Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Based on an oral subacute study with the substance registered, the NOAEL is considered to be 1000 mg/kg bw/day.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline Study
Qualifier:
according to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Japanese Guidelines for Screening, Toxicity Testing of Chemicals: Testing Methods for new Substances, enacted July 13, 1974, amended December 5, 1986
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: RCC Ltd, Laboratory Animal Services, CH-4414 Fullinsdorf / Switzerland
- Age at study initiation: 6 weeks
- Housing: in groups of five in Makrolon type-4 cages with wire mesh tops and standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, ad libitum
- Water (e.g. ad libitum): Community tap water from Itingen, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Humidity (%): 30-70%
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
The dose formulations were prepared weekly. The substance was weighed into a glass beaker on a tared Mettler balance and the vehicle added. The mixure was prepared using a megnetic stirrer and stored at room temperature.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The mean concentrations of the homogeneity samples taken during pretest preparations were found to be 106.7%, 94.3%, and 99.6% of the nominal concentrations of dose group 2 (5.0 mg/mL), dose group 3 (20.0 mg/mL), and dose group 4 (100 mg/mL), respectively.
The mean concentrations of the homogeneity samples were found to be 97.4%, 102.7%, and 104.7% of the nominal concentrations of dose group 2 (5.0 mg/mL), dose group 3 (20.0 mg/mL), and dose group 4 (100 mg/mL), respectively.
The individual concentrations varied in the range from -6% to +3% of the mean concentrations. Therefore, the test item was found to be homogeneously distributed in the vehicle.
Duration of treatment / exposure:
28 days
Frequency of treatment:
7 days/week
Remarks:
Doses / Concentrations:
0, 50, 200, 1000 mg/kg bw/day
Basis:
nominal in water
No. of animals per sex per dose:
Control group: 10 males and 10 females
Low-dose group: 5 males and 5 females
Mid-dose group: 5 males and 5 females
High-dose group: 10 males and 10 females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Derived from a 5-day pre-test
- Rationale for selecting satellite groups: % males and 5 females from the control and the high-dose group were further observed for a recovery period of 2 weeks.
- Post-exposure recovery period in satellite groups: 2 weeks
Positive control:
no
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily (days 1-3: twice daily)

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 6 weeks (recovery groups: after 6 weeks)
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- Parameters checked in table 1 were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after 6 weeks (recovery groups: after 6 weeks)
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- Parameters checked in table 1 were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: after 6 weeks (recovery groups: after 6 weeks)
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes a
- Parameters checked in table 1 were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: during week 4
- Dose groups that were examined:
- Battery of functions tested: grip strength / locomotor activity
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Statistics:
The following statistical methods were used to analyze the grip strength, locomotor activity, body weight, organ weights and ratios, as well as:
- The Dunnett-test (many to one t-test) based on a pooled variance estimate were applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
- The Steel-test (many-one rank test) were applied instead of the Dunnett-test when the data can not be assumed to follow a normal distribution.
- Fisher's exact-test were applied to the macroscopic findings.
The following statistical methods were used for statistical analysis of clinical laboratory data:
- Quantitative data were analyzed by a one-way analysis of variance (ANOVA) when the variances are considered homogeneous according to Bartlett. Alternatively, if the variances are considered to be heterogenous (p≤0.05), a non-parametric Kruskal-Wallis test was used. Treated groups were compared to the control groups using Dunnett's test if the ANOVA was significant at the 5% level and by Dunn's test in the case of a significant Kruskal-Wallis test (p≤0.05).

Clinical signs:
no effects observed
Description (incidence and severity):
All animals survived until scheduled necropsy. No test item-related findings of toxicological relevance were noted during cage-side and clinical observations at any dose level. Rats treated with 200 and 1000 mg/kg/day showed red discoloration of the feces
Mortality:
no mortality observed
Description (incidence):
All animals survived until scheduled necropsy. No test item-related findings of toxicological relevance were noted during cage-side and clinical observations at any dose level. Rats treated with 200 and 1000 mg/kg/day showed red discoloration of the feces
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Description (incidence and severity):
No test item-related differences in the hematology parameters were noted after four weeks treatment or after two weeks recovery.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
No test item-related differences in the clinical biochemistry parameters were noted after four weeks treatment or after two weeks recovery.
Urinalysis findings:
no effects observed
Description (incidence and severity):
No test item-related differences of toxicological relevance were noted in the urinalysis parameters after four weeks treatment or after 2 weeks recovery.
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
No test item-related findings of toxicological relevance were noted at any dose level during the functional observational battery performed at week 4.
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
No test item-related changes in mean absolute or relative organ weights were noted after four weeks of treatment or after 2 weeks of recovery.
Gross pathological findings:
no effects observed
Description (incidence and severity):
At the end of the treatment and following recovery period, no test item-related gross lesions were observed.
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
At the end of the treatment and following recovery period, no test item-related microscopic lesions were observed.
Histopathological findings: neoplastic:
no effects observed
Description (incidence and severity):
At the end of the treatment and following recovery period, no test item-related gross lesions were observed.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Dose descriptor:
NOEL
Effect level:
200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Critical effects observed:
not specified
Conclusions:
Based on the results of this study, 1000 mg/kg bw/day of RED LF 6382/18L-R was established as the no-observed-adverse-effect-level (NOAEL).
Executive summary:

In order to assess the toxicity after repeated oral application of the registration substance a valid subacute oral gavage study was performed. Oral administration of the test substance to Wistar rats at doses of 50, 200 and 1000 mg/kg/day (vehicle: water) resulted in no deaths, no clinical signs of toxicological relevance during daily or weekly observations or during the functional observation battery (week 4), no effects upon food consumption or body weight, no toxicologically relevant differences in hematology, clinical biochemistry or urine analysis, no changes in mean absolute or relative organ weights, and no macroscopcal or microscopcal changes. Based on the results of this study, 1000 mg/kg/day is considered to be the NOAEL of the registration substance.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Study reliability of 1.

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

In order to assess the toxicity after repeated oral application of the registration substance a valid 28 -day subacute oral gavage study was performed. Oral administration of the test substance to Wistar rats at doses of 50, 200 and 1000 mg/kg/day (vehicle: water) resulted in no deaths, no clinical signs of toxicological relevance during daily or weekly observations or during the functional observation battery (week 4), no effects upon food consumption or body weight, no toxicologically relevant differences in hematology, clinical biochemistry or urine analysis, no changes in mean absolute or relative organ weights, and no macroscopically or microscopically changes. Based on the results of this study, 1000 mg/kg/day is considered to be the NOAEL of the registration substance.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
The selected study was performed under GLP and in accordance with OECD TG 407. No other studies are available.

Justification for classification or non-classification

Resulting from the oral subacute toxicity study on the substance registered, the test substance is not to be classified as to its repeated dose toxicity properties.