Registration Dossier

Administrative data

Description of key information

Key value for chemical safety assessment

Carcinogenicity: via oral route

Endpoint conclusion
Endpoint conclusion:
no study available

Carcinogenicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
carcinogenicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1975 to 1979
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Taken from publically available data. The study is well documented and contains suitable information for use in the assessment. The full literature report is appended for reference.
Qualifier:
no guideline followed
Principles of method if other than guideline:
CFC-12 was tested by inhalation on Sprague-Dawley rats and Swiss Mice using whole body exposure. The animals were exposed by inhalation, 4 hours daily, 5 days weekly, for 104 weeks (rats: Exp. BT 601, 601 bis) and 78 weeks (mice: Exp. BT 602).
All the animals were kept under control until spontaneous death. The status and behaviour of the animals were controlled three times daily. The animals were submitted to clinical examination for gross changes every 2 weeks. The animals were weighed every 2 weeks during treatment, and then every 8 weeks. Full necropsy was performed on all the animals. The housing and the diet of the animals were the same highly standardized ones adopted in the BT Experimental Unit during the last 15 years before the beginning of this experiment.
GLP compliance:
not specified
Species:
other: mouse & rat
Strain:
other: Swiss mice & Sprague-Dawley rats
Sex:
male/female
Details on test animals and environmental conditions:
Male and female Sprague-Dawley rats 9 (Exp. BT 601), and 13 (Exp. BT 601 bis,) weeks old at the start of the experiment.
Male and female Swiss mice 8 weeks old at the start of the experiment (Exp. BT 602).
The animals were of the breed currently used in the BT Experimental Unit for more than 15 years.
All the animals were kept under control until spontaneous death.
The status and behaviour of the animals were controlled three times daily.
The housing and the diet of the animals were the same highly standardized ones adopted in the BT Experimental Unit during the last 15 years before the beginning of this experiment.
Route of administration:
inhalation
Type of inhalation exposure (if applicable):
whole body
Vehicle:
unchanged (no vehicle)
Details on exposure:
The chambers for inhalation exposure were made of stainless steel, with two glass doors, and they measure 135 x 98 x 65 cm. The volume of each chamber is 860 litres. Continuous air flow provided 12-15 air changes per hour. Before its introduction the air was filtered, and the chamber arrangement was such that air flowed from one part of the chamber to the other without recirculation. The internal pressure was about 1 mm Hg less than that of the room where the chambers were situated to avoid any possible contamination of the outside environment.
FC 12 was tested in rats with two experiments each: BT 601 and BT 601 bis (CFC-12). Experiment BT 601 bis included 60 animals (30 males and 30 females). Experiment BT 601 was started concurrently in rats of the same litters. Experiment BT 601 bis was started concurrently in rats of the same litters after experiment BT 601. At the start of the project, the animals of experiments BT 601 bis were scheduled for interim sacrifice for clinical laboratory analyses and histopathology. Since it was then decided not to perform such exams, all the animals of the experiments were treated and examined in the same way, and therefore are considered as a whole.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The concentrations in air were checked by continuous gas chromatographic monitoring-no further details referenced within the publication.
Duration of treatment / exposure:
The animals were exposed by inhalation, 4 hours daily, for 104 weeks (rats: Exp. BT 601, 601 bis) and 78 weeks (mice: Exp. BT 602).
Frequency of treatment:
5 days weekly,
Post exposure period:
Not specified within the publication.
Remarks:
Doses / Concentrations:
0, 1000, 5000 ppm
Basis:
nominal conc.
No. of animals per sex per dose:
Mouse: 420 total (210 male & 210 female)
Rat: 660 total (330 male & 330 female)
Control animals:
yes, concurrent no treatment
Details on study design:
Detailed in table form, attached under any other information.
Positive control:
Not specified
Observations and examinations performed and frequency:
The status and behaviour of the animals were controlled three times daily.
The animals were submitted to clinical examination for gross changes every 2 weeks.
The animals were weighed every 2 weeks during treatment, and then every 8 weeks.
Sacrifice and pathology:
Systematic and standardized histopathologic examinations were performed on each animal on the subcutaneous lymph nodes, brain and cerebellum, Zymbal glands, interscapular brown fat, salivary glands, Harderian glands, tongue, thymus and mediastinal lymph nodes, lungs, diaphragm, liver, kidneys, adrenals, spleen, mesenteric lymph nodes, stomach, various segments of the intestine, bladder, uterus, gonads, bone marrow smear, and any other organs with pathologic lesions.
Full necropsy was performed on all the animals.
Other examinations:
Not specified
Statistics:
log rank test
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
The tumors most frequently expected in the rats of the strain used, on the basis of the literature and of the historical controls of the BT Experimental Unit, are mammary tumors (benign and malignant), leukemias, pheochromocytomas and pheochromoblastomas. Moreover, a variety of other miscellaneous tumors are also observed.
In mice, the tumors most frequently expected on the basis of the literature and of the historical controls of the BT Experimental Unit, are mammary carcinomas (in females), pulmonary tumors and leukemias. Moreover, a variety of other miscellaneous tumors are also observed.
FC12: Rats (Exp. BT 601 + 601 bis): Survival and body weight; No important differences related to the exposure to the test compound were found.
Carcinogenicity; No noticeable differences, related to treatment, were found in the incidence of total benign and malignant, and malignant tumors, and of the most frequent expected or rare tumors.
FC12: Mice (Exp. BT 602): Survival and body weight; The survival rate varied from group to group, was generally lower in control than in treated groups, and showed statistically significant differences after various periods. The treatment did not affect the body weight.
Carcinogenicity; A higher number of the following tumors was observed in treated groups: total number of tumors in males and females, dose-related in males; pulmonary adenomas in males and females, at 5000 ppm; and leukemias in males, at 5000 and 1000 ppm, and in females at 1000
ppm.
Since the above-mentioned tumors were generally age-related, a statistical method was used, namely the log rank test, which allows for such variations. When such a test was used, no statistically significant increases in total number of tumors, pulmonary adenomas, and leukemias was found in treated male or female mice in comparison with control animals.
Relevance of carcinogenic effects / potential:
No carcinogenic effects were noted within the study.
Dose descriptor:
NOEC
Effect level:
5 000 ppm
Based on:
not specified
Sex:
male/female
Basis for effect level:
other: Mouse & Rat data
Remarks on result:
other: Effect type: carcinogenicity (migrated information)

Dichlorodifluoromethane (FC12): Total Tumors in Rats (Exp. BT 601 + 601 bis)

Group No.

Concentration (ppm)

Animals

Percentage of Animals Bearing Tumors

No. of Malignant Tumors per 100 Animals

Sex

No. at Start

TBMTa

MTb

I

5000

M

90

30.0

10.0

10.0

F

90

80.0

20.0

20.0

M + F

180

55.0

15.0

15.0

II

1000

M

90

46.7

17.8

17.8

F

90

78.9

26.7

26.7

M + F

180

62.8

22.2

22.2

III

0 (Control)

M

150

34.0

16.7

17.3

F

150

82.7

28.7

31.3

M + F

300

58.3

22.7

24.3

aTotal benign and malignant tumors.

bMalignant tumors.

 

Dichlorodifluoromethane (FC12): Mammary Tumors, Leukemias, Pheochromocytomas and Pheochromoblastomas in Rats (Exp. BT 601 + 601 bis)

Group No.

Concentration (ppm)

Animals

Percentage of Animals Bearing Tumors

Sex

No. at Start

Mammary Tumors

 

BMTa

MTb

Leukemiasc

Pheochromocytomas

Pheochromoblastomas

I

5000

M

90

12.2

2.2

3.3

5.5

-

F

90

74.4

10.0

1.1

1.1

-

M + F

180

43.3

6.1

2.2

3.3

-

II

1000

M

90

15.5

1.1

7.8

12.2

1.1

F

90

68.9

14.4

3.3

1.1

-

M + F

180

42.2

7.8

5.5

6.7

0.5

III

0 (Control)

M

150

12.7

4.0

6.0

4.0

-

F

150

72.0

13.3

5.3

3.3

-

M + F

300

42.3

8.7

5.7

3.7

-

aBenign and malignant tumors.

bMalignant tumors.

cThe term “leukemia” includes a variety of hemolymphoreticular neoplastic diseases at different sites.

 

Dichlorodifluoromethane (FC12): Total Tumors in Mice (Exp. BT 602)

Group No.

Concentration (ppm)

Animals

Percentage of Animals Bearing Tumors

No. of Malignant Tumors per 100 Animals

Sex

No. at Start

TBMTa

MTb

I

5000

M

60

25.0

11.7

11.7

F

60

21.7

10.0

10.0

M + F

120

23.3

10.8

10.8

II

1000

M

60

15.0

6.7

6.7

F

60

25.0

23.3

23.3

M + F

120

20.0

15.0

15.0

III

0 (Control)

M

90

10.0

5.5

5.5

F

90

16.7

10.0

11.1

M + F

180

13.3

7.8

8.3

aTotal benign and malignant tumors.

bMalignant tumors.

 

Dichlorodifluoromethane (FC12): Mammary Carcinomas, Pulmonary Tumors and Leukenias in Mice (Exp. BT 602)

Group No.

Concentration (ppm)

Animals

Percentage of Animals Bearing Tumors

Sex

No. at Start

Mammary Carcinomas

Pulmonary Tumors

Leukemiasa

Adenomas

Adenocarcinomas

I

5000

M

60

-

8.3

-

6.7

 

 

F

60

-

5.0

-

10.0

 

 

M + F

120

-

6.7

-

8.3

II

1000

M

60

-

5.0

-

6.7

 

 

F

60

3.3

1.7

-

20.0

 

 

M + F

120

1.7

3.3

.

13.3

III

0 (Control)

M

90

-

3.3

-

3.3

 

 

F

90

1.1

2.2

-

8.9

 

 

M + F

180

0.5

2.8

-

6.1

aThe term “leukemia” includes a variety of hemolymphoreticular beoplastic diseases at different sites.

 

Dichlorodifluoromethane (FC12): Liver Angiosarcomas in Rats

Experiment and Compound

Group No.

Concentration (ppm)

Animals

Percentage of Animals Bearing Liver Angiosarcomas

Sex

No. at Start

BT 601 + BT 601 bis (FC12)

I

5000

M

90

-

F

90

3.3

M + F

180

1.7

II

1000

M

90

-

F

90

1.1

M + F

180

0.5

III

0 (Control)

M

150

0.7

F

150

0.7

M + F

300

0.7

Conclusions:
Under the experimental conditions, all three compounds failed to show any carcinogenic effects.
Executive summary:

The propellant chlorofluorocarbon dichlorodifluoromethane (CFC-12)was administered by inhalation at a concentration of 5000, 1000 and 0 ppm, 4 hours daily, 5days weekly, for 104 and 78 weeks, to rats and mice, respectively.

The animals were kept under observation until spontaneous death.

No unexpected treatment-related tumors were observed in rats exposed to CFC-12. No important differences in the incidence of total benign and malignant tumors and of frequently occurring spontaneous tumors were observed in the rats exposed to CFC-12.

The incidence of all tumors and of some particular frequently occurring spontaneous tumors in mice showed a tendency to increase in animals exposed toCFC-12. The increased incidence was usually observed in one sex and was not always dose-related, possibly due toalonger survival of the treated mice compared with controls.

Under the experimental conditions, and on the basis of statistical analysis and of the oncological evaluation of the results, CFC-12 failed to show carcinogenic effects.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEC
24 725 mg/m³
Study duration:
chronic
Species:
rat
Quality of whole database:
Study is considered to be appropriate for use as it presents well documented results in an appropriate format, despite not being to GLP.
Calculation based on 5000 ppm * 120.91 /24.45 = 24725 mg/m3 where 24.45 is the molar volume as reported into the ACGIH ("2010 TLVs and BEIs, Based on the Documentation of the Threshold LImit Values for Chemical Substances and Physical Agents & Biological Exposure Indices") for conversion ppm vs mg/m3.

Additional information

A carcinogenicity study on the substance was conducted. Dichlorodifluoromethane (CFC-12)was administered by inhalation at a concentration of 5000, 1000 and 0 ppm, 4 hours daily, 5days weekly, for 104 and 78 weeks, to rats and mice, respectively.

As a result of this study, no unexpected treatment-related tumors were observed in rats exposed. No important differences in the incidence of total benign and malignant tumors and of frequently occurring spontaneous tumors were observed in the rats exposed to the tested propellant. The incidence of all tumors and of some particular frequently occurring spontaneous tumors in mice showed a tendency to increase in animals exposed to CFC-12.The increased incidence was usually observed in one sex and was not always dose-related, possibly due to a longer survival of the treated mice compared with controls.

 

Under the experimental conditions, and on the basis of statistical analysis and of the oncological evaluation of the results, CFC-12 failed to show carcinogenic effects.

 


Justification for selection of carcinogenicity via inhalation route endpoint:
Well documented literature paper available.

Justification for classification or non-classification

The above study has been ranked reliability 2 according to the Klimish et al system. This ranking was deemed appropriate because the study was not conducted to GLP and is 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 Dangerous Substance Directive (67/548/EEC) and the CLP Regulation (EC No 1272/2008). No classification for mutagenic effects is therefore required.