Propylidynetrimethyl trimethacrylate

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From January 01 to March 22, 2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

mammalian erythrocyte micronucleus test

Test material

Test material information

Constituent 1

Reference substance name:

Propylidynetrimethyl trimethacrylate

EC Number:

221-950-4

EC Name:

Propylidynetrimethyl trimethacrylate

Cas Number:

3290-92-4

Molecular formula:

C18H26O6

IUPAC Name:

2,2-bis[(methacryloyloxy)methyl]butyl methacrylate (non-preferred name)

Test material form:

other: Clear to slightly turbid yellowish liquid

Details on test material:

- Name of test material (as cited in study report): Trimethylolpropantrimethacrylate (TMPTMA)
- Physical state: Clear to slightly turbid yellowish liquid

- Storage condition of test material: At room temperature

Test animals

Species:

mouse

Strain:

NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: HARLAN WINKELMAN, Borchen, Germany
- Age at study initiation: 23-33 g
- Assigned to test groups randomly: Yes
- Housing: Housed in a group of 5 mice/sex in Macaroon Type III cage
- Diet (e.g. ad libitum): Pelleted standard diet (ALTROMIN, Lage/Lippe, Germany)
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25
- Humidity (%): 55 ± 10
- Photoperiod (hours dark / hours light): 12 hours dark/12 hours light

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: CMC (carboxymethyl cellulose)
- Justification for choice of solvent/vehicle: Nontoxic to animals
- Source: SIGMA, Germany
- Amount of vehicle (if gavage or dermal): 33.3 mL/kg bw
- Lot/batch no. (if required): 36H0738

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Test material was prepared and diluted with carboxymethylcellulose.

Duration of treatment / exposure:

24 hours (in all test groups) and 48 hours (in 2000 mg/kg bw group)

Frequency of treatment:

Once

Post exposure period:

Not applicable

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 200, 600 and 2000 mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

Five

Control animals:

yes, concurrent vehicle

Positive control(s):

CPA (cyclophosphamide)
- Source: SIGMA, Germany
- Analytical purity: At least 98%
- Route of administration: i.p.
- Doses / concentrations: 30 mg/kg bw
- Volume administered: 10 mL/kg bw
- Lot/batch no.: 087H0207

Examinations

Tissues and cell types examined:

- Femora bones were removed for marrow extraction and the prepared slides were examined for polychromatic erythrocytes (PCEs), normochromatic erythrocytes (NCEs) and total erythrocytes

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: Dose selection was based on a preliminary range-finding test conducted on 3 mice/sex/dose at 2000 mg/kg bw. No toxic signs were recorded.

TREATMENT AND SAMPLING TIMES: Femora were removed for marrow extraction from single animals in each treatment and control group at 24 or 48 hours after exposure.

DETAILS OF SLIDE PREPARATION: Bone marrow cells extracted, preparations smeared on slides and air-dried. The slides were stained with May-Grunwald solution/ Giemsa and coded.

METHOD OF ANALYSIS: Slides were scanned using Olympus microscopes to determine the frequency of micronuclei in 2000 polychromatic erythrocytes (PCEs) per animal. In addition, PCE:NCE ratio was determined in the same sample and expressed as NCE/1000 PCEs.

Evaluation criteria:

- Criteria for a positive response: Detection of either a statistically significant dose related increase in the number of micronucleated PCEs or a reproducible statistically significant positive response for at least one of the test points.
- Test article that does not induce both of these responses is considered negative.
- Both biological and statistical significance should be considered together.

Statistics:

Statistical significance at the 5% level (p < 0.05) was evaluated by means of the non-parametric Mann-Whitney test.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- Dose: 2000 mg/kg bw
- Clinical signs of toxicity in test animals: No

RESULTS OF DEFINITIVE STUDY
- Frequency of micronuclei in polychromatic erythrocytes and PCE:NCE ratio: See table 1

Any other information on results incl. tables

Table 1: Micronucleus Assay - summary table

 

Treatment	Dose	Harvest time (hours)	% Micronucleated PCEs (mean of 2000 PCEs per animal)				Ratio PCE:NCE (mean)
			males		females		males	females
			all	small	all	small
Vehicle control	Carboxymethylcellulose 33.3 mL/kg bw	24	0.16	0.14	0.16	0.14	1.14	1.35
Positive control	Cyclophosphamide 30 mg/kg bw	24	1.01	0.94	0.98	0.92	1.35	1.45
Test Article	200 mg/kg bw	24	0.18	0.13	0.14	0.11	1.19	0.99
	600 mg/kg bw	24	0.09	0.07	0.12	0.09	0.86	0.93
	2000 mg/kg bw	24	0.12	0.11	0.1	0.09	0.89	1.11
		48	0.16	0.16	0.12	0.12	0.58	1.19

Applicant's summary and conclusion

Conclusions:

Under the test conditions, trimethylolpropantrimethacrylate is not considered as mutagenic in the mouse bone marrow micronucleus test.

Executive summary:

In a bone marrow micronucleus test, performed according to OECD guideline 474 and in compliance with GLP, groups of NMRI mice (5/sex/dose) were given a single oral (gavage) dose of trimethylolpropantrimethacrylate in carboxymethylcellulose at doses of 0, 200, 600 and 2000 mg/kg bw. Bone marrow was extracted after 24 hours (in all test groups) or 48 hours (in 2000 mg/kg bw group) of exposure and the prepared slides were scanned to determine the frequency of micronuclei in 2000 polychromatic erythrocytes (PCEs) for each animal. In addition, PCE:NCE ratio was determined in the same sample and expressed as NCE/1000 PCEs. A preliminary range-finding test was also conducted on 3 mice/sex/dose at 2000 mg/kg bw in which no toxic signs were recorded.

 

No statistically significant increases in the frequency of micronucleated PCEs were observed at any dose levels. PCE:NCE ratio was slightly affected at a dose level of 2000 mg/kg bw at 24 and 48 hours indicating slight cytotoxicity of the test material. Positive control (cyclophosphamide, 30 mg/kg bw) induced the appropriate response.

 

Under the test conditions, trimethylolpropantrimethacrylate is not considered as mutagenic in the mouse bone marrow micronucleus test.