Registration Dossier

Administrative data

Endpoint:
sub-chronic toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and OECD guideline study, available as unpublished report, no restrictions, fully adequate for assessment.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report Date:
1993

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)
Qualifier:
according to
Guideline:
EPA OTS 798.2450 (90-Day Inhalation Toxicity)
Qualifier:
according to
Guideline:
other: Japanese Ministry of International Trade and Industry Guidelines for Subchronic Toxicity Studies (No. 1: 1974, amended 1987)
Qualifier:
according to
Guideline:
EPA OPP 82-4 (90-Day Inhalation Toxicity)
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
FM-200 was received from Great Lakes Chemical Corporation, El Dorado, Arkanzas
Cylinder Lot numbers: 93-200-172C (WIL Log No.2292E), 93-200-174A (WIL Log No. 2292C), 93-200-174B (WIL Log No. 2292C), 93-200-174C (WIL Log No.2292E), 93-200-176A (WIL Log No.2292E), 93-200-176B (WIL Log No.2292E), 93-200-177A (WIL Log No.2292E), 93-200-179A (WIL Log No.2292E)
Dates of receipt: 07-07-93 (WIL Log No.2292E) and 06-30-93 (WIL Log No. 2292C)
Storage: in original cylinders at ambient conditions
Purity: 99.9%

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: CRL:CD BR rats, Charles River Laboratories, Inc., Portadge, Michigan
- Age at receipt: approx. 31 days
- Age at study initiation: approx. 6 weeks old
- Weight at study initiation: 154-211 g (males), 125-158 g (females)
- Housing: individually, in clean, wire-mesh cages, suspended above cage boards
- Diet: Purina Certified Rodent Chow, #5002, ad libitum, except of fasting period before blood collection and during exposure periods
- Water: automatic watering system, ad libitum, except during exposure periods
- Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS
- Temperature : 65-76 F
- Humidity (%): 26-70%
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
inhalation: gas
Type of inhalation exposure:
whole body
Vehicle:
other: unchanged (no vehicle)
Remarks on MMAD:
MMAD / GSD: Not applicable
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 0.5 m3 stainless steel and glass whole body exposure chamber
- Temperature, humidity, pressure in air chamber: 22 +/- 2 C, 40-70%,
- Air flow rate: 12-15 chamber flows per hour
- Treatment of exhaust air: treatment of the exhaust air consisted of drawing the air through an activated charcoal bed, a HEPA filter, and a water-spray fume scrubber.

TEST ATMOSPHERE
- Brief description of analytical method used: gas chromatography
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Exposure concentrations within each chamber were measured by gas chromatography at least at the beginning, at the intermediate time, and at the end of each daily exposure period. Mass air flow, temperature, humidity and oxygen content were monitored continuously and recorded at least every 30 min. The test material was analysed by gas chromatography with a Thermal Conductivity Detector (TCD).
Duration of treatment / exposure:
13 weeks (at least 65 exposures)
Frequency of treatment:
6 hours/day, 5 days/week
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
139370, 348420 and 731690 mg/m3 (20000, 50000 and 105000 ppm)
Basis:
other: target concentration
Remarks:
Doses / Concentrations:
20001, 50324 and 104865 ppm
Basis:
analytical conc.
Remarks:
Doses / Concentrations:
18231, 46286 and 78167 ppm
Basis:
nominal conc.
No. of animals per sex per dose:
12/sex/dose
Control animals:
yes
Details on study design:
Rationale for animal assignment: computer randomization program, based on body weight stratification in a block design

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: yes
- Time schedule: twice daily, in the morning and in the afternoon

DETAILED CLINICAL OBSERVATIONS: yes
- Time schedule: weekly, starting one week prior to test article administion until just before the necropsy. In addition, the clinical condition of the animals was monitored during the exposure periods.

BODY WEIGHT: Yes
- Time schedule for examinations: weekly, starting one week prior to test article administion, with final body weight recorded at the day of necropsy

FOOD CONSUMPTION: Yes
Food consumprion was measured weekly, starting one week prior to test article administion. Food consumption for each animal was calculated as g/animal/day for the corresponding body weights intervals

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: prior to study initiation and in week 12
- Dose groups that were examined: all

HAEMATOLOGY: Yes
- Time schedule for collection of blood: weeks 3 and 12
- Animals fasted: Yes
- Parameters examined: total leukocyte count, erythrocyte count, hemoglobin, hematocrit, mean corpuscular volume, mean corpusrular hemoglobin concentration, platelet count, platelet estimate, RBC morphology, differential leykocyte count (performed on the control and 100,000 ppm groups only, included unsegmented neutrophil, lymphocyte, monocyte, eosinophil, basophic, segmented neutrophil).

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: week 3 and 12
- Animals fasted: Yes, overnight
- Parameters examined: albumin, A/G ratio, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, bilirubin, blood urea nitrogen, calcium, chloride, cholesterol, creatinine.

SERUM CHEMISTRY: Yes
- Time schedule for collection of blood: week 3 and 12
- Parameters examined: glucose, urea nitrogen, creatinine, total protein, albumin, albumin/globulin ratio (A/G), calcium, phophorus, chloride, total bilirubin, gamma glutamyltransferase, serum aspartate aminitransferase, serum alanine aminitransferase, serum alkaline phosphatase, sodium, potassium, total cholesterol.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Included examination of the external surface, all orifices and the cranial, thoracic, abdominal and pelvic cavities, including viscera.
The following organ weights were determined: adrenals, brain, lidneys, liver, lungs (prioir to inflation with fixative), ovaries and testes. Paired organs were weighed together.

HISTOPATHOLOGY: Yes
Histopathological examination was performed on the following tissues: adrenals, aorta, bone with marrow, bone marrow smear, brain, eyes with optic nerve, gastrointestinal tract, heart, kidneys, liver, lungs, mesenteric lymph node, mammary gland, ovaries with oviducts, pancreas, sciatic peripheral nerve, pituitary, prostate, salivary glands, seminal vesicles, skeletal muscle, skin, spinal cord, spleen, testes with epididymides, thymus, thyroids, trachea, urinary bladder, uterus with vagina, all gross lesions.
Statistics:
Two-tailed tests for minimum significance levels of 1% and 5% comparing the treatment groups to the vehicle control group by sex. All means were presented with standard deviations and the numbers of sampling units used to calculate the means. Body weight, body weight change, food consumption, clinical lavoratory and absolute and relative organ weights were subjected to a one-way analysis of variance, followed by Dunnetts test.

Results and discussion

Results of examinations

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):
no effects observed
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
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:
All animals survived until the termination of the study. Soft stool and red material around the nose were observed in both treated and control groups. Non-dose related effects such as hair loss and scabbing on the forelimbs, and occasional salivation and dried yellow genital matting were observed in the treated group at a similar incidence to the control group.
There was no difference found in the mean body weight or body weight gains between the control and the treated groups. No treatment-related effects on food consumption were observed in any dose group. There were small differences in food consumption in higher dose females compared with the control group; however, this occurred sporadically and was not considered to be dose or time related.
There were no treatment-related changes in the haematology parameters observed at any dose level.
There were no treatment-related changes in serum chemistry parameters observed at any dose level. Slight differences between the treated and the control groups in mean aspartate aminotransferase, gamma glutamyl transferase, glucose, total bilirubin, calcium and potassium were observed in a non dose-related manner. Some of the differences were also inconsistent between sexes and therefore not considered to be of toxicological importance.
Ophthalmological examinations did not show pathological lesions indicative of toxic effects, in any dose groups.
The organ weight values of the treated groups were comparable with the control group. No treatment-related gross lesions were observed in any dose group. Haemorrhagic thymus gland and clear fluid uterus contents were observed in both treated and control groups; single animals demonstrated small kidneys and mottled lungs.
Histological findings revealed lymphocyte infiltration and splenic hemosiderosis in both treated and control groups. Germinal epithelium degeneration of the testes, a small benign glioma and a splenic cyst was present in one animal in both treated and control groups. Tubular mineralisation of the renal cortico-medullary junctions and alveolar oedema was observed in a non dose-related manner.

Effect levels

Dose descriptor:
NOAEC
Effect level:
731 690 mg/m³ air
Sex:
male/female
Basis for effect level:
other: No effects were observed at the highest concentration tested.

Target system / organ toxicity

Critical effects observed:
not specified

Applicant's summary and conclusion