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Description of key information

A  valid NOAEL of 200 mg/kg/day was established in a guideline conform 28 days oral toxiyity study with rats. Marked to moderate hairloss  and a 10 %  body weight loss were the most prominet effect observed in this study.

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was according to Japanese MHW-MITI Regulations and in compliance with GLP. The Guideline used is similar to OECD guideline 407.
Reference:
Composition 0
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Test material information:
Composition 1
Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River UK (Margate, Kent, England)
- Age at study initiation: ca. 4 weeks
- Weight at study initiation: 75 - 84 g
- Housing: groups of 5 in polypropylene cages
- Diet (e.g. ad libitum): ad libitum(except when urine was collected and overnight befor blood sampling) Diet type: LAD-1, Biosur, Manea, Cambridgeshire, England
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21°C (18-25°C)
- Humidity (%): 55 % (40-70 % )
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 h
Route of administration:
oral: gavage
Vehicle:
maize oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
Formulations of the test substance were prepared for adminitration as a series of graded concentrations in maize oil to provide the required dosages at a constant volume-dosage of 5 ml/kg. All formulations were prepared freshly each day

VEHICLE
- Justification for use and choice of vehicle (if other than water): solubility

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Duplicate samples form each formulation prepared on day 1 and 26 were analyzed.
Duration of treatment / exposure:
28 days and 14 days recovery period
Frequency of treatment:
once daily by gavage
Remarks:
Doses / Concentrations:
0, 40, 200, 1000 mg/kg
Basis:
other: oral gavage
No. of animals per sex per dose:
0 and 1000 mg/kg: 10 /sex
40, 200 mg/kg: 5/sex
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: pilot study (3 males and 3 female were gavaged 200, 500 and 100 mg/kg in maze oil for 7 days)
- Post-exposure recovery: 14 days
Positive control:
not applicable
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

CLINICAL OBSERVATIONS: Yes
- Time schedule: daily
- Parameters examined: Observations were recorded

BODY WEIGHT: Yes
- Time schedule for examinations: Weighing was on day 0, 3, 7, 10, 14, 17, 21, 24, 27

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
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 4 weeks treatment and after 2 weeks recovery
- Anaesthetic used for blood collection: No data
- Animals fasted: Yes
- How many animals: 55
- Parameters examined: PCV, Hb, RBC, WBC, platelet, neutrophils, lymphocytes, esinophils, basophils, monocytes, reticulocytes, protrombine time, blood cell microscopy

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after 4 weeks treatment and after 2 weeks recovery
- Animals fasted: Yes
- How many animals: 55
- Parameters examined: AP, ALT, AST, gamma GT, GGT, urea, creatinine, glucose, bilirubin, triglycerides, cholesterol, proteins ,Na, K, Cl, Ca, P,


URINALYSIS: Yes
- Time schedule for collection of urine: after 3 weeks treatment and after 1 weeks recovery
- Metabolism cages used for collection of urine: Yes / No / No data
- Animals fasted: at urine overnight collection
- Parameters examined: pH, specific gravity, protein content, glucose, ketones, bilirubin, urobilinogen, nitrite, blood, sediment (leucocytes, Erytrocytes, eppithelial cells, etc.)


NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (results see below)
HISTOPATHOLOGY: Yes (results see below)
Other examinations:
no
Statistics:
Different methods were used depending on the data to be compared: Student's t-test, Bartlett's test, Behrens-Fisher test, Dunnett's Test, Fisher Exact test.
Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
effects observed, treatment-related
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY
Two female rats treated at 200 mg/kg/day died on Day 5. The deaths were considered to be incidental to treatment with Rapi-cure CHVE. There was no other death (failure of gavage dosing).
Salivation at dosing was observed from Day 4 onwards in most rats treated at 1000 mg/kg/day, less frequently in animals treated at 200 mg/kg/day and on two occasions in those treated at 40 mg/kg/day. Hair loss was observed from Day 4 onwards in the majority of animals treated at 1000 mg/kg/day (9/10 females; 4/10 males). The sign persisted during the recovery period.

BODY WEIGHT AND WEIGHT GAIN
The body weight gain of rats receiving 1000 mg/kg/day was about inferior to that of controls during the treatment period (p<0.001 males; <0.05 females). The body weight of rats receiving 1000 mg/kg/day was about 10% less to that of controls.

FOOD EFFICIENCY
Food consumption and food utilisation efficiency were considered to have been unaffected by treatment with Rapi-cure CHVE.

HAEMATOLOGY
The packed cell volumes, haemoglobin concentrations and erythrocyte numbers of animals treated at 1000 mg/kg/day were slightly but significantly lower than those of the controls. In addition, neutrophil numbers of rats treated at 1000 mg/kg/day were slightly higher than those of the controls (p<0.05). Most of the differences were still observed after the two weeks recovery period.

CLINICAL CHEMISTRY
The plasma alkaline phosphatase activities were higher and sodium and chloride concentrations lower in rats treated at 1000 mg/kg/day than in the control animals. These differences were not apparent two weeks after cessation of treatment. Urea (male) ALT and gamma GT (females) were also higher slightly but significantly higher at 1000 mg/kg/day. These differences, however, numerically small and were not considered toxicologically relevant.

URINALYSIS
Urinary volumes of rats treated at 1000 mg/kg/day and of male rats treated at 200 mg/kg/day were higher than those of the controls. The specific gravity of the urine of males treated at 200 or 1000 mg/kg/day was slightly lower than that of the controls. Urinary protein concentrations of male rats treated at 1000 mg/kg/day were marginally lower than those of the male controls and blood was detected in two males and one female treated at this dosage. Blood was also detected in these animals in the recovery period.


ORGAN WEIGHTS
After four weeks of treatment, kidney weights of males and females treated at 1000 mg/kg/day and of males treated at 200 mg/kg/day were slightly higher than those of the control animals. The liver weights of animals treated at 1000 mg/kg/day were higher and those of animals treated at 200 mg/kg/day slightly higher than the liver weights of the control animals. Two weeks after cessation of treatment, liver and kidney weights relative to bodyweight of male rats treated at 1000 mg/kg/day were still slightly higher than those of the male controls.

GROSS PATHOLOGY
There was no organ macroscopic change which was attributed to treatment with Rapi-cure CHVE.

HISTOPATHOLOGY: NON-NEOPLASTIC
There were no histopathological changes in test animals which were considered to be substance related and of toxicological significance.
Dose descriptor:
NOAEL
Effect level:
200 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: overall effects clinical signs; body weight; organ weights; other: hair loss
Critical effects observed:
not specified
Executive summary:

0, 40, 200 and 1000 mg/kg/day Rapi-cure CHVE was administered by gavage to male and female CD rats in a guideline conform 28 days study. The study was according to Japanese MHW-MITI Regulations and in compliance with GLP. The Guideline used is similar to OECD guideline 407. The study included a 14 days recovery period. Marked to moderate hair loss was observed in 9/10 female and 4 /10 males at 1000 mg/kg. An increase of alkaline phosphatase, ALT and gamma GT and increased absolute kidney and liver weights were observed. Effects were numerically small but significant. Beyond this a reduction of the bodyweight about 10 % in comparison to the control was observed. Effects were still present at the end of the recovery period.

A  oral NOAEL of 200 mg/kg was established in this study.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
200 mg/kg bw/day
Study duration:
subacute
Species:
rat

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

Mode of Action Analysis / Human Relevance Framework

Additional information

0, 40, 200 and 1000 mg/kg/day Rapi-cure CHVE was administered by gavage to male and female CD rats in a guideline conform 28 days study. The study was according to Japanese MHW-MITI Regulations and in compliance with GLP. The Guideline used is similar to OECD guideline 407. The study included a 14 days recovery period. Salivation after dosing was the most prominent clinical sign. Marked to moderate hair loss was observed in 9/10 female and 4 /10 males at 1000 mg/kg. An increase of alkaline phosphatase, ALT and gamma GT and increased absolute kidney and liver weights were observed. Effects were significant but numerically small and were not considered toxicologically relevant.. Beyond this a reduction of the bodyweight about 10 % in comparison to the control was observed. Effects were still present at the end of the recovery period.


Repeated dose toxicity: via oral route - systemic effects (target organ) other: all gross lesions and masses

Justification for classification or non-classification

Classification for repeated dose toxicity is not warranted according to the criteria of EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.