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Toxicological information

Repeated dose toxicity: inhalation

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Administrative data

Endpoint:
sub-chronic toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference
Reference Type:
publication
Title:
Effects on experimental animals of long-term inhalation of trichloroethylene, carbon tetrachloride, 1,1,1-trichloroethane, dichlorodifluoromethane, and 1,1-dichloroethylene.
Author:
Prendergast JA, Jones RA, Jenkins LJ Jr, Siegel J.
Year:
1967
Bibliographic source:
Toxicol Appl Pharmacol., vol. 10, no. 2, p. 270-89.

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
- Principle of test: groups of animals were exposed to dichlorodifluoromethane daily for 30 exposures, or as a continuous exposure for 90 days.
- Short description of test conditions:
- Parameters analysed / observed:
GLP compliance:
no
Remarks:
Prior to GLP guidelines
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
Dichlorodifluoromethane
EC Number:
200-893-9
EC Name:
Dichlorodifluoromethane
Cas Number:
75-71-8
Molecular formula:
CCl2F2
IUPAC Name:
dichlorodifluoromethane
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Matheson Gas Company, East Rutherford, New Jersey


Test animals

Species:
other: rats, guinea-pigs, monkeys, rabbits, dogs
Strain:
other: as stated in the publication: 15 Sprague-Dawley or Long-Evans, 15 Hartley guinea-pigs, 3 squirrel monkeys, 3 New-Zealand rabbits, 2 beagle dogs
Sex:
not specified
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: no data
- Age at study initiation: no data
- Weight at study initiation: 192 g (range: 145-244g) at start of the repeated exposure study; 213 g (range: 165-262g) at the start of the continuous exposure study
- Housing:
- Diet : not specified
- Water : ad libitum
- Acclimation period:

DETAILS OF FOOD AND WATER QUALITY: commercial dry chow.

ENVIRONMENTAL CONDITIONS : (exposure chambers)
- Temperature (°C): 75 to 80°F (23 to 26.7°C)
- Humidity (%): 50%
- Air changes (per hr): 1.23 m3/ minute
- Photoperiod (hrs dark / hrs light): no data

IN-LIFE DATES: no data

Administration / exposure

Route of administration:
inhalation: gas
Type of inhalation exposure:
whole body
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: chamber (no details)
- Source and rate of air: no data
- Method of conditioning air: a stream of pure gaseous dichlorodifluoromethane from a high-pressure cylinder was metered through a rotameter into a mixing bottle and carried into the chember where it was diluted to the atrget concentration with air.
- Temperature, humidity, pressure in air chamber: 75-80°F (23.6-26.6 °C), 50% humidity, negative pressure at 2.0 inches if water
- Air flow rate: 1.25 m3/minute
- Air change rate: no data
- Treatment of exhaust air: no data

TEST ATMOSPHERE
- Brief description of analytical method used: continous monitoring of chamber concentrations by infrared analysis.
- Samples taken from breathing zone: no data

VEHICLE (if applicable)
- Justification for use and choice of vehicle: no data
- Composition of vehicle: air
Analytical verification of doses or concentrations:
yes
Remarks:
Infrared analysis
Details on analytical verification of doses or concentrations:
Infrared analysis. A stream of chamber air was drawn through a variable pathlength gas cell fitted to an infrared spectrophotometer. The instrument was locked on the analytical wavelength selected for the contaminant and the percent transmission was continuously recorded. The contaminant level was then read from a graph of concentration vs percent transmission previously prepared by vaporizing known amounts of the test material in the gas cell.
Chemical analysis were performed daily and correlated with nominal input data.
Duration of treatment / exposure:
- repeated exposure study: 30 exposures
- continuous exposure study: 90 days
Frequency of treatment:
- Repeated exposures: Daily 8 hours/day, 5 days/week for a total of 30 exposures
- Continuous exposure for 90-day, except for feeding the animals and servicing the chambers (< 2% of the total chamber time)
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/m³ air
Dose / conc.:
4 136 mg/m³ air
Remarks:
30 repeated exposures (Mean +/- SD 87)
Dose / conc.:
3 997 mg/m³ air
Remarks:
90-day continuous exposure (Mean +/- SD 113)
No. of animals per sex per dose:
15 rats per dose (sex not specified)
Control animals:
yes, concurrent no treatment
Positive control:
no

Examinations

Observations and examinations performed and frequency:
DETAILED CLINICAL OBSERVATIONS: yes
All animals were routinely examined for signs of toxicity, marked alterations in behaviour, physical appearance, respiration pattern, locomotor activity and prostration.

BODY WEIGHT: Yes
- Time schedule for examinations: prior to exposure, at monthly intervals and at the termination of the study.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: no data
- Anaesthetic used for blood collection: Not specified
- Animals fasted: Not specified
- How many animals: Not specified
- Parameters examined : Total and differential Leukocyte counts, Hemoglobin concentration, Hematocrit, before and after the exposure

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood:
- Animals fasted: Yes / No / Not specified
- How many animals:
- Parameters examined:
* Reduced nicotinamide adenine dinucleotide (NADH), reduced nicotinamide adenine dinucleotide phosphate ( NADPH), activities of succinic dehydrogenase ( SDH ), lactic dehydrogenase ( LDH), isocitric dehydrogenase ( ICD), glucose-6-phosphate dehydrogenase ( GGPD), and p-hydroxybutyric dehydrogenase (B-OHBD).
* Alkaline phosphatase activity
* Serum glutamic-pyruvic transaminase levels
* Serum urea nitrogen concentrations
* Liver lipids

URINALYSIS: Not specified

Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes : heart, lung, liver, spleen, kidney

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
Repeated exposure: No signs of toxicity were observed in the rat survivors.
Continuous exposure: no visible signs of toxicity
Mortality:
mortality observed, treatment-related
Description (incidence):
Rats: (no details on day)
1/15 in the repeated exposure group
2/15 in the continuous exposure group
7/304 controls

Guinea-pigs:
1/15 in the continuous exposure group.

No mortality observed in the other groups and species.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
* Repeated exposure: no effect reported
* continuous exposure: No effects reported in rats. Body weights of the rabbits and guinea-pigs were depressed.
Food consumption and compound intake (if feeding study):
not examined
Ophthalmological findings:
not examined
Clinical biochemistry findings:
not specified
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
No details
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
* Repeated exposure: several rats and guinea pigs, had varying degrees of lung congestion.
Other organs appeared normal.
No effects were reported in rats, but guinea-pigs showed focal necrosis of the liver, but which could not be defintely attributed to the exposure.

* Continuous exposure: high incidence of varying degrees of lung congestion in rats, rabbits, monkeys, guinea-pigs.
In guinea-pigs, submassive necrosis of the liver was noted in the surviving guinea-pigs.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
* Repeated exposure: non-specific interstitial inflammatory changes in the lungs of both treated and control animals.
- Several guinea-pigs showed focal necrosis or fatty infiltration of the liver.
- one monkey had heavy pigment deposits in the liver, spleen, kidney.

* Continuous exposure: non-specific interstitial inflammatory changes in the lungs (all species)
- one guinea-pig showed a focal giant cell pneumonitis. All guinea-pig liver sections: slight to extensive fatty infliltration of the hepatic cells; several sections with focal or submassive necrosis of the liver. It could not be determined if this toxic response was due to the continuous exposure or to the higher susceptibility of the guinea-pigs, already noted in other studies.
Histopathological findings: neoplastic:
not specified

Effect levels

Key result
Dose descriptor:
NOAEC
Effect level:
ca. 3 997 mg/m³ air (nominal)
Based on:
test mat.
Sex:
not specified

Target system / organ toxicity

Key result
Critical effects observed:
no

Applicant's summary and conclusion