Registration Dossier

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Remarks:
7-day range-finding tolerability study
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
24 October 2008 to 29 January 2009
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:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Guideline:
other: In-house 7-day repeated oral dose protocol
Principles of method if other than guideline:
In-house protocol. The notified chemical in the vehicle, corn oil, was administered orally by gavage once daily for 7 consecutive days to 3 groups of test animals at the dose levels of 100, 300 and 1,000 mg/kg bw/day. A concurrent control group received vehicle only on a comparable regimen. The dose volume was 5 mL/kg bw for all groups. Following 7 days of dose administration, all test animals were euthanized for gross necropsies.

During the study, all test animals were observed twice daily for mortality and moribundity. Clinical examinations were performed daily at the time of dosing and approximately 1 and 4 hours post-dosing and detailed physical examinations were performed weekly. Individual body weight and food consumption were recorded on study Days 0 and 7.
GLP compliance:
no
Remarks:
The study was not commissioned with compliance with REACH as a goal, rather the study was commissioned during early product development.
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Lot no. Q151-170
- purity: 99.0%

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Details on species / strain selection:
This species and strain of animal is recognized as appropriate for subchronic toxicity studies. The Sprague Dawley rat was selected because it is a widely used strain for which significant historical control data are available.
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Inc., Raleigh, NC
- Females nulliparous and non-pregnant: yes
- Age at study initiation: approximately 8 weeks old at the initiation of dose administration
- Weight at study initiation: 281 g to 322 g for males and from 182 g to 233 g for females at the initiation of dosing
- Housing: housed individually
- Diet: ad libitum throughout the study
- Water: ad libitum throughout the study
- Acclimation period: 14-day acclimation/pretest period

DETAILS OF FOOD AND WATER QUALITY:
Food: PMI Nutrition International, LLC, Certified Rodent LabDiet® 5002 meal
Water: Reverse osmosis-treated (on-site) drinking water

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.3°C to 21.5°C
- Humidity (%): 40.4% to 49.9%
- Air changes (per hr): a minimum of 10 fresh air changes per hour
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 29 October 2008 To: 5 November 2008

Administration / exposure

Route of administration:
oral: gavage
Details on route of administration:
The dose volume for all groups was 5 mL/kg.
Vehicle:
corn oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: The test article formulations were volume/volume (test article/vehicle) mixtures. The test article formulations were prepared daily as single formulations for each dosage level, divided into aliquots for daily dispensation and stored at room temperature. The test article formulations were stirred continuously throughout the preparation, sampling and dose administration procedures. Due to the nature of the vehicle, the pH was not measured.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The accuracy and stability of the test substance at 3 concentrations was confirmed using High Pressure Liquid Chromatography (HPLC). The results indicated that the three formulations used for dose levels 100, 300, and 1000 mg/kg bw were within +/- 10% of the target concentrations. Also the traces revealed a clear peak for the test substance and no incipient peaks.
Duration of treatment / exposure:
once daily for 7 consecutive days
Frequency of treatment:
once daily for 7 consecutive days
Doses / concentrationsopen allclose all
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
300 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
5/sex/dose
Control animals:
yes, concurrent vehicle
Details on study design:
The notified chemical in the vehicle, corn oil, was administered orally by gavage once daily for 7 consecutive days to 3 groups of test animals at the dose levels of 100, 300 and 1,000 mg/kg bw/day. A concurrent control group received vehicle only on a comparable regimen. The dose volume was 5 mL/kg bw for all groups. Following 7 days of dose administration, all test animals were euthanized for gross necropsies.
Positive control:
None

Examinations

Observations and examinations performed and frequency:
During the study, all test animals were observed twice daily for mortality and moribundity. Clinical examinations were performed daily at the time of dosing and approximately 1 and 4 hours post-dosing and detailed physical examinations were performed weekly. Individual body weight and food consumption were recorded on study Days 0 and 7.
Sacrifice and pathology:
A gross necropsy was conducted for all animals. The necropsies included examination of the external surface, all orifices, and the cranial, thoracic, abdominal and pelvic cavities, including viscera.
Other examinations:
None
Statistics:
All statistical tests were performed using appropriate computing devices or programs.

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Test substance related clinical observations noted in the 1,000 mg/kg bw/day group as early as study Day 0 and throughout 7-day dosing period included impaired muscle coordination and/or impaired equilibrium in both sexes of the test animals with higher frequency in females. Hypoactivity, decreased respiration rate and prostration were noted in females on study Day 0. These effects did not persist to the 4-hour post-dosing observation on study Days 0 to 6 for males and study Days 4 to 6 for females. These effects were considered by the study authors as adverse since they persisted throughout the 7-day dosing period at approximately 1 hour post-dosing. No significant clinical observations were recorded in the 100 and 300 mg/kg bw/day groups.
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
A test substance related effect on body weight was noted in test substance treated animals, showing a trend towards slightly lower body weight gains. However, this effect was not considered by the study authors as adverse.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
There were no test substance-related effects on food consumption.
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:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
No significant macroscopic findings noted.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not examined

Effect levels

Key result
Dose descriptor:
NOAEL
Remarks:
7 days
Effect level:
300 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
gross pathology

Target system / organ toxicity

Critical effects observed:
yes
Lowest effective dose / conc.:
1 000 mg/kg bw/day (nominal)
Organ:
not specified
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
not specified

Applicant's summary and conclusion

Conclusions:
Based on the results of this study, adverse toxicity of the test substance administered orally (gavage) to rats for 7 consecutive days was observed at 1000 mg/kg/day as evidenced by clinical observations noted following dosing throughout the 7-day dosing period. The test article was well tolerated in male and female rats at 100 and 300 mg/kg/day.
Executive summary:

The test substance in the vehicle, corn oil, was administered orally by gavage once daily for 7 consecutive days to 3 groups (Groups 2-3) of rats. Dosage levels were 100, 300 and 1000 mg/kg/day. A concurrent control group (Group 1) received the vehicle on a comparable regimen. The dose volume was 5 mL/kg for all groups. Each group (Groups 1-4) consisted of 5 animals/sex. Following 7 days of dose administration, all rats were euthanized.

 

All animals were observed twice daily for mortality and moribundity. Clinical examinations were performed daily at the time of dosing and approximately 1 and 4 hours post-dosing, and detailed physical examinations were performed weekly. Individual body weight and food consumption were recorded on study days 0 and 7. All carcasses were discarded following examination and collection of gross lesions.

 

All animals survived to the scheduled necropsy. There were no test article-related effects on food consumption and there were no significant macroscopic findings. There were no significant clinical observations in the 100 and 300 mg/kg/day groups.

 

Test article-related clinical observations noted in the 1000 mg/kg/day group as early as study day 0 and throughout the 7-day dosing period included impaired muscle coordination and/or impaired equilibrium in males and females; hypoactivity, decreased respiration rate, and prostration were also noted in females on study day 0. These effects did not persist to the 4-hour post-dose observation on study days 0-6 for males and study days 4-6 for females. These clinical observations were considered adverse, especially since they persisted throughout the 7-day dosing period at approximately 1-hour post-dosing.

 

A test article-related effect on body weight included a trend towards slightly lower body weight gains noted in all test article-treated groups compared to the control group. However, the changes in body weights were not of a magnitude to be considered adverse.

 

Based on the results of this study, adverse toxicity of the test substance administered orally (gavage) to rats for 7 consecutive days was observed at 1000 mg/kg/day as evidenced by clinical observations noted following dosing throughout the 7-day dosing period. The test article was well tolerated in male and female rats at 100 and 300 mg/kg/day.