Registration Dossier

Administrative data

Description of key information

NOAEL = 1000 mg/kg bw/d (OECD 407, GLP, K, rel. 1)

NOAEL = 1000 mg/kg bw/day (OECD 408, GLP, K, rel.1) - read-across to Cyclohexane-1,4-dicarboxylic acid

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15th July 1999 to 14th November 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Version / remarks:
(adopted 27 July 1995)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
Version / remarks:
Commission Directive 96/54/EC
Deviations:
no
Principles of method if other than guideline:
not applicable
GLP compliance:
yes (incl. certificate)
Remarks:
Date of inspection: 23rd March 1998 Date of signature: 21st July 1998
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Ltd, Margate, Kent, United Kingdom.
- Age at study initiation: Approximately 5 to 7 weeks old.
- Weight at study initiation: 178 - 223 g (male) and 159 - 194 g (female).
- Fasting period before study: No data.
- Housing: The animals were housed in groups of 5 by sex in polypropylene grid-floor cages suspended over trays lined with absorbent paper.
- Diet (e.g. ad libitum): A pellet diet (Rat and Mouse SQC Expanded Diet No.1, Special Diets Services Limited, Witham, Essex, United Kingdom). Ad libitum.
- Water (e.g. ad libitum): Mains water. Ad libitum.
- Acclimation period: 11 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2ºC.
- Humidity (%): 55 ± 15%.
- Air changes (per hr): ≥ 15 changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours continuous light and 12 hours darkness.

IN-LIFE DATES: Day 0 (the day of dosing) to Day 28.
Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: The test material was prepared at 7.5, 75 and 500 mg/ml, as a solution in Arachis oil BP. The stability and homogeneity of the test material formulations were determined at the test laboratory. The formulations to be stable for at least 14 days. Formulations were therefore prepared weekly and stored at approximately 4ºC in the dark.

DIET PREPARATION
Not applicable.

VEHICLE
- Justification for use and choice of vehicle (if other than water): The stability and homogeneity of the test material formulations were determined at the test laboratory. The samples used were test material extracted with acetonitrile with final theoretical concentration of approximately 0.01 mg/ml. The formulations were found to be stable for at least 14 days.
- Concentration in vehicle: 7.5, 75 and 500 mg/ml.
- Amount of vehicle (if gavage): 2 ml/kg.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Summary; The concentration of ST 03 C 99 in the test material formulations was determined by gas chromatography (GC) using an external standard technique.

Samples; The test material formulations were extracted with acetonitrile with final theoretical concentration of approximately 0.01 mg/ml.

Standards; Standard solutions of test material were prepared in acetonitrile at a nominal concentration of 0.01 mg/ml.

Procedure; The sample and standard solutions were analysed by GC.

Homogeneity determinations; It was apparent by visual inspection that the test material formulations were homogeneous.

Stability determinations; The test material formulations were sampled and analysed initially and then after storage at 4ºC in the dark for 14 days.

Verification of test material formulation concentrations; The test material formulations were sampled and analysed within 3 days of preparation.

Results; Mean concentration found was in the range of 90 – 102% of nominal concentrations. The results showed the formulation to be stable for at least 14 days.
Duration of treatment / exposure:
28 days
Frequency of treatment:
Once daily
Dose / conc.:
15 mg/kg bw/day (actual dose received)
Dose / conc.:
150 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
5 animals per sex per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were chosen based on the results of the range-finding study performed. See the summary of the range-finding study in 'any other information on materials and methods incl. tables'.

Positive control:
No
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS / DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Immediately before dosing, and 1 and 5 hours after dosing during the working week. Animals were observed immediately before dosing, and 1 hour after dosing at weekends.
- Cage side observations checked: Overt signs of toxicity, ill-health and behavioural change.

BODY WEIGHT: Yes
- Time schedule for examinations: Individual bodyweights were recorded on Day 0 (the day before the sart of treatment) and on Day 7, 14, 21 and 28. Bodyweights were also recorded at terminal kill.

FOOD CONSUMPTION: Yes
Weekly food consumption was recorded for each cage group.

FOOD EFFICIENCY: Yes
- Food efficiency (group mean bodyweight gain / group mean food consumption) was calculated.

WATER CONSUMPTION: Yes
- Time schedule for examinations: Water intake was observed daily for each cage group by visual inspection of the water bottles for any overt changes.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY AND CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood:
Haematological and blood chemical investigations were performed at the end of the study (Day 28). Blood samples were obtained from the lateral tail vein. Where necessary repeat samples were obtained by cardiac puncture prior to necropsy on Day 29.
- Anaesthetic used for blood collection: Not reported.
- Animals fasted: No.
- How many animals: All animals from each test and control group.
- Parameters checked:
(Haematology) Hb, RBC, Hct, MCH, MCV, MCHC, WBC, APTT, Neut, Lymph, Mono, Eos, Bas, CT and PLT.
(Chinical Chemistry) Urea, Glucose, Tot.Prot, Albumin, A/G, Na+, K+, Cl-, Ca++, P, ASAT, ALAT, AP, Creat, Chol and Bili.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations:
Behavioural assessment: Prior to the start of treatment and on Days 6, 13, 20 and 27.
Functional Performance Tests: On Days 24 and 27.
Sensory Reactivity: On Day 27.
- Dose groups that were examined: All animals.
- Parameters examined:
Behavioural assessment: Detailed individual clinical observations were performed for each animal using a purpose-built arena. The following parameters were observed:
Gait, hyper/hypothermia, tremors, skin colour, twitches, respiration, convulsions, palpebral closure, bizarre/abnormal/stereotypic behaviour, urination, salivation, defecation, pilo-erection, transfer arousal, exophthalmia, tail elevation and lachrymation.
Functional Performance Tests: Motor Activity. Twenty purpose-built 44 infra-red beam automated activity monitors were used to assess motor activity. Animals of one sex were tested at each occasion and were randomly allocated to the activity monitors. The tests were performed at approximately the same time each day, under similar laboratory conditions. The evaluation period was sixteen hours for each animal. The percentage of time each animal was active and mobile was recorded for the overall time period and also during the final 20% of thetrial period (considered to be the asymptotic period).

Forelimb/Hindlimb Grip Strength. An automated grip strength meter was used. Each animal was allowed to grip the proximal metal bar of the meter with its forepaws. The animal was pulled by the base of the tail until its grip was broken. The animal was drawn along the trough of the meter by the tail until its hind paws gripped the distal metal bar. The animal was pulled by the base of the tail until its grip was broken. A record of the force required
to break the grip of each animal was made. Three consecutive trials were performed for each animal.

Sensory Reactivity: Each animal was individually assessed for sensory reactivity to auditory, visual and proprioceptive stimuli. The following parameters were observed:
Grasp response, touch escape, vocalisation, pupil reflex, toe pinch, startle reflex, tail pinch, blink reflex and finger approach.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
All animals were killed by intravenous overdose of sodium pentobarbitone (Sagatal, 60 mglml; May and Baker Limited, Dagenham, Essex, United Kingdom), followed by exsanguination on completion of the dosing period. All animals were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded.
Organ weights:
The following organs, removed from animals that were killed at the end of the study, were dissected free from fat and weighed before fixation.
Adrenals, liver, brain, ovaries, epididymides, spleen, heart, testes, kidneys and thymus.

HISTOPATHOLOGY: Yes
Samples of the following tissues were preserved from all animals and preserved in buffered 10% formalin.
Adrenals, aorta (thoracic), bone & bone marrow (femur including stifle joint), bone & bone marrow (sternum), brain (including cerebrum, cerebellum and pons), caecum, colon, duodenum, epididymides, eyes, gross lesions, heart, ileum, jejunum, kidneys, liver, lungs (with bronchi. Lungs were inflated to approximately normal inspiratory volume with buffered 10% formalin before immersion in fixative), lymph nodes (cervical and mesenteric), muscle (skeletal), oesophagus, ovaries, pancreas, pituitary, prostate, rectum, salivary glands (submaxillary), sciatic nerve, seminal vesicles, skin (hind limb), spinal cord (cervical), spleen, stomach, testes, thymus, thyroid/parathyroid, trachea, urinary bladder and uterus.

All tissues were despatched to the external facility for processing. The tissues (except aorta, bone & bone marrow, eyes, muscle, oesophagus, pancreas, pituitary, salivary glands and skin) from all control and 1000 mg/kg/day group animals were prepared as paraffin blocks, sectioned at nominal thickness of 5 μm and stained with haematoxylin and eosin for subsequent microscopic examination. Any macroscopically observed lesions were also processed together with the liver and spleen from all 15 and 150 mg/kg/day dose group animals.
Other examinations:
None
Statistics:
Haematology, blood chemistry and absolute and bodyweight relative organ weights, weekly bodyweight gain and quantitative functional performance and sensory reactivity data were assessed for dose response relationships by linear regression analysis, followed by one way analysis of variance (ANOVA) incorporating Levene’s test for homogeneity of variance. Where variances were shown to be homogenous, pairwise comparisons were conducted using Dunnett’s test. Where Levene’s test showed unequal variances the data were analysed using non-parametric methods: Kruskal-Wallis ANOVA and Mann-Whitney ‘U’ test.

Probability values (p) were calculated as follows:
p < 0.001 ***
p < 0.01 **
p < 0.05 *
p ≥ 0.05 (not significant)
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:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Other effects:
not specified
Details on results:
CLINICAL SIGNS AND MORTALITY: No death. No clinically observable signs of toxiicty detected.
Increased salivation was observed which often accompanies the oral administration of a test material formulation and is usually attributed to an unpleasant tasting or locally irritant formulation rather than an indication of systemic toxicity.

BODY WEIGHT AND WEIGHT GAIN: No adverse effects seen.
Females treated with 1000 mglkglday showed a slight but statistically significant reduction in bodyweight gain during Week 3 ofthe study When compared with controls. The level of significance achieved was minimal (p<0.05) and in view of the superior bodyweight gains noted at this dose level during the first half of the study, this isolated reduction was considered to be of no toxicological consequence.

FOOD CONSUMPTION AND FOOD EFFICIENCY: No adverse effects were detected.

WATER CONSUMPTION: No adverse effects were detected.

HAEMATOLOGY: No treatment-related changes were seen.

CLINICAL CHEMISTRY: No treatment-related changes were seen.
Males treated with 1000 mg/kg bw/day showed a statistically significant reduction in plasma glucose when compared with controls but in the absence of any degenerative liver changes or effects on food consumption, this reduction was considered to be of no toxicological importance. Males from this treatment group also showed an increase in plasma creatinine concentration when compared with controls. There was no concomitant change in plasma urea concentration amongst these animals nor any histopathological evidence to support a view of renal dysfunction and, in isolation, this intergroup difference was considered to be of no toxicological significance.
Females treated with 1000 mg/kg bw/day showed a statistically significant reduction in plasma potassium concentration together with an increase in plasma inorganic phosphorus concentration when compared with controls. In the absence of any imbalances in other plasma electrolytes or any evidence of microscopic renal change, these intergroup differences were considered to be of no toxicological importance.

NEUROBEHAVIOUR: There were no intergroup differences in sensory reactivity detected during the study.
Statistical analysis of functional performance revealed a significant increase in forelimb grip strength among 1000 mg/kg bw/day females when compared with controls but in the absence of any changes in the other functional performance parameters measured, this isolated intergroup difference was considered to be of no toxicological significance.

ORGAN WEIGHTS: No treatment-related changes were seen.

GROSS PATHOLOGY: No macroscopic abnormalities were seen.

HISTOPATHOLOGY: No treatment-related changes were seen.
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no toxicologically significant effects were observed
Key result
Critical effects observed:
no

none

Conclusions:
Repeated oral administration of the test material, ST 03 C 99, to rats by gavage for a period of 28-consecutive days at dose levels of up to 1000 mg/kg bw/day produced no toxicologically significant effects in the parameters measured. The NOAEL for this substance was considered to be 1000 mg/kg bw/day.
Executive summary:

Introduction, materials and methods

The study was designed to investigate the systemic toxicity of the test material. It complies with the requirements for notification of a new chemical substance in the EC and follows the testing method described in Commission Directive 96/54/EC (Method B7) and OECD Guidelines for Testing of Chemicals No. 407 "Repeated Dose 28 Day Oral Toxicity Study in Rodents" (Adopted 27 July 1995).

The test material was administered by gavage to three groups, each of five male and five female Sprague-Dawley CrI:CD®BR strain rats, for twenty-eight consecutive days, at dose levels of 15, 150 and 1000 mg/kg bw/day. A control group of five males and five females was dosed with vehicle alone (Arachis oil BP). Clinical signs, functional observations, bodyweight development and food and water consumption were monitored during the study. Haematology and blood chemistry were evaluated for all animals at the end of the study. All animals were subjected to a gross necropsy examination and histopathological evaluation of selected tissues from 1000 mg/kg bw/day and control animals was performed.

Results

There were no deaths during the study. No clinically observable signs of toxicity were detected during the study.

No intergroup differences were observed during Behavioural Assessments, Functional Performance Tests and Sensory Reactivity Assessments.

No adverse effect on bodyweight development, food consumption and water intake was observed during the study.

No treatment-related haematological changes, organ weight changes, macroscopic abnormalities or microscopic abnormalities were detected.

Clinical Chemistry: High-dose males showed increased plasma creatinine and reduced plasma glucose, while the females showed reduced potassium and increased inorganic phosphorous. Apart from the difference in creatinine, the differences were not reflected in the lower dose groups or animals of the other sex. While not statistically significant, the creatinine level for high-dose females was higher than for the controls, and in all treated animals, very small non-significant increases relative to controls were seen. While there is an appearance of a dose-response for creatinine concentration, the differences are at most barely significant, and no corresponding change in plasma urea or kidney histopathology was seen to indicate kidney dysfunction.

Conclusions

Repeated oral administration of the test material, ST 03 C 99, to rats by gavage for a period of twenty-eight consecutive days at dose levels of up to 1000 mg/kg bw/day produced no toxicologically adverse significant changes in the parameters measured. The "No Observed Adverse Effect Level" (NOAEL) was therefore considered to be 1000 mg/kg bw/day.

This study is considered as acceptable and satisfies the requirement for sub-acute repeated dose toxicty endpoint.

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
02 August 2005 to 21 May 2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Version / remarks:
Adopted on 21 September 1998
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.26 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Version / remarks:
Adopted on 30 September 1996
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Details on species / strain selection:
Recognised by the international guidelines as the recommended test system
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS (HanRcc:WlST)
- Source: RCC Ltd, Laboratory Animal Services, CH-4414 Fullinsdorf, Switzerland.
- Age at delivery: 6 weeks
- Weight at acclimatization: Males: 135-160 g (mean 148 g); Females: 116-133 g (mean 125 g)
- Housing: Animals were housed in groups of five in Makrolon type-4 cages with wire mesh tops and standardized softwood bedding (Lignocel Schill AG, CH-4132 Muttenz, Switzerland)
- Diet: Pelleted standard Provimi Kliba 3433 (batch no. 39/05, 63/05) rat maintenance diet (Provimi Kliba AG, CH-4303, Kaiseraugst, Switzerland) was available ad libitum
- Water: Community tap-water from Itingen was available ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 30-70%
- Air changes: Air-conditioned with 10-15 air changes per hour
- Photoperiod: 12-hour fluorescent light/12-hour dark cycle with music during the light period.
Route of administration:
oral: gavage
Vehicle:
other: PEG 300
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
Test substance was weighed into a glass beaker on a tared Mettler balance and the vehicle polyethylene glycol 300 (PEG 300) added. The mixtures were prepared using a magnetic stirrer and stored at room temperature (15-25 °C).
Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.
The dose formulations were prepared daily.

VEHICLE
- Concentration in vehicle: 20, 60 and 200 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg bw/day
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Concentration, homogeneity and stability (after 4 hours and 7 days) of the dose formulations were determined in samples taken before experimental start. Analyses were performed according to a HPLC analytical method.
Concentration and homogeneity of the dose formulations were determined in samples taken monthly during the treatment period.
Duration of treatment / exposure:
Duration of treatment: 91/92 days
Duration of recovery: 28 days
Frequency of treatment:
Daily
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
Main group: 10 animals/sex/dose
Recovery group: 5 animals/sex/dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Rationale for animal assignment: Computer-generated random algorithm
- Post-exposure recovery period in satellite groups: Additional 5 rats per sex and group were used at 0 and 1000 mg/kg bw/day. These animals were treated for 90 days and then allowed a 28-day treatment-free recovery period.
Positive control:
Not applicable
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Observations for mortality/viability were recorded twice daily.
The animals were observed for clinical signs once daily before commencement of administration and once daily during the treatment and recovery periods.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly
The animals were observed in their home cages, outside their home cages in a standard arena and in the hand. These observations were performed in random sequence once before commencement of administration and once weekly (weeks 1-12 and 14-16) thereafter.

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were recorded weekly during the acclimatization, treatment and recovery periods and before necropsy, using an on-line electronic recording system consisting of a Mettler balance connected to the RCC computer.

FOOD CONSUMPTION: Yes
- Food consumption was recorded once during the pretest period and weekly thereafter, using an on-line electronic recording system consisting of a Mettler balance connected to the RCC computer.

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Eye examinations were performed at acclimatization in all animals (toxicity and recovery groups), during Week 13 in control and high dose animals (toxicity and recovery groups), and at Week 17 in control and high dose animals (recovery groups).
- Ophthalmoscopic examinations of both eyes of all animals were performed after the application of a mydriatic solution using an ophthalmoscope.

HAEMATOLOGY AND CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: After 13 weeks (toxicity and recovery groups) and after 17 weeks (recovery groups)
- Anaesthetic used for blood collection: Yes; Blood samples for haematology and clinical biochemistry were collected from all animals under light isoflurane anaesthesia
- Animals fasted: Yes; animals were fasted in metabolism cages for ca. 18 hours before blood sampling but allowed access to water ad libitum.
- Blood samples were drawn from the retro-orbital plexus using a microhematocrit glass capillary tube.
- Parameters checked:
Haematology: Erythrocyte count, Hemoglobin, Hematocrit, Mean corpuscular volume, Red cell volume distribution width, Mean corpuscular hemoglobin, Mean corpuscular hemoglobin concentration, Hemoglobin concentration distribution width, Platelet (thrombocyte) count, Reticulocvte count, Reticulocyte maturity index, Methemoglobin, Heinz bodies, Total leukocyte count, Differential leukocyte count, Coagulation: Thromboplastin time and Activated partial thromboplastin time
Clinical chemistry: Glucose, Urea, Creatinine, Bilirubin (total), Cholesterol (total), Triglycerides, Phospholipids, Aspartate aminotransferase, Alanine aminotransferase, Lactate dehydrogenase, Glutamate dehydrogenase, Creatinine kinase, Alkaline phosphatase, Gamma-glutamyl-transferase, Sodium, Potassium, Chloride, Calcium, Phosphorus inorganic, Protein (total), Albumin, Globulin, Albumin/Globulin ratio

URINALYSIS: Yes
- Time schedule for collection of urine: After 13 weeks (toxicity and recovery groups) and after 17 weeks (recovery groups)
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes; Urine was collected during the 18-hour fasting period into a specimen vial.
- Parameters checked: Volume (18 hours), Specific gravity (relative density), Osmolality, Color, Appearance, pH, Nitrite, Protein, Glucose, Ketones, Urobilinogen, Bilirubin, Blood

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Weeks 13 (toxicity and recovery groups) and 17 (recovery groups)
- Battery of functions tested: Grip strength / motor activity
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Necropsy:
After 13 weeks (toxicity groups) and after 17 weeks (recovery groups)
- All animals were weighed and necropsied. Descriptions of all macroscopic abnormalities were recorded. All animals surviving to scheduled necropsy and all moribund animals were anaesthetised by intraperitoneal injection of sodium pentobarbitone and killed by exsanguination.

HISTOPATHOLOGY: Yes
- Samples of the following tissues and organs were collected from all animals at necropsy and fixed in neutral phosphate buffered 4% formaldehyde solution:
Adrenal glands*, Aorta*, Bone (sternum, femur incl. joint), Bone marrow (femur)*, Brain (4 levels)*, Cecum*, Colon*, Duodenum*, Epididymides (fixed in Bouin’s solution)*, Esophagus*, Exorbital lacrimal glands, Eyes with optic nerve (fixed in Davidson's solution)*, Harderian gland (fixed in Davidson's solution), Heart*, Ileum with Peyer's patches*, Jejunum with Peyer's patches*, Kidneys*, Larynx, Lacrimal gland (exorbital), Liver*, Lungs (infused with formalin)*, Lymph nodes - mesenteric, mandibular*, Mammary gland area*, Nasal cavity (turbinates), Ovaries*, Pancreas*, Pituitary gland*, Prostate gland*, Rectum*, Salivary glands - mandibular, sub lingual, Sciatic nerve*, Seminal vesicles*, Skeletal muscle, Skin*, Spinal cord - cervical, mid thoracic, lumbar*, Spleen*, Stomach*, Testes (fixed in Bouin's solution)*, Thymus*, Thyroid (incl. parathyroid gland, if possible)*, Tongue*, Trachea*, Urinary bladder (infused formalin)*, Uterus*, Vagina*, Gross lesions*
Histotechnique: All organ and tissue samples were processed, embedded and cut at an approximate thickness of 2 to 4 micrometers, and stained with hematoxylin and eosin.
Histopathology: Slides of all organs and tissues indicated as “*” (above) that were collected at scheduled sacrifice from the animals of control and high-dose groups were examined.
As possible test item-related morphologic changes were detected in the organs of high-dose animals, the same organs (liver (females only), ovaries, uterus, vagina, and thymus (males only from animals of the mid- and low-dose groups were examined.
Other examinations:
Organ weights: Brain, Heart, Liver, Thyroids with parathyroids, Thymus, Kidneys, Adrenals, Uterus, Spleen, Testes, Epididymides, Ovaries
The organ to terminal body weight ratios as well as organ to brain weight ratios were determined.
The determination of the terminal body weight was performed immediately prior to necropsy.
Statistics:
The following statistical methods were used to analyze the grip strength, locomotor activity, food consumption, body weight clinical laboratory data, 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 were assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex. Alternatively, the t-test was used for grip strength and locomotor activity.
- The Steel-test (many-one rank test) were applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
- Fisher's exact-test was applied to the ophthalmoscopic findings.
Clinical signs:
no effects observed
Description (incidence and severity):
- No test item-related clinical signs of toxicological relevance were noted at any dose level during daily clinical observations or during weekly detailed clinical observations (Weeks 1-12 and 14-16).

General cage side observations (daily):
- Daily clinical observations showed soft feces in rats at all dose levels This finding is commonly associated with the use of PEG 300 as vehicle and is considered to be a passive finding rather than one of systemic toxicity. Pale feces were noted in male and female rats treated with 1000 mg/kg bw/day. In the absence of any concomitant findings which could be related to this change, this was considered to be due to the light-colored test item and is of no toxicological relevance. Soft and pale feces both resolved during the recovery period.
- All other clinical signs noted during daily observations (such as crusts, alopecia, salivation, kinked tail, half-closed eyes, exophthalmia, rales, etc) were considered to be typical background changes and are of no toxicological relevance.

Detailed clinical observations (weekly):
- A small number of typical background findings were noted in males and females. These changes included bedding consumption, ruffled fur, rales, localized erythema or alopecia, exophthalmic (bilateral) and/or miosis (bilateral). None of the observed findings was considered to be of toxicological relevance.
- During the post-treatment weekly detailed clinical observations (weeks 14-16), observations included soft and/or pale feces, rales, paresis, salivation, exophthalmia (bilateral) and/or ptosis (bilateral).
Mortality:
no mortality observed
Description (incidence):
- No test item-related deaths occurred.
- One control male was killed for ethical reasons during the recovery phase of the study. The poor condition of this rat was found to be due to a kidney nephroblastoma. One control female rat died during the treatment period as a result of dosing error.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
- Reductions in mean body weight noted in rats treated with 1000 mg/kg bw/day were considered to be a slight test item-related effect. The reductions noted in females were more clearly expressed than the reductions noted in males.
- In males treated with 1000 mg/kg bw/day, marginally lower mean body weights were noted (ca. 5-7%) during the treatment period, beginning around day 36 of treatment. This was also reflected in the mean body weight gain values. The body weights and body weight gain increased slightly during the recovery period.
- In females treated with 1000 mg/kg bw/day, reductions in mean body weight attained statistical significance on day 43 (p<0.05), day 50 (p<0.05), day 64 (p<0.05), day 71 (p<0.01), day 78 (p<0.05) and day 91 (p<0.05) of treatment. The reductions in mean body weight gain attained statistical significance throughout the treatment period (p<0.01 or p<0.05), with the exception of day 57. Lower mean body weights persisted during the recovery period in females treated previously with 1000 mg/kg bw/day, attaining statistical significance on days 3 and 15 (p<0.05). Mean body weight gain was also lower than control females but the differences did not attain statistical significance.
- The transient significant reductions in mean body weight which were noted in females treated with 100 mg/kg bw/day (days 36 and 43 (p<0.05)) were considered to be incidental. Similarly, the transient significant reductions in mean body weight gain (days 8, 36 and 43 (p<0.01or p<0.05)) were considered to be incidental.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
- The mean daily food consumption and mean relative daily food consumption of the test item treated rats were unaffected during the treatment and recovery period.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
no effects observed
Description (incidence and severity):
- No test item-related ophthalmoscopic findings were noted at any dose level after 13 weeks of treatment or 4 weeks of recovery.
Haematological findings:
no effects observed
Description (incidence and severity):
- No test item-related differences in the haematological parameters were noted during the treatment and recovery in males or females at any dose level.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
- No test item-related changes of toxicological relevance were noted during the treatment and recovery in males or females at any dose level.
Urinalysis findings:
no effects observed
Description (incidence and severity):
- Male and female rats treated with 1000 mg/kg bw/day had significantly elevated urine output (p<0.05) when compared with the respective controls. In females at this dose level, significant reductions in relative density (p<0.05), osmolality (p<0.01) and ketones (P<0.05) were noted, whereas males had significantly lower pH (p<0.05) and leukocytes (P<0.05) than the control males. Of these changes, only the elevated urine output was considered to be a marginal test item-related effect.
- After the 4-week recovery period, the urinalysis parameters of the control and previously test item-treated rats were similar.
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
- No test item-related clinical signs of toxicological relevance were noted during functional observational battery (Week 13 of treatment or Week 4 of recovery) at any dose level.
- A small number of typical background findings were noted in males and females. These changes included bedding consumption, ruffled fur, localized erythema or alopecia, exophthalmia (bilateral), paresis, eye discoloration and/or miosis (bilateral). None of the observed findings was considered to be of toxicological relevance.
Grip Strength: No test item-related differences in the mean fore- and hindlimb grip strength values were noted at any dose level when assessed during week 13 of treatment and week 4 of recovery.
Locomotor activity: No test item-related effects upon locomotor activity were noted after 13 weeks of treatment or 4 weeks of recovery.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
After 13 weeks:
- In males treated at 1000 mg/kg bw/day, statistically significant reductions of the mean spleen-to-brain weight ratio (p<0.05) were noted. This finding was accompanied by reduced mean absolute thymus weights (p<0.05), reduced thymus-to-body weight ratio (not significant) and reduced thymus-to-brain weight ratio (p<0.05). The reduction in mean and absolute thymus weights correlated with microscopical findings (lymphoid depletion). These findings were considered to be related to the test item.
- The mean kidney-to-body weight ratio of males treated with 1000 mg/kg bw/day was significantly elevated (p<0.01) when compared to the controls, but without microscopical correlation. This difference was considered to be unrelated to the test item.
- The significant difference noted in mean brain-to-body weight ratio of males treated with 1000 mg/kg bw/day was considered to be incidental, as no correlating microscopic finding was found. The significant reduction in heart-to-brain weight ratio (p<0.01) noted in males treated with 1000 mg/kg bw/day was also considered to be incidental; no correlating microscopic findings were found.
- In females treated at 1000 mg/kg bw/day, slightly reduced mean absolute and relative ovary weights (not significant) were noted, together with reduced mean absolute and relative uterus weights (p<0.05 or p<0.01).
- The significantly elevated liver-to-body weight ratio (p<0.01) noted in these females was considered to be a test item-related adaptive change.
- In males treated at 300 mg/kg bw/day, no test item-related changes in mean absolute or relative organ weights were ascertained.
- In females treated with 300 mg/kg bw/day, reduced mean absolute and relative uterus weights (p<0.05) were considered to be test item-related changes. The significantly elevated liver-to-body weight ratio (p<0.05) noted in these females was considered to be test item-related adaptive change.
- In males treated with 100 mg/kg bw/day, the mean absolute testes weight and mean testes-to-body weight ratio was significantly elevated (p<0.05) when compared with the controls. This finding was considered to be incidental, as no correlating microscopic finding was found. In females treated at 100 mg/kg bw/day, no test item-related changes in mean absolute or relative organ weights were ascertained.
After 17 Weeks:
- In males treated previously with 1000 mg/kg bw/day, test item-related differences in the spleen and thymus were reversible after the recovery period.
- In females treated previously with 1000 mg/kg bw/day, effects on mean uterus weights were reversible after the recovery period (not statistically significant, very wide normal range). Test item-related adaptive differences in the liver were reversible after the recovery period.
- The incidental elevation of mean brain-to-body weight ratio noted in males treated with 1000 mg/kg bw/day (p<0.05) persisted after the recovery period. Females had a marginal but significant increase in the mean brain-to-body weight ratio (p<0.05) after the recovery period. Neither of these differences was accompanied by any microscopical changes and both were considered to be incidental.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
After 13 Weeks:
- After 13 weeks of treatment period, liquid contents in the cecum were recorded at necropsy in 10/10 males and in 4/10 females treated with 1000 mg/kg bw/day. One control male and female and one female (300 mg/kg bw/day) also had liquid contents in the cecum.
- In one control female rat which died during the treatment period, the lung was not collapsed and was discolored. Foamy fluid was released from the bronchi.
- The few other findings recorded, such as pelvic dilation of the kidneys or dark red foci in the thymus were regularly distributed among control and treatment groups, and were considered to be within the range of normal background changes which may be seen in rats of this strain and age in 13-week studies. These were considered incidental and to reflect the usual individual variability.

After 17 Weeks:
After 4 weeks of recovery, there were no macroscopic findings related to the treatment with the test item. One male of control group was killed for ethical reasons during the recovery period, the right kidney had a nodule which also adhered to the right adrenal gland.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
After 13 Weeks:
- At the end of the treatment period, the following test item-related microscopic lesions were observed:
Liver: After 13 weeks of treatment period, minimal or slight centrilobular hypertrophy of the hepatocytes was observed in all females treated with 1000 mg/kg bw/day.
Thymus: After 13 weeks of treatment, a minimal or slight lymphoid depletion was seen in 2/10 males treated with 300 mg/kg bw/day and in 6/10 males treated with 1000 mg/kg bw/day.
- Liquid contents in the cecum recorded at necropsy in all males and some females treated with 1000 mg/kg bw/day (as well as one control male and female, and one female treated with 300 mg/kg bw/day) did not correlate microscopically as the cecum morphology did not differ between control and test item-treated animals. Microscopically the gastro-intestinal tract was without test item-related findings.
- One control female rat which died during the treatment period, the microscopical findings correlated with edema and congestion, and were considered to confirm that the animal died from a dosing error.
- The remaining microscopic findings were regularly distributed among the groups and were within the range and severity of spontaneous background lesions that may be observed in rats of this strain and age in this laboratory. They were considered to be of no toxicological significance.

After 17 Weeks:
- At the end of the recovery period, the liver and thymus morphology returned to normal. One control male was killed for ethical reasons during the recovery phase of the study due to a kidney nephroblastoma. This rare tumor occurs spontaneously in young rats of this strain.
Histopathological findings: neoplastic:
not examined
Other effects:
not specified
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects observed
Critical effects observed:
no

None

Conclusions:
Under the test conditions, the NOAEL is 1000 mg/kg bw/day in rats.
Executive summary:

In a repeated dose toxicity study performed in accordance with OECD test guideline No. 408 and in compliance with GLP, test substance was administered daily by oral gavage to groups of HanRcc:WlSTrats (10/sex/dose) at dose levels of 100, 300 and 1000 mg/kg bw/day for 90 days. A similarly constituted control group received the vehicle, PEG 300 only. Additional 5 rats per sex and group were used at 0 and 1000 mg/kg bw/day. These animals were treated for 90 days and then allowed a 28-day treatment-free recovery period. During the study, clinical observations, functional observational battery, grip strength and locomotor activity, body weight, food consumption, ophthalmoscopy, haematology, coagulation, blood chemistry, urinalysis, organ weights, macropathology and histopathology investigations were undertaken.

                            

No test item-related deaths occurred. No test item-related clinical signs of toxicological relevance were noted at any dose level during daily clinical observations or during weekly detailed clinical observations (Weeks 1-12 and 14-16). No test item-related clinical signs of toxicological relevance were noted during functional observational battery (Week 13 of treatment or Week 4 of recovery) at any dose level. No test item-related differences in the mean fore- and hindlimb grip strength values were noted at any dose level when assessed during week 13 of treatment and week 4 of recovery. No test item-related effects upon locomotor activity were noted after 13 weeks of treatment or 4 weeks of recovery.

 

Transient reductions in body weight and body weight gains was noted for males and females at 100 mg/kg bw/day resulting in slightly reduced body weights and body weight gains at the end of the exposure (-4.4%/-4.4% for males and -4.5%/-6.5%). The mean body weights and mean body weight gain of the rats treated with 300 mg/kg bw/day were considered to be unaffected by treatment. Slightly reduced mean body weights and body weight gains were noted in males rats treated with 1000 mg/kg bw/day (-5% and -7.6% at D91, respectively). These changes were more distinct in the females (-6.6% and -16.9% at D91, respectively). During the recovery period, mean body weights and body weight gain was similar to controls in males. In females, body weights and body weight gains were also improved at the end of the recovery period, even if complete recovery was not observed (-4.1% and -7.4%, respectively). Based on the absence of dose-relationship, on the limited severity and on the reversibility, these effects are not considered to be adverse. The mean daily food consumption and mean relative daily food consumption of the test item treated rats were unaffected during the treatment and recovery period.

No test item-related ophthalmoscopic findings were noted at any dose level after 13 weeks of treatment or 4 weeks of recovery. No test item-related differences in the hematological parameters were noted during the treatment and recovery in males or females at any dose level. No test item-related changes of toxicological relevance were noted during the treatment and recovery in males or females at any dose level. Male and female rats treated with 1000 mg/kg bw/day had significantly elevated urine output during treatment when compared to the controls. Urine output reverted to control levels during recovery therefore this finding was not considered to be adverse.

 After 13 weeks, at 1000 mg/kg bw/day males showed reduction in mean spleen-to-brain weight ratios accompanied by reduced mean absolute thymus weights, reduced thymus-to-body weight ratio and reduced thymus-to-brain weight ratio. The reduction in mean and absolute thymus weights correlated with microscopical findings (lymphoid depletion). The test item-related differences noted in the spleen and thymus of males were fully reversible after 4 weeks of recovery and are therefore not considered to be adverse.

At 1000 mg/kg bw/day, females showed slightly reduced mean absolute ovary weights as well as reduced ovary-to-body weight ratios and ovary-to-brain weight ratios. Statistical significance was not achieved and the effect was no longer apparent after the recovery period. The effect on ovary are therefore not considered to be adverse.

At 1000 and 300 mg/kg bw/day, females had reduced mean absolute uterus weights, reduced uterus-to-body weight and reduced uterus-to-brain weights. The effect was however not clearly dose-related at 13 weeks and reversibility occurred after the recovery period in the 1000 mg/kg bw/day group. Indeed, after 4 weeks of recovery, no statistical difference was noted and all individual values were within or very near to the control range. In the absence of histopathology correlates in the uterine tissue, the effects are considered to be spurious chance differences and are not considered to be averse.

Elevated liver-to-body weight ratio was observed in all females treated with 1000 mg/kg bw/day and was accompanied by minimal or slight centrilobular hypertrophy of the hepatocytes. All the findings were reversible after the recovery period and were considered to be adaptive change.

At 300 mg/kg bw/day in males and at 100 mg/kg bw/day in both sexes, no test item-related changes of toxicological relevance were ascertained.

During necropsy, after 13 weeks of treatment, liquid cecal contents were noted in all males and four females treated with 1000 mg/kg bw/day. This macroscopic change did not correlate with any microscopic change in the cecum morphology and was not considered to be adverse.

 

Based on the results of the study, the NOAEL is 1000 mg/kg bw/day in rats.

The test material is not classified for damage to organs through prolonged oral dose repeated exposure according to the criteria of the Annex VI of the Regulation (EC) No 1272/2008 (CLP).

This study is considered as acceptable and satisfies the requirement for sub-chronic repeated dose toxicty endpoint.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
The sub-acute oxicity study performed on the registered substance, and the sub-chronic toxicity study conducted on the supporting substance are both GLP-compliant and of high quality (Klimisch score = 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

Repeated dose toxicity: oral

The sub-acute toxicity of Safepharm (2000, rel. 1) was considered to be the key study for the sub-acute repeated dose toxicity endpoint. In this study performed according to the OECD test guideline No. 407 and in compliance with GLP, the test material diluted in arachis oil was administered by gavage to rats at 0, 15, 150 or 1000 mg/kg bw/day for 28 days.

There were no deaths during the study. No clinically observable signs of toxicity were detected during the study.

No intergroup differences were observed during Behavioural Assessments, Functional Performance Tests and Sensory Reactivity Assessments.

No adverse effect on bodyweight development, food consumption and water intake was observed during the study.

No treatment-related haematological changes, organ weight changes, macroscopic abnormalities or microscopic abnormalities were detected.

High-dose males showed increased plasma creatinine and reduced plasma glucose, while the females showed reduced potassium and increased inorganic phosphorous. Apart from the difference in creatinine, the differences were not reflected in the lower dose groups or animals of the other sex. While not statistically significant, the creatinine level for high-dose females was higher than for the controls, and in all treated animals, very small non-significant increases relative to controls were seen. While there is an appearance of a dose-response for creatinine concentration, the differences are at most barely significant, and no corresponding change in plasma urea or kidney histopathology was seen to indicate kidney dysfunction.

Based on these observations, the "No Observed Adverse Effect level" (NOAEL) was considered to be 1000 mg/kg bw/day.

The sub-chronic toxicity of RCC (2006, rel.1) conducted on the supporting substance, Cyclohexane-1,4-dicarboxylic acid, was considered to be the key study for the sub-chronic repeated dose toxicity endpoint. In this study performed in accordance with OECD test guideline No. 408 and in compliance with GLP, test substance diluted on PEG 300 was administered daily by oral gavage to groups of HanRcc:WlSTrats (10/sex/dose) at dose levels of 0, 100, 300 and 1000 mg/kg bw/day for 90 days. Additional 5 rats per sex and group were used at 0 and 1000 mg/kg bw/day. These animals were treated for 90 days and then allowed a 28-day treatment-free recovery period.                             

No test item-related deaths occurred. No test item-related clinical signs of toxicological relevance were noted at any dose level during daily clinical observations or during weekly detailed clinical observations (Weeks 1-12 and 14-16). No test item-related clinical signs of toxicological relevance were noted during functional observational battery (Week 13 of treatment or Week 4 of recovery) at any dose level. No test item-related differences in the mean fore- and hindlimb grip strength values were noted at any dose level when assessed during week 13 of treatment and week 4 of recovery. No test item-related effects upon locomotor activity were noted after 13 weeks of treatment or 4 weeks of recovery.

Transient reductions in body weight and body weight gains was noted for males and females at 100 mg/kg bw/day resulting in slightly reduced body weights and body weight gains at the end of the exposure (-4.4%/-4.4% for males and -4.5%/-6.5%). The mean body weights and mean body weight gain of the rats treated with 300 mg/kg bw/day were considered to be unaffected by treatment. Slightly reduced mean body weights and body weight gains were noted in males rats treated with 1000 mg/kg bw/day (-5% and -7.6% at D91, respectively). These changes were more distinct in the females (-6.6% and -16.9% at D91, respectively). During the recovery period, mean body weights and body weight gain was similar to controls in males. In females, body weights and body weight gains were also improved at the end of the recovery period, even if complete recovery was not observed (-4.1% and -7.4%, respectively). Based on the absence of dose-relationship, on the limited severity and on the reversibility, these effects are not considered to be adverse. The mean daily food consumption and mean relative daily food consumption of the test item treated rats were unaffected during the treatment and recovery period.

No test item-related ophthalmoscopic findings were noted at any dose level after 13 weeks of treatment or 4 weeks of recovery. No test item-related differences in the hematological parameters were noted during the treatment and recovery in males or females at any dose level. No test item-related changes of toxicological relevance were noted during the treatment and recovery in males or females at any dose level. Male and female rats treated with 1000 mg/kg bw/day had significantly elevated urine output during treatment when compared to the controls. Urine output reverted to control levels during recovery therefore this finding was not considered to be adverse.

 After 13 weeks, at 1000 mg/kg bw/day males showed reduction in mean spleen-to-brain weight ratios accompanied by reduced mean absolute thymus weights, reduced thymus-to-body weight ratio and reduced thymus-to-brain weight ratio. The reduction in mean and absolute thymus weights correlated with microscopical findings (lymphoid depletion). The test item-related differences noted in the spleen and thymus of males were fully reversible after 4 weeks of recovery and are therefore not considered to be adverse.

At 1000 mg/kg bw/day, females showed slightly reduced mean absolute ovary weights as well as reduced ovary-to-body weight ratios and ovary-to-brain weight ratios. Statistical significance was not achieved and the effect was no longer apparent after the recovery period. The effect on ovary are therefore not considered to be adverse.

At 1000 and 300 mg/kg bw/day, females had reduced mean absolute uterus weights, reduced uterus-to-body weight and reduced uterus-to-brain weights. The effect was however not clearly dose-related at 13 weeks and reversibility occurred after the recovery period in the 1000 mg/kg bw/day group. Indeed, after 4 weeks of recovery, no statistical difference was noted and all individual values were within or very near to the control range. In the absence of histopathology correlates in the uterine tissue, the effects are considered to be spurious chance differences and are not considered to be averse.

Elevated liver-to-body weight ratio was observed in all females treated with 1000 mg/kg bw/day and was accompanied by minimal or slight centrilobular hypertrophy of the hepatocytes. All the findings were reversible after the recovery period and were considered to be adaptive change.

At 300 mg/kg bw/day in males and at 100 mg/kg bw/day in both sexes, no test item-related changes of toxicological relevance were ascertained.

During necropsy, after 13 weeks of treatment, liquid cecal contents were noted in all males and four females treated with 1000 mg/kg bw/day. This macroscopic change did not correlate with any microscopic change in the cecum morphology and was not considered to be adverse.

Based on the results of the study, the NOAEL was considered to be 1000 mg/kg bw/day in rats.

Justification for classification or non-classification

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No 1272/2008.

Self-classification:

Based on the available data, no self-classification is proposed regarding the specific target organ toxicity after oral dose-repeated exposure according to the Regulation (EC) No. 1272/2008 (CLP).

There were no data regarding the dermal and inhalation route.