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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

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

For the assessment of dodecyl methacrylate there are peer reviewed data of a review available and 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.

 

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.

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.

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

Genetic toxicity in vivo

Description of key information

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 Dodecyl methacrylate) received negative results. The absence of a mutagenic potential of Dodecyl methacrylate in vitro was supported by peer reviewed data of the review (Johannsen et al., 2008). Genetic toxicity data in vivo: Mouse micronucleus test, OECD474: negative (Klimisch score: 1, GLP, (RCC-CCR, 1989))

Link to relevant study records
Reference
Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09-July-1989 - 31-Aug.- 1989
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
OECD 474, GLP.
Qualifier:
according to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Deviations:
no
Principles of method if other than guideline:
Pre-Experiment for Toxicity
A preliminary study on acute toxicity was performed with 2 animals per sex under identical conditions as in the mutagenicity study concerning:
starvation period, animal strain; vehicle; route, frequency, and volume of administration.
The animals were treated orally with a single dose of 5000 mg/kg bw in 1% CMC with the test item and examined for acute toxic symptoms.
GLP compliance:
yes
Type of assay:
micronucleus assay
Specific details on test material used for the study:
- Supplier: Evonik Röhm GmbH, D-64293 Darmstadt, Germany
Species:
mouse
Strain:
NMRI
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BRL Tierfarm, Füllinsdorf, Switzerland
- Number of animals. 84 (42 males/42 females)
- Age at study initiation: minimum 11 weeks
- Weight at study initiation: approximately 30 g
- Assigned to test groups randomly: yes
- Fasting period before study: 18 hours before treatment
- Housing: single
- Diet: pelleted standard diet,ad libitum
- Water: tap water, ad libitum
- Acclimation period: minimum 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ±3°C
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs artifical light
Route of administration:
oral: gavage
Vehicle:
- Vehicle(s)/solvent(s) used: 1% CMC (carboxymethyl cellulose)
- Justification for choice of solvent/vehicle: vehicle was chosen to its relative non-toxicity for animals. All animals received a single standard volume
of 10 mL/kg body weight orally.
- Concentration of test material in vehicle:
- Supplier: Fluka; SIGMA-Aldrich Vertiebs-GmbH, 82041 Deisenhofen, Germany
- Lot/batch no.: no data
- Catalogue no.: 21902
- Purity: no data
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
On the day of the experiment, the test substance was formulated in 1% CMC. All animals received a single standard volume of 10 ml/kg body weight
orally.

DIET PREPARATION
- Rate of preparation of diet (frequency): no data
- Mixing appropriate amounts with (Type of food): no data
- Storage temperature of food: no data
Duration of treatment / exposure:
24h, 48h and 72 hours
Frequency of treatment:
single dose
Remarks:
Doses / Concentrations:
5000 mg/kg suspended in Carboxymethylcellulose (1 %)
Basis:
nominal conc.
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Positive control(s):
CPA; cyclophosphamide
- Supplier: SERVA, Heidelberg, Germany
- Purity: commercial grade
- Dissolved in: physiological saline
- Route of administration: orally, once
- Doses / concentrations: 40 mg/kg bw
Volume administered: 10 mL/kg bw

The stability of CPA at room temperature is sufficient. At 20 °C only 1% of CPA is hydrolysed per day in aqueous solution.
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION:
It is generally recommended to use the maximum tolerated dose or the highest dose that can be formulated and administered reproducibly or
the highest dose that can be formulated and administered reproducibly.
The maximum tolerated dose level is determined to be the dose that causes toxic reactions without having major effects on survival within 72
hours.
The volume to be administered should be compatible with physiological space available.

TREATMENT AND SAMPLING TIMES:

Treatment:
Approximately 18 hours before treatment the animals received no food but water ad libitum. At the beginning of the treatment the animals (including
the controls) were weighed and the individual volume to be administered was adjusted to the animals body weight. The animals received the test item, the vehicle or the positive control substance once. Twelve animals, six males and six females, were treated per dose group and sampling time.
Sampling of bone marrow was done 24, 48 and 72 hours after treatment, respectively.


DETAILS OF SLIDE PREPARATION:

Preparation of the Animals:
The animals were sacrificed by cervical dislocation. The femora were removed, the epiphyses were cut off and the marrow was flushed out with fetal
calf serum, using a syringe. The cell suspension was centrifuged at 1000 rpm for 5 minutes and the supernatant was discarded. A small
drop of the resuspended cell pellet was spread on a slide. The smear was air-dried and then stained with May-Grünwald (Kindler, D-79110 Freiburg,
Germany/Giemsa. Cover slips were mounted with EUKITT (KINDLER, D-79110 Freiburg, Germany). At least one slide was made from each bone
marrow sample.

METHOD OF ANALYSIS:
Analysis of Cells:
Evaluation of the slides was performed using NlKON microscopes with 100x oil immersion objectives. At least 1000 polychromatic erythrocytes (PCE)
were analysed per animal for micronuclei. To describe a cytotoxic effect the ratio between polychromatic and normochromatic erythrocytes was
determined in the same sample and expressed in normochromatic erythrocytes per 1000 erythrocytes. The analysis was performed with coded
slides.
Ten animals (5 males, 5 females) per test group were evaluated as described. The remaining animal of each test group was evaluated in case an
animal had died in its test group spontaneously or due to gavage error.
Evaluation criteria:
A test article is classified as mutagenic if it induces either a statistically significant dose-related increase in the number of micronucleated
polychromatic erythrocytes or a reproducible statistically significant positive response for at least one of the test points.
A test article producing neither a statistically significant dose-related increase in the number of micronucleated polychromatic erythrocytes nor a
statistically significant and reproducible positive response at anyone of the test points is considered nonmutagenic in this system.
This can be confirmed by means of the nonparametric Mann-Whitney test.
However, both biological and statistical significance should be considered together.
mntire
Statistics:
Statistical methods (nonparametric Mann-Whitney test) will be used as an aid in evaluating the results. However, the primary point of consideration
is the biological relevance of the results.
Sex:
male/female
Genotoxicity:
negative
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RESULTS OF RANGE-FINDING STUDY (see Remarks on results including tables and figures)

In a pre-experiment 4 animals (2 males, 2 females) received orally a single dose of 5000 mg/kg b.w., Dodecylmethacrylate
suspended in CMC (1%). The volume administered was 10 ml/kg b.w..
All treated animals expressed toxic reactions: reduction of spontaneous activity, eyelid closure and apathy. One female expressed pilo erection.

Summary of results test group                     dose mg/kg bw              sampling              PCEs with              range                            PCE/NCE                                                                                time (hours)        micronuclei        ------------------------------------------------------------------------------------------------------------------------------ suspending agent                     0                            24                     0.09%                 0 - 2                           1000 / 404                        test article                            5000                         24                    0.06%                  0 - 3                           1000 / 511 cyclophosphamide                40                           24                    1.06%                 3 - 27                           1000 /935         suspending agent                     0                            48                     0.14%                  1 - 4                           1000 / 688                      test article                            5000                          48                     0.03%                  0 - 2                          1000 / 900     suspending agent                     0                            72                      0.09%                 0 - 3                           1000 / 676                      test article                            5000                          72                      0.07%               0 - 2                           1000 / 670 -------------------------------------------------------------------------------------------------------------------------------

Conclusions:
Interpretation of results: negative
During the study decribed and under the experimental conditions reported, Dodecylmethacrylate did not induce micronuclei. This was determined by the micronucleus test with bone marrow cells of the mouse (OECD guideline 474).
Therefore, Dodecylmethacrylate is considered to be non-mutagenic in this micronucleus assay.
Executive summary:

In a NMRI mouse bone marrow micronucleus assay, 5 animals/male/female/dose were treated orally with Dodecylmethacrylate (98%, stabilizied) at a dose of 0, 5000 mg/kg bw. The test article was suspended in Carboxymethylcellulose (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 accceptable. This study satisfies the requirement for Test Guideline OECD 474 for in vivo cytogenetic mutagenicity 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 conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vivo:

For the assessment of dodecyl methacrylate there are peer reviewed data of a review available and 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.

In vivo there is one micronucleus study with dodecyl methacrylate 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.

 

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.

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.

 

In vivo Micronucleus test

In a NMRI mouse bone marrow micronucleus assay, 5 animals/male/female/dose were treated orally with 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, dodecylmethacrylate 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.

These results are representative for Dodecyl 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.



Justification for selection of genetic toxicity endpoint
There is one reliable (Klimisch score: 1, GLP) in vivo study with Dodecyl methacrylate.
In three different reliable (Klimisch score: 1, with GLP) in vitro genetic toxicity studies Isodecyl methacrylate (structurally related substance of Dodecyl methacrylate) received negative results.

Justification for classification or non-classification

Based on the reliable available data on Dodecyl methacrylate and Isodecyl methacrylate (structurally related substance of Dodecyl methacrylate), Dodecyl 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.