Poly[oxy(methyl-1,2-ethanediyl)], α-hydro-ω-hydroxy-, ether with 2-ethyl-2-(hydroxymethyl)-1,3-propanediol (3:1), 2-propenoate

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 November 02, 1998 to January 16, 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

no

GLP compliance:

yes

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

Propylidynetrimethanol, ethoxylated, esters with acrylic acid

EC Number:

500-066-5

EC Name:

Propylidynetrimethanol, ethoxylated, esters with acrylic acid

Cas Number:

28961-43-5

Molecular formula:

n.a.

IUPAC Name:

Poly(oxy-1,2-ethanediyl), .alpha.-hydro-.omega.-[(1-oxo-2- propenyl)oxy]-, ether with 2-ethyl-2-(hydroxymethyl)-1,3-propanediol (3:1)

Constituent 2

Reference substance name:

2-ethyl-2-[[(1-oxoallyl)oxy]methyl]-1,3-propanediyl diacrylate

EC Number:

239-701-3

EC Name:

2-ethyl-2-[[(1-oxoallyl)oxy]methyl]-1,3-propanediyl diacrylate

Cas Number:

15625-89-5

Molecular formula:

C15H20O6

IUPAC Name:

2,2-bis(prop-2-enoyloxymethyl)butyl prop-2-enoate

Test material form:

liquid

Specific details on test material used for the study:

Identification: PURIFIED TMPEOTA
Test Substance: Code PRO-2687
Chemical name (IUPAC): ETHOXYLATED TRIMETHYLOL PROPANE TRIACRYLATE
Description: Clear slightly yellow liquid
Batch: CLX 598/1
Purity: treated as 100% pure
Test substance storage: In refrigerator in the dark
Stability under storage conditions: Stable
Expiry date: 12 May 1999

Test animals

Species:

mouse

Strain:

NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test animals
- Source: Charles River, Sulzfeld, Germany
- Age at study initiation: 5-7 weeks old
- Weight at study initiation: 27.2-34.8
- Housing: housed in labelled polycarbonate cages containing purified sawdust
- Diet (e.g. ad libitum): standard pelleted laboratory animal diet; ad libitum
- Water (e.g. ad libitum):tap-water; ad libitum

Environmental conditions
- Temperature: 21 ± 3°C
- Humidity: 30-70%.
- Air changes (per hr): 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours dark per day

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

- Vehicle(s)/solvent(s) used: corn oil
- Concentration of test substance in vehicle: 250mg/10mL, 125mg/10mL and 62.5mg/10mL

Details on exposure:

The mice received a single intraperitoneal injection of a maximum tolerated (high), of an intermediate and of a low dose of the test substance. The route of administration was chosen to maximize the chance of the test substance to reach the target tissue. The dosing volume was 10 mL/kg bw.

Duration of treatment / exposure:

Single dose

Frequency of treatment:

Once

Post exposure period:

- 24h and 48h: treatment groups
- 48h: positive conrol

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (actual dose received)

Remarks:

nominal concentration

Doses / concentrations 2

Dose / conc.:

62.5 mg/kg bw/day (actual dose received)

Remarks:

nominal concentrations

Doses / concentrations 3

Dose / conc.:

125 mg/kg bw/day (actual dose received)

Remarks:

nominal concentrations

Doses / concentrations 4

Dose / conc.:

250 mg/kg bw/day (actual dose received)

Remarks:

nominal concentrations

No. of animals per sex per dose:

5 (two groups per dosage)

Control animals:

yes, concurrent vehicle

Positive control(s):

- positive control: cyclophosphamide (CPA)
- Route of administration: intraperitoneal
- Doses / concentrations: 50mg/kg bw

Examinations

Tissues and cell types examined:

- Bone marrow
- The number of micronucleated polychromatic erythrocytes was counted in 2000 polychromatic erythrocytes.

Details of tissue and slide preparation:

The slides were automatically stained using the "Wright-stain-procedure" in an "Ames" HEMA-tek slide stainer (Miles, Bayer Nederland B.V.). The dry slides were dipped in xylene before they were embedded in MicroMount and mounted with a coverslip.

Evaluation criteria:

- A test substance is considered positive in the micronucleus test if:
It Induced a biologically as well as a statistically significant (Wilcoxon Rank Sum Test; two-sided test at P < 0.05) increase in the frequency of micronucleated polychromatic erythrocytes (at any dose or at any sampling time) in the combined data for both sexes or in the data for male or female groups separately.
- A test substance is considered negative in the micronucleus test if:
None of the tested concentrations or sampling times showed a statistically significant (P < 0.05) increase in the incidence of micronucleated polychromatic erythrocytes neither in the combined data for both sexes nor in the data for male or female groups separately.

Statistics:

yes, e.G, Wilcoxon Rank Sum Test; two-sided test at P < 0.05

Results and discussion

Test results

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Remarks:

lethargic and rough coat; no abnormalities at 62.5 mg/kg bw

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: toxicity signs (at 125 and 250 mg/kg bw): lethargic behaviour and rough coat

Remarks:

no abnormalities at 62.5 mg/kg bw

Additional information on results:

- No increase in the frequency of micronucleated polychromatic erythrocytes was observed in the polychromatic erythrocytes of the bone marrow of the animals treated with the test substance. The animals of the groups that received 250 mg/kg bw showed slight decreases in the ratio of polychromatic to normochromatic erythrocytes at the 48 hours sampling time compared to the ratio observed in the animals treated with 62.5 mg/kg bw. This result reflected a toxic effect of the test substance on the erythropoiesis, implying that it reached the bone marrow cells of the mouse.
- Several toxicity signs such as lethargic behaviour and rough coat were observed in mice following the administration of 125.0 and 250.0 mg/kg bw. No abnormalities was recorded in the groups that had received 62.5 mg/kg bw test substance.

Applicant's summary and conclusion

Conclusions:

Under the study conditions, the test substance was considered to be not clastogenic in mouse (in vivo Mammalian Erythrocyte Micronucleus Test).

Executive summary:

A study was conducted to determine the in vivo toxicity of the test substance according to OECD Guideline 474 (in vivo Mammalian Erythrocyte Micronucleus Test), in compliance with GLP. Six groups each comprising 5 male and 5 female NMRI mice, received a single intraperitoneal injection of 10 mL/kg bw test substance. Two groups were dosed with 250 mg/kg bw, two groups were dosed with 125 mg/kg bw and two groups were dosed with 62.5 mg/kg bw. One group treated with the vehicle alone (corn oil) or with cyclophosphamide (CPA) at 50 mg/kg bw served as negative and positive controls, respectively. The tissue examinated was the bone marrow. The number of micronucleated polychromatic erythrocytes was counted in 2000 polychromatic erythrocytes. Mortality and toxicity signs were recorded daily. No increase in the frequency of micronucleated polychromatic erythrocytes was observed in the polychromatic erythrocytes of the bone marrow of the animals treated with the test substance. The animals of the groups that received 250 mg/kg bw showed slight decreases in the ratio of polychromatic to normochromatic erythrocytes at the 48 hours sampling time compared to the ratio observed in the animals treated with 62.5 mg/kg bw. This result reflected a toxic effect of the test substance on the erythropoiesis, implying that it reached the bone marrow cells of the mouse. Several toxicity signs such as lethargic behaviour and rough coat were observed in mice following the administration of 125.0 and 250.0 mg/kg bw. No abnormalities was recorded in the groups that had received 62.5 mg/kg bw test substance. Under the study conditions, the test substance was considered to be not clastogenic in mouse (Bertens, 2001).