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Toxicological information

Genetic toxicity: in vivo

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Administrative data

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:
1998-03-25 to 1998-05-03
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1998
Report Date:
1998

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Version / remarks:
draft version 1997
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)
Version / remarks:
29 December 1992
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): 1,4-Butanediol dimethacrylate

Test animals

Species:
mouse
Strain:
NMRI
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Winkelmann, Borchen, Germany
- Age at study initiation: no data
- Weight at study initiation: 26 – 36 g
- Assigned to test groups randomly: yes
- Fasting period before study: no data
- Housing: 5 animals of identical sex per cage
- Diet (e.g. ad libitum): pellet standard diet (Altromin)
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25°C
- Humidity (%): 55+/-10°C
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light):12/12

IN-LIFE DATES: no data

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
- Vehicle(s)/solvent(s) used: cotton seed oil
- Justification for choice of solvent/vehicle: relatively nontoxic to the animals
- Concentration of test material in vehicle:
- Amount of vehicle (if gavage or dermal): 16.7 mL/kg bw
- Lot/batch no. (if required): 27H0534
- Purity: no data
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
dilution in cotton seed oil; single standard volume of 16.7 mL/kg bw
Duration of treatment / exposure:
The animals received the test item once. Sampling of the bone marrow was carried out on animals 24 and 48 h after treatment.
Frequency of treatment:
single dose
Post exposure period:
The animals were sacrificed 24 and 48 hours after treatment.
Doses / concentrationsopen allclose all
Dose / conc.:
200 mg/kg bw/day (actual dose received)
Remarks:
24 h interval
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
Remarks:
24 h interval
Dose / conc.:
2 000 mg/kg bw/day (actual dose received)
Remarks:
24 h interval
Dose / conc.:
2 000 mg/kg bw/day (actual dose received)
Remarks:
48 h interval
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Positive control(s):
cyclophosphamide
- Justification for choice of positive control(s): appropriate reference mutagen
- Route of administration: single administration i.p.
- Doses / concentrations: 30 mg/kg bw in 0.9% NaCl (10 mL/kg bw)

Examinations

Tissues and cell types examined:
bone marrow erythrocytes (polychromatic erythrocytes)
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION:
dose selection based on acute toxicity pre-test: at 2000 mg/kg bw no toxic signs were found two lower dose groups (200 and 1000 mg/kg bw) were selected for 24 h treatment interval

DETAILS OF SLIDE PREPARATION:
- removal of femura
- bone marrow was flushed out with 1 mL FCS followed by resuspension of cells and centrifugation at 1500 rpm for 10 min
- cells were resuspended in FCS and smeared on slides
- air drying of slides, staining with May-Grünwald/Giemsa
- at least one slide prepared from each bone marrow sample

METHOD OF ANALYSIS:
- 2000 polychromatic erythrocytes were analysed per animal for micronuclei
- ratio between polychromatic erythrocytes (PCE) and normochromatic erythrocytes (NCE) was determined to assess cytotoxicity (expressed as NCE per 1000 PCE)
- analysis carried out with coded slides
Evaluation criteria:
A test item is classified mutagenic if it induces either a statistically significant dose related increase in the number of micronucleated PCE or a reproducible statistically significant positive result for at least one test point.
Statistics:
non-parametric Mann-Whitney test

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
yes
Positive controls validity:
valid
Additional information on results:
RESULTS OF RANGE-FINDING STUDY
- Dose range: 2000 mg/kg bw
- Solubility: no data
- Clinical signs of toxicity in test animals: after 1 hour reduced spontaneous activity
- Other: only pre-test for toxicity, no further analysis

RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): no induction of micronuclei over background (see table); the mean values of micronuclei observed after treatment with the test item (0.11% to 0.15%) were in the same range as the negative control groups (0.10% to 0.13%)
- Ratio of PCE/NCE (for Micronucleus assay): slight effect on PCE/NCE ration by treatment with test item at a concentration of 2000 mg/kg bw at 24 and 48 h indicating slight cytotoxic properties
- Statistical evaluation: no significance found for 1000 mg/kg bw (24 h), 2000 mg/kg bw (24 h), 2000 mg/kg bw (48 h); lowest dose of 200 mg/kg bw (24 h) not tested for significance, as values were equal to or lower than negative control

Any other information on results incl. tables

group sex mean MN/2000 PCE %MN relative to N.C. PCE/NCE
N.C. m 2 0.1 1.0 1000/808.2
P.C. m 34.6 1.73 17.3 1000/1045.6
test item 2000mg/kg, 24 h m 2.6 0.13 1.3 1000/995.2
test item 2000mg/kg, 48 h m 3 0.15 1.5 1000/1011
test item 1000mg/kg, 24 h m 2.4 0.12 1.2 1000/868
test item 200mg/kg, 24 h m 2.2 0.11 1.1 1000/820.2
N.C. f 2.6 0.13 1.0 1000/844.2
P.C. f 38.8 1.94 14.9 1000/1055.8
test item 2000mg/kg, 24 h f 2.4 0.12 0.9 1000/972.8
test item 2000mg/kg, 48 h f 2.6 0.13 1.0 1000/1016.8
test item 1000mg/kg, 24 h f 2.8 0.14 1.1 1000/884.4
test item 200mg/kg, 24 h f 2.4 0.12 0.9 1000/828.6

N.C. = negative control

P.C. = positive control

m = male

f = female

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
In this study under the experimental conditions reported, 1,4-BDDMA did not induce micronuclei over background as determined by the micronucleus test in bone marrow cells of the mouse.
Executive summary:

In a NMRI mouse bone marrow micronucleus assay according to OECD guideline 474, draft version 1997, 5 animals per sex per dose were treated by oral gavage (single dose) with 1,4-BDDMA (93.9% a.i.) at doses of 0, 200, 1000 and 2000 mg/kg bw. Bone marrow cells were harvested at 24 h (200, 1000 and 2000 mg/kg dose groups) and 48 h (2000 mg/kg dose group) post-treatment. The vehicle was cotton seed oil.

There were no signs of toxicity during the study based on mortality or clinical signs. A slight effect on the ratio of polychromatic to normochromatic erythrocytes was observed in the 2000 mg/kg dose group 24 and 48 h post-treatment indicating a possible weak toxic effect to the bone marrow. There was no significant increase in the frequency of micronucleated polychromatic erythrocytes in bone marrow after any treatment time. 1,4-BDDMA was tested at an adequate dose evel which included the guideline limit dose of 2000 mg/kg bw. The positive control induced the appropriate response.

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