3-chloropropene

Toxicity to reproduction

Administrative data

Endpoint:

fertility, other

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: - scientifically sound study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1981

Report date:

1981

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

male mice were sacrificed 5 wks after the last of 5 consecutive inhalative treatments and sperm was extracted from the cauda epididymes fixed, stained and microscopically analyzed for abnormalities

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

3-chloropropene

EC Number:

203-457-6

EC Name:

3-chloropropene

Cas Number:

107-05-1

Molecular formula:

C3H5Cl

IUPAC Name:

3-chloroprop-1-ene

Details on test material:

- Name of test material (as cited in study report): 3-chloropropene
- Physical state: clear, colourless liquid
- Analytical purity: 98 %
- Impurities (identity and concentrations): not reported
- Purity test date: not reported
- Lot/batch No.: Batch No. 17835
- Supplier: Aldrich Chemical Company Limited (18 March 1980)
- Expiration date of the lot/batch: not reported
- Stability under test conditions: not reported, but expected to be stable
- Storage condition of test material: retained in the dark under ambient conditions in the company dispensary until used
- Other:

Test animals

Species:

mouse

Strain:

B6C3F1

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (U.S.A.)
- Age at study initiation: 10 - 12 wks
- Weight at study initiation: not reported
- Fasting period before study: not reported
- Housing: not reported
- Diet (e.g. ad libitum): Spratts-Spillers No. 1
- Water (e.g. ad libitum): not reported
- Acclimation period: not reported


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.5 - 26, average 22
- Humidity (%): 30 - 68, average 50
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

inhalation: vapour

Type of inhalation exposure (if applicable):

whole body

Vehicle:

unchanged (no vehicle)

Details on exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: chambers constructed of glass and steel, 1.5 m³ inner size
- Method of holding animals in test chamber: caged individually
- Source and rate of air: 12 airchanges/h, Compressed air was supplied py means of 2 Broomwade compressors (Type CAR31) fitted with automatic pressure control switches. These supplied filtered, conditioned, oil-free compressed air for subsequent dilution of test atmospheres.
- Method of conditioning air: see above
- Temperature, humidity, pressure in air chamber: not reported
- System of generating particulates/aerosols: The high level atmosphere was produced by bubbling dry, oxygen-free nitrogen (BOC Limited) through a liquid reservoir of 3-chloropropene contained in a glass, gas washing or Drechel bottle immersed in a temperature controlled water bath at 1 °C. The low level atmosphere was generated by inserting a 100 µL glass syringe, through a rubber septum, into a glass T piece and feeding a constant volume of 3-chloropropene into a stream of nitrogen gas. The micro syringe was activated by a syringe driver. The nitrogen/3-chloropropene vapour mixture so generated was ducted through 7/16" ID stainless steel piping to a glass mixing vessel and diluted with filtered, compressed air. The resulting mixture of 3-chloropropene/air was ducted through 7/8" stainless steel piping to the top of the exposure chamber. The required atmospheric concentrations within the exposure chambers were maintained by finely regulating the flow of nitrogen and diluting air into the mixing vessels, by means of adjustable flow meters.
- Air flow rate: not reported in detail,
- Air change rate: 12 airchanges/h were maintained
- Treatment of exhaust air: Test atmospheres were exhausted from the exposure chambers using a Gast extract pump. Contaminated air extracted from
the exposure chamber was 'scrubbed' using methylated spirits/water treatment. It was then diluted in the building exhaust air before discharging to the external atmosphere. The exposure chambers were maintained under slight negative pressure (variable, but normally 2-3 cm water) to minimise any possible leakage of test material into the working environment.


TEST ATMOSPHERE
- Brief description of analytical method used: The atmospheres within the exposure chambers were analysed by infra-red spectroscopy using Miran-1A Portable Gas Analysers (Foxboro/Wilks Inc).
- Samples taken from breathing zone: yes


VEHICLE (if applicable)
- no vehicle used

Details on mating procedure:

not applicable, no mating occured, only sperm was analyzed

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The atmospheres within the exposure chambers were analysed by infra-red spectroscopy using Miran-1A Portable Gas Analysers (Foxboro/Wilks Inc). This type of instrument is a single beam, variable wavelength spectrometer, scanning the infra-red spectrum between 2.5 and 14.5 µm. It is equipped with a gas cell having a variable pathlength of between 0.75 and 21.75 m. Samples of the chamber air were continuously pumped (5 l/min) through stainless steel sample lines of 3/8'' ID, to the gas cell of the analyser. The concentration was measured and relayed to a chart recorder (Servoscribe RE 541) to provide a permanent record of the chamber concentrations.
The infra-red gas analyser used to monitor chamber atmospheres of 3-chloropropene were calibrated each day before vapour generation commenced using a closed loop calibration system.

Duration of treatment / exposure:

7 h

Frequency of treatment:

on five consecutive days

Details on study schedule:

not applicable

Doses / concentrations

Remarks:

Doses / Concentrations:
1 and 25 ppm
Basis:
analytical conc.

No. of animals per sex per dose:

10

Control animals:

yes, sham-exposed

Details on study design:

- Dose selection rationale: not reported
- Rationale for animal assignment (if not random): Empty cages were placed on racks and, upon receipt of the animals, starting with the male rats, a transporting box was opened and a rat placed in the first cage. A second rat was removed from the same transport box and placed in the second c8ge and so on until all the cages designated for the male rats each contained one animal.

Positive control:

yes, 200 mg/Kg ethyl methanesulphonate was administered orally by gavage to the rodents at a constant dose volume of 10 mL/kg at on 5 consecutive days.

Examinations

Parental animals: Observations and examinations:

not applicable

Oestrous cyclicity (parental animals):

not applicable

Sperm parameters (parental animals):

Parameters examined in males:
sperm morphology:
The following types of sperm were not scored:
(1) separated tails and heads.
(2) clumps of sperm.
(3) sperm orientated so that the hook could not be
seen.
(4) sperm partially masked by any remaining stain
droplets.
Otherwise, sperm were scored and placed in one of the
following categories:
I Normal
II Abnormal
A. hook upturned or elongated.
B. banana-shaped head.
c. amorphous hoad.
D. abnormal tail (sharp, 1800 angle or tight
coiling only).
E. miscellaneous tthese were specified in footnotes, could include multiple tails, double heads, twisted neck, filamentous midplece, enlarged mid-piece, plier type).

Litter observations:

not applicable

Postmortem examinations (parental animals):

not applicable

Postmortem examinations (offspring):

not applicable

Statistics:

The data were transformed using the Freeman-Tukey transformation for proportions. A one-sided t test was used on the transformed values. This analysis was performed on (a) total abnormal cells and (b) each of the abnormal categories A-E.

Reproductive indices:

not applicable

Offspring viability indices:

not applicable

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

not examined

Body weight and weight changes:

not examined

Food consumption and compound intake (if feeding study):

not examined

Organ weight findings including organ / body weight ratios:

not examined

Histopathological findings: non-neoplastic:

not examined

Other effects:

not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

not examined

Details on results (P0)

There were no increases in the frequencies of abnormal sperm in any of the categories examined following exposure to 3-chloropropene atmospheres. (see table 1)
There were small increases in aberrant sperm frequencies in Categories C and D (amorphous head and folded tail, respectively) with the positive control substance indicating the suitability of the test system.

Effect levels (P0)

1

Dose descriptor:

NOAEL

Effect level:

> 25 ppm

Sex:

male

Basis for effect level:

other: no effects were seen at this level

Remarks on result:

not determinable

Remarks:

no NOAEL identified

2

Dose descriptor:

NOEL

Effect level:

25 ppm

Sex:

male

Basis for effect level:

other: no effects were seen at this level

Results: F1 generation

General toxicity (F1)

Clinical signs:

not examined

Mortality / viability:

not examined

Body weight and weight changes:

not examined

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Histopathological findings:

not examined

Details on results (F1)

not applicable

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

- table 1: abnormality scoring

Dose Group	Number normal	Number Abnormal						Percent Abnormal
		A	B	C	D	E	Total	A	B	C	D	E	Total i
Air Control, 7 h/day	9590	10	15	183	51	151	410	0.10	0.15	1.83	0.51	1.51	4.10
1 ppm, 7 h/day	9626	11	19	132	77	135	374	0.11	0.19	1.32	0.77	1.35	3.74
25 ppm, 7 h/day	9636	6	18	148	52	140	364	0.06	0.15	1.48	0.52	1.40	3.64
EMS, 200 mg/kg/day	9495	16	20	1228	110	131	505	0.16	0.20	2.28	1.10	1.13	5.05

A: hook upturned or elongated.

B: banana-shaped head.

C: amorphous hoad.

D: abnormal tail (sharp, 1800 angle or tight coiling only).

E: miscellaneous tthese were specified in footnotes, could include multiple tails, double heads, twisted neck, filamentous midplece, enlarged mid-piece, plier type).

EMS: ethyl methanesulphonate

Applicant's summary and conclusion

Conclusions:

A sperm abnormality test was conducted with 3-chloropropene in male mice 5 wks after the last of 5 consecutive inhalative treatments.
No adverse effects were seen with both exposure concentrations.

Executive summary:

In the present study a sperm abnormality test was conducted with 3-chloropropene in male mice 5 wks after the last of 5 consecutive inhalative treatments (7 h; 1, 25 ppm). No adverse effects were seen with both exposure concentrations. As the exposure concentrations are very low even for a subacute exposure no clear conclusion can be drawn on the genotoxicity in vivo.

Mice were killed 5 weeks from the last day of dosing by neck dislocation, sperm was extracted from the cauda epididymes and smears were analyzed microscopically for abnormalities.