Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

There is no study available for tetradecyl methacrylate but for structural analogue substances Isodecyl methacrylate CAS: 29964 -84 -9.

Read across from structural analogue substance Isodecyl methacrylate CAS: 29964-84-9:

For the assessment of tetradecyl methacrylate no data available but data of the structurally related substance isodecyl methacrylate (three in vitro studies).

One reverse mutation assay in bacteria with isodecyl methacrylate, one chromosome aberration micronucleus test and one HPRT test with the structurally related substance isodecyl methacrylate are available.

Bacterial reverse mutation assay

No study is available on dodecyl methacrylate but there is a study with the structurally related substance isodecyl methacrylate.

In a reverse gene mutation assay in bacteria according to OECD guideline 471 Salmonella typhimurium TA98, TA100, TA1535, TA1537 and TA102 strains were exposed to isodecyl methacrylate in THF at concentrations up to 5000 µg/plate in the presence and absence of mammalian metabolic activation. The positive controls induced the appropriate responses in the corresponding strains. There was no evidence of induced mutant colonies over background.

 

In vitro Chromosome aberration test in human lymphocytes

No study is available on dodecyl methacrylate but there is a study with the structurally related substance isodecyl methacrylate.

 

In a chromosome aberration test in human lymphocytes according to OECD guideline 473 (adopted May 26, 1983), human lymphocyte cultures were exposed to Isodecyl methacrylate (98.9%) in THF with and without metabolic activation (S9 mix).

The following concentrations were evaluated:

Experiment I:

22 hrs prep. interval, 4 h treatment without metabolic activation: 14.5, 25.4, 44.4, 77.7, 136.0, 238.1, 416.7, 729.1, 1276.0, 2233 µg/ml

22 hrs prep. interval, 4 h treatment with metabolic activation: 14.5, 25.4, 44.4, 77.7, 136.0, 238.1, 416.7, 729.1, 1276.0, 2233 µg/ml

Experiment II:

22 hrs prep. interval, 22 h treatment without metabolic activation: 44.4, 77.7, 136.0, 238.1, 416.7, 729.1, 1276.0, 2233 µg/ml

22 hrs prep. interval, 4 h treatment with metabolic activation: 44.4, 77.7, 136.0, 238.1, 416.7, 729.1, 1276.0, 2233 µg/ml

In Experiment I, visible precipitation of the test item in the culture medium was observed at 136.0 µg/ml and above in the absence and presence of S9 mix. In addition, precipitation occurred in Experiment II, in the absence of S9 mix, at 136.0 µg/ml and above and in the presence of S9 mix at 416.7 µg/ml and above. No relevant increase in the osmolarity or pH value was observed. In Experiment I, in the absence and presence of S9 mix, and in Experiment II, in the presence of S9 mix, no cytotoxicity was observed up to the highest applied concentration. In Experiment II, in the absence of S9 mix, a single clearly reduced mitotic index was observed at the highest dose evaluated for cytogenetic damage.

Isodecyl methacrylate was tested up to cytotoxic concentration.

HPRT-test in mammalian cells

No study is available on dodecyl methacrylate but there is a study with the structurally related substance isodecyl methacrylate.

Isodecyl methacrylate was assessed for its potential to induce gene mutations at the HPRT locus using V79 cells of the Chinese hamster according to OECD 476.

The assay was performed in two independent experiments with identical experimental procedures, using two parallel cultures each. The first main experiment was performed with and without liver microsomal activation and a treatment period of 4 h. The second experiment was performed with a treatment period of 24 hours in the absence of metabolic activation and 4 hours in the presence of metabolic activation.

The cell cultures were evaluated at the following concentrations:

Experiment I:

without S9 mix:  0.1; 0.3; 0.5; 1.0; and 2.0 µg/ml

with S9 mix: 37.5; 75; 150; 300; and 1200 µg/ml

 

Experiment II:

without S9 mix: 18.8 ;37.5; 75.0; 150; and 600 µg/ml

with S9 mix: 37.5; 75.0; 150; 300; and 600 µg/ml

In both experiments of this study (with and without S9 mix) the range of the solvent controls was from 5.7 up to 24.0 mutants per 106cells; the range of the groups treated with the test item was from 3.3 up to 34.1 mutants per 106cells.

 

EMS(150 µg/mL in experiment I and 75 µg/mL in experiment II) and DMBA (2.0 µg/mL) were used as positive controls and showed a distinct increase in induced mutant colonies. This showed the sensitivity of the test system and the activity of the S9 mix.

 

In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells.

 

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
27-03-2008 - 08-04-2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
OECD 471, GLP. Study according to relevant guideline.
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
adopted May 26, 1983
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Specific details on test material used for the study:
- Name of test material (as cited in study report): Isodecyl methacrylate
- Supplier: Evonik Röhm GmbH, Darmstadt, Germany
- Substance type: organic
- Physical state at room temperature: liquid
- Expiration date of the lot/batch: Aug 26, 2008
- Stability under test conditions: Stability in water: Several days at room temperature, refrigerated and in the freezer
- Storage condition of test material: At room temperature
Species / strain / cell type:
other: TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Metabolic activation system:
S9-Mix fraction of Aroclor 1254-induced, male Sprague-Dawley rats and male Syrian hamster livers
Test concentrations with justification for top dose:
Main experiment:
-S9 mix; 0; 33; 100; 333; 1000; 2500; 5000 µg/plate
+S9 mix; 0; 33; 100; 333; 1000; 2500; 5000 µg/plate

According to the results of the pre-experiment the concentrations applied in the main experiments were chosen.
The maximum concentration was 5000.0 µg/plate. The concentration range include two logarithmic decades. Six adequately spaced concentrations
were tested. Two independent experiments were performed.


Each chemical was tested initially at half-log dose intervals up to a dose that elicited toxicity, or to a dose immediately below one which was toxic in
the preliminary toxicity test.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Tetrahydrofuran (THF)
- Justification for choice of solvent/vehicle: The solvent was chosen because of its solubility properties and its relative nontoxicity to the bacteria.
Untreated negative controls:
yes
Remarks:
Concurrent untreated and solvent control: DMSO were performed
Negative solvent / vehicle controls:
yes
Remarks:
vehicle: THF
True negative controls:
yes
Negative solvent / vehicle controls:
yes
Positive controls:
yes
Remarks:
for TA 1535, TA 100
Positive control substance:
sodium azide
Remarks:
sodium azide (purity: >= 99.0%, supplier: Serva, D-69042 Heidelberg, Germany) dissolved in aqua dest.; concentration: 10 µg/plate
Positive controls:
yes
Remarks:
for TA 1537, TA 98
Positive control substance:
other: 4-nitro-o-phenylenediamine, without metabolic activation
Remarks:
4-nitro-o-phenylenediamine (purity: > 99.9%, supplier: Sigma, D-82041 Deisenhofen, Germany) dissolved in DMSO; concentration: 10 µg/plate
Positive controls:
yes
Remarks:
for TA 102
Positive control substance:
methylmethanesulfonate
Remarks:
methylmethanesulfonate (purity: > 99.0%, supplier: Merck-Schuchardt, D-85662 Hohenbrunn, Germany) dissolved in aqua dest.; concentration: 1.0 µl/plate Migrated to IUCLID6: without metabolic activation
Positive controls:
yes
Remarks:
for TA 1535, TA 1537, TA 98, TA 100, TA 102
Positive control substance:
other: 2-aminoanthracene, with metabolic activation
Remarks:
2-aminoanthracene (purity: > 99%, supplier: MERCK, D-64293 Darmstadt, Germany) dissolved in DMSO; concentration: 2.5 µg/plate (10 µg/plate in TA 102)
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation assay as described by Haworth et al. 1983, with some differences.

- Salmonella typhimurium strains were obtained from Dr. Bruce Ames (University of California, Berkeley, U.S.A.) and were stored as recommended (Maron and Ames, 1983).
- Cultures were grown overnight with shaking at 37 °C in Oxoid No. 2 broth, and their phenotypes were analyszed prior to their use for mutagenicity assays.

Test conditions:
System of testing:
-Metabolic activation system: S9 from rat liver, induced with phenobarbital and 5,6-benzoflavone.
Administration:
-Number of replicates: 2
-Plate per test: 3
-Application: pre-incubation
Evaluation criteria:
EVALUATION OF RESULTS
The generally accepted conditions for the evaluation of the results are:

- corresponding background growth on both negative controls and test plates
- normal range of spontaneous reversion rates.

Range of spontaneous reversion frequencies*
------------------------------------------------------------------------------------
TA 135 TA 1537 TA 98 TA 100 TA 102
10 - 29 5 - 28 15 - 57 77 - 189 121 - 293

* These values refer to the negative control without metabolic activation and represent our historical control range in 1993.

According to international guidelines a statistical evaluation of the results is recommended. However, no evaluated statistical procedure can be
recommended for analysis of data from the bacterial assays at this time.
A test article is considered positive if either a dose related and reproducible increase in the number of revertants or a significant and reproducible
increase for at least one test concentration is induced.
A test article producing neither a dose related and reproducible increase in the number of revertants nor a significant and reproducible positive
response at any one of the test points is considered non-mutagenic in this system.

A significant response is described as follows:
A test article is considered mutagenic if in strain TA 100 and TA 102 the number of reversions is at least twice as high and in strains TA 1535,
TA 1537, and TA 98 it is at least three times higher as compared to the spontaneous reversion rate. Also, a dose-dependent and reproducible
increase in the number of revertants is regarded as an indication of possibly existing mutagenic potential of the test article regardless whether the
highest dose induced the above described enhancement factors or not.
Statistics:
According to OECD guideline 471, a statistical analysis of the data is not mandatory.
Key result
Species / strain:
other: Salmonella typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Conclusions:
Interpretation of results: negative

In conclusion, it can be stated that during the decribed mutagenicity test and under the experimental conditions reported, the test item did not
induce gene mutations by base pair changes or frameshifts in the genome of the strains tested.
Therefore, Isodecyl methacrylate has to be judged as nonmutagenic in the presence and absence of mammalian metabolic activation
according to the Ames test results.
Executive summary:

In a reverse gene mutation assay in bacteria (Ames test), strains TA1535, TA1537, TA98, TA100, and TA102 of Salmonella typhimurium were exposed to Isodecyl methacrylate at concentrations of up to 5000 µg/plate in the presence and absence of mammalian metabolic activation S9 -mix. 

No toxic effects occurred in the test groups with and without metabolic activation.The plates incubated with the test article showed normal background growth up to ug/plate with and without S9-mix in all strains used. No substantial increases in revertant colony numbers of any of the five tester strains were observed following treatment with at any concentration level, either in the presence or absence of metabolic activation (S9 -mix). There was also no tendency to higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance.

Appropriate reference mutagens were used as positive controls.

The positive controls induced the appropriate responses in the corresponding strains.

 

There was no evidence of induced mutant colonies over background.

Therefore, Isodecyl methacrylate is considered to be non-mutagenic in this Salmonella typhimurium reverse mutation assay.

 

This reliable and valid study satisfies the requirement for tests for in vitro mutagenicity (bacterial reverse gene mutation) data.

 

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Study period:
10-03-2008 - 30-04-2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
OECD 473, GLP. Study according to relevant guideline.
Qualifier:
according to guideline
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Version / remarks:
adopted May 26, 1983 and EEC Directive 92/69, L 383 A, Annex V, B 10, dated December 29, 1992
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
in vitro mammalian chromosome aberration test
Specific details on test material used for the study:
- Name of test material (as cited in study report): Isodecyl methacrylate
- Supplier: Evonik Röhm GmBH, Darmstadt, Germany
- Substance type: organic
- Physical state at room temperature: liquid
- Expiration date of the lot/batch: Aug 26, 2008
- Stability under test conditions: Stability in water: Several days at room temperature, refrigerated and in the freezer
- Storage condition of test material: At room temperature
Species / strain / cell type:
lymphocytes: human, short-term culture
Details on mammalian cell type (if applicable):
Blood samples were obtained from healthy donors not receiving medication.
Metabolic activation:
with and without
Metabolic activation system:
Mammalian rat liver microsomal fraction induced with phenobarbital / ß-naphthoflavone (S9 mix)
Test concentrations with justification for top dose:
Experiment I: 22 hrs prep. interval, 4 hrs exposure period: without S9 mix: 14.5; 25.4; 44.4; 77.7; 136.0; 238.1; 416.7; 729.1; 1276.0; 2233 µg/ml
22 hrs prep. interval, 4 hrs exposure period: with S9 mix: 14.5; 25.4; 44.4; 77.7; 136.0; 238.1; 416.7; 729.1; 1276.0; 2233 µg/ml




Experiment II: 22 hrs prep. interval, 22 hrs exposure period: without S9 mix: 44.4; 77.7; 136.0; 238.1; 416.7; 729.1; 1276.0; 2233 µg/ml
22 hrs prep. interval, 4 hrs exposure period: with S9 mix: 44.4; 77.7; 136.0; 238.1; 416.7; 729.1; 1276.0; 2233 µg/ml



Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Tetrahydrofuran (THF) (E. Merck, D-64293 Darmstadt, Germany; purity: >= 99.5%)
- Justification for choice of solvent/vehicle: The solvent was chosen according to its solubility properties and its relatrive non-toxicity to the cells.
The final concentration of DMSO in the culture medium was 0.5 % (v/v).
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
concurrent negative and solvent controls
True negative controls:
no
Positive controls:
yes
Remarks:
dissolved in: nutrient medium
Positive control substance:
ethylmethanesulphonate
Remarks:
Final concentration: 825 µg/ml Migrated to IUCLID6: without metabolic activation
Positive controls:
yes
Remarks:
dissolved in: nutrient medium
Positive control substance:
cyclophosphamide
Remarks:
Final concentration: 15 µg/ml in Experiment I and 30 µg/ml in Experiment II Migrated to IUCLID6: with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium


DURATION
- Preincubation period: 50 - 80 hrs
- Exposure duration: 4 h (with S9 mix); 22 hours (without S9 mix)




SPINDLE INHIBITOR (cytogenetic assays): colcemid
STAIN (for cytogenetic assays): Giemsa (Merck, D-64293 Darmstadt, Germany)


NUMBER OF REPLICATIONS: 2


NUMBER OF CELLS EVALUATED: 100 metaphases were scored


DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; in addition the number of polyploid cells in 250 metaphase cells was scored (% polyploid metaphases)
Evaluation criteria:
ACCEPTABILITY OF THE ASSAY
The chromosomal aberration assay is considered acceptable if it meets the following criteria:

a) The number of aberrations found in the negative and/or solvent controls falls within the range of historical laboratory control
data: 0.00 % - 4.00 %.
b) The positive control substances should produce significant increases of the number of cells with structural chromosomal aberrations.


EVALUATION OF RESULTS

A test article is classified as mutagenic if it induces reproducibly either a significant concentration-related increase in the number of structural
chromosomal aberrations or a significant and reproducible positive response for at least one of the test points.

A test article producing reproducibly neither a significant concentration-related increase in the number of structural chromosomal aberrations nor
a significant and reproducibly positive response at any one of the test points is considered nonmutagenic in this system.

Statistical significance was confirmed by means of the Fisher's exact test (p < 0.05).

However, both biological and statistical significance should be considered together.
Statistics:
Statistical significance at the five per cent level (p < 0.05) was evaluated by means of the Fisher's exact test. Evaluation was performed only for cells
carrying aberrations exclusive gaps.
Key result
Species / strain:
lymphocytes: human, short-term culture
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
ADDITIONAL INFORMATION ON CYTOTOXICITY:
Cytotoxicity was only observed in Experiment II, in the absence of S9 mix, where a single clearly reduced mitotic index was observed at the highest dose
evaluated for cytogenetic damage
Remarks on result:
other: strain/cell type: Human Lymphocytes
Conclusions:
Interpretation of results: negative

In the Chromosomal aberration test with human lymphocytes in vitro according to OECD 473 in the presence and absence of
mammalian metabolic activation structural chromosomal aberrations were not observed when tested up to cytotoxic
and/or precipitationg concentrations.
Therefore, the test substance is considered to be non-mutagenic under the experimental conditions reported.
Executive summary:

The test item Isodecyl methacrylate, dissolved in THF, was assessed for its potential to induce structural chromosomal aberrations in human lymphocytes in vitro in two independant experiments.

 

The study design was performed as follows:

 

Experiment I: 22 hrs preparation interval, 4 hrs exposure period without and with S9 mix

                      

Experiment II: 22 hrs prep. interval, 22 hrs exposure period without S9 mix and 22 hrs prep. interval, 4 hrs exposure period with S9 mix.

 

In each experimental group, two parallel cultures were analysed. Per culture 100 metaphase plates were scored for structural chromosomal aberrations, except for the positive controls in Experiment I, in the presence of S9 mix and Experiment II, in the absence of S9 mix, where 50 metaphase plates were scored due to strong clastogenic effects.

 

The highest applied concentration in Experiment I (2333.0 µg/ml of the test item, approx. 9.9 mM) was chosen with regard to the solubility of the test item in an appropriate solvent (THF) and with respect to the current OECD Guideline 473.

 

Dose selection of the cytogenetic experiment was performed considering the toxicity data and the occurrence of test item precipitation in accordance with OECD Guideline 473.

In Experiment I, visible precipitation of the test item in the culture medium was observed at 136.0 µg/ml and above in the absence and presence of S9 mix. In addition, precipitation occurred in Experiment II, in the absence of S9 mix, at 136.0 µg/ml and above and in the presence of S9 mix at 416.7 µg/ml and above. No relevant increase in the osmolarity or pH value was observed. In Experiment I, in the absence and presence of S9 mix, and in Experiment II, in the presence of S9 mix, no cytotoxicity was observed up to the highest applied concentration. In Experiment II, in the absence of S9 mix, a single clearly reduced mitotic index was observed at the highest dose evaluated for cytogenetic damage.

 

In both independent experiments, neither a statistically significant nor a biologically relevant increase in the number of cells carrying structural chromosomal aberrations was observed after treatment with the test item.

 

No relevant increase in the frequencies of polyploid metaphases was found after treatment with the test item as compared to the frequencies of the controls.

 

Appropriate mutagens were used as positive controls. They induced statistically significant increases (p < 0.05) in cells with structural chromosome aberrations.

 

 

In conclusion, it can be stated that under the experimental conditions reported, the test item did not induce structural chromosomal aberrations as determined by the chromosome aberration test in human lymphocytes in vitro.

 

Therefore, the test substance is considered to be non-clastogenic under the experimental conditions reported.

                          

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

Endpoint:
in vitro gene mutation study in mammalian cells
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11-03-2008 - 12-06-2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
OECD 476, GLP. Study according to relevant guideline.
Qualifier:
according to guideline
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
Version / remarks:
adopted July 21, 1997
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of assay:
mammalian cell gene mutation assay
Specific details on test material used for the study:
- Name of test material (as cited in study report): Isodecyl methacrylate
- Supplier: Evonik Röhm GmBH, Darmstadt, Germany
- Substance type: organic
- Physical state at room temperature: liquid
- Expiration date of the lot/batch: Aug 26, 2008
- Stability under test conditions: Stability in water: Several days at room temperature, refrigerated and in the freezer
- Storage condition of test material: At room temperature
Species / strain / cell type:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Metabolic activation system:
Mammalian liver microsomal fraction S9 mix
Test concentrations with justification for top dose:
Experiment I:
without S9 mix: 0.1; 0.3; 0.5; 1.0; and 2.0 µg/ml
with S9 mix: 37.5; 75; 150; 300; and 1200 µg/ml

Experiment II:
without S9 mix: 18.8 ;37.5; 75.0; 150; and 600 µg/ml
with S9 mix: 37.5; 75.0; 150; 300; and 600 µg/ml
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Tetrahydrofuran (THF)
- Justification for choice of solvent/vehicle: relative non-toxicity towards the cells and solubility properties of the test item
Negative solvent / vehicle controls:
yes
Remarks:
concurrent solvent controls (THF)
Positive controls:
yes
Positive controls:
yes
Positive control substance:
ethylmethanesulphonate
Remarks:
Purity > 98 % ; Supplier :ACROS Organics, Geel, Belgium
Positive controls:
yes
Positive control substance:
7,12-dimethylbenzanthracene
Remarks:
Purity 99.5 %; Supplier: Merck, Darmstadt, Germany Migrated to IUCLID6: with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in suspension

DURATION
- Exposure duration: Experiment I: 4 hours with and without metabolic activation
Experiment II: 4 hours with and 24 hours without metabolic activation. The experimental parts of the second experiment with
and without metabolic activation were performed in two separate experiments (experiment II and IIA) for technical reasons. The
results are combined and reported as experiment II.



NUMBER OF CELLS EVALUATED: The stained colonies with more than 50 cells were counted. In doubt the colony size was checked with a preparation
microscope (Nikon, 40407 Düsseldorf, Germany).
Evaluation criteria:
Acceptability of the Assay
The gene mutation assay is considered acceptable if it meets the following criteria:
- the numbers of mutant colonies per 10exp+6 cells found in the negative and/or solvent controls fall within the laboratory historical control data
range of 2001 – 2006.
- the positive control substances must produce a significant increase in mutant colony frequencies (Historical data).
- the cloning efficiency II (absolute value) of the negative and/or solvent controls must exceed 50 %.

Evaluation of Results
A test item is regarded as positive if it induces either a concentration-related increase of the mutant frequency or a reproducible and positive
response at one of the test points.
A test item producing neither a concentration- related increase of the mutant frequency nor a reproducible positive response at any of the test
points is considered non-mutagenic in this system.
A positive response is described as follows:
A test item is regarded as mutagenic if it reproducibly induces a mutation frequency that is three times above the spontaneous mutation frequency
at least at one of the concentrations in the experiment.
The test item is regarded as mutagenic if there is a reproducible concentration-related increase of the mutation frequency. Such evaluation may be
considered also in the case that a threefold increase of the mutant frequency is not observed.
However, in a case by case evaluation this decision depends on the level of the corresponding solvent control data. If there is by chance a low
spontaneous mutation rate in the range normally found ( mutants per 10exp+6 cells) a concentration-related increase of the mutations
within this range has to be discussed. The variability of the mutation rates of solvent controls within all experiments of this study was also taken into consideration.
Statistics:
Statistical Analysis
A linear regression (least squares) was performed to assess a possible dose dependent increase of mutant frequencies using SYSTAT®11 (SYSTAT
Software, Inc., 501, Canal Boulevard, Suite C, Richmond, CA 94804, USA) statistics software. The number of mutant colonies obtained for the
groups treated with the test item were compared to the solvent control groups. A trend is judged as significant whenever the p-value (probability
value) is below 0.05. However, both, biological and statistical significance were considered together.
Key result
Species / strain:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
in forward gene mutations in mammalian cells
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
at 1200 µg/mL with S9 mix; at 1.0 µg/mL and above in Experiment I and 37.5 µg/mL in Experiment II without S9 mix
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid

Results and Discussion

Isodecyl methacrylate was assessed for its potential to induce gene mutations at the HPRT locus using V79 cells of the Chinese hamster.

The assay was performed in two independent experiments with identical experimental procedures, using two parallel cultures each. The first main experiment was performed with and without liver microsomal activation and a treatment period of 4 h. The second experiment was performed with a treatment period of 24 hours in the absence of metabolic activation and 4 hours in the presence of metabolic activation.

The cell cultures were evaluated at the following concentrations:

Experiment I:

without S9 mix:  0.1; 0.3; 0.5; 1.0; and 2.0 µg/ml

with S9 mix: 37.5; 75; 150; 300; and 1200 µg/ml

 

Experiment II:

without S9 mix: 18.8 ;37.5; 75.0; 150; and 600 µg/ml

with S9 mix: 37.5; 75.0; 150; 300; and 600 µg/ml

Phase separation of the test item was observed at 300 µg/mL and above in the first experiment with metabolic activation and at 150 µg/mL and above in the second experiment without metabolic activation. In the second experiment with metabolic activation phase separation was noted at 300 µg/mL and above.

Relevant toxic effects indicated by a relative cloning efficiency 1 below 50 % occurred at 1.0 µg/mL and above in the first experiment without metabolic activation and at 1200.0 µg/mL and above with metabolic activation. In the second experiment toxic effects as described above occurred at 37.5 µg/mL without metabolic activation and at 1200 µg/mL with metabolic activation. The striking difference of toxic concentrations with and without metabolic activation is probably based on protein or lipid binding effects. In the presence of metabolic activation the protein and lipid concentration is higher due to the S9-mix added.This fact is furthermore supported by the considerably less severe cytotoxicity following 24 h treatment without metabolic activation. During long term exposure 10 % FCS have to be added increasing the protein and lipid concentration of the medium.

No relevant and reproducible increase in mutant colony numbers/106cells was observed in the main experiments up to the maximum concentration.

The induction factor reached or exceeded the threshold of three times the corresponding solvent control in experiment I at 37.5 µg/mL in the first culture with metabolic activation and at the same concentration in the first culture in experiment II without metabolic activation. However, both effects were judged as biologically irrelevant fluctuations since no increase was observed at higher concentrations or in the parallel cultures under identical conditions. Furthermore, the effects were not dose-dependent as indicated by the lacking statistical significance.

In both experiments of this study (with and without S9 mix) the range of the solvent controls was from 5.7 up to 24.0 mutants per 106cells; the range of the groups treated with the test item was from 3.3 up to 34.1 mutants per 106cells.

 

EMS(150 µg/mL in experiment I and 75 µg/mL in experiment II) and DMBA (2.0 µg/mL) were used as positive controls and showed a distinct increase in induced mutant colonies. This showed the sensitivity of the test system and the activity of the S9 mix.

 

Conclusion

 

In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells.

 

Therefore, Isodecyl methacrylate is considered to be non-mutagenic in this HPRT assay.

 

Conclusions:
Interpretation of results: negative

In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells.
Therefore, Isodecyl methacrylate is considered to be non-mutagenic in this HPRT assay.
Executive summary:

Isodecyl methacrylate was assessed for its potential to induce gene mutations at the HPRT locus using V79 cells of the Chinese hamster.

The assay was performed in two independent experiments with identical experimental procedures, using two parallel cultures each. The first main experiment was performed with and without liver microsomal activation and a treatment period of 4 h. The second experiment was performed with a treatment period of 24 hours in the absence of metabolic activation and 4 hours in the presence of metabolic activation.

The cell cultures were evaluated at the following concentrations:

Experiment I:

without S9 mix:  0.1; 0.3; 0.5; 1.0; and 2.0 µg/ml

with S9 mix: 37.5; 75; 150; 300; and 1200 µg/ml

 

Experiment II:

without S9 mix: 18.8 ;37.5; 75.0; 150; and 600 µg/ml

with S9 mix: 37.5; 75.0; 150; 300; and 600 µg/ml

Phase separation of the test item was observed at 300 µg/mL and above in the first experiment with metabolic activation and at 150 µg/mL and above in the second experiment without metabolic activation. In the second experiment with metabolic activation phase separation was noted at 300 µg/mL and above.

Relevant toxic effects indicated by a relative cloning efficiency 1 below 50 % occurred at 1.0 µg/mL and above in the first experiment without metabolic activation and at 1200.0 µg/mL and above with metabolic activation. In the second experiment toxic effects as described above occurred at 37.5 µg/mL without metabolic activation and at 1200 µg/mL with metabolic activation. The striking difference of toxic concentrations with and without metabolic activation is probably based on protein or lipid binding effects. In the presence of metabolic activation the protein and lipid concentration is higher due to the S9-mix added.This fact is furthermore supported by the considerably less severe cytotoxicity following 24 h treatment without metabolic activation. During long term exposure 10 % FCS have to be added increasing the protein and lipid concentration of the medium.

 

No relevant and reproducible increase in mutant colony numbers/106cells was observed in the main experiments up to the maximum concentration.

 

The induction factor reached or exceeded the threshold of three times the corresponding solvent control in experiment I at 37.5 µg/mL in the first culture with metabolic activation and at the same concentration in the first culture in experiment II without metabolic activation. However, both effects were judged as biologically irrelevant fluctuations since no increase was observed at higher concentrations or in the parallel cultures under identical conditions. Furthermore, the effects were not dose-dependent as indicated by the lacking statistical significance.

In both experiments of this study (with and without S9 mix) the range of the solvent controls was from 5.7 up to 24.0 mutants per 106cells; the range of the groups treated with the test item was from 3.3 up to 34.1 mutants per 106cells.

 

EMS(150 µg/mL in experiment I and 75 µg/mL in experiment II) and DMBA (2.0 µg/mL) were used as positive controls and showed a distinct increase in induced mutant colonies. This showed the sensitivity of the test system and the activity of the S9 mix.

 

Conclusion

 

In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells.

 

Therefore, Isodecyl methacrylate is considered to be non-mutagenic in this HPRT assay.

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

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

Genetic toxicity in vivo

Description of key information

There is no study available for tetradecyl methacrylate but for structural analogue substances Isodecyl methacrylate CAS: 29964 -84 -9 and Dodecyl methacrylate CAS: 142 -90 -5.

Read across from structural analogue substances Isodecyl methacrylate and Dodecyl methacrylate.

No single key study has been selected for in vitro genetic toxicity; instead, all available studies have been used in a weight-of-evidence approach. Read-across in vitro: In three different reliable (Klimisch score: 1, with GLP) in vitro genetic toxicity studies Isodecyl methacrylate (structurally related substance of Tetradecyl methacrylate) received negative results. The absence of a mutagenic potential of Tetradecyl methacrylate in vitro was supported by peer reviewed data from Dodecyl methacrylate of the review (Johannsen et al., 2008). Genetic toxicity data in vivo with test substance Isodecyl methacrylate: Mouse micronucleus test, OECD 474: negative (Klimisch score: 1, GLP, (RCC-CCR, 1989))

Additional information

Read across from structurally related substances isodecyl methacrylate CAS: 29964 -84 -9 and monomer mix ((methacrylic acid ester of an alcohol mixture with an average C-number of 14,8) CAS:

For the assessment of tetradecyl methacrylate there are data of the structurally related substances isodecyl methacrylate (three in vitro studies) and monomer mix ((methacrylic acid ester of an alcohol mixture with an average C-number of 14,8) two in vitro studies) available.

One reverse mutation assay in bacteria, one chromosome aberration test and one HPRT test with the structurally related substance isodecyl methacrylate are available. With monomer mix as test substance one reverse mutation assay in bacteria and one chromosome aberration test in V79 cells are available.

In vivo there is one micronucleus study with structural analogue dodecyl methacrylate available.

 

Bacterial reverse mutation assay

No study is available on tetradecyl methacrylate but there are two studies on the structurally related substances isodecyl methacrylate and monomer mix.

In a reverse gene mutation assay in bacteria according to OECD guideline 471 Salmonella typhimurium TA98, TA100, TA1535, TA1537 and TA102 strains were exposed to isodecyl methacrylate in THF at concentrations up to 5000 µg/plate in the presence and absence of mammalian metabolic activation. The positive controls induced the appropriate responses in the corresponding strains. There was no evidence of induced mutant colonies over background (Klimisch score: 1, GLP, (RCC-CCR, 2008)).

In a reverse gene mutation assay in bacteria according to OECD guideline 471 Salmonella typhimurium TA98, TA100, TA1535, TA1537 and TA102 strains were exposed to monomer mix in THF at concentrations up to 3000 µg/plate in the presence and absence of mammalian metabolic activation. The positive controls induced the appropriate responses in the corresponding strains. There was no evidence of induced mutant colonies over background (Klimisch score: 1, GLP, (RCC-CCR, 2005)).

 

In vitro Chromosome aberration test in human lymphocytes

No study is available on tetradecyl methacrylate but there is a study on the structurally related substances isodecyl methacrylate.

 

In a chromosome aberration test in human lymphocytes according to OECD guideline 473 (adopted May 26, 1983), human lymphocyte cultures were exposed to Isodecyl methacrylate (98.9%) in THF with and without metabolic activation (S9 mix).

The following concentrations were evaluated:

Experiment I:

22 hrs prep. interval, 4 h treatment without metabolic activation: 14.5, 25.4, 44.4, 77.7, 136.0, 238.1, 416.7, 729.1, 1276.0, 2233 µg/ml

22 hrs prep. interval, 4 h treatment with metabolic activation: 14.5, 25.4, 44.4, 77.7, 136.0, 238.1, 416.7, 729.1, 1276.0, 2233 µg/ml

Experiment II:

22 hrs prep. interval, 22 h treatment without metabolic activation: 44.4, 77.7, 136.0, 238.1, 416.7, 729.1, 1276.0, 2233 µg/ml

22 hrs prep. interval, 4 h treatment with metabolic activation: 44.4, 77.7, 136.0, 238.1, 416.7, 729.1, 1276.0, 2233 µg/ml

In Experiment I, visible precipitation of the test item in the culture medium was observed at 136.0 µg/ml and above in the absence and presence of S9 mix. In addition, precipitation occurred in Experiment II, in the absence of S9 mix, at 136.0 µg/ml and above and in the presence of S9 mix at 416.7 µg/ml and above. No relevant increase in the osmolarity or pH value was observed. In Experiment I, in the absence and presence of S9 mix, and in Experiment II, in the presence of S9 mix, no cytotoxicity was observed up to the highest applied concentration. In Experiment II, in the absence of S9 mix, a single clearly reduced mitotic index was observed at the highest dose evaluated for cytogenetic damage.

Isodecyl methacrylate was tested up to cytotoxic concentration.

 

In the absence and presence of S9 mix neither a statistically nor a biologically relevant increase in the number of cells carrying structural chromosome aberrations were observed after treatment with the test item (Klimisch score: 1, GLP, (RCC-CCR, 2008))

In vitro Chromosome aberration test in Chinese hamster V79 cells

No study is available on tetradecyl methacrylate but there is a study with the structurally related substance, monomer mix (details on composition see above).

 

In a chromosome aberration test in Chinese hamster V79 cells according to OECD guideline 473 (adopted July 21, 1997), human lymphocyte cultures were exposed to monomer mix dissolved in THF with and without metabolic activation (S9 mix).

The following concentrations were evaluated:

Experiment I:

18 hrs prep. interval, 4 h treatment without and with metabolic activation: 5.9, 11.7, 23.4, 46.9, 93.8, 187.5 (p), 375.0 (p), 750.0 (p), 1500.0 (p), 3000.0 (p) µg/ml (p): Precipitation occurred

Experiment II:

18 hrs prep. interval, 18 h treatment without metabolic activation: 46.9, 93.8, 187.5 (p), 375.0 (p), 750.0 (p), 1500.0 (p), 3000.0 (p µg/ml

28 hrs prep. interval, 28 h treatment without metabolic activation: 23.4, 46.9, 93.8, 187.5 (p), 375.0 (p), 750.0 (p), 1500.0 (p), 3000.0 (p) µg/ml

Experiment II: 28 hrs prep. interval, 4 hrs exposure period: with S9 mix: 23.4, 46.9, 93.8, 187.5 (p), 375.0 (p), 750.0 (p) µg/ml

(p): Precipitation occurred

No clear toxic effects indicated by reduced mitotic indices or cell numbers of below 50% were observed up to the highest required test item concentration. Therefore, concentrations at the border of test item solubility in culture medium were scored for cytogenetic damage.

 

In both independent experiments, no biologically relevant increase in the number of cells carrying structural chromosomal aberrations was observed after treatment with the test item. However, in Experiments I and II, in the absence of S9 mix, statistically significant increases (2 % - 2.5 % aberrant cells, exclusive gaps) were observed, but were within our historical control range (0.0 - 4.0 % aberrant cells, exclusive gaps) and are regarded as being biologically irrelevant.

 

No relevant increase in the frequencies of polyploid metaphases was found after treatment with the test item as compared to the frequencies of the controls.

 

In the absence and presence of S9 mix neither a statistically nor a biologically relevant increase in the number of cells carrying structural chromosome aberrations were observed after treatment with the test item (Klimisch score: 1, GLP, (RCC-CCR, 2005))

HPRT-test in mammalian cells

No study is available on tetradecyl methacrylate but there is a study on the structurally related substance isodecyl methacrylate.

Isodecyl methacrylate was assessed for its potential to induce gene mutations at the HPRT locus using V79 cells of the Chinese hamster according to OECD 476.

The assay was performed in two independent experiments with identical experimental procedures, using two parallel cultures each. The first main experiment was performed with and without liver microsomal activation and a treatment period of 4 h. The second experiment was performed with a treatment period of 24 hours in the absence of metabolic activation and 4 hours in the presence of metabolic activation.

The cell cultures were evaluated at the following concentrations:

Experiment I:

without S9 mix:  0.1; 0.3; 0.5; 1.0; and 2.0 µg/ml

with S9 mix: 37.5; 75; 150; 300; and 1200 µg/ml

 

Experiment II:

without S9 mix: 18.8 ;37.5; 75.0; 150; and 600 µg/ml

with S9 mix: 37.5; 75.0; 150; 300; and 600 µg/ml

In both experiments of this study (with and without S9 mix) the range of the solvent controls was from 5.7 up to 24.0 mutants per 106cells; the range of the groups treated with the test item was from 3.3 up to 34.1 mutants per 106cells.

 

EMS(150 µg/mL in experiment I and 75 µg/mL in experiment II) and DMBA (2.0 µg/mL) were used as positive controls and showed a distinct increase in induced mutant colonies. This showed the sensitivity of the test system and the activity of the S9 mix.

 

In conclusion it can be stated that under the experimental conditions reported the test item did not induce gene mutations at the HPRT locus in V79 cells.

 

In vivo Micronucleus test

In a NMRI mouse bone marrow micronucleus assay, 5 animals/male/female/dose were treated orally with structural analogue dodecyl methacrylate (98%, stabilizied) at a dose of 0, 5000 mg/kg bw according to OECD 474. The test article was suspended in carboxymethyl cellulose (1%). This suspending agent was used as negative control. The volume administered orally was 10 ml/kg b.w.. 24 h, 48 h and 72 h after a single application of the test article the bone marrow cells were collected for micronuclei analysis. Ten animals (5 males, 5 females) per test group were evaluated for the occurrence of micronuclei. 1000 polychromatic erythrocytes (PCE) per animal were scored for micronuclei.

To describe a cytotoxic effect due to the treatment with the test article the ratio between polychromatic and normochromatic erythrocytes (NCE) was determined in the same sample and reported as the number of NCE per 1000 PCE.

The following dose level of the test article was investigated:

24 h, 48 h, and 72 h preparation interval: 5000 mg/kg b.w..

In a pre-experiment this dose level was estimated to be the maximum attainable dose. The animals expressed toxic reactions. After treatment with the test article the ratio between PCEs and NCEs was not affected as compared to the corresponding negative controls thus indicating no cytotoxic effects.

In comparison with the corresponding negative controls there was no enhancement in the frequency of the detected micronuclei at any preparation interval after application of the test article.

An appropriate reference mutagen was used as positive control which showed a distinct increase of induced micronucleus frequency. 

This study is classified as acceptable. It satisfies the requirement for Test Guideline OECD 474 for in vivo cytogenetic mutagenicity data.

Therefore, Dodecyl methacrylate is considered to be non-mutagenic in this micronucleus assay.

Conclusion:

The absence of a mutagenic potential was demonstrated for gene mutations as well as chromosome mutations for isodecyl methacrylate and monomer mix.

These results are representative for tetradecyl methacrylate as isodecyl methacrylate represents a worst-case in terms of bioavailability due to its molecular size, lipophilicity and water solubility.

A negative mouse micronucleus tests with Dodecyl methacrylate supports the absence of a mutagenic potential in vivo.



Short description of key information:
There is no study available for tetradecyl methacrylate. But there are serveral studies available for structural analogue substances dodecyl methacrylate and isodecyl methacrylate as well as Monomer Mix (Methacrylic acid ester of an alcohol mixture with an average C-number of 14.8).
No single key study has been selected for in vitro genetic toxicity; instead, all available studies have been used in a weight-of-evidence approach.
Read-across in vitro:
In three different reliable (Klimisch score: 1, with GLP) in vitro genetic toxicity studies with Isodecyl methacrylate (structurally related substance of tetradecyl methacrylate) received negative results. Furthermore there are two additional in vitro studies with Monomer mix (Klimisch score: 1, GLP) which received negative results as well.

Genetic toxicity data in vivo:
Mouse micronucleus test, OECD474 with structural analogue dodecyl methacrylate: negative (Klimisch score: 1, GLP, (RCC-CCR, 1989))

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Based on the reliable available data on structural analogue substances dodecyl methacrylate, isodecyl methacrylate and monomer mix (structurally related substances of tetra decyl methacrylate), tetradecyl methacrylate does not need to be classified for mutagenicity according to the criteria given in regulation (EC) 1272/2008 (CLP (EU-GHS) or the former European directive on classification and labelling 67/548/EEC. Thus, no labelling is required.