell/dish, 40 cell/observer). From the results, an appearance ratio was calculated of cells with structural abnormality against those deemed to be polyploid. Inducibility of chromosome abnormality was comprehensively evaluated by referring to the Fisher’s exact test result (see Statistics) and from the biological perspectives.
- Statistics:
- A significance test (Fisher’s exact test, one-sided) of the appearance ratio was conducted for the test substance treatment, positive control, and negative control for 6-hour and 24-hour tests. Cochran-Armitage's trend test, one-sided was performed for 6-hour tests with and without metabolic activation.
- Key result
- Species / strain:
- other: Hamster lung (CHI/IU) cells
- Remarks:
- 6-hour treatment
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Key result
- Species / strain:
- other: Hamster lung (CHI/IU) cells
- Remarks:
- 24-hour treatment
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Following chromosome analysis, no statistically significant increase in the number of polyploid cells or the number of cells with structural abnormalities was reported.
- Conclusions:
- An in vitro mammalian chromosome aberration experiment using hamster lung (CHI/IU) cells found that 2-iso-butoxyethanol did not bring about significant inhibition of cell proliferation, nor were abnormalities in the chromosomal DNA such as structural aberration and polyploid cell reported from exposure to 2-iso-butoxyethanol. The registered substance can be concluded to be non-mutagenic under the experimental conditions and it does not require classification (CLP Regulation (EC) No. 1272/2008).
- Executive summary:
To establish the potential genetic toxicity of 2-iso-butoxyethanol, an in vitro mammalian chromosome aberration experiment with hamster lung (CHI/IU) cells was performed according to Japanese Methods of Testing New Chemical Substances (2011) and the Japanese Standard Concerning Testing Laboratories Implementing Tests for New Chemical Substances (2011), which is similar / equivalent to OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test). The test aimed to establish whether 2-iso-butoxyethanol had mutagenic potential.
An intial test to examine the potential cell proliferation inhibition at 0.019 - 1.2 mg/ml 2-iso-butoxyethanol was used to determine the test concentrations . Four test doses ranging from 0.15 mg/ml to 1.2 mg/ml (common ratio: 2) were used in the chromosomal aberration test. Three conditions were arranged, including a 6-hour treatment with and without metabolic activation and a 24-hour continuous treatment without metabolic activation. The S9 mix was derived from the liver of 7-week old Sprague-Dawley male rats. Mitomycin was selected as a positive control for the short-term and 24-hour treatment without S9 mix at 15 µl/dish (final concentration:0.1µl/ml) and 12.5µl/dish (final concentration:0.05 µl/ml) respectively. Cyclophosphamide was selected as a positive control to the short-term treatment with S9 mix at 30 µl/dish (final concentration: 10 µl/ml). Distilled water for injection was added at a level of 10 vol% per dish in the negative control.
A significance test (Fisher’s exact test, p<0.01, one-sided) of the appearance ratio, calculated from the frequency of cells with structural abnormalities and polyploid cell, was conducted among the test substance, positive control, and negative control groups. The potential of 2-iso-butoxyethanol to induce chromosome abnormality was comprehensively evaluated by referring to the Fisher’s exact test result and from biological perspectives.
Cell proliferation inhibition was found not to exceed 50% in all experimental conditions at up to 1.2 mg/ml 2-iso-butoxyethanol. In addition, there was no statistically significant increase in the number of cells that exhibited structural abnormalities or polyploidy. The positive and negative controls were valid. Subsequently, it has been determined conclusively that 2-isobutoxyethanol did not inhibit growth nor micronucleus abnormality in CHL/IU hamster cells and, therefore, the registered substance is non-mutagenic under the experimental conditions.
Referenceopen allclose all
In all test bacteria, colony numbers in the test material conditions were less than double the colony numbers of the negative control in both the presence and absence of metabolic activation. No bacterial contamination was detected in the test substance solution or S9 mix. Mutagenicity of the positive control was detected in all of the test bacteria. Furthermore, the value of the positive and negative controls was within the range of background data (average ±3 x standard deviation).
Examination for the inhibition of cell proliferation revealed a decline in cell proliferation to 79 % in the 24-hour continuous treatment. However, cell proliferation inhibition did not exceed 50 % in all experimental conditions with 1.2 mg/ml test substance. At the start and end of the treatment, precipitation was not identified with bare eyes in the culture solution in all test substance groups. Based on the obtained result, chromosome analysis was carried out for the conditions described below:
- Short-term treatment without the S9 mix: 0.15, 0.30, 0.60, 1.2mg/ml (common ratio: 2)
- Short-term treatment with the S9 mix: 0.15, 0.30, 0.60, 1.2mg/ml (common ratio: 2)
- 24-hours of continuous treatment: 0.15, 0.30, 0.60, 1.2mg/ml (common ratio: 2).
At the start and end of the treatment, precipitation was not identified in the culture solution in the all test substance groups. However, precipitation was identified with bare eyes during the experiment involving the negative control and 0.15 mg/ml groups of short-term treatment without the S9 mix. The experiment of the short-term treatment without the S9 mix was cancelled and re-examination was conducted as below.
- Short-term treatment with the S9 mix (re-examination): 0.15, 0.30, 0.60, 1.2 mg/ml (common ratio: 2)
At the start and end of the treatment, precipitation was not identified with bare eyes in the culture solution in all test substance groups. Mitotic index analysis indicated that the highest analysable concentration was 1.2 mg/ml in the experiment. Therefore, the groups described below were used for chromosome analysis.
- Short-term treatment without the S9 mix: 0.30, 0.60, 1.2mg/ml
- Short-term treatment with the S9 mix: 0.30, 0.60, 1.2mg/ml
- 24-hours of continuous treatment: 0.30, 0.60, 1.2mg/ml
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
A Klimisch score of 1 (reliable without restriction) has been assigned to the endpoint conclusion attained from the bacterial chromosomal aberration experiment (Reference 1). This reliability level has been recommended as the study was performed according to the Japanese Methods of Testing New Chemical Substances (2011) and Standard Concerning Testing Laboratories Implementing Tests for New Chemical Substances (2011) without deviation, which is similar / equivalent to Good Laboratory Practise (GLP) and OECD Guideline 471 (Bacterial Reverse Mutation Assay). The experiment detailed in Reference 2 was performed in line with Japanese Methods of Testing New Chemical Substances (2011) and Standard Concerning Testing Laboratories Implementing Tests for New Chemical Substances (2011), which is similar / equivalent to OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test). No deviation from the protocol was reported. As the test was not compliant with Good Laboratory Practise (GLP), the endpoint conclusion is suitable for regulatory use with some restriction and has been assigned a Klimisch score of 2.
Justification for classification or non-classification
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.
