Registration Dossier

Administrative data

Description of key information

A study similar to OECD TG 408 was performed to assess the cumulative toxicity of the test item. Groups of 10 male and 9 or 10 female F344/N rats were given 0, 3000, 6000, 12000, 25000, or 50000 ppm test item (equivalent to average daily doses of approximately 200, 400, 800, 1600, or 3500 mg test item/kg body weight to males and 200, 400, 800, 1700, or 3600 mg/kg to females) in feed for 13 weeks. Based on the results of this study, the NOAEL was 800 mg/kg bw/day for males and females (reference 7.5.1-1).

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
28 April 1987 - 29 July 1987
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
GLP compliance:
yes
Remarks:
Food and Drug Administration Good Laboratory Practice Regulations (21 CFR, Part 58)
Limit test:
no
Species:
rat
Strain:
Fischer 344
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Taconic Farms (Germantown, NY)
- Age at study initiation: 6 weeks
- Weight at study initiation: 132 g (males) and 109 g (females)
- Housing: Polycarbonate suspended cages (Lab Products, Inc., Rochelle Park, NJ), changed twice per week
- Diet: NIH-07 open formula meal diet (Zeigler Brothers, Inc., Gardners, PA), available ad libitum, changed weekly
- Water: Tap water (Bethesda municipal supply) via automatic watering system (Edstrom Industries, Inc., Waterford, WI), available ad libitum, changed every 2 weeks
- Acclimation period: 12 to 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23
- Humidity (%): 50-67
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
DIET PREPARATION
- Rate of preparation of diet: The dose formulations for all studies were prepared weekly by mixing test item with feed
- Mixing appropriate amounts with feed: A test item premix was prepared by hand and was then blended with feed in a Patterson-Kelly twin-shell blender for 15 minutes using an intensifier bar for the initial 5 minutes.
- Storage temperature of food: Formulations were stored in double plastic bags at temperatures at or below -20° C for up to 3 weeks.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Stability was monitored during the 13-week using high-performance liquid chromatography. No degradation of the bulk chemical was detected. Homogeneity was confirmed and the stability of the dose formulation was confirmed for at least 3 weeks when stored protected from light at room temperature. All of the dose formulations analyzed during the were within 10% of the target concentrations.
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
continuously via feed
Dose / conc.:
0 ppm
Dose / conc.:
3 000 ppm
Remarks:
equivalent to average daily doses of approximately 200 mg/kg bw/day
Dose / conc.:
6 000 ppm
Remarks:
equivalent to average daily doses of approximately 400 mg/kg bw/day
Dose / conc.:
12 000 ppm
Remarks:
equivalent to average daily doses of approximately 800 mg/kg bw/day
Dose / conc.:
25 000 ppm
Remarks:
males: equivalent to average daily doses of approximately 1600 mg/kg bw/day
females: equivalent to average daily doses of approximately 1700 mg/kg bw/day
Dose / conc.:
50 000 ppm
Remarks:
males: equivalent to average daily doses of approximately 3500 mg/kg bw/day
females: equivalent to average daily doses of approximately 3600 mg/kg bw/day
No. of animals per sex per dose:
10 males and 9 or 10 females
Control animals:
yes, plain diet
Positive control:
no
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

BODY WEIGHT: Yes
- Time schedule for examinations: animals were weighed initially, weekly, and at the end of the studies

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
Feed consumption was recorded weekly by cage.

WATER CONSUMPTION AND COMPOUND INTAKE: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Blood was collected from the retroorbital sinus of rats in the clinical pathology groups on days 5 and 21 and of core study rats at the end of the study for hematology and clinical chemistry.
- Anaesthetic used for blood collection: Not specified
- Animals fasted: Not specified
- How many animals: all
- Parameters checked: hematocrit, hemoglobin, erythrocyte and reticulocyte counts, mean cell volume, mean cell hemoglobin, mean cell hemoglobin concentration, platelet count, and total leukocyte counts and differentials. Clinical chemistry: urea nitrogen, creatinine, total protein, albumin, alanine aminotransferase, alkaline phosphatase, creatine kinase, sorbitol dehydrogenase, and bile acids.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Blood was collected from the retroorbital sinus of rats in the clinical pathology groups on days 5 and 21 and of core study rats at the end of the study for hematology and clinical chemistry.
- Animals fasted: Not specified
- How many animals: all
- Parameters checked: urea nitrogen, creatinine, total protein, albumin, alanine aminotransferase, alkaline phosphatase, creatine kinase, sorbitol dehydrogenase, and bile acids.

URINALYSIS: Yes

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Necropsy was performed on all core study rats . Organs weighed were brain, heart, right kidney, liver, lung, right testis, and thymus.

HISTOPATHOLOGY: Yes

Complete histopathology was performed on 0 and 50,000 ppm rats . In addition to gross lesions and tissue masses, the tissues examined included: adrenal gland, brain, clitoral gland (rats), esophagus, femur (including marrow), heart, gallbladder (mice), large intestine (cecum, colon, and rectum), small intestine (duodenum, jejunum, and ileum), kidney, liver, lungs (and mainstem bronchi), lymph nodes (mandibular and mesenteric), mammary gland, nose, ovaries, pancreas, parathyroid gland, pituitary gland, preputial gland (rats), prostate gland, salivary gland, skin, spleen, stomach (including forestomach and glandular stomach), testes (with epididymis and seminal vesicle), thymus, thyroid gland, trachea, urinary bladder, and uterus. In rats, the liver (females), lung (males), and ovaries were examined to a no effect level. In mice, the bone marrow and spleen (males) were examined to a no effect level.
Other examinations:
At the end of the study, sperm samples were collected from male rats (core study) in the 0, 12,000, 25,000, and 50,000 ppm groups for sperm morphology evaluations. The parameters evaluated included sperm density, morphology, and motility. The right cauda, right epididymis, and right testis were weighed. Vaginal fluid samples were collected for up to 7 consecutive days prior to the end of the studies from female rats (core study) in the 0, 12,000, 25,000, and 50,000 ppm groups. The parameters evaluated were relative frequency of estrous stages and estrous cycle length.
Statistics:
Williams' or Dunnett's test
Clinical signs:
no effects observed
Mortality:
no mortality observed
Description (incidence):
All rats survived to the end of the study.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
The final mean body weight of the 50,000 ppm females and the body weight gains of the 25,000 and 50,000 ppm females were significantly lower than those of the controls. The final mean body weights and body weight gains of all other exposed groups were similar to those of the controls.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
Feed consumption by exposed groups was similar to that by the controls. Dietary levels of 3,000, 6,000, 12,000, 25,000, or 50,000 ppm test item resulted in average daily doses of approximately 200, 400, 800, 1,600, or 3,500 mg test item/kg body weight to males and 200, 400, 800, 1,700, or 3,600 mg/kg to females.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
The few differences in the hematology parameters were sporadic and were not considered to be chemical related.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
The few differences in the clinical chemistry parameters were sporadic and were not considered to be chemical related.
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
The absolute and relative liver weights of 25,000 and 50,000 ppm males and the relative liver weights of 12,000 ppm males and of 25,000 and 50,000 ppm females were significantly greater than those of the controls.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No chemical-related gross lesions were observed.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No chemical-related microscopic lesions were observed.
Histopathological findings: neoplastic:
not examined
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
The percentage of motile sperm in the 12,000 ppm males was significantly greater than that in the controls, but no other significant differences in sperm morphology or vaginal cytology were observed between exposed and control groups.
Key result
Dose descriptor:
NOEL
Effect level:
6 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
organ weights and organ / body weight ratios
Remarks on result:
other: Corresponding to 400 mg/kg bw/d.
Key result
Dose descriptor:
NOAEL
Effect level:
12 000 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
organ weights and organ / body weight ratios
Remarks on result:
other: Corresponding to 800 mg/kg bw/d.
Critical effects observed:
not specified
Conclusions:
Based on the results of this study, the NOAEL was 12000 ppm (800 mg/kg bw/day) for males and females.
Executive summary:

A study similar to OECD TG 408 was performed to assess the cumulative toxicity of the test item when administered daily to rats in feed for a period of 13 weeks.

Groups of 10 male and 9 or 10 female F344/N rats were given 0, 3000, 6000, 12000, 25000, or 50000 ppm test item (equivalent to average daily doses of approximately 200, 400, 800, 1600, or 3500 mg test item/kg body weight to males and 200, 400, 800, 1700, or 3600 mg/kg to females) in feed for 13 weeks. Additional groups of 10 male and 10 female rats designated for clinical pathology evaluations were also given 0, 3000, 6000, 12000, 25000, or 50000 ppm test item in feed until day 21.

All core study rats survived to the end of the study. The final mean body weight of the 50000 ppm females and the mean body weight gains of the 25000 and 50000 ppm females were significantly lower than those of the controls. The final mean body weights and mean body weight gains of all other exposed groups were similar to those of the controls. There was no cathartic action or any other clinical finding attributed to exposure to the test item. The few differences in the haematology and clinical chemistry parameters were sporadic and were not considered being chemical related. The percentage of motile sperm in the 12000 ppm males was significantly greater than that in the controls, but no other significant differences in sperm morphology or vaginal cytology between exposed and control groups were observed. Absolute and relative liver weights of 25000 and 50000 ppm males were significantly greater than those of the controls. No chemical-related gross or microscopic lesions were observed.

Based on the results of this study, the NOAEL was 12000 ppm (800 mg/kg bw/day) for males and females.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
800 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
similar to OECD TG 408

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

A study similar to OECD TG 408 was performed to assess the cumulative toxicity of the test item when administered daily to rats in feed for a period of 13 weeks.

Groups of 10 male and 9 or 10 female F344/N rats were given 0, 3000, 6000, 12000, 25000, or 50000 ppm test item (equivalent to average daily doses of approximately 200, 400, 800, 1600, or 3500 mg test item/kg body weight to males and 200, 400, 800, 1700, or 3600 mg/kg to females) in feed for 13 weeks. Additional groups of 10 male and 10 female rats designated for clinical pathology evaluations were also given 0, 3000, 6000, 12000, 25000, or 50000 ppm test item in feed until day 21.

All core study rats survived to the end of the study. The final mean body weight of the 50,000 ppm females and the mean body weight gains of the 25,000 and 50,000 ppm females were significantly lower than those of the controls. The final mean body weights and mean body weight gains of all other exposed groups were similar to those of the controls. There was no cathartic action or any other clinical finding attributed to exposure to the test item. The few differences in the haematology and clinical chemistry parameters were sporadic and were not considered being chemical related. The percentage of motile sperm in the 12,000 ppm males was significantly greater than that in the controls, but no other significant differences in sperm morphology or vaginal cytology between exposed and control groups were observed. Absolute and relative liver weights of 25,000 and 50,000 ppm males were significantly greater than those of the controls. No chemical-related gross or microscopic lesions were observed.

Based on the results of this study, the NOAEL was 12000 ppm for males and females.

In a 14-day oral toxicity study the test item was administered daily to rats via feed. Groups of five male and five female F344/N rats were given 0, 6250, 12500, 25000, 50000 or 100000 ppm test item in feed for 14 days. Dietary levels of 6250, 12500, 25000, 50000, or 100000 ppm test item resulted in average daily doses of approximately 500, 1,000, 2,000, 4,500, or 10,500 mg test item/kg body weight to males and 500, 1000, 2000, 4000, or 11000 mg/kg to females. All rats survived to the end of the study. The final mean body weights of all exposed groups of rats were similar to those of the controls. No chemical-related gross or microscopic lesions were observed.

Based on the results of this study, no test item related adverse effects have been observed up to 11000 mg/kg bw/day (100000 ppm).

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. The substance is not considered to be classified for repeated dose toxicity according to Regulation (EC) No 1272/2008 (CLP), as amended for the fourteenth time in Regulation (EU) 2020/217.