Registration Dossier

Administrative data

Key value for chemical safety assessment

Toxic effect type:
concentration-driven

Effects on fertility

Description of key information

No adverse effects were reported regarding reproductive parameters and/or reproductive organs based on tests conducted by the oral (3-generation study) and/or inhalation routes (2-year repeated dose toxicity study). There was no dominant lethal effect indicating no specific concern for male reproduction.

Link to relevant study records

Referenceopen allclose all

Endpoint:
three-generation reproductive toxicity
Remarks:
based on test type
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Not specified
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
The primary report was not available for review but the data has been reviewed by the World Health Organisation (WHO) experts who have agreed that no effects or dominant lethality was found at either dose level tested.
Qualifier:
no guideline followed
Principles of method if other than guideline:
In a three-generation, oral gavage study in rats using CFC-12 (in corn oil) at average doses of 15 and 150 mg/kg per day. Limited details available but the data has been reviewed by the World Health Organisation (WHO) experts.
GLP compliance:
not specified
Remarks:
likely pre-dates the GLP standards
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
Charles River CD
Sex:
male/female
Details on test animals or test system and environmental conditions:
Not specified
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
In a three-generation, oral gavage study in rats using CFC-12 (in corn oil) at average doses of 15 and 150 mg/kg per day
Details on mating procedure:
Not specified
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
Not specified
Duration of treatment / exposure:
Not specified
Frequency of treatment:
Not specified
Details on study schedule:
Not specified
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
15 mg/kg bw/day (nominal)
Dose / conc.:
150 mg/kg bw/day (nominal)
No. of animals per sex per dose:
50 /sex/dose
Control animals:
yes, concurrent vehicle
Details on study design:
Male and female rats were given Freon 12 in doses of 15 or 150 mg/kg by intubation. The rats were then bred and evaluated for fertility, corpora lutea, implantation sites, resorption sites, and number of live fetuses per litter.

Groups of 50 males and 50 females of the F1a generation remained in the test for 2 years, with an interim killing at 1 year.
Animals received CFC-12 in corn oil or corn oil alone daily by gavage for 6 weeks, and 5 times/week thereafter.

Dominant Lethal assay, part of a reproductive toxicity study:
The F0 animals of the reproduction study were mated to initiate an F1b litter. Animals were treated with the test substance gavage until pregnancies were terminated in mid-term to give a dominant lethal assay
Positive control:
Not specified (not required)
Parental animals: Observations and examinations:
Not specified
Oestrous cyclicity (parental animals):
Not specified
Sperm parameters (parental animals):
Not specified
Litter observations:
Not specified
Postmortem examinations (parental animals):
Not specified
Postmortem examinations (offspring):
Not specified
Statistics:
Not specified
Reproductive indices:
Fertility, corpora lutea, implantation sites, resorption sites, and number of live fetuses per litter.
Offspring viability indices:
Number of live fetuses per litter.
Clinical signs:
not specified
Mortality:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Organ weight findings including organ / body weight ratios:
not specified
Histopathological findings: non-neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
No effects on fertility index (percentage of matings resulting in pregnancy), no effects on gestation index (% of pregnancies resulting in birth of liver litters).
No dominant lethality was found at either dose level.
Key result
Dose descriptor:
NOEL
Effect level:
> 150 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Remarks:
(limited details available)
Clinical signs:
no effects observed
Description (incidence and severity):
No overt signs of toxicity.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
no significant differences between treated and control groups in survival and post-natal survival (survival after 4 days or 21 days)
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
in F1a parental generation (treatment started at 6 weeks of age): Body weight gain was depressed in the high-dose groups, particularly among the females.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
Slight decline in food efficiency in high dose females, compared to controls
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
No effects reported on fertility index of the second parental generation
No details provided on the unpublished study report reviewed by WHO.
Key result
Dose descriptor:
NOAEL
Effect level:
> 150 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Remarks:
(limited details available)
Key result
Critical effects observed:
no
Clinical signs:
not specified
Description (incidence and severity):
No effects on viability index (percentage of rats born that survived four days)
No effects on lactation index (percentage of rats alive at 4 days that survived to be weaned at 21 days)
Body weight and weight changes:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
no effects observed
Description (incidence and severity):
No dominant lethality was found at either dose level, indicating no chromosomal damage to germ cells.
Behaviour (functional findings):
not specified
No dominant lethality was found at either dose level.
Key result
Dose descriptor:
NOEC
Generation:
F1
Effect level:
>= 150 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Remarks:
No effects reported on the reproductive parameters of the F1 generation and no effects on postnatal survival
Key result
Critical effects observed:
no
Key result
Reproductive effects observed:
no
Conclusions:
No dominant lethality was found at either dose level.
The substance is not considered to be classified for fertility effects.
Executive summary:

In a 3-generation oral study using CFC12 (in corn oil) at average doses of 15 and 150 mg/kg per day, Sherman (1974) found no adverse effects in reproductive capability as measured by the fertility index, gestation index, viability index and lactation index. No classification is applicable.

Endpoint:
extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
the study does not need to be conducted because a two- (or multi-) generation reproductive toxicity study is available
Justification for type of information:
JUSTIFICATION FOR DATA WAIVING
Available data in multi-generation experimental study did not indicate adverse effects on reproduction. There was no dominant lethal effect in rats. In chronic studies in rats and mice, no histopathological lesions were reported in the reproductive organs (uterus and gonads) that were examined.
The use of this substance is regulated (Regulation (EC) No. 1005/2009) with limited volumes allowed for the listed use. The process is fully automated and operated remotely, and technical measures are set up to prevent and minimise potential emissions as reported in the CSR.
Existing information in the dataset support a low toxicity profile of the substance, with no specific alerts for fertility or reproduction parameters. There are no consumer uses. As use volumes are controlled and regularly lowered by the Regulation, additional animal tests are not considered necessary
Reason / purpose for cross-reference:
data waiving: supporting information
Reason / purpose for cross-reference:
data waiving: supporting information
Reason / purpose for cross-reference:
data waiving: supporting information
Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

In a 3-generation oral study using CFC12 (in corn oil) at average doses of 15 and 150 mg/kg per day, Sherman (1974) found no adverse effects in reproductive capability as measured by the fertility index, gestation index, viability index and lactation index. 

This study has been reviewed by the World Health Organisation (WHO) and international experts have agreed that no effects or dominant lethality was found at either dose level tested. 

Effects on developmental toxicity

Description of key information

No effects were observed on fetal development in rats or rabbits exposed by the oral or inhalation routes.

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
Not specified
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
Study report, not conducted to GLP but well documented with methodology and results included.
Qualifier:
no guideline followed
Principles of method if other than guideline:
See "details on exposure" and "details on test animals and environmental conditions" below.
GLP compliance:
not specified
Remarks:
prior to GLP standards
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Remarks:
Charles River-CD
Details on test animals or test system and environmental conditions:
Seventy-eight pregnant (primigravida) albino rats (Charles River-CD) were received from Charles Rivers Breeding Laboratories in two shipments one day apart. From the time of arrival at Haskell Laboratory through the fifth day of gestation, the animals were housed in suspended stainless steel wire cages, assigned to three groups, and fed Ground Purina Laboratory Chow (GPLC) supplemented with 1% corn oil (CO).
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
On day six of the gestation period, all rats in all groups were given the following diet: 74.25% GPLC + 0.25% CO + 25% Casein + 0.5% vitamin mix. One group, the control, was given 2.0ml of corn oil each day by intragastric intubation for 10 consecutive days; the other two groups were given 2.0ml of Haskell No. 6966 (0.2% solution of Freon 12 in corn oil) or Haskell No. 6967 (2.0% solution of Freon 12 in corn oil), respectively, for the same period of time, i.e., through day 15 of gestation. From day 16 of the gestation period to the time of sacrifice, the control and test rats received their initial basal diet (GPLC + 1% CO).
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
Not specified
Details on mating procedure:
Not specified
Duration of treatment / exposure:
10 days (gestation day 6 to gestation day 15)
Frequency of treatment:
Daily
Duration of test:
To day 21 of gestation period.
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
16.6 mg/kg bw/day (nominal)
Dose / conc.:
170.9 mg/kg bw/day (nominal)
No. of animals per sex per dose:
78 rats in total.
Group I (0) - 25
Group II (16.6) - 26
Group III (170.9) - 27
Control animals:
yes, concurrent vehicle
Details on study design:
The animals were sacrificed by chloroform inhalation on the twenty-first day of gestation. Pregnancy was dated by counting as day one the morning on which a copulation plug was found. An incision was made in the abdominal wall and both ovaries and uterine horns were promptly removed, inspected and weighed. The uterus was carefully opened and the foetuses removed. The following observations and measurements were made:
1. Number and location of live foetuses, dead foetuses, and resorption sites in the respective uterine horns.
2. Body weight and crown-rump length of all live foetuses.
3. External examination of all foetuses, from head to tail, under Macro Scope (Ednakite 2½ x) for gross abnormalities.
About two-thirds of the foetuses from each litter were preserved in 95% alcohol for subsequent maceration in 1% aqueous KOH, clearing and staining with Alizarin Red S, and examination to detect skeletal abnormalities. The remaining foetuses were fixed in Bouin’s fluid for free-hand razor-blade sectioning (Wilson Method) and examination for visceral and neural anomalies under the dissecting microscope.
Maternal examinations:
Not specified
Ovaries and uterine content:
An incision was made in the abdominal wall and both ovaries and uterine horns were promptly removed, inspected and weighed. The uterus was carefully opened and the foetuses removed.
Fetal examinations:
Number and location of live foetuses, dead foetuses, and resorption sites in the respective uterine horns. Body weight and crown-rump length of all live foetuses. External examination of all foetuses, from head to tail, under Macro Scope (Ednakite 2½ x) for gross abnormalities.
Statistics:
Not specified
Indices:
Not specified
Historical control data:
The data was also compared with the standard values established for Charles River-CD (ChR-CD) rats.
Clinical signs:
no effects observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Details on maternal toxic effects:
Maternal toxic effects: no effects

Body Weight: The oral administration of Freon 12 to pregnant rats during the period of organogenesis had no adverse effect on body weight gain
Food Consumption: There were no meaningful differences among the control and test groups of rats with respect to food intake during any of the time periods measured.
Pregnancy Outcome : The uteri of all females in all groups were free from gross abnormalities. There was no difference in the numbers of implantation sites, viable foetuses and resorption sites per female between the control group and those given Freon 12.
Key result
Dose descriptor:
NOAEL
Effect level:
>= 171 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
clinical signs
food consumption and compound intake
mortality
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects: no effects

Details on embryotoxic / teratogenic effects:
Pregnancy Outcome and Fetal Development: The uteri of all females in all groups were free from gross abnormalities. There was no difference in the numbers of implantation sites, viable foetuses and resorption sites per female between the control group and those given Freon 12. Likewise, there was no significant difference in mean weight and length of foetuses. The smaller weight and length in the control group were due to sacrifice of several females on day earlier (20 days). The data was also compared with the standard values established for Charles River-CD (ChR-CD) rats and were found to be in good agreement.
Malformations and Anomalies: No major gross, skeletal or soft tissue abnormalities were detected. Maternal feeding of Freon 12 did not influence the fetal development. The incidence and type of other minor anomalies were compared with those of the control group and with the standard values for ChR-CD rats. They were found to be similar.
Key result
Dose descriptor:
NOAEL
Effect level:
> 170 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Abnormalities:
no effects observed
Key result
Developmental effects observed:
no

TABLE I – AVERAGE BODY WEIGHTS OF PREGNANT RATS RECEIVING FREON 12 DURING ORGANOGENESIS

Group Characteristics

No. of Pregnant Rats

No. of Rats Received

Average Body Weight in Grams on Day of Gestation Period

6

16

21

2 ml corn oil (control)

22/25

199

269

337

2 ml corn oil containing 0.2% Freon 12 (test)

20/26

203

278

345

2 ml corn oil containing 2.0% Freon 12 (test)

25/27

196

271

346

 

TABLE II – AVERAGE FOOD CONSUMPTION DATA OF PREGNANT RATS RECEIVING FREON 12 DURING ORGANOGENESIS

Group Characteristics

No. of Pregnant Rats

No. of Rats Received

Average Daily Food Consumption (g/rat/day)

Control Diet Prior to Day 6

Diet During Dosing Day 6-16

Control Diet Days 16-21

2 ml corn oil (control)

22/25

17.1

17.0

21.0

2 ml corn oil containing 0.2% Freon 12 (test)

20/26

18.7

19.3

22.7

2 ml corn oil containing 2.0% Freon 12 (test)

25/27

17.4

17.8

23.1

 

TABLE III – AVERAGE DOSE OF FREON 12 RECEIVED BY PREGNANT RATS DURING ORGANOGENESIS

Group Characteristics

Average Body Weight In Grams During Day 6-16

Average Dose of Freon 12 (mg/kg/day)

2 ml corn oil (control)

234

-

2 ml corn oil containing 0.2% Freon 12 (test)

241

16.6

2 ml corn oil containing 2.0% Freon 12 (test)

234

170.9

 

TABLE IV – EFFECT OF ORAL ADMINISTRATION OF FREON 12 TO PREGNANT RATS IN THE OUTCOME OF PREGNANCY AND ON VARIOUS PARAMETERS OF FETAL DEVELOPMENT

 

Groups (mg/kg/day of Freon 12)

Standard Values (Range) for Charles River (CD) Rats (1)

Group I (0)

Group II (16.6)

Group III (170.9)

Number Females Pregnant

22/25 (88.0%)

20/26 (76.9%)

25/27 (92.6%)

90% (62-100)

Number Implantation Sites

201

186

236

 

Number Implantation Sites / Pregnant Female

9.13

9.30

9.44

10.9 (8.6-14.1)

Number Live Fetuses

197

179

231

 

Number Dead Fetuses

0

0

0

 

Number of Live Fetuses/Litter

8.95

8.95

9.24

10.2 (8.2-12.7)

Number Females Showing Complete Resorption

0

0

0

0.3% (0-5.9)

Number Females Showing Premature Birth

0

0

0

0.1% (0-5.9)

Number Females Showing Partial Resorption (Excluding Complete Resorptions)

4 (18.2%)

4 (20.0%)

3 (12.0%)

40.6% (10.5-77.8)

Total Number Resorbed

4 (2.0%)

7 (3.8%)

5 (2.1%)

6.0% (1.2-12.4)

Mean Fetus Weight (g)

3.35

4.19

3.93

3.69 (2.90-4.22)

Mean Fetus Crown-Rump Length (cm)

2.98

3.52

3.36

 

(1) Personal communication; unpublished data obtained from Charles River Breeding Laboratories, Wilmington, Massachusetts for period 1964 -1969.  

 

TABLE V – GROSS EXTERNAL, SKELETAL AND SOFT TISSUE ABNORMALITIES FOUND IN FETUSES FROM MOTHERS ADMINISTERED FREON 12 DURING FETAL DEVELOPMENT

Abnormalities Found

Groups (mg/kg/day of Freon 12)

Standard Values (Range) of Abnormalities for ChR-CD Rats (1)

Group I (0)

Group II (16.6)

Group III (170.9)

1. Gross External

Number Fetuses Examined

197

179

231

32915

a. Small, stunted

1 (0.5%)

0 (0%)

4 (1.73%)

0.3% (0-3.5)

2. Skeletal

Number Fetuses Examined

12

117

154

7127

a. Xiphisternum and one or more sternebrae unoissified

10 (7.7%)

10 (8.5%)

17 (11.0%)

11% (5-30)

b. With 14 rib(s)

15 (11.7%)

9 (7.6%)

12 (7.8%)

21% (6.36)

c. Vertebral bipartite centra

1 (0.8%)

1 (0.9%)

0 (0%)

2% (0-4)

3. Soft Tissue (Visceral and Neural)

Number Fetuses Examined

68

*

77

 

a. Found normal

68

-

77

 

(1) Personal communication; unpublished data obtained from Charles River Breeding Laboratories, Wilmington, Massachusetts for period 1964 -1969.

* Not examined.

Conclusions:
Maternal feeding of CFC-12 was not embryotoxic under the conditions of this experiment. CFC-12 was not teratogenic.
Executive summary:

The present study was undertaken, as part of a comprehensive toxicological program, to evaluate the embryotoxic and teratogenic potential, if any, of CFC-12 when it was administered by intragastric intubation to pregnant rats during the period of fetal organogenesis. 

Gastric intubation of dichlorodifluoromethane (CFC-12) to pregnant rats from day six through day fifteen of their gestation period, at average dose levels of 16.6 and 170.9 mg/kg/day, had no effect on the food intake and the gain in body weight of the mothers.

No clinical signs of toxicity were observed in any of the treated animals. The outcome of pregnancy, measured by the number of implantation sites, resorptions and live foetuses per litter, was not adversely affected by the treatment. The treatment did not affect the embryonal development as measured by weight and crown-rump length of the foetuses. The absence of any major external, skeletal, and soft tissue abnormalities in live foetuses and a similar incidence of spontaneous minor anomalies in the control and treated groups indicate that oral administration for CFC-12 was not teratogenic.

CONCLUSIONS

The absence of any effect on the number of implantation sites, resorptions, live foetuses per litter and mean weight and crown-rump length of the foetuses indicate that maternal feeding of CFC-12 was not embryotoxic under the conditions of this experiment.

Likewise, the absence of any major abnormalities in live foetuses and a similar incidence of spontaneous minor anomalies in the control and treated groups indicate that CFC-12 was not teratogenic.

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
Not specified
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Qualifier:
no guideline followed
Principles of method if other than guideline:
Summary only.
GLP compliance:
not specified
Limit test:
yes
Species:
rat
Strain:
Wistar
Details on test animals or test system and environmental conditions:
pregnant females
Route of administration:
inhalation
Type of inhalation exposure (if applicable):
not specified
Vehicle:
air
Details on exposure:
Not specified
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
Animals inseminated
Duration of treatment / exposure:
Rats: gestation day 4 to 16
Frequency of treatment:
2 hours per day.
Duration of test:
Half of the Rats were sacrificed on day 20 of gestation, the rest were allowed to deliver
Dose / conc.:
200 000 ppm (nominal)
Remarks:
eq. to 20% concentration of a mixture of CFC-11 (10%) and CFC-12 (90%) in air
(i.e., approx. 180000 ppm CFC-12)
No. of animals per sex per dose:
20 female rats
Control animals:
not specified
Details on study design:
Inseminated Wistar albino rats were administered a mixture of 90% CFC-12 (dichlorodifluoromethane) and 10% Freon 11 by inhalation for 2hr/day. Pregnant rats were exposed on days 4 to 16 of gestation. The mixture was administered in a 20% concentration (200,000 ppm).
Maternal examinations:
DETAILED CLINICAL OBSERVATIONS: no data

BODY WEIGHT: Yes
Ovaries and uterine content:
Not specified
Fetal examinations:
Yes, but no details provided
Clinical signs:
not specified
Mortality:
not specified
Body weight and weight changes:
no effects observed
Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
not specified
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not specified
Details on maternal toxic effects:
Maternal toxic effects: no effects
Key result
Dose descriptor:
NOEC
Effect level:
> 200 000 ppm (nominal)
Based on:
test mat.
Remarks:
(mixture of 10% CFC-11 and 90% CFC-12)
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
No effects on survival at birth
Changes in sex ratio:
not specified
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
no effects observed
Description (incidence and severity):
No effects on survival after 1 and 4 weeks
External malformations:
no effects observed
Description (incidence and severity):
No details on examinations performed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects: no effects reported.
Limited details available on teratogenicity examination.
Key result
Dose descriptor:
NOEC
Effect level:
>= 200 000 ppm
Based on:
test mat.
Basis for effect level:
other: No embryotoxicity or fetotoxicity observed at that concentration
Remarks on result:
not determinable due to absence of adverse toxic effects
Abnormalities:
no effects observed
Description (incidence and severity):
(limited details available)
Key result
Developmental effects observed:
no

No indications of any embryotoxic, fetotoxic, or teratogenic changes were found when dams were sacrificed and foetuses removed on day 20 of gestation (rats).

Conclusions:
No indications of any embryotoxic, fetotoxic, or teratogenic changes were found in the offspring of rats exposed during gestation days 4 to 16.
Executive summary:

In studies of inseminated Wistar albino rats, Paulet et al. (1974) administered a mixture of 90% CFC-12 (dichlorodifluoromethane) and 10% Freon 11 by inhalation for 2hr/day. Rats were exposed on days 4 to 16 of gestation. The mixture was administered in a 20% concentration (200,000 ppm). No indications of any embryotoxic, fetotoxic, or teratogenic changes were found when dams were sacrificed and foetuses removed on day 20 of gestation (rats). No classification is applicable.

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
Not specified
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
WHO assessment.
Principles of method if other than guideline:
Summary only.
GLP compliance:
not specified
Limit test:
yes
Species:
rabbit
Strain:
not specified
Details on test animals or test system and environmental conditions:
Not specified
Route of administration:
inhalation
Type of inhalation exposure (if applicable):
not specified
Vehicle:
air
Details on exposure:
Not specified
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
Not specified
Duration of treatment / exposure:
Rabbits: Exposure on days 5-20 of gestation
Frequency of treatment:
2 h/day
Duration of test:
Not specified
Dose / conc.:
200 000 ppm (nominal)
Remarks:
of a mixture of 10% CFC-11 and 90% CFC-12
eq. to 20% concentration of a mixture of CFC-11 (10%) and CFC-12 (90%) in air (i.e., approx. 180000 ppm CFC-12)
No. of animals per sex per dose:
Not specified
Control animals:
not specified
Details on study design:
Not specified
Maternal examinations:
Not specified
Ovaries and uterine content:
Not specified
Fetal examinations:
Not specified
Statistics:
Not specified
Indices:
Not specified
Historical control data:
No data
Details on maternal toxic effects:
Maternal toxic effects:no data
Remarks on result:
not determinable
Remarks:
no details available on maternal toxicity
Abnormalities:
not specified
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No evidence of embryotoxicity, fetotoxicity, or teratogenicity was seen when rabbits were sacrificed at 30 days of gestation. In addition, offspring of the dams that were allowed to deliver naturally showed no evidence of toxicity relative to survival or growth.
Dose descriptor:
NOAEC
Effect level:
>= 200 000 ppm
Based on:
test mat.
Sex:
not specified
Remarks on result:
not determinable due to absence of adverse toxic effects
Remarks:
no embryotoxicity, fetotoxicity or teratogenicity at the highest concentration tested
Abnormalities:
no effects observed
Developmental effects observed:
no
Conclusions:
In conclusion, CFC-12 did not show any evidence of reproductive or developmental toxicity in rabbits.
Executive summary:

In a study of a mixture (10% CFC-11 and 90% CFC-12), groups of rats and rabbits were exposed by inhalation on days 4-16 (rats) or days 5-20 (rabbits) of gestation for 2 h/day at a concentration of 989 g/m3(200 000 ppm). No evidence of embryotoxicity, fetotoxicity, or teratogenicity was seen when rats and rabbits were sacrificed at 20 days (rats) or 30 days (rabbits) of gestation. In addition, offspring of the dams that were allowed to deliver naturally showed no evidence of toxicity relative to survival or growth (US EPA, 1983).

 

In conclusion, CFC-12 did not show any evidence of reproductive or developmental toxicity.

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
Not specified
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
WHO assessment.
Principles of method if other than guideline:
Summary only.
GLP compliance:
not specified
Limit test:
yes
Species:
rat
Strain:
not specified
Details on test animals or test system and environmental conditions:
Not specified
Route of administration:
inhalation
Type of inhalation exposure (if applicable):
not specified
Vehicle:
air
Details on exposure:
Not specified
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
Not specified
Duration of treatment / exposure:
Rats: Exposure on days 4-16 of gestation
Frequency of treatment:
2 h/day
Duration of test:
Not specified
Dose / conc.:
200 000 ppm (nominal)
Remarks:
of a mixture of 10% CFC-11 and 90% CFC-12
eq. to 20% concentration of a mixture of CFC-11 (10%) and CFC-12 (90%) in air (i.e., approx. 180000 ppm CFC-12)
No. of animals per sex per dose:
Not specified
Control animals:
not specified
Details on study design:
Not specified
Maternal examinations:
Not specified
Ovaries and uterine content:
Not specified
Fetal examinations:
Not specified
Statistics:
Not specified
Indices:
Not specified
Historical control data:
No data
Details on maternal toxic effects:
Maternal toxic effects:no data
Remarks on result:
not determinable
Remarks:
no details available on maternal toxicity
Abnormalities:
not specified
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects: no effects

Details on embryotoxic / teratogenic effects:
No evidence of embryotoxicity, fetotoxicity, or teratogenicity was seen when rats and rabbits were sacrificed at 20 days (rats) or 30 days (rabbits) of gestation. In addition, offspring of the dams that were allowed to deliver naturally showed no evidence of toxicity relative to survival or growth.
Dose descriptor:
NOAEC
Effect level:
>= 200 000 ppm
Based on:
test mat.
Sex:
not specified
Remarks on result:
not determinable due to absence of adverse toxic effects
Remarks:
No reported effects on embryotoxicity, fetotoxicity, fetal malformations
Abnormalities:
no effects observed
Developmental effects observed:
no
Conclusions:
In conclusion, CFC-12 did not show any evidence of reproductive or developmental toxicity in rats.
Executive summary:

In a study of a mixture (10% CFC-11 and 90% CFC-12), groups of rats and rabbits were exposed by inhalation on days 4-16 (rats) or days 5-20 (rabbits) of gestation for 2 h/day at a concentration of 989 g/m3(200 000 ppm). No evidence of embryotoxicity, fetotoxicity, or teratogenicity was seen when rats and rabbits were sacrificed at 20 days (rats) or 30 days (rabbits) of gestation. In addition, offspring of the dams that were allowed to deliver naturally showed no evidence of toxicity relative to survival or growth (US EPA, 1983).

 

In conclusion, CFC-12 did not show any evidence of reproductive or developmental toxicity.

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
Not specified
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Qualifier:
no guideline followed
Principles of method if other than guideline:
Summary only.
GLP compliance:
not specified
Limit test:
yes
Species:
rabbit
Strain:
not specified
Remarks:
albino rabbits
Details on test animals or test system and environmental conditions:
pregnant females
Route of administration:
inhalation
Type of inhalation exposure (if applicable):
not specified
Vehicle:
air
Details on exposure:
Not specified
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
Animals inseminated
Duration of treatment / exposure:
Rabbits: gestation day 5 to 20
Frequency of treatment:
2 hours per day.
Duration of test:
Half of the Rabbits were sacrificed on day 30 of gestation, the rest were allowed to deliver
Dose / conc.:
200 000 ppm (nominal)
Remarks:
eq. to 20% concentration of a mixture of CFC-11 (10%) and CFC-12 (90%) in air
(i.e., approx. 180000 ppm CFC-12)
No. of animals per sex per dose:
10 female rabbits
Control animals:
not specified
Details on study design:
Inseminated albino rabbits were administered a mixture of 90% CFC-12 (dichlorodifluoromethane) and 10% Freon 11 by inhalation for 2hr/day, on days 5 to 20. The mixture was administered in a 20% concentration (200,000 ppm).
Maternal examinations:
DETAILED CLINICAL OBSERVATIONS: no data

BODY WEIGHT: Yes
Ovaries and uterine content:
Not specified
Fetal examinations:
Yes, but no details provided
Clinical signs:
not specified
Mortality:
not specified
Body weight and weight changes:
no effects observed
Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
not specified
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not specified
Details on maternal toxic effects:
Maternal toxic effects: no effects
Key result
Dose descriptor:
NOEC
Effect level:
>= 200 000 ppm (nominal)
Based on:
test mat.
Remarks:
(mixture of 10% CFC-11 and 90% CFC-12)
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
No effects on survival at birth
Changes in sex ratio:
not specified
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
no effects observed
Description (incidence and severity):
No effects on survival after 1 and 4 weeks
External malformations:
no effects observed
Description (incidence and severity):
No details on examinations performed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects: no effects reported.
Limited details available on teratogenicity examination.
Key result
Dose descriptor:
NOEC
Effect level:
>= 200 000 ppm
Based on:
test mat.
Basis for effect level:
other: No observed embryotoxicity, fetotoxicity, teratogenicity observed at that concentration
Remarks on result:
not determinable due to absence of adverse toxic effects
Abnormalities:
no effects observed
Description (incidence and severity):
(limited details available)
Key result
Developmental effects observed:
no

No indications of any embryotoxic, fetotoxic, or teratogenic changes were found when dams were sacrificed and foetuses removed on day 30 of gestation (rabbits).

Conclusions:
No indications of any embryotoxic, fetotoxic, or teratogenic changes were found in the offspring of rabbits exposed during gestation days 5 to 20.
Executive summary:

In studies with inseminated albino rabbits, Paulet et al. (1974) administered a mixture of 90% CFC-12 (dichlorodifluoromethane) and 10% Freon 11 by inhalation for 2hr/day. Rabbits were exposed on days 5 to 20. The mixture was administered in a 20% concentration (200,000 ppm). No indications of any embryotoxic, fetotoxic, or teratogenic changes were found when dams were sacrificed and foetuses removed on day 30 of gestation in rabbits. No classification is applicable.

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
Not specified
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
Taken from publically available data, and is considered accurate based on the registrants experience of the substance and the source as the WHO.
Reason / purpose for cross-reference:
reference to same study
Qualifier:
no guideline followed
Principles of method if other than guideline:
Summary only.
GLP compliance:
not specified
Remarks:
Not likely (prior to GLP standards)
Limit test:
yes
Species:
rat
Strain:
other: Charles River
Details on test animals or test system and environmental conditions:
Not specified
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
Not specified
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
Not specified
Duration of treatment / exposure:
Duration 10 days (dosed on days 6-15 of gestation).
Frequency of treatment:
Daily
Dose / conc.:
16.6 mg/kg bw/day (nominal)
Dose / conc.:
179 mg/kg bw/day (nominal)
No. of animals per sex per dose:
Groups of 25 to 27 pregnant rats used within the study.
Control animals:
yes, concurrent vehicle
not specified
Details on study design:
No data
Maternal examinations:
Not specified
Ovaries and uterine content:
Not specified
Fetal examinations:
Not specified
Statistics:
Not specified
Indices:
Not specified
Historical control data:
Not specified
Details on maternal toxic effects:
Maternal toxic effects:no data
Dose descriptor:
NOAEL
Effect level:
>= 179 mg/kg bw/day
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Remarks:
no reported maternal toxicity
Abnormalities:
not specified
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Neither dose induced any evidence of embryotoxicity or teratogenicity.
Dose descriptor:
NOAEL
Effect level:
> 179 mg/kg bw/day
Based on:
test mat.
Sex:
not specified
Remarks on result:
not determinable due to absence of adverse toxic effects
Remarks:
no reported embryotoxicity or teratogenicity
Abnormalities:
no effects observed
Developmental effects observed:
no
Conclusions:
In conclusion, CFC-12 did not show any evidence of reproductive or developmental toxicity.
Executive summary:

In a study, groups of 25 to 27 pregnant Charles River rats were given CFC-12 in corn oil by gavage at doses of 16.6 or 179 mg/kg per day on days 6-15 of gestation. Neither dose induced any evidence of embryotoxicity or teratogenicity (Sherman, 1974).

 

In conclusion, CFC-12 did not show any evidence of reproductive or developmental toxicity. No classification is applicable.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no adverse effect observed
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

Several studies conducted in the past in rats and rabbits were available and assessed as a weight of evidence because they did not fulfill the current standards and available documentation was limited. However the studies reports cited here were reviewed by international experts as part of the International Program on Chemical Safety and summarised in the Environmental Health Criteria document No.113: Fully halogenated chlorofluorocarbons (WHO/IPCS, 1990).

http://www.inchem.org/documents/ehc/ehc/ehc113.htm#PartNumber:7

 

One of the most relevant studies was undertaken, as part of a comprehensive toxicological program, to evaluate the embryotoxic and teratogenic potential of CFC-12 when administered by intragastric intubation to pregnant rats during the period of fetal organogenesis. 

Gastric intubation of dichlorodifluoromethane (CFC-12) to pregnant rats from day six through day fifteen of their gestation period, at average dose levels of 16.6 and 170.9 mg/kg/day, had no effect on the food intake and the gain in body weight of the mothers.

No clinical signs of toxicity were observed in any of the treated animals. The outcome of pregnancy, measured by the number of implantation sites, resorptions and live foetuses per litter, was not adversely affected by the treatment. The treatment did not affect the embryonal development as measured by weight and crown-rump length of the foetuses. The absence of any major external, skeletal, and soft tissue abnormalities in live foetuses and a similar incidence of spontaneous minor anomalies in the control and treated groups indicate that oral administration for CFC-12 was not teratogenic.

 

CONCLUSIONS

The absence of any effect on the number of implantation sites, resorptions, live foetuses per litter and mean weight and crown-rump length of the foetuses indicate that maternal feeding of CFC-12 was not embryotoxic under the conditions of this experiment.

Likewise, the absence of any major abnormalities in live foetuses and a similar incidence of spontaneous minor anomalies in the control and treated groups indicate that CFC-12 was not teratogenic.

 

Other supporting studies:

In studies of inseminated Wistar albino rats and albino rabbits, Paulet et al. (1974) administered a mixture of 90% CFC-12 (dichlorodifluoromethane) and 10% Freon 11 by inhalation for 2hr/day. Rats were exposed on days 4 to 16 of gestation, and rabbits on days 5 to 20. The mixture was administered in a 20% concentration (200,000 ppm). No indications of any embryotoxic, fetotoxic, or teratogenic changes were found when dams were sacrificed and foetuses removed on day 20 of gestation (rats) or day 30 of gestation (rabbits). No classification is applicable.

 

In a study of a mixture (10% CFC-11 and 90% CFC-12), groups of rats and rabbits were exposed by inhalation on days 4-16 (rats) or days 5-20 (rabbits) of gestation for 2 h/day at a concentration of 989 g/m3(200 000 ppm). No evidence of embryotoxicity, fetotoxicity, or teratogenicity was seen when rats and rabbits were sacrificed at 20 days (rats) or 30 days (rabbits) of gestation. In addition, offspring of the dams that were allowed to deliver naturally showed no evidence of toxicity relative to survival or growth (US EPA, 1983). (possibly the same study by Paulet et al).

 

In a study, groups of 25 to 27 pregnant Charles River rats were given CFC-12 in corn oil by gavage at doses of 16.6 or 179 mg/kg per day on days 6-15 of gestation. Neither dose induced any evidence of embryotoxicity or teratogenicity (Sherman, 1974).

 

In conclusion, CFC-12 did not show any evidence of reproductive or developmental toxicity. No classification is applicable.

Justification for classification or non-classification

The above studies have all been ranked reliability 2 according to the Klimish et al system. This ranking was deemed appropriate because the studies were not conducted to GLP and are not in compliance with agreed protocols. However these are considered appropriate for use in assessment purposes in accordance with the criteria listed in Annex XI. As such, it is deemed appropriate to apply a weight of evidence approach based on similarity and animal welfare grounds.

The above results triggered no classification under the CLP Regulation (EC No 1272/2008). No classification for reproductive or developmental effects is therefore required.

Additional information