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Description of key information

In a subacute study in rats main test substance-related changes were degeneration/necrosis in the myocardial cell and in myocardial cell around the vein in both sexes of the 40 mg/kg bw/d group. Therefore the NOAEL is 10 mg/kg bw/d under the conditions of the 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
Study period:
13 April - 20 August 2012
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
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories Japan, Inc. (Hino Breeding Center)
- Age at study initiation: 6 weeks
- Weight at study initiation (day 1): 202.4 - 228.0 g (males), 131.8 - 167.2 g (females)
- Fasting period before study: 18-23 h before scheduled necropsy
- Housing: single housing in stainless-steel cages
- Diet: Autoclave-sterilized pellet diet (CRF-1, Oriental Yeast Co, Ltd.)
- Water: Well water admixed with NaClO (free residual chlorine level: about 2 ppm), ad libitum (except during motor activity measurements)
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.5 - 24.8
- Humidity (%): 39.5 - 65.0
- Air changes (per hr): 10-20
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 5 April 2012 To: 11 May 2012 (dosing group), 25 May 2012 (recovery group)
Route of administration:
oral: gavage
Vehicle:
other: olive oil containing cremophor (10%)
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
Formulations of 2 and 8 mg/mL were prepared every 7 days. Test substance was weighed and transferred to a beaker. About 50% of the total volume of the vehicle was added to the beaker and the dosing formulation was stirred with a magnetic stirrer. After stirring, the dosing formulation was transferred to the mesuring cylinder. The beaker and stirring bar were washed with the vehicle, and the washing was transferred tot he measuring cylinder. The final volume was adjusted by adding a proper quantity of the vehicle to the required concentrations of 2 and 8 mg/mL.

VEHICLE
- Concentration in vehicle: 2 and 8 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg bw
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Homogeneity and stability of the test substance preparations were confirmed with an analytical GC method according to an internal protocol. The relative standard deviation (RSD) of analytical values (concentration) was within the criteria (actual RSD: 0.2% to 0.3%, criteria: not more than 10%), and measured concentration (mean) was within the criteria (actual concentration 98.7% to 100.5% of nominal concentrations, criteria: nominal concentration ± 10%)
Duration of treatment / exposure:
28 days
Frequency of treatment:
Once daily
Remarks:
Doses / Concentrations:
2, 10, 40 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
Main study: 10 (control, mid and high dose), 5 (low dose)
Recovery period: 5 (control, mid and high dose), 0 (low dose) animals from the main study each
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: In two 7-day screening studies animals were dosed with 50, 250, and 100 mg/kg bw/day (5 animals per sex and dose) or 1, 5, and 20 mg/kg bw/day (3 male animals per dose). Death and moribund animals (both sexes) were found in the 250 and 1000 mg/kg bw dose group. High AST and liver and kidney weights were observed in both sexes of the 50 mg/kg bw/day group. 1 male of this group showed discoloration of the heart, dark reddish macule of the lung, low values of body weight, food consumption and reticulocytes, and high values of platelets and monocytes. Effects on body weight and food consumption was also observed in 1 female of the 50 mg/kg bw dose group. Discoloration of the heart was also observed in dead animals of the 250 and 1000 mg/kg bw/day groups. No treatment-related changes were observed in the 1, 5, and 20 mg/kg bw/day groups. Thus 40 mg/kg bw/day was set as the high dose level in the main study expecting that there would be no death or some toxicity change.

- Post-exposure recovery period in satellite groups: 14 days
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily (dosing period), once daily (recovery period)
- Cage side observations checked: mortality and general signs of toxicity

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once before start of dosing and once a week during the dosing and recovery periods.

BODY WEIGHT: Yes
- Time schedule for examinations: Days 1, 8, 15, 22 and 28 (during dosing period), Days 29, 36, and 42 during recovery period

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: 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: No

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No


HAEMATOLOGY: Yes
- Time schedule for collection of blood: Days 29 and 43 (dosing and recovery period, respectively)
- Anaesthetic used for blood collection: Yes (sodium pentobarbital, 30 mg/kg bw)
- Animals fasted: Yes (18.5-23 h)
- How many animals: all animals
- Parameters examined: leukocytes, erythrocytes, hemoglobin, hematocrit, MCV, MCH, MCHC, reticulocyte ratio and count, platelets, Differential leukocyte ratio and count, prothrombin time (PT), Activated partial thromboplastin time (APTT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Days 29 and 43 (dosing and recovery period, respectively)
- Animals fasted: Yes (18.5-23 h)
- How many animals: all animals
- Parameters examined: total protein, albumin and albumin/globulin ratio, AST, ALT, gamma-Glutamyltranspeptidase, alkaline phosphatase, total bilirubin, total bile acids, total cholesterol, triglycerides, glucose, blood urea nitrogen, creatinine, calcium, inorganic phosphorus, sodium, potassium, chloride

URINALYSIS: Yes
- Time schedule for collection of urine: Day 27
- Metabolism cages used for collection of urine: No
- Animals fasted: Yes (only access to water)
- Parameters examined: pH, Protein, Glucose, ketone body, occult blood

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule: all animals, once in week 4 of the dosing period
- Dose groups that were examined: all animals
- Battery of functions tested: sensory activity / grip strength/ motor activity
Sacrifice and pathology:
SACRIFICE
- All animals were euthanized after blood sampling by exsanguination and were subjected to necropsy

GROSS NECROPSY
Necropsy of all males and females of the control and 40 mg/kg bw/day group was performed.
The following organs/tissues were collected and examined:
Brain (cerebrum, cerebellum and pons), heart, ovaries (females), liver, uterus (females), kidneys, adrenal glands, prostate, seminal vesicles with coagulating glands as a whole (males), thymus, urinary bladder, thyroid (incl. parathyroid), spleen, mammary glands, all gross lesions, eye, lymphnodes (submaxillary), submaxillary gland, parathyroid, stomach, duodenum, jejunum, ileum (including Peyer´s patches), cecum, colon, rectum. mesenteric lymph node, pancreas, testis (males), epididymides, vagina, femur, sternum, bone marrow (femur and sternum), M. biceps femoris, peripheral nerve (e.g. sciatic nerve), pituitary gland, spinal cord, trachea, lungs (including bronchus)


HISTOPATHOLOGY / ORGAN WEIGHTS
All full histopathology was carried out on the organs and tissues of all males and females of the control and 40 mg/kg bw/day group. Examination of heart, lung, and liver was performed in all animals of all dose groups (end of dosing period) and in all animals of the control and high dose group (end of recovery period)

The following tissues were prepared for microscopic examination:
With weight determination:
brain (cerebrum, cerebellum and pons), heart, ovaries (females), liver, uterus (females), kidneys, adrenal glands, prostate, seminal vesicles with coagulating glands as a whole (males), thymus, urinary bladder, thyroid (incl. parathyroid), spleen, mammary glands, all gross lesions

Without weight determination:
Eye, lymphnodes (submaxillary), submaxillary gland, parathyroid, stomach, duodenum, jejunum, ileum (including Peyer´s patches), cecum, colon, rectum. mesenteric lymph node, pancreas, testis (males), epididymides, vagina, femur, sternum, bone marrow (femur and sternum), M. biceps femoris, peripheral nerve (e.g. sciatic nerve), pituitary gland, spinal cord, trachea, lungs (including bronchus)
Statistics:
Statistical analysis was performed with a computer system (MiTOX-PPL, Mitsui Zosen Systems Research Inc.). For the numeral data, Bartlett´s test was performed to compare variances among groups (significance level: 5%). When variance of data was homogenous, Dunnett´s multiple comparison test was performed to compare with the control group. When variance of data was heterogenous, Steel´s multiple comparison test was performed to compare with the control group. For results of urinalysis with reagent strip, Steel´s multiple comparison test was performed after the grades were converted into numeric values. Mann-Whitney´s U test was used for the histopathological findings after the grades were converted into numeric values. In all cases, levels of p<0.01 (1%) and p<0.05 (5%) were considered to be significant and two-tailed test was used.
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 examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
40 mg/kg bw: high values of monocytes in both sexes at the end of the dosing period (non-adverse). Several non-adverse and not treatment-related effects were observed in all dose groups.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
40 mg/kg bw: high values of AST (both sexes), ALT (males), and inorganic phosphorus (males) and low levels of glucose (males) (non-adverse). Several non-adverse and not treatment-related effects were observed in the 10 and 40 mg/kg bw/day dose groups.
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
40 mg/kg bw: high values or tendencies of absolute and relative heart (both sexes) and kidney weights (females) after dosing and recovery period (adverse)
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
40 mg/kg bw: degeneration/necrosis (myocardial cell) and inflammatory cellular infiltration in the myocardium(both sexes, adverse) after dosing period, fibrosis in the myocardium after the recovery period (both sexes, adverse), effects on lung and liver
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
No treatment-related changes were observed.

BODY WEIGHT AND WEIGHT GAIN
No treatment-related changes were observed.

FOOD CONSUMPTION
No treatment-related changes were observed.

HAEMATOLOGY
40 mg/kg bw/day: high values of monocytes in both sexes at the end of the dosing period. High value of platelets and low values of basophils and the ratio were noted (males, end of recovery period)
10 mg/kg bw/day: high value of platelets (males, end of recovery period), prolongations of PT and PTT (females) (within mean±2SD of historical control data)
2 mg/kg bw/day: high values of eosinophil ratio males at the end of the dosing period (no dose-response correlation).

CLINICAL CHEMISTRY
40 mg/kg bw: High values of AST (both sexes), ALT (males), and inorganic phosphorus (males) and low levels of glucose (males) (end of dosing period, non-adverse). High values of ALP and sodium were noted in the males (within mean±2SD of historical control data). High sodium (males), potassium, and chlorine (males) values at the end of the recovery period (within mean±2SD of historical control data).
10 mg/kg bw/day: high potassium values (within mean±2SD of historical control data), high A/G ratio (males, no dose-response correlation)

URINALYSIS
No treatment-related changes were observed.

NEUROBEHAVIOUR
No treatment-related changes were observed.

ORGAN WEIGHTS
40 mg/kg bw: high values or tendencies of absolute and relative heart (both sexes) and kidney weights (females) after dosing and recovery period (adverse)

GROSS PATHOLOGY
No treatment-related changes were observed.

HISTOPATHOLOGY: NON-NEOPLASTIC
40 mg/kg bw:
Heart:degeneration/necrosis (myocardial cell) and inflammatory cellular infiltration in the myocardium(5/5 males, 5/5 females) after dosing period, fibrosis in the myocardium after the recovery period (1/5 males, 1/5 females)
Lung: degeneration/necrosis (myocardial cell around the vein) and inflammatory cellular infiltration in the myocardium around the vein (2/5 males, 2/5 females)
Liver: mild hypertrophy of the centrilobular hepatocytes (2/5 males, 3/5 females)
10 mg/kg bw/day: mild and focal inflammatory cellular infiltration in the myocardium (2/5 males, 1/5 females) at the end of the dosing period and in 1/5 males after the recovery period
Control group: mild and focal inflammatory cellular infiltration in the myocardium (3/5 males) at the end of the recovery group

HISTOPATHOLOGY: NEOPLASTIC
No findings were reported.

HISTORICAL CONTROL DATA
Historical control data were provided and used for discussion for endpoints where data have been observed that were outside the range of the actual control group.
Dose descriptor:
NOAEL
Effect level:
10 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Degeneration/necrosis in the myocardial cell and in myocardial cells around the vein at 40 mg/kg bw/day.
Dose descriptor:
LOAEL
Effect level:
40 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Degeneration/necrosis in the myocardial cell and in myocardial cells around the vein at 40 mg/kg bw/day.
Critical effects observed:
not specified

Table 1: Histopathological findings in males after the dosing period.

Organ and findings

 

Dose groups

 

 

 

 

 

Control

2 mg/kg bw/day

10 mg/kg bw/day

40 mg/kg bw/day

Liver

Hypertrophy, hepatocyte, centrilobular

0/5

0/5

0/5

2/5 (mild)

 

Cellular infiltration, inflammatory, focal

2/5 (mild)

0/5

2/5 (mild)

3/5 (mild)

Lung

Degeneration/necrosis, myocardial cell, vein, focal

0/5

0/5

0/5

2/5 (mild)

 

Cellular infiltration, vein, myocardium, inflammatory, focal

0/5

0/5

0/5

2/5 (mild)

Heart

Degeneration/necrosis, myocardial cell, diffuse

0/5

0/5

0/5

5/5 (mild)**

 

Cellular infiltration, myocardium, inflammatory, focal

0/5

0/5

2/5 (mild)

0/5

 

Cellular infiltration, myocardium, inflammatory, diffuse

0/5

0/5

0/5

5/5 (mild)**

 

Fibrosis, myocardium, diffuse

0/5

0/5

0/5

1/5 (mild)

**: p<0.01 (significantly different from control)


Table 2: Histopathological findings in females after the dosing period. 

Organ and findings

 

Dose groups

 

 

 

 

 

Control

2 mg/kg bw/day

10 mg/kg bw/day

40 mg/kg bw/day

Liver

Vacuolation, hepatocyte, periportal

0/5

1/5 (mild)

0/5

0/5

 

Hypertrophy, hepatocyte, centrilobular

0/5

0/5

0/5

3/5 (mild)

 

Cellular infiltration, inflammatory, focal

1/5 (mild)

0/5

0/5

0/5

Lung

Degeneration/necrosis, myocardial cell, vein, focal

0/5

0/5

0/5

2/5 (mild)

 

Cellular infiltration, vein, myocardium, inflammatory, focal

0/5

0/5

0/5

2/5 (mild)

 

Cellular infiltration, inflammatory, focal

0/5

1/5 (mild)

0/5

0/5

Heart

Degeneration/necrosis, myocardial cell, diffuse

0/5

0/5

0/5

5/5 (mild)**

 

Cellular infiltration, myocardium, inflammatory, focal

0/5

0/5

1/5 (mild)

0/5

 

Cellular infiltration, myocardium, inflammatory, diffuse

0/5

0/5

0/5

5/5 (mild)**

 

Fibrosis, myocardium, diffuse

0/5

0/5

0/5

1/5 (mild)

**: p<0.01 (significantly different from control)


 

Table 3: Histopathological findings in males and females after the recovery period.

Organ and findings

 

Dose groups

 

 

 

 

Control

10 mg/kg bw/day

40 mg/kg bw/day

Liver

Necrosis, hepatocyte, focal

Males: 0/5

Males: 0/5

Males: 1/5 (mild)

 

Cellular infiltration, inflammatory, focal

Males: 1/5 (mild) Females: 0/5

Males: 1/5 (mild) Females: 1/5 (mild)

Males: 2/5 (mild) Females: 1/5 (mild)

Lung

 

0/5

0/5

0/5

Heart

Cellular infiltration, myocardium, inflammatory, focal

Males: 3/5 (mild)

Males: 1/5 (mild)

Males: 1/5 (mild)

 

Fibrosis, myocardium, diffuse

Males: 0/5

Females: 0/5

Males: 0/5

Females: 0/5

Males: 4/5 (mild)*

Females: 3/5

*: p<0.05 (significantly different from control)


 

Table 4: Selected organ weights of males and females after the dosing and recovery period.

Absolute organ weight (relative organ weight)

 

Dose groups

 

 

 

 

 

Control

2 mg/kg bw/day

10 mg/kg bw/day

40 mg/kg bw/day

after dosing period

males

 

 

 

 

 

Heart

1.43±0.15 (0.38±0.03)

1.36±0.21 (0.37±0.03)

1.39±0.09 (0.37±0.03)

1.61±0.06 (0.42±0.01)

 

Liver

12.01±0.8 (3.21±0.09)

11.83±1.39 (3.23±0.17)

12.26±0.87 (3.25±0.20)

13.38±1.49 (3.49±0.21)

 

Females

 

 

 

 

 

Heart

0.81±0.05 (0.38±0.03)

0.84±0.09 (0.41±0.03)

0.87±0.10 (0.40±0.04)

0.99±0.08** (0.51±0.04)**

 

Liver

6.54±0.31 (3.11±0.14)

6.48±0.82 (3.16±0.21)

7.25±0.64 (3.33±0.22)

7.31±1.07 (3.72±0.29)**

after recovery period

males

 

 

 

 

 

Heart

1.52±0.15 (0.36±0.02)

-

1.68±0.15 (0.38±0.05)

1.57±0.10 (0.37±0.02)

 

Liver

13.25±1.34 (3.14±0.20)

-

12.75±1.13 (2.87±0.17)

12.88±1.02 (2.98±0.20)

 

Females

 

-

 

 

 

Heart

0.88±0.04 (0.38±0.03)

-

0.85±0.11 (0.37±0.05)

1.03±0.10 (0.44±0.05)

 

Liver

6.71±0.57 (2.91±0.06)

-

6.62±1.21 (2.88±0.12)

7.35±0.20* (3.14±0.16)*

*: p<0.05 (significantly different from control)

**: p<0.01 (significantly different from control)

 

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
10 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The available information comprises an adequate and reliable study (Klimisch score 1), and is thus sufficient to fulfil the standard information requirements set out in Annex VIII, 8.6, of Regulation (EC) No 1907/2006.

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

3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl 2-chloropropenoic acid ester (CAS 96383-55-0) was tested in a subacute repeated dose oral toxicity according to OECD 407 and in compliance with GLP in Crl:CD(SD) rats (Saitoh, 2012). Ten animals per sex and dose were treated with 10 and 40 mg/kg bw/d or vehicle only (corn oil containing 10% cremophor). Each 5 of the animals were part of the main group (28-day treatment) or recovery group (28-day treatment plus 14-day recovery period). Furthermore, 5 animals per sex were additionally treated with 2 mg/kg bw/d as part of the main group.

In two 7-day screening studies animals were dosed with 50, 250, and 1000 mg/kg bw/day (5 animals per sex and dose) or 1, 5, and 20 mg/kg bw/day (3 male animals per dose). Death and moribund animals (both sexes) were found in the 250 and 1000 mg/kg bw dose group. High AST and liver and kidney weights were observed in both sexes of the 50 mg/kg bw/day group. 1 male of this group showed discoloration of the heart, dark reddish macule of the lung, low values of body weight, food consumption and reticulocytes, and high values of platelets and monocytes. Effects on body weight and food consumption was also observed in 1 female of the 50 mg/kg bw dose group. Discoloration of the heart was also observed in dead animals of the 250 and 1000 mg/kg bw/day groups. No treatment-related changes were observed in the 1, 5, and 20 mg/kg bw/day groups. Thus 40 mg/kg bw/day was set as the high dose level in the main study expecting that there would be no death or changes in toxicity.

In the main study, no mortality or clinical signs of toxicity were observed throughout the study period. No effects on body weight or food consumption were observed. In the 40 mg/kg bw/d group high values of monocytes were observed in both sexes at the end of the dosing period only. These changes were considered to be caused by injury to myocardial cell. Furthermore, high value of platelets and low values of basophils and the ratio were noted (males, end of recovery period). In the mid dose group high value of platelets (males, end of recovery period) and prolongations of PT and PTT (females) were observed. These effects were judged to be not treatment-related as they were within the mean ±2 SD of historical control data.

High values of AST (both sexes), ALT (males), and inorganic phosphorus (males) and low levels of glucose (males) were observed at the end of the dosing period, only. Further changes of blood chemistry parameters were judged to be not treatment-related as there was no dose-response relationship or the values were within the mean ±2 SD of historical control data. Urinalysis revealed no changes. No relevant effects were observed in any of the parameters of the functional observation battery before and at the end of the treatment period.

In the high dose group high values or tendencies of absolute and relative heart (both sexes) and kidney weights (females) after dosing and recovery period were observed. Necropsy revealed no findings. But after histopathology of the heart degeneration/necrosis (myocardial cell) and inflammatory cellular infiltration in the myocardium (5/5 males, 5/5 females) were observed after the dosing period, and fibrosis in the myocardium (1/5 males, 1/5 females) was observed after the recovery period in the high dose group. In the 10 mg/kg bw/d group mild and focal inflammatory cellular infiltration in the myocardium (2/5 males, 1/5 females) was observed at the end of the dosing period and in 1/5 males after the recovery period. Mild and focal inflammatory cellular infiltration in the myocardium (3/5 males) was also observed in the control group at the end of the recovery period. Degeneration/necrosis in the myocardial cell is the change suggested the injury to the myocardium, and inflammatory cellular infiltration and fibrosis are the repair reaction to the injury.

Furthermore, effects on the lung as degeneration/necrosis (myocardial cell around the vein) and inflammatory cellular infiltration in the myocardium around the vein (2/5 males, 2/5 females) were observed in the high dose group. These effects were considered to be similar to the change observed in the heart as the myocardium extends along the vein from the heart continuously in rats. In addition, mild hypertrophy of the centrilobular hepatocytes (2/5 males, 3/5 females) was observed in the high dose group. These effects were judged to be a biological adaptive response as the change was not observed at the end of the recovery period and was not accompanied by impaired changes such as necrosis etc.

As a result, main test substance-related changes were degeneration/necrosis in the myocardial cells and in myocardial cells around the vein in both sexes of the 40 mg/kg bw/d group. Therefore the NOAEL was estimated to be 10 mg/kg bw/d under the conditions of the study.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:

The reliable GLP compliant OECD Guideline study was chosen.

Repeated dose toxicity: via oral route - systemic effects (target organ) cardiovascular / hematological: heart

Justification for classification or non-classification

The available data on repeated dose toxicity of the test substance do meet the criteria for classification as "STOT RE Cat. 2 (H373)" according to Regulation (EC) 1272/2008 or "Xn (R48/22)" according to Directive 67/548/EEC.

Regarding Regulation (EC) 1272/2008 the criteria for classification as STOT after repeated exposure require toxic effects, occurring at doses >30 to ≤300 mg/kg bw after oral exposure (study duration 28 days). The LOAEL, indicating toxic effects, was 40 mg/kg bw in the 28 day study. Thus, the criteria for classification as STOT after repeated exposure are met.

Regarding Directive 67/548/EEC the criteria for classification based on serious damage after repeated exposure require toxic effects, occurring at doses ≤150 mg/kg bw after oral exposure (study duration 28 days). The LOAEL, indicating toxic effects, was 40 mg/kg bw in the 28 day study. Thus, the test substance fulfills the criteria for classification for serious damage after repeated exposure.