Propane-1,3-diol

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

ACC PDO panel assigned reliability of 1. PDO from chemical process.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: EEC Directive 92/69/EEC Part B: Methods for the Determination of Toxicity, B.12. Micronucleus Test

GLP compliance:

yes

Type of assay:

mammalian erythrocyte micronucleus test

Test material

Test material information

Constituent 1

Reference substance name:

Propane-1,3-diol

EC Number:

207-997-3

EC Name:

Propane-1,3-diol

Cas Number:

504-63-2

Molecular formula:

C3H8O2

IUPAC Name:

propane-1,3-diol

Details on test material:

- Name of test material (as cited in study report): 1,3-propanediol, trimethylene glycol
- Physical state: Clear, colourless liquid.
- Analytical purity: >= 99.8%
- Stability under test conditions: The stability of the test substance under the given storage conditions was guaranteed by the sponsor throughout the experimental period.
- Storage condition of test material: Tightly closed container under nitrogen at room temperature.

Test animals

Species:

mouse

Strain:

other: Hsd/Win: NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan-Winkelmann GmbH
- Age at study initiation: approximately 6 weeks
- Weight at study initiation: males: 30-36 g; females: 25-30 g
- Assigned to test groups randomly: yes (computer generated random numbers)
- Fasting period before study: 16 hours before treatment.
- Housing: Macrolon cages, type II, 1 animal per cage.
- Diet: ad libitum
- Water:ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0-21.5°C
- Humidity (%): 50-58%
- Photoperiod (hrs dark / hrs light): 12 hours light, 12 hours dark

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: The test material was freshly diluted with aqua ad iniectabilia (Ampuwa, batch No. EL 1009, supplied by Fresenius AG, D-61343 Bad Homburg v d Hohe)
- Administration volume: 10.0 mL/kg b.w.

Frequency of treatment:

single administration

Post exposure period:

24 and 48 hours

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

1 470 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

2 150 mg/kg bw/day (nominal)

No. of animals per sex per dose:

Main test: 28 (14 males and 14 females) at 2150 mg/kg bw
Repetition Test: 12 (6 males and 6 females) each at 1000, 1470 and 2150 mg/kg bw

Control animals:

other: Main Test: Physiological saline solution; 12 males + 12 females. Repetition Test: 6 males + 6 females.

Positive control(s):

Main Test positive control: cyclophosphamide (31.6 mg/kg bw) in physiological saline solution; 6 males + 6 females.
Repeat Test positive control: cyclophosphamide (31.6 mg/kg bw) in physiological saline solution; 6 males + 6 females.

Examinations

Tissues and cell types examined:

clinical observation; bone marrow

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: Range finding study identified that 4640 mg/kg bw caused severe toxicity symptoms and death, and at 3160 mg/kg bw distinct toxicity symptoms but no death was observed. Test guideline considers 2000 mg/kg bw to be the maximum single (limit) dose to be administered.

DETAILS OF SLIDE PREPARATION: Mice were sacrificed by CO2 overdose. Both femurs removed from each mouse and the bone marrow cells flushed into a labeled centrifuge tube with approx. 1.5 mL of fetal calf serum. Centrifuged at approx. 180 x g for 5 minutes. Supernatant serum was discarded and bone marrow cells suspended upon a thin layer of serum. A small drop of the marrow serum suspension was smeared on a slide and allowed to dry overnight.

METHOD OF ANALYSIS: Microscopical evaluation; 1000 polychromatic erythrocytes per animal were scored for the incidence of polychromatic erythrocytes with micronuclei. Calculation of PCE/NCE, based on 1000 erythrocytes (PCE+NCE) scored per slide.

Evaluation criteria:

If a test material produced neither a statistically significant and reproducible positive response nor a dose related statistically significant response at any one of the test points compared to the negative control group, it is considered non-mutagenic in this system.

Statistics:

POISSON test

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Remarks:

non-mutagenic

Toxicity:

no effects

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Under the experimental conditions reported, 1,3-Propanediol is considered not to induce chromosomal mutations in mice by damage to the chromosomes or the miotic apparatus at 24 or 48 hour intervals after the animals had received a single oral dose of 2150 mg/kg body weight. However, a weak clastogenic effect cannot be totally excluded due to a statistically significant increase in micronucleated polychromatic erythrocytes at the 48 hour sampling time when calculations were performed for males and females combined. According to the test guideline, a second test (repetition test) was performed to examine the reproducibility of the statistically significant response at the 48 hour sampling time in males and females combined and to assess a possible dose relation in the increase of micronucleated polychromatic erythrocytes. These responses could not be verified during the repetition test using two additional dose levels and is therefore considered to be an incidental finding.

Therefore, 1,3-Propanediol is non-mutagenic in the reported in vivo mouse micronucleus test.

Any other information on results incl. tables

	Mean PCEs with MN (Males)		Mean PCEs with MN (Females)
Main Test	24-Hours	48-Hours	24-Hours	48-Hours
Negative Control	1.4	1.2	1.4	1.1
2150 mg/kg	1.0	2.2	1.8	1.5
Positive Control	13.8		14.7
Repetition Test
Negative Control		2.0		2.0
1000 mg/kg		0.8		1.4
1470 mg/kg		1.0		1.0
2150 mg/kg		2.0		1.2
Positive Control		6.6		6.3

Applicant's summary and conclusion

Conclusions:

1,3-Propanediol is non-mutagenic in the reported in vivo mouse micronucleus test.

Executive summary:

This in vivo micronucleus test in mice was performed to assess the potential mutagenic activity, induced by 1,3-propanediol through damage to the chromosomes of to the mitotic apparatus. The main test was performed with three groups, the negative control, the positive control, and the test material group (2150 mg/kg). Half the mice were sacrificed at 24 hours after treatment and the remaining mice were sacrificed 48 hours after treatment. Bone marrow was removed from the femurs for examination. All animals of the positive control group were sacrificed 24 hours after administration. One thousand polychromatic erythrocytes (PCE) per animal were scored for micronuclei. The ratio of polychromatic to normochromatic erythrocytes (NCE) was used to assess the toxicity of the test material. No test substance-related clinical observations were observed in the mice. No mortality was observed. Under the experimental conditions reported, 1,3-Propanediol is considered not to induce chromosomal mutations in mice by damage to the chromosomes or the miotic apparatus at 24 or 48 hour intervals after the animals had received a single oral dose of 2150 mg/kg body weight. However, a weak clastogenic effect cannot be totally excluded due to a statistically significant increase in micronucleated polychromatic erythrocytes at the 48 hour sampling time when calculations were performed for males and females combined. 

 

According to the test guideline, a second test (repetition test) was performed to examine the reproducibility of the statistically significant response at the 48 hour sampling time in males and females combined and to assess a possible dose relation in the increase of micronucleated polychromatic erythrocytes. Dose levels of 1000, 1470, and 2150 were used in the repetition test. The responses of the first test could not be verified during the repetition test using two additional dose levels and is therefore considered to be an incidental finding. Therefore, 1,3-Propanediol is non-mutagenic in the reported in vivo mouse micronucleus test.