Registration Dossier
Registration Dossier
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EC number: 943-011-4 | CAS number: -
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Negative results in Salmonella typhimurium TA 98, TA 100, TA102, TA 1535
and TA 1537, with and without metabolic activation (OECD 471, GLP).
Negative results in mammalian chromosomal aberration test with Chinese
hamster lung cells (OECD 473, GLP).
Negative results in mammalian cell gene mutation tests using mouse
lymphoma L5178Y cells, with and without metabolic activation (OECD 476,
GLP).
The hazard assessment is based on the data currently available. New studies with the registered substance and/or other member substances of the glycol esters category will be conducted in the future. The finalised studies will be included in the technical dossier as soon as they become available and the hazard assessment will be re-evaluated accordingly.
For further details, please refer to the category concept document attached to the category object (linked under IUCLID section 0.2) showing an overview of the strategy for all substances within the glycol esters category.
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:
- 19 Jan - 5 Feb 2015
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Amt für Arbeitsschutz - Arbeitnehmerschutz, Behörde für Gesundheit und Verbraucherschutz, Hamburg, Germany
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- his operon
- 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:
- cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of male rats treated with Aroclor 1254
- Test concentrations with justification for top dose:
- 31.6, 100, 316, 1000, 3160 and 5000 µg/plate with and without metabolic activation
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: ethanol (concentration in vehicle: 10 ng/mL or 20 ng/mL)
- Justification for choice of solvent/vehicle: Due to the limited solubility of the test substance in water and DMSO, ethanol was selected as vehicle. - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- ethanol
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: Sodium Azide (SA), 2-Nitrofluorene (2-NF), 9-Aminoacridine (9-AA), Mitomycin C (MC), Benzo(a)pyrene (BaP), 2-Aminoanthracene (2-AA)
- Remarks:
- -S9: 2-NF (10 µg/pl in DMSO; TA 98); SA (10 µg/pl in water; TA 100 and TA 1535); 9-AA (100 µg/pl in ethanol; TA 1537), MC (10 µg/pl in water; TA 102); +S9: 2-AA (2 µg/pl in DMSO; TA 100 and TA 1535); BaP (10 µg/pl in DMSO; TA 98, TA 102 and TA 1537)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION:
First experiment: in agar (plate incorporation)
Second experiment: pre-incubation
DURATION
- Preincubation period: 20 min (second experiment)
- Exposure duration: 48 - 72 h (first and second experiment)
NUMBER OF REPLICATIONS: triplicates each in two independent experiments
DETERMINATION OF CYTOTOXICITY
- Method: reduction in the number of spontaneous revertants by at least 50%, a clearing or diminution of the background lawn or by the degree of survival of the treated cultures - Evaluation criteria:
- A test item is considered to have mutagenic potential, if a significant (p ≤ 0.05) increase of revertant colonies per plate is determined in both experiments (increase factor ≥ 2 for TA 98, TA 100, TA 1535 and TA 1537 or increase factor ≥ 1.5 for TA 102) in at least one strain or a concentration-related increase over the range tested is observed.
Positive results have to be reproducible and the histidine independence of the revertants has to be confirmed by streaking random samples on histidine-free agar plates.
Biological relevance of the results should be considered first.
A test item for which the results do not meet the above mentioned criteria is considered as non-mutagenic in the AMES test. - Statistics:
- Mean values and standard errors were calculated.
Statistical significance was determined via U-test according to MANN and WHITNEY (increase in revertants). In case that a concentration-related increase over the range tested was observed, a Spearman's rank correlation coefficient was applied. - Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- in the preincubation test at 5000 µg/plate with and without S9 mix
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium 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
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- in the preincubation test at 5000 µg/plate with and without S9 mix
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- in the plate incorporation test at 5000 µg/plate with and without S9 mix and in the preincubation test at 5000 µg/plate with S9 mix
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: The test item was fully soluble in stock concentrations of 10 and 20 mg/mL. Stock dilutions with higher concentrations (31.6 and 50 mg/mL or 63.2 and 100 mg/mL (corresponding to final concentrations of 3160 or 5000 µg/plate)) were emulsions. Precipitation was noted at 5000 µg/plate in all strains.
- Other: For the positive control substances, no respective solvent control was included. However, due to the range of the mutagenic response, a comparison to the vehicle control ethanol is considered as acceptable.
RANGE-FINDING/SCREENING STUDIES:
A preliminary test was performed in tester strain TA 100 to determine cytotoxcity. Concentrations ranging from 0.316 to 5000 µg/plate were tested in a plate incorporation test without and with metabolic activation. No signs of cytotoxicity were noted in any concentration. Precipitation was observed at 5000 µg/plate. Based on the results of the preliminary study, 5000 µg/plate was chosen as maximum dose for the main study.
COMPARISON WITH HISTORICAL CONTROL DATA:
The mutant frequencies of the solvent controls were within the range of historical control data for any strain. Furthermore, the results of the positive control cultures were within the range of the historical data.
ADDITIONAL INFORMATION ON CYTOTOXICITY:
To prevent cytotoxicity of the solvent ethanol in the preincubation test, the volume of the test item and negative control was reduced from the generally employed 100 µL to 50 µL per plate as the preincubation test is more sensitive than the plate incorporation test. - Conclusions:
- Interpretation of results: negative
- Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Comparable to guideline study with acceptable restrictions (no discussion of results and no historical control data).
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- no discussion of results and no historical control data.
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- no discussion of results and no historical control data.
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Ministerium für Umwelt, Raumordnung und Landwirtschaft des Landes Nordrhein-Westfalen, Düsseldorf, Germany
- Type of assay:
- in vitro mammalian chromosome aberration test
- Target gene:
- Not applicable
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- - Type and identity of media:
MEM medium supplemented with 2 mM L-Glutamine and
- 4% (v/v) fetal calf serum (MEM4) or
- 0% (v/v) fetal calf serum (MEM0)
- 100 IU/mL penicillin/streptomycin
During exposure to the test substancewith S9 mix, MEM0 medium was used and replaced by MEM4 after 3 h after test substance administration. - Additional strain / cell type characteristics:
- other: modal chromosome number of 22 and a cell cycle length of approx. 16.5 h
- Metabolic activation:
- with and without
- Metabolic activation system:
- cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254
- Test concentrations with justification for top dose:
- 18h treatment: 10, 40 and 80 µg/mL (without metabolic activation)
18h treatment: 10, 60, 80 and 100 µg/mL (with metabolic activation)
28h treatment: 80 µg/mL (without metabolic activation)
28h treatment: 100 µg/mL (with metabolic activation) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: ethanol
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- Remarks:
- MEM4 medium (resp. MEM0 medium in the test with S9 mix) containing 1% (v/v) ethanol
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: cyclophosphamide, 3 and 4 µg/mL in MEM0 medium, +S9; mitomycin C, 0.03 and 0.04 µg/mL in MEM4 medium, -S9
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 18 and 28 h
- Fixation time (start of exposure up to fixation or harvest of cells): 18 h treatment: 18 h; 28 h treatment: 28 h
STAIN (for cytogenetic assays): Giemsa
NUMBER OF REPLICATIONS: 2 replications each in one (28 h treatment) or two (18 h treatment) independent experiments
NUMBER OF CELLS EVALUATED: 100 per culture
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index of 1000 cells per slide
OTHER EXAMINATIONS:
- Determination of polyploidy: yes
- Determination of endoreduplicated cells: yes - Evaluation criteria:
- The test chemical is to be considered clastogenic in this assay if:
- it induces chromosomal aberrations (excl. gaps) in a statistically significant manner in one or more concentrations
- the induced proportion of aberrant cells at such test substance concentrations exceeds the normal range of the test system
- positive results can be verified in an independent experiment. - Statistics:
- The number of aberrant cells in each replica was used to establish acceptable homogeneity between replicates by means of a binomial dispersion test ( Richardson, C., Williams, D. A., Allen, J. A., Amphlett, G., Chanter, D. O. and Phillips, B. Analysis of Data from In Vitro Cytogenetic Assays, in: Statistical Evaluation of Mutagenicity Test Data, Kirkland, D. J., (ed) Cambridge University Press, Cambridge, pp. 41-64., 1990). The proportion of cells that was treated with the test substance and harboured structural aberrations (excl. gaps) was compared with the corresponding proportion of the negative controls in the Chi-square test. Probability values of p < 0.05 were accepted as statistically significant.
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- systematic influence of the test compound which led to a reduction in the mitotic index from 10 µg/mL.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: The test substance was soluble in ethanol, in MEM4 medium containing 1% ethanol. The solubility limit of the test substance was determined to be 100 µg/mL (homogeneous emulsion).
- In the first test without metabolic activation the negative controls exhibited only a mitotic index of 2.0%. Therefore, test 1 without metabolic activation was completely repeated with the same doses and named test #1a. - Conclusions:
- Interpretation of results: negative
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- 17 Jun 2010 - 17 Aug 2010
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Remarks:
- GLP-Guideline study, tested with the source substance Fatty acids, C16-18, esters with ethylene glycol (CAS 91031-31-1). According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (Dec 2012)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- TK locus
- Species / strain / cell type:
- mouse lymphoma L5178Y cells
- Details on mammalian cell type (if applicable):
- - Type and identity of media: RPMI 1640 Hepes buffered medium (Dutch modification) containing penicillin/streptomycin, sodium pyruvate and L-glutamin amd 10% (v/v) heat-inactivated horse serum (24 h exposure); for 3 h exposure only 5% (v/v) heat-inactivated horse serum were included. Selective medium consisted of the basic medium and 20% (v/v) heat-inactivated horse serum and 5 µg/mL trifluorothymidine (TFT).
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically "cleansed" against high spontaneous background: yes - Metabolic activation:
- with and without
- Metabolic activation system:
- co-factor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Phenobarbital/β-naphtoflavone
- Test concentrations with justification for top dose:
- Experiment 1:
With and without S9-mix: 0.1, 0.3, 1.0, 3.0, 10.0, 33.0, 100.0 and 333.0 µg/mL (3 h)
Experiment 2:
Without S9-mix: 3.0, 10.0, 33.0, 100.0, 125.0, 140.0 and 175.0 µg/mL (24 h)
With 12% (v/v) S9-mix: 0, 0.1, 0.3, 1.0, 3.0, 10.0, 33.0, 100.0 and 333.0 µg/mL (3 h) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: cyclophosphamide, 7.5 µg/mL, with S9; methylmethanesulfonate, 15 µg/mL and 5 µg/mL without S9, for 3 h and 24 h treatment, respectively
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 3 and 24 h
- Expression time (cells in growth medium): 48 h
SELECTION AGENT (mutation assays): 5 µg/mL trifluorothymidine
- Selection time (if incubation with a selection agent): 11-12 days
NUMBER OF REPLICATIONS: 2 replications each in 5 independent experiments
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency, relative total growth, relative survival, suspension growth, relative suspension growth, growth rate - Evaluation criteria:
- A mutation assay was considered acceptable if it met the following criteria:
a) The absolute cloning efficiency of the solvent controls is between 65 and 120%. An acceptable number of surviving cells (10E6) could be analysed for expression of the TK mutation.
b) The spontaneous mutation frequency in the solvent control is ≥ 50E-06 and ≤ 170E-06.
c) The growth rate (R) over the 2-day expression period for the negative controls should be between 8 and 32 (3 h treatment) and between 32 and 180 (24 h treatment).
d) The mutation frequency of MMS should not be below 500E-06, and fro CP not below 700E-06.
The global evaluation factor (GEF) has been identified by the IWTG as the mean of the negative/solvent mutation frequeny (MF) distribution plus one standard deviation. For the micro well version of the assay the GEF is 126.
A test substance is considered positive (mutagenic) in the mutation assay if it induces a MF of more than the MF of the controls + 126 in a dose-dependent manner. An observed increase should be biologically relevant and will be compared with the historical control data range.
A test substance is considered equivocal (questionable) in the mutation assay if no clear conclusion for positive or negative result can be made after an additional confirmation study.
A test substance is considered negative (not mutagenic) in the mutation assay if: a) None of the tested concentrations reaches a mutation frequency of the MF of the controls + 126. b) The results are confirmed in an independently repeated test. - Species / strain:
- mouse lymphoma L5178Y cells
- 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:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility and precipitation: The test substance precipitated in the exposure medium at concentrations of 100 µg/mL and above. The test substance was tested beyond the limit of the solubility to obtain adequate cytotoxicity data, the concentration used as the highest test substance concentration for the dose range ending test was 333 µg/mL.
RANGE-FINDING/SCREENING STUDIES:
In the absence of S9-mix, the relative suspension growth was 66% at the test substance concentration of 333 µg/mL compared to the relative suspension growth of the solvent control. In the presence of S9-mix, no toxicity in the relative suspension growth was observed up to and including the highest test substance concentration of 333 µg/mL compared to the suspension growth of the solvent control.
COMPARISON WITH HISTORICAL CONTROL DATA: The spontaneous mutation frequencies in the solvent treated control cultures were within the ranges of the historical controls. - Conclusions:
- Interpretation of results: negative
Referenceopen allclose all
Table 2. Results of the plate incorporation test
With or without S9-Mix |
Test substance concentration (μg/plate) |
Mean number of revertant colonies per plate ± standard deviation |
||||
Base-pair substitution type |
Frameshift type |
|||||
TA 100 |
TA 102 |
TA 1535 |
TA 98 |
TA 1537 |
||
– |
0 (100 µL/plate) |
173.0 ± 17.3± |
277.0 ± 21.8 |
26.3 ± 2.3 |
29.0 ± 3.6 |
7.7 ± 2.1 |
– |
31.6 |
152.3 ± 20.2 |
265.0 ± 11.4 |
25.3 ± 1.5 |
30 ± 7.9 |
4.3 ± 1.2 |
– |
100 |
158.0 ± 6.0 |
267.3 ± 7.6 |
28.0 ± 3.6 |
26.7 ± 9.0 |
4.3 ± 1.2 |
– |
316 |
134 ± 21.3 |
268.3 ± 7.8 |
29.7 ± 6.7 |
30.0 ± 3.6 |
4.7 ± 2.1 |
– |
1000 |
160 ± 8.9 |
269.3 ± 7.8 |
37.3 ± 2.9 |
29.7 ± 0.6 |
5.0 ± 3.5 |
– |
3160 |
164.3 ± 2.5 |
262.0 ± 14.4 |
28.0 ± 8.9 |
29.3 ± 1.2 |
5.3 ± 0.6 |
– |
5000 |
170p ± 2.0 |
247.0p ± 2.0 |
27.7p ± 3.5 |
33.3p ± 9.0 |
2.7p ± 0.6 |
– |
Positive controls |
SA |
MC |
SA |
2NF |
9AA |
Mean No. of colonies/plate ± SD |
1023.3 ± 50.0 |
1119.3 ± 7.0 |
130.7 ± 17.6 |
116.7 ± 6.0 |
61 ± 3.6 |
|
+ |
0 (100 µL/plate) |
146.3 ± 15.1 |
266.7 ± 24.8 |
26.7 ± 4.0 |
40.7 ± 6.7 |
6.7 ± 2.9 |
+ |
31.6 |
121.7 ± 0.6 |
251.0 ± 1.0 |
27.7 ± 1.2 |
36.0 ± 5.2 |
6.0 ± 1.0 |
+ |
100 |
116.7 ± 7.0 |
255.3 ± 4.2 |
32.3 ± 6.4 |
30.3 ± 0.6 |
7.3 ± 1.5 |
+ |
316 |
113.7 ± 6.4 |
282.0 ± 2.6 |
29.7 ± 3.1 |
29.0 ± 1.7 |
5.0 ± 1.0 |
+ |
1000 |
109.3 ± 5.0 |
221.3 ± 98.2 |
26.3 ± 1.2 |
27.0 ± 1.0 |
4.3 ± 0.6 |
+ |
3160 |
164.0 ± 5.2 |
272.0 ± 3.6 |
25.3 ± 2.5 |
27.7 ± 2.9 |
4.7 ± 1.5 |
+ |
5000 |
148.0p ± 22.5 |
270.7p ± 3.1 |
25.3p ± 1.2 |
32p ± 2.0 |
2.3p ± 0.6 |
+ |
Positive controls |
2AA |
BaP |
2AA |
BaP |
BaP |
Mean No. of colonies/plate ± SD |
986.3± 2.1 |
1081.7± 10.5 |
125.3 ± 18.0 |
120.7± 3.5 |
62.7± 0.6 |
|
SA = Sodium Azide
MC = Mitomycin C
2NF = 2-Nitrofluorene
9AA = 9 -Aminoacridine
2AA = 2 -Aminoanthracene
BaP = Benzo(a)pyrene
SD = standard deviation
p = precipitate
Table 3. Results of the preincubation test
With or without S9-Mix |
Test substance concentration (μg/plate) |
Mean number of revertant colonies per plate ± standard deviation |
||||
Base-pair substitution type |
Frameshift type |
|||||
TA 100 |
TA 102 |
TA 1535 |
TA 98 |
TA 1537 |
||
– |
0 (50 µL/plate) |
147.7± 17.2 |
263.0 ± 1.0 |
19.3 ± 3.8 |
31.0 ± 8.0 |
6.0 ± 1.0 |
– |
31.6 |
147.3 ± 6.1 |
279.7 ± 11.6 |
20.3 ± 1.2 |
26.0 ± 2.0 |
7.3 ± 2.9 |
– |
100 |
159.3 ± 3.8 |
280.3 ± 9.0 |
25.3 ± 4.0 |
24.7 ± 4.2 |
8.3 ± 1.2 |
– |
316 |
137.0 ± 3.0 |
282.7 ± 8.0 |
19.7 ± 2.1 |
26.0 ± 2.0 |
5.7 ± 1.2 |
– |
1000 |
137.0 ± 3.6 |
283.7 ±3.2 |
19.0 ± 1.0 |
23.3 ± 1.5 |
6.7 ± 1.5 |
– |
3160 |
117.0 ± 114.0 |
260.0 ± 16.5 |
23.7 ± 1.5 |
39.0 ± 3.5 |
5.7 ± 0.6 |
– |
5000 |
103.0p ± 2.0 |
245.3p ± 0.6 |
8.7p ± 0.6 |
14.0p ± 2.0 |
3.3p ± 2.3 |
– |
Positive controls |
SA |
MC |
SA |
2NF |
9AA |
Mean No. of colonies/plate ± SD |
882.7 ± 16.8 |
1057.7 ± 57.0 |
147.7 ± 4.0 |
164.7 ± 8.5 |
67.3 ± 0.6 |
|
+ |
0 (50 µL/plate) |
134.3 ± 4.2 |
271.3 ± 3.1 |
20.7 ± 3.8 |
29.0 ± 4.4 |
7.3 ± 1.2 |
+ |
31.6 |
128.0 ± 1.7 |
276.0 ± 12.8 |
27.0 ± 1.0 |
32.0 ± 1.0 |
7.0 ± 2.6 |
+ |
100 |
134.7 ± 8.1 |
271.7 ± 0.6 |
26.3 ± 0.6 |
32.3 ± 0.6 |
5.3 ± 2.1 |
+ |
316 |
140.7 ± 30.7 |
268.3 ± 2.1 |
28.7± 2.1 |
30.3 ± 2.1 |
7.7 ± 0.6 |
+ |
1000 |
138.3 ± 1.5 |
277.0 ± 13.9 |
26.3 ± 0.6 |
28.0 ± 1.7 |
7.7 ± 0.6 |
+ |
3160 |
122.0 ± 9.6 |
272.3 ± 1.5 |
27.0 ± 1.0 |
28.7± 0.63 |
4.0 ± 1.0 |
+ |
5000 |
102.0p ± 2.6 |
244.0p ± 1.0 |
8.0p ± 1.0 |
13.7p ± 1.2 |
2.0p ± 0.0 |
+ |
Positive controls |
2AA |
BaP |
2AA |
BaP |
BaP |
Mean No. of colonies/plate ± SD |
889.3 ± 12.7 |
1089.0 ± 14.7 |
15.03 ± 1.5 |
168.3 ± 3.2 |
61.3 ± 10.8 |
|
SA = Sodium Azide
MC = Mitomycin C
2NF = 2-Nitrofluorene
9AA = 9 -Aminoacridine
2AA = 2 -Aminoanthracene
BaP = Benzo(a)pyrene
SD = standard deviation
p = precipitate
Table 1. Summary of data obtained in experiment #1a.
Test item |
Concentration |
Mitotic Index |
Aberrant cells in % |
|
|
in µg/mL |
in % mean |
with gaps |
without gaps |
Exposure period 18 h, without S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
10.7 |
2.0 |
0.0 |
MMC |
0.03 |
2.1 |
24.6 |
18.5* |
MMC |
0.04 |
2.9 |
40.0 |
31.9* |
Test substance |
10 |
8.0 |
3.5 |
1.5 |
40 |
6.7 |
3.0 |
0.5 |
|
80 |
5.3 |
5.0 |
0.0 |
|
Exposure period 18 h, with S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
7.0 |
6.5 |
4.0 |
CP |
3.0 |
2.7 |
45.0 |
34.0* |
Test substance |
10 |
6.7 |
3.5 |
1.5 |
60 |
6.1 |
4.5 |
1.0 |
|
100 |
5.7 |
4.0 |
2.0 |
|
MMC: Mitomycin C; CP: Cyclophosphamide (positive controls)
*: significant, no statistical evaluation
Table 2. Summary of data obtained in experiment #2.
Test item |
Concentration |
Mitotic Index |
Aberrant cells in % |
|
|
in µg/mL |
in % mean |
with gaps |
without gaps |
Exposure period 18 h, without S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
7.9 |
6.0 |
3.5 |
MMC |
0.03 |
3.4 |
23.5 |
14.0* |
Test substance |
10 |
7.5 |
4.0 |
0.5 |
40 |
8.0 |
9.5 |
3.0 |
|
80 |
5.8 |
5.5 |
0.5 |
|
Exposure period 18 h, with S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
6.3 |
8.0 |
2.0 |
CP |
3.0 |
1.6 |
42.0 |
33.3* |
Test substance |
10 |
6.5 |
5.5 |
0.0 |
60 |
5.7 |
4.5 |
1.5 |
|
100 |
6.7 |
6.5 |
1.5 |
|
Exposure period 28 h, without S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
7.9 |
3.5 |
0.0 |
MMC |
0.03 |
4.4 |
43.5 |
32.5* |
Test substance |
80 |
5.8 |
4.5 |
0.5 |
Exposure period 28 h, with S9 mix |
||||
Control, Ethanol |
1.0% (v/v) |
9.2 |
6.5 |
1.5 |
CP |
3.0 |
6.0 |
36.5 |
27.5* |
Test substance |
100 |
8.4 |
3.5 |
1.0 |
MMC: Mitomycin C; CP: Cyclophosphamide (positive controls)
*: significant, no statistical evaluation
Table 1: Cytotoxic and mutagenic responses
Treatment |
Concentration [µg/mL] |
Cloning efficiency [%] |
Relative total growth [%] |
Mutation frequency x 10-6
|
||
total |
small colonies |
large colonies |
||||
3 h treatment without S9-mix |
||||||
Solvent control (mean) |
-- |
86 |
100 |
61 |
42 |
19 |
Test substance |
0.1 |
101 |
114 |
56 |
29 |
25 |
0.3 |
101 |
108 |
52 |
28 |
23 |
|
1.0 |
118 |
140 |
66 |
48 |
16 |
|
3.0 |
91 |
109 |
63 |
46 |
15 |
|
10.0 |
95 |
108 |
66 |
32 |
33 |
|
33.0 |
97 |
104 |
76 |
46 |
28 |
|
100.0* |
91 |
97 |
81 |
32 |
46 |
|
333.0* |
102 |
77 |
64 |
48 |
14 |
|
MMS |
15 |
60 |
45 |
685 |
521 |
118 |
3 h treatment with 8% (v/v) S9-mix |
||||||
Solvent control (mean) |
-- |
91 |
100 |
67 |
36 |
29 |
Test substance |
0.1 |
108 |
111 |
74 |
47 |
25 |
0.3 |
89 |
94 |
71 |
45 |
24 |
|
1.0 |
108 |
113 |
64 |
42 |
20 |
|
3.0 |
98 |
107 |
78 |
53 |
23 |
|
10.0 |
95 |
100 |
87 |
54 |
29 |
|
33.0 |
108 |
96 |
55 |
32 |
22 |
|
100.0* |
98 |
89 |
83 |
60 |
21 |
|
333.0* |
94 |
92 |
63 |
23 |
39 |
|
CP |
7.5 |
60 |
32 |
1074 |
829 |
144 |
24 h treatment without S9-mix |
||||||
Solvent control (mean) |
-- |
115 |
100 |
51 |
26 |
23 |
Test substance |
3 |
135 |
92 |
58 |
42 |
14 |
10 |
137 |
109 |
51 |
39 |
11 |
|
33 |
133 |
85 |
71 |
32 |
35 |
|
100* |
110 |
30 |
100 |
35 |
60 |
|
125* |
118 |
31 |
70 |
31 |
36 |
|
140* |
118 |
20 |
103 |
51 |
47 |
|
175* |
102 |
9 |
134 |
53 |
72 |
|
MMS |
5 |
101 |
73 |
865 |
463 |
233 |
3 h treatment with 12% (v/v) S9-mix |
||||||
Solvent control (mean) |
-- |
109 |
100 |
72 |
47 |
23 |
Test substance |
0.1 |
110 |
100 |
73 |
49 |
21 |
0.3 |
123 |
117 |
72 |
47 |
23 |
|
1.0 |
104 |
105 |
82 |
52 |
27 |
|
3.0 |
97 |
104 |
94 |
62 |
29 |
|
10.0 |
115 |
126 |
87 |
57 |
26 |
|
33.0 |
107 |
108 |
85 |
59 |
23 |
|
100.0* |
131 |
123 |
80 |
51 |
26 |
|
333.0* |
97 |
83 |
92 |
64 |
24 |
|
CP |
7.5 |
70 |
59 |
979 |
621 |
221 |
*precipitation of test substance in the exposure medium
MMS = methylmethanesulfonate
CP = cyclophosphamide
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
The hazard assessment is based on the data currently available. New studies with the registered substance and/or other member substances of the glycol esters category will be conducted in the future. The finalised studies will be included in the technical dossier as soon as they become available and the hazard assessment will be re-evaluated accordingly.
For further details, please refer to the category concept document attached to the category object (linked under IUCLID section 0.2) showing an overview of the strategy for all substances within the glycol esters category.
Genetic toxicity in bacteria (Ames)
CAS 1323-39-3/29013-28-3
The in-vitro genetic toxicity of Octadecanoic acid, monoester with 1,2-propanediol (CAS 1323-39-3) and Palmitic acid, monoester with propane-1,2-diol (CAS 29013-28-3) was assessed in a bacterial reverse mutation assay (Ames test) according to OECD 471 and in compliance with (Laboratory of Pharmacology and Toxicology, 2015). The mutagenic potential of the test substance was assessed in S. typhimurium strains TA 98, TA 100, TA 102, TA 1535 and TA 1537 at concentrations up to 5000 µg/plate in 2 independent experiments, including the plate incorporation and preincubation method. Precipitates were visible at the highest dose tested. The test substance did not induce an increase in reversions in any of the S. typhimurium strains with or without metabolic activation. Cytotoxicity was observed at 5000 µg/plate in TA 1537 (plate incorporation test), TA 98 and TA 1535 (preincubation test) with and without metabolic activation and in TA 1537 with metabolic activation (preincubation test). The vehicle and positive controls were valid and lay within the range of historical control data.
Furthermore, Propylene Glycol Monostearate was tested for mutagenicity in a series of different in vitro microbial assays with and without metabolic activation. Bacterial reverse mutation assays performed with the S. typhimurium strains TA 1535, TA 1537 and TA 1538 and suspension tests with S. cerevisiae D4 were negative (Litton Bionetics, 1975; Cosmetic Ingredient Review, 1983).
Genetic toxicity (cytogenicity) in mammalian cells in vitro
CAS 853947-59-8
An in vitro mammalian chromosome aberration test was conducted with C8-C10-1,3-Butandiolester in accordance with OECD guideline 473 under GLP conditions (Hüls AG, 1997). The induction of structural chromosome aberrations was evaluated in vitro in Chinese hamster lung fibroblasts (V79) cells, incubated for 18 and 28 h with and without a metabolic activation system (S9-mix from rats treated with Aroclor 1245). Concentrations of 10-100 µg/mL (18 h incubation) and 80 and 100 µg/mL (28 h incubation) of the test substance in the vehicle ethanol were applied. The solubility limit of the test substance in the vehicle ethanol in the culture medium was determined to be 100 µg/mL. In the first experiment without metabolic activation, the negative controls exhibited a mitotic index of 2.0% only and the experiment was therefore repeated. Thereafter, the negative as well as the positive controls showed the expected results and were within the range of historical control data. The frequency of polyploidy cells with and without metabolic activation was within the expected range (< 10%). In the experiments both with and without metabolic activation, a systematic influence of the test substance was observed, which led to a reduction in the mitotic index. No statistically or biologically significant increase in the incidence of chromosome aberrations was observed.
Therefore, under the conditions of the study, C8-C10-1,3-Butandiolester did not show clastogenic activity in this chromosomal aberration test with and without metabolic activation performed in Chinese hamster lung fibroblasts in vitro.
Genetic toxicity (mutagenicity) in mammalian cells in vitro
CAS 91031-31-1
Mutagenic properties of Fatty acids, C16-18, esters with ethylene glycol were characterized in an in vitro mammalian cell gene mutation study according to OECD guideline 476 under GLP conditions (NOTOX B.V., 2010). Gene mutations in the thymidine kinase locus were investigated in L5178Y mouse lymphoma cells in the presence and absence of a metabolic activation system (Phenobarbital/β-naphtoflavone-induced rat liver S9). In the first experiment, cells were exposed for 3 h to test substance at concentrations of 0.1-333 µg/mL (in DMSO) with and without metabolic activation. Concentrations of the second experiment without metabolic activation for an exposure time of 24 h ranged from 3-175 µg/mL and with metabolic activation (3 h; 12% S9-mix) from 0.1-333 µg/mL. The vehicle and positive controls in the study showed the expected results and were within the range of historical control data. No cytotoxicity was observed up to the precipitating concentration of 100 µg/mL and up to 333 µg/mL, respectively. There was no significant increase in the number of forward mutations at the thymidine kinase locus of L5178Y mouse lymphoma cells treated with the test material, neither in the presence nor in the absence of a metabolic activation system. Under the conditions of the study, Fatty acids, C16-18, esters with ethylene glycol did not show gene mutation activity in this test performed in L5178Y mouse lymphoma cells in vitro.
Conclusion on genetic toxicity
The available data do not provide evidence that the target or source substances exhibit mutagenic or clastogenic properties in bacteria or mammalian cells. Therefore, no properties for genetic toxicity are expected for Octadecanoic acid, monoester with 1,2-propanediol (CAS 1323-39-3) and Palmitic acid, monoester with propane-1,2-diol (CAS 29013-28-3).
Justification for classification or non-classification
Based test substance-specific data and on the analogue read-across approach, the available data on genetic toxicity do not meet the classification criteria according to Regulation (EC) 1272/2008, and are therefore conclusive but not sufficient for 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.
