2-hydroxyethyl methacrylate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well conducted study, carried out by the Hatano research Institute, Food and Drug Safety Center (Japan), Guideline study, GLP; german translation available, only study summary is written English.

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Reference 2

Reference Type:

publication

Title:

Relevance of chemical structure and cytotoxicity to the induction of chromosome aberrations based on the testing results of 98 high production volume industrial chemicals.

Author:

Kusakabe H, Yamakage K, Wakuri S, Sasaki K, Nakagawa Y, Watanabe M, Hayashi M, Sofuni T, Ono H, Tanaka N

Year:

2002

Bibliographic source:

Mutation Research 517 (1-2): 187-198

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

2-hydroxyethyl methacrylate

EC Number:

212-782-2

EC Name:

2-hydroxyethyl methacrylate

Cas Number:

868-77-9

Molecular formula:

C6H10O3

IUPAC Name:

2-hydroxyethyl methacrylate

Test material form:

liquid

Specific details on test material used for the study:

Supplier: Nippon Shokubai
Batch No. 5P05LA
Purity: 97.6 %
Ethylenglycol dimethacrylate: 0.2 - 0.3 %
Diethylenglycol monomethacrylate: 2.0 -2.5%
Stabilzer: 50 ppm Hydrochinon monomethylether

Method

Species / strain

Species / strain / cell type:

other: Chinese hamster lung (CHL/IU) cells

Cytokinesis block (if used):

The cytotoxic effect of the test substance on CHL/IU cells was determined using a monolayer cell density meter (Monocellater TM, manufactured by Olympus Kogaku Kogyo (AG)) by determining the growth rate for each group.
The cell growth was compared control group with /neutral/ medium. It was found that with both continuous and short-term medication, the cytotoxic did not exceed 50% at all concentrations used.

Metabolic activation:

with and without

Metabolic activation system:

Rat liver S9-mix

Test concentrations with justification for top dose:

-S9 mix (continuous treatment): 0, 0.16, 0.33, 0.65, 1.3 mg/ml; -S9-mix and +S9-mix(short-term treatment): 0, 0.33, 0.65 and 1.3 mg/ml

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: solubility tests

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: -S9 mix, Mitomycin C; +S9 mix, Cyclophosphamide S-9: Rat liver, induced with phenobarbital and 5,6-benzoflavone

Details on test system and experimental conditions:

Test duration: continous treatment: 24-hours                
short-term treatment: 6-hours
Plates/test: 2

Evaluation criteria:

A substance is considered clastogenic if: - any dose level shows
a statistically signicant increase in aberration-bearing
cells - the increase is over historical controls - the increase is present in both replicates

Statistics:

yes

Results and discussion

Test results

Species / strain:

other: Chinese hamster lung (CHL/IU) cells

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

other: Toxicity was not observed up to 0.65 mg/ml in continuous and short-term treatment with or without S9-mix.

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

Structural chromosomal aberrations (including gap) were induced under the following conditions: 
24 h continuous treatment (0.65 and 1.3 mg/ml: mid and high concentrations, 10.0 and 70.6 %, respectively); 
48 h continuous treatment (0.16 - 0.65 mg/ml: all concentrations, 6.0 - 84.0 %);
short-term treatment with an exogenous metabolic activation system (1.3 mg/ml: high concentration, 13.0 %). 
Polyploidy was induced under the following conditions: the 48 h continuous treatment (0.65 mg/ml, 3.25 %); 
short-term treatment with an exogenous metabolic activation system (0.65 mg/ml: mid concentration, 1.25 %); 
short-term treatment without the metabolic activation system (0.33 and 1.3 mg/ml: low and high concentrations, 0.88 and 6.13 %, respectively). 
However, a trend test showed no dose-dependency for the  polyploidy with short-term treatment and the metabolic activation system.
Lowest concentration producing cytogenetic effects in vitro:
Without metabolic activation (continuous treatment): 0.16 mg/ml (clastogenicity)                           
0.65 mg/ml (polyploidy)
Without metabolic activation (short-term treatment):  0.33 mg/ml (polyploidy)
With metabolic activation (short-term treatment): 1.3 mg/ml (clastogenicity)      
0.65 mg/ml (polyploidy)

Any other information on results incl. tables

Result table see under attachments

Applicant's summary and conclusion

Conclusions:

Interpretation of results:
positive

Structural chromosomal aberrations (including gap) were induced under the conditions of the study.

Executive summary:

HEMA has been evaluated for its ability to induce chromosomal aberrations in mammalian cells in culture (MHW 1997). Kusakabe et al. (2002) evaluated the clastogenic potential of HEMA along with a large number of other substances in Chinese hamster lung cells in culture, exposed to concentrations up to 1.3 mg/ml. HEMA was reported to induce structural chromosome aberrations following 6-hour exposure of cells but only in the presence of S9 at 1.3 mg/ml. Continuous exposure of cells for 24 or 48 hours without S9 also caused an elevated incidence of chromosome aberrations (from 0.16 mg/ml for the 48-hour exposure and from 0.65 mg/ml for the 24-hour exposure). Polyploidy was reported after both short-term treatment and 48-hour continuous treatment exposures. However, no dose-dependency was observed for polyploidy in the short-term treatment with metabolic activation. These effects were found at exposure levels without cytotoxicity or at concentrations which caused <50% cell death (no toxicity up to 0.65 mg/ml). For careful interpretation of these results two publications by Fujita et al (2016) should be considered. If the cytotoxicity index relative cell count (RCC) is replaced with a new index, RICC or RPD (relative increase in cell count/relative population doubling), the result was identified as being possibly false positive.