Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
9 December 2003 - 7 March 1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was conducted according to OECD guideline 412 and under GLP conditions.
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1994
Report date:
1994
Reference Type:
publication
Title:
Twenty-eight day nose-only inhalation toxicity study of resorcinol bis-diphenylphosphate (Fyrolflex RDP) in rats
Author:
Henrich, R.T., Johnson, W.D., Rajendran, N., Freudenthal, R.I., Tomlinson, M.J., Aranyi, C.
Year:
2000
Bibliographic source:
International Journal of Toxicology, 19: 223-231 (2000)

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)
Deviations:
yes
Remarks:
in addition, MNSE activity was determined to study neurotoxicity
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction mass of 3-[(diphenoxyphosphoryl)oxy]phenyl triphenyl 1,3-phenylene bis(phosphate) and tetraphenyl 1,3-phenylene bis(phosphate)
EC Number:
701-337-2
Cas Number:
not available
Molecular formula:
C30H24O8P2
IUPAC Name:
Reaction mass of 3-[(diphenoxyphosphoryl)oxy]phenyl triphenyl 1,3-phenylene bis(phosphate) and tetraphenyl 1,3-phenylene bis(phosphate)
Details on test material:
- Name of test material (as cited in study report): Fyrolflex RDP
- Physical state: Liquid
- Analytical purity: Confidential information
- Composition of test material, percentage of components: Confidential information
- Purity test date: Confidential information
- Lot/batch No.: Confidential information

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories
- Age at study initiation: 8-9 weeks
- Weight at study initiation:
Males: 196-233 g
Females: 167-212 g
- Housing: According to guideline
- Diet (e.g. ad libitum): Ad libitum, rodent chow
- Water (e.g. ad libitum): Ad libitum, tap water
- Acclimation period: 3 days (of 15 days quaratine)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-26
- Humidity (%): 32-60
- Air changes (per hr): 15 +/- 2
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
other: unchanged (no vehicle)
Remarks on MMAD:
MMAD / GSD: By Quartz Crystal Microbalance (QCM)
MMAD: 1.39-1.70 um
GSD: 1.67-2.04 um
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 52-port nose-only inhalation exposure chambers
- Method of holding animals in test chamber: Holding tubes
- Source and rate of air: Regulated compressed air source
- System of generating particulates/aerosols: nebulizers and regulated compressed air source
- Air flow rate: Adjusted to generate aerosol with particle size of <3 um MMAD
- Treatment of exhaust air: Moved through an inertial trap and a particulate filter by a ring compressor and exhausted outside the building

TEST ATMOSPHERE
- Brief description of analytical method used:
Aerosol mass concentraiton: Gravimetric analysis (filter), light scattering method (sensors) and chemical analysis (filter)
Aerosol particle size: Quartz Crystal Microbalance (QCM)
- Samples taken from breathing zone: no, only from filters and sensors
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Gravimetry and chemical analysis
Duration of treatment / exposure:
4 weeks
Frequency of treatment:
6 hours/day 5 days/week
Doses / concentrations
Remarks:
Doses / Concentrations:
0.1, 0.5, 2.0 mg/l
Basis:
nominal conc.
No. of animals per sex per dose:
Low- and mid-dose group: 10
High-dose group: 20
Control animals:
other: filtered air
Details on study design:
- Rationale for animal assignment: Random (constrained by body weight)
- Post-exposure recovery period in satellite groups: 60 days
- Section schedule rationale: Random
Positive control:
Not relevant

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily
- Cage side observations: mortality and moribundity

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least weekly

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes, weekly

HAEMATOLOGY: Yes
- Time schedule for collection of blood: 7 days prior to exposure
- Anaesthetic used for blood collection: Yes (CO2 or sodium pentobarbital at termination)
- Animals fasted: Yes (overnight before termination)
- How many animals: All
- Parameters checked:
APTT
Erythrocyte count and morphology
Hematocrit
Hemoglobin concentration
Leukocyte count (total and differential)
Mean corpuscular hemoglobin
Mean corpuscular hemoglobin concentration
Mean corpuscular volume
Platelet count
Prothrombin time
Reticulocyte count

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood:
- Animals fasted: Yes (overnight before termination)
- How many animals: All
- Parameters checked:
Albumin
Albumin/globulin ratio (calculated)
Bilirubin (total)
Blood urea nitrogen
Calcium
Chloride
CPK
Cholesterol
Cholinesterase (plasma)
Cholinesterase (erythrocyte)
Creatinine
Gamma glutamyl transpeptidase
Globulin
Glucose
LDH
Monocyte non-specific esterase
Phosphorus
Potassium
Serum alanine aminotransferase
Serum alkaline phosphatase
Serum aspartase aminotransferase
Sodium
Total protein
Triglycerides
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, examination of external surface and all orifices; external surfaces of brain and spinal cord; organs and tissues of cranial, thoracic, abdominal and pelvic cavities and neck; remainder of the carcass

ORGAN WEIGHTS: Yes, see below:
Liver
Kidney
Lungs
Spleen
Adrenal glands
Heart
Brain
Testes with epididymides or ovaries
Pituitary
Thymus
Thyroid/parathyroids

At recovery only lung weight determined

HISTOPATHOLOGY: Yes, microscopic examination of tissue of control and high-dose group (+ tissues with lesions and target organs from low- and mid-dose group):
Adrenal glands
Brain
Buccal mucosa
Epididymides
Esophagus
Eyes
Heart
Kidneys
Larynx
Liver
Lungs
Nose
Ovaries
Pancreas
Pituitary gland
Spinal cord
Spleen
Testes
Thymus
Thyroid gland
Trachea

Gross lesions
Target organs
Other examinations:
Not relevant
Statistics:
ANOVA followed by Dunnett's test for comparison of multiple groups with a single control group

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
not examined
Details on results:
BODY WEIGHT AND WEIGHT GAIN
Mean body weight significantly decreased in high-dose males in weeks 1-3, and did not recover during the first 5 weeks of recovery. No effect were found in females, exept for a significant increase in mean body weight for high-dose females in week 4 of recovery.

FOOD CONSUMPTION
Food consumption significantly decreased in high dose males during weeks 1 and 3 of exposure

HEMATOLOGY
Low-dose female rats had significantly increased total erythrocyte counts, hemoglobin and hematocrit.

CLINICAL CHEMISTRY
No toxicologically significant effects on any clinical chemistry parameter during exposure.

Mean plasma cholinesterase activity levels significantly decreased in high-dose males and mid- and high-dose females at end of exposure period. Significant decrease was still present in high-dose females at end of recovery. No effects on erythrocyte cholinesterase levels.

MNSE activity levels increased in low- and mid-dose rats at end of exposure period. In high-dose animals this activity was comparable to controls, not indicating a dose related effect.

ORGAN WEIGHTS
Dose-related increase in absolute and relative lung weights in test group, significantly in mid and high dose animals at end of exposure period. This persisted in high-dose animals during recovery.
Mean absolute liver weights significantly increased in high-dose females, and mean relative liver weight significantly increased in mid- and high-dose females and high-dose males after exposure period.

GROSS PATHOLOGY
All animals of high-dose group had confluent white foci in the lungs. This effect was still apparent after recovery and not seen in the control group animals.

HISTOPATHOLOGY: NON-NEOPLASTIC
Alveolar histiocytosis observed in all mid- and high-dose animals at end of exposure period. According to the athors, these changes are consistent with a response to foreign material (water insoluble liquid aerosol) and they were not considered to be a response to chronic injury. Chronic foreign body inflammation was observed in high-dose animals at end of recovery.

Effect levels

Dose descriptor:
NOEL
Effect level:
0.1 mg/L air (nominal)
Sex:
male/female
Basis for effect level:
other: Lung effects, decrease in plasma cholinesterase (females)

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Not relevant

Applicant's summary and conclusion

Conclusions:
Under the conditions of this study, a No Observed Effect Level (NOEL) of 0.1 mg/L was established for the inhalation of Fyroflex RDP in rats. This NOEL was based on the presence of lung effects, which persisted after recovery.
Executive summary:

This study was performed to determine the ability of Fyroflex RDP to induce toxicity in rats by inhalation. The study was conducted according to OECD guideline 412 and under GLP conditions. Additionally, the Monocyte NonSpecific Esterase (MNSE) activity was evaluated to study neurotoxicity.

 

No deaths or toxicologically significant clinical signs were noted. In males of the high-dose group body weight and body weight gain was decreased. This returned to normal after recovery. Plasma cholinesterase activity was decreased in high-dose males and mid- and high-dose females, which persisted for high-dose females after recovery. No inhibition of erythrocyte cholinesterase acitivy levels was seen. MNSE activity levels were increased in low- and mid-dose rat, however, in high-dose animals this activity was comparable to that of controls. A dose-related increase in absolute and relative lung weights was seen in the test group, and was significant in mid- and high-dose animals. This persisted in high-dose animals during recovery. Mean absolute liver weights were significantly increased in high-dose females, and mean relative liver weight significantly increased in mid- and high-dose females and high-dose males. All animals of the high-dose group had confluent white foci in the lungs. This effect was still apparent after recovery and alveolar histiocytosis was observed in all mid- and high-dose animals. Chronic foreign body inflammation was observed in the high-dose animals at end of recovery.

Under the conditions of this study, a No Observed Effect Level (NOEL) of 0.1 mg/L was established for the inhalation of RDP in rats. This NOEL was based on the presence of lung effects, which persisted after recovery.