Registration Dossier

Administrative data

Description of key information

REACH_NOAEL = 1000 mg/kg bw/d | rat (male/female) | OECD 407 | #key study#

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

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:
Feb - Nov 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Deviations:
yes
Remarks:
see below "Any other information on results"
Qualifier:
according to guideline
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
Deviations:
yes
Remarks:
see below "Any other information on results"
GLP compliance:
yes (incl. QA statement)
Remarks:
Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
PREPARATION OF THE ANIMALS
Prior to the start of the treatment period a detailed clinical observation outside the home cage was made. Animals showed no pathological signs before the first administration. Before the first administration all animals to be used for the study were weighed and assigned to the experimental groups with achieving a most homogenous variation in body weight throughout the groups of males and females, respectively (randomisation will be performed with IDBS Workbook 10.1.2 software).

ANIMALS
Species/strain: healthy Wistar rats, Crl: WI(Han) (Full Barrier)
Source: Charles River, 97633 Sulzfeld, Germany
Sex: male and female; the female animals were non-pregnant and nulliparous.
Age at the start of the treatment period: males: 6-7 weeks old, females: 6-7 weeks old.
Body weight at the allocation of the animals to the experimental groups:
males: 136 - 162 g (mean: 147.50 g, ± 20 % = 118.00 – 177.00 g); females: 130 - 151 g (mean: 143.10 g, ± 20 % = 114.48 – 171.72 g)
The animals were derived from a controlled full-barrier maintained breeding system (SPF). According to the German Act on Animal Welfare the animals were bred for experimental purposes.
This study was performed in an AAALAC-accredited laboratory. According to German animal protection law, the study type has been reviewed and accepted by local authorities. Furthermore, the study has been subjected to Ethical Review Process and was authorised by the Bavarian animal welfare administration.

HUSBANDRY
- Full barrier in an air-conditioned room
- Temperature: 22 ± 3 °C
- Relative humidity: 55 ± 10 %
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice
- Free access to tap water, sulphur acidified to a pH of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept in groups of 5 animals / sex / group / cage in IVC cages (type IV, polysulphone cages) on Altromin saw fibre bedding
- Certificates of food, water and bedding are filed for two years at BSL Munich and afterwards archived at Eurofins Munich
- Adequate acclimatisation period (at least 5 days) under laboratory conditions
Route of administration:
oral: gavage
Vehicle:
corn oil
Remarks:
Manufacturer: Sigma-Aldrich Batch No.: MKCD1021 / MKCC9871 Physical State: liquid Storage Conditions: room temperature Expiry Date: 30 March 2018 (MKCD1021), 24 April 2018 (MKCC9871)
Details on oral exposure:
Experimental Groups and Doses
According to the results of a previous dose range finding study (BSL Munich Study No. 178287, non GLP) and in consultation with the sponsor the following doses were selected for the 3 dose groups (LD = low dose, MD = medium dose, HD = high dose).

C 0 mg/kg bw
LD 100 mg/kg bw
MD 300 mg/kg bw
HD 1000 mg/kg bw

The animals were treated with the test item formulation or vehicle on 7 days per week for a period of 28 days.
The highest dose level was chosen with the aim of inducing toxic effects, but not death or severe suffering. Thereafter, a descending sequence of dose levels was selected with a view to demonstrate any dose-related response and a NOAEL.
The animals in the control group were handled in an identical manner to the test group subjects and received the vehicle using the same volume as used for the high dose group.

Administration of Doses
The test item formulation or vehicle was administered at a single dose to the animals by oral gavage. The application volume for all groups was 4 mL/kg body weight. For each animal the individual dosing volume was calculated on the basis of the body weight most recently measured.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Dose Formulation Analysis
Before beginning of the treatment period, formulation samples were prepared and analysed in order to obtain knowledge about stability and homogeneity of the test item in the selected vehicle at Eurofins Munich as part of a separate GLP study (Eurofins Munich Study No. 178290).
Prestart homogeneity investigation were included on the samples collected from various levels (top, middle and bottom) of high dose and low dose groups.
Since the test item was shown to be homogenous according to Eurofins Study No. 178290, samples were only analysed from the middle of prepared formulations from all dose groups and from the control group in study week 1 and 3 of the treatment (8 samples).
Each sample taken during the study was retained in duplicate (sample A, sample B, each of at least 5 mL). The A-samples were analysed at Eurofins Munich (Eurofins Munich Study Phase No. 178291) and until then stored under appropriate conditions based on available stability data. The B-samples were retained at -15 to -35 °C at BSL Munich (test facility) and discarded after completion of the final study report.
The phase plan was amended to this study plan. The results were reported in an analytical phase report which was attached in the report of this study.
Duration of treatment / exposure:
The animals were treated with the test article or vehicle for a period of 28 days.
Frequency of treatment:
daily
Dose / conc.:
0 other: mg/kg bw/day
Dose / conc.:
100 other: mg/kg bw/day
Dose / conc.:
300 other: mg/kg bw/day
Dose / conc.:
1 000 other: mg/kg bw/day
No. of animals per sex per dose:
40 animals (20 males and 20 females) were included in the study (5 male and 5 female animals per group).
Control animals:
yes, concurrent vehicle
Details on study design:
The aim of this study was to assess the possible health hazards which could arise from repeated exposure of Sa 190 via oral administration to rats over a period of 28 days.
The test item was administered daily in graduated doses to 3 groups of test animals, one dose level per group for a treatment period of 28 days. Animals of an additional control group were handled identically as the dose groups but received corn oil, the vehicle used in this study. The 4 groups comprised of 5 male and 5 female Wistar rats.
During the period of administration, the animals were observed precisely each day for signs of toxicity. Animals that died were examined macroscopically and at the conclusion of the test, surviving animals were sacrificed and observed macroscopically.
Body weight and food consumption were measured weekly.
At the conclusion of the treatment period, all animals were sacrificed and subjected to necropsy. The wet weight of a subset of tissues was taken and a set of organs/tissues was preserved. A full histopathological evaluation of the tissues was performed on high dose and control animals.
Observations and examinations performed and frequency:
Body Weight and Food Consumption
The body weight was recorded once before the assignment to the experimental groups, on the first day of administration and weekly during the treatment period. Food consumption was measured weekly during the treatment period.

Clinical Observations
All animals were observed for clinical signs during the entire treatment period of 28 days.
General clinical observations were made at least once a day, preferably at the same time each day and considering the peak period of anticipated effects after dosing. The health condition of the animals was recorded. Twice daily all animals were observed for morbidity and mortality except on weekends and public holidays when observations were made once daily.
Detailed cage side observations considering spontaneous activity, lethargy, recumbent position, convulsions, tremors, apnoea, asphyxia, vocalisation, diarrhoea, changes in skin and fur, eyes and mucous membranes (salivation, discharge), piloerection and pupil size were made outside the home cage in a standard arena once before the first administration and once in weeks 3 and 4.
Ophthalmological examination, using an ophthalmoscope was made on all animals before the first administration and in the last week of the treatment period.

Functional Observations
Once before the first exposure and once in the fourth week of exposure multiple detailed behavioural observations were made outside the home cage using a functional observational battery of tests. These tests were conducted in all animals.


Sacrifice and pathology:
Haematology
Haematological parameters were examined at the end of the treatment prior to or as part of the sacrifice of the animals.
After overnight fasting of male animals, blood from the abdominal aorta was collected in EDTA-coated tubes. Blood from female animals was collected in a non-fasted condition. The following haematological parameters were examined: haematocrit value, haemoglobin content, red blood cell count, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, reticulocytes, platelet count, white blood cells, neutrophils, lymphocytes, monocytes, eosinophils, basophils and large unstained cells.

Blood Coagulation
Coagulation parameters were examined at the end of the treatment prior to or as part of the sacrifice of the animals.
After overnight fasting of male animals, blood from the abdominal aorta was collected in citrate tubes. Blood from female animals was collected in a non-fasted condition. The following coagulation parameters were examined: prothrombin time and activated partial thromboplastin time.

Clinical Biochemistry
Parameters of clinical biochemistry were examined at the end of the treatment prior to or as part of the sacrifice of the animals.
After overnight fasting of male animals, blood from the abdominal aorta was collected in serum separator tubes. Blood from female animals was collected in a non-fasted condition.
The following parameters of clinical biochemistry were examined: alanine aminotransferase, aspartate-aminotransferase, alkaline phosphatase, creatinine, total protein, albumin, urea, total bilirubin, total bile acids, total cholesterol, glucose, sodium and potassium.
Additionally, at necropsy serum samples of all animals were retained at the end of the treatment period and stored at -20° C for a possible evaluation of test item-related effects on the pituitary-thyroid axis and thyroid hormones (if requested). The samples were discarded at finalisation of the study report.

Urinalysis
A urinalysis was performed with samples collected from all animals prior to or as part of the sacrifice of the animals.
The following parameters were measured using qualitative indicators (Henry Schein Urine Stripes URI 10SL): specific gravity, nitrite, pH-value, protein, glucose, ketone bodies, urobilinogen, bilirubin, erythroctes and leukocytes. Additionally, urine colour/ appearance were recorded. 

Pathology
One day after the last administration (study day 29) all surviving animals of the treatment period were sacrificed using anesthesia (ketamine and xylazin) and subjected to a detailed gross necropsy which includes careful examination of the external surface of the body, all orifices and the cranial, thoracic and abdominal cavities and their contents.

Organ Weight
The wet weight of the following organs of all sacrificed animals was recorded as soon as possible: liver, uterus with cervix, kidneys, thymus, adrenals, thyroid/ parathyroid glands, testes, spleen, epididymides, brain, prostate, seminal vesicles and coagulating glands, pituitary gland, ovaries and heart. Paired organs were weighed together. Organ weights of animals found dead or euthanised for animal welfare reasons were not recorded.
The following tissues from all animals were preserved in 4 % neutral-buffered formaldehyde except eyes, testes and epididymides which were fixed in Modified Davidson’s fixative for approximately 24 hours before they were transferred to 70 % ethanol:
adrenal glands, all gross lesions, aorta, brain (incl. medulla/pons, cerebellar and cerebral cortex), caecum, colon, duodenum, epididymides, eyes with optic nerve and Harderian gland, femur with knee joint, heart, ileum (including Peyer´s patches), jejunum, kidneys, liver, lungs, lymph nodes (mandibular), lymph nodes (mesenteric and axillary), mammary gland area (male and female), oesophagus, ovaries, oviducts, pancreas, parathyroid glands, pituitary, prostate and seminal vesicles with coagulating glands as a whole, rectum, salivary glands (sublingual, submandibular), sciatic nerve, skeletal muscle, skin, spinal cord (cervical, thoracic and lumbar segments), spleen, sternum (with bone marrow), stomach, testes, thymus, thyroid gland, tongue, trachea, ureters, urinary bladder and uterus with cervix and vagina.
All animals found dead and/or intercurrently euthanised for animal welfare reasons were subjected to a gross necropsy and the organs preserved for a histopathological examination.

Histopathology
The following organs were examined histopathologically after preparation of paraffin sections and haematoxylin-eosin staining for the animals of the groups 1 and 4 sacrificed at the end of the treatment period and any animal found dead or euthanised before the planned day of sacrifice:
adrenal glands, all gross lesions, brain (incl. medulla/pons, cerebellar and cerebral cortex), caecum, colon, duodenum, epididymides, eyes with optic nerve and Harderian gland, heart, ileum (including Peyer´s patches), jejunum, kidneys, liver, lungs, lymph nodes (mesenteric and axillary), oesophagus, ovaries, prostate and seminal vesicles with coagulating glands as a whole, rectum, sciatic nerve, skeletal muscle, spinal cord (cervical, thoracic and lumbar segments), spleen, sternum (with bone marrow), stomach, testes, thymus, thyroid gland, trachea, urinary bladder and uterus with cervix and vagina.
These examinations were extended to animals of all other dosage groups for treatment-related changes that were observed in the high dose group. Only organs and tissues of the other dosage groups showing changes in the high dose group were examined.
Any gross lesion macroscopically identified was examined microscopically in all animals. Discoloration possibly due to the test item was evaluated in the organs of all dose groups.
The histological processing of tissues to microscope slides was performed at the GLP-certified contract laboratory AnaPath Services, Hammerstrasse 49, 4410 Liestal, Switzerland (test site for tissue processing). The histopathological evaluation was performed at the GLP-certified contract laboratory AnaPath GmbH, Buchsweg 56, 4625 Oberbuchsiten, Switzerland (test site for histopathology). The Study phase from test site 1 and 2 was performed in compliance with the Swiss Ordinance relating to Good Laboratory Practice adopted 18 May 2005 [SR 813.112.1] (Status as of 01 December 2012. Blocking, embedding, cutting, H&E staining and scientific slide evaluation were performed according to the corresponding SOP’s of the test sites.
The principal investigator for histopathological tissue processing sent all raw data (including blocks, slides, paper raw data, statement of compliance and quality assurance statement) to the study director.
The principal investigator for histopathological evaluation provided the histopathology results to the study director by e-mail and sent a pathology phase report to the study director upon the completion of the study.
Statistics:
A statistical assessment of the results of the body weight, food consumption, parameters of haematology, blood coagulation and clinical biochemistry and absolute and relative organ weights were performed for each gender by comparing values of dosed with control animals using either a parametric one-way ANOVA and a post-hoc Dunnett Test or a non-parametric Kruskal-Wallis Test and a post-hoc Dunn’s Test, based on the results of homogeneity and normality tests. These statistics were performed with Ascentos 1.3.4 software or GraphPad Prism V.6.01 software (p<0.05 is considered as statistically significant).
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
see Details on results
Mortality:
mortality observed, non-treatment-related
Description (incidence):
see Details on results
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
see Details on results
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
see Details on results
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Description (incidence and severity):
see Details on results
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
see Details on results
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
see Details on results
Urinalysis findings:
no effects observed
Description (incidence and severity):
see Details on results
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
see Details on results
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
see Details on results
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
see Details on results
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
see Details on results
Histopathological findings: neoplastic:
no effects observed
Description (incidence and severity):
see Details on results
Other effects:
not examined
Details on results:
Mortality
A single female animal no. 29 of the LD group was euthanized in a bad health condition on study day 19. Pericardial inflammation, pleural inflammation and mediastinal inflammation at the thymus indicated gavage error.
Besides, no mortality occurred during the treatment period of this study.

Clinical Observations
Prior to euthanization of animal no. 29 of the LD group the following clinical symptoms were noted on the same day: reduced spontaneous activity, piloerection, half eyelid closure, hyperaesthesia, abnormal breathing and nasal discharge.
Moving the bedding and slight salivation were observed in all animals of the high dose group from the second treatment week onwards. Moving the bedding was also observed transiently in a single male animal of the MD group. These findings indicate slight local reactions after oral gavaging.
Mild clinical signs of alopecia or crust observed at the forelimbs, tail and upper back of a single control animal noted on study day 27 were of little relevance and are not assumed to have an impact on the results of the study.
Besides, there were no clinical signs of toxicity in this study. No abnormalities were seen during once-weekly detailed clinical observations in weeks 3 and 4.

Functional Observation Battery
In males and females, no relevant effects were observed in any of the parameters of the functional observation battery before and at the end of the treatment period when compared with the controls. There were no biologically relevant differences in body temperature between the groups.

Body Weight Development
Sa 190 had no effect on body weight development in this study. During and at the end of the treatment period there were no considerable differences in body weights between dose groups and control group.
A slight but statistically significantly higher body weight gain of male animals of the LD group is not assumed to be biologically relevant.

Food Consumption
Sa 190 had no effect on food consumption in this study. Food consumption was generally lower in treatment week 3 when compared to the other weeks. A reason is unclear. During the overall treatment period no considerable differences in food intake were observed between dose group and control group.

Haematology and Blood Coagulation
Sa 190 had no effect on haematological parameters and coagulation parameters determined at the end of the treatment period.
Very slightly but statistically significantly higher rate of reticulocytes found in male animals of the MD group are not considered toxicologically relevant as means were in the normal range of historical control data.
White blood cells were slightly higher in males of the MD and HD groups. As the increase was not dose-dependent, means were within historical control data and female animals were not affected, it is not assumed to be toxicologically relevant.
A slightly but statistically significantly higher rate of large unstained cells in male animals of the MD group is not assumed to be toxicologically relevant.
A very slight but statistically significantly longer aPTT was noted in male animals of the MD group. Due to the slightness and as means were within the range of historical control data this is not considered toxicologically relevant.
None of the statistically significantly different parameters described above were correlated with abnormalities in any of the other parameters examined in this study.

Clinical Biochemistry
Sa 190 had no effect on parameters of clinical biochemistry determined at the end of the treatment period.
Total protein was slightly and statistically significantly higher in serum of male but not female animals of the HD group when compared to controls. As values were within the normal range of historical control data this is not assumed to be toxicologically relevant.

Urinalysis
At the end of the treatment period no consistent differences were noted in urinary parameters between dose groups and control group of this study. A tendency towards lower pH values in male animals of MD and HD groups is not considered toxicologically relevant as it was not associated with histopathological findings.

Pathology
The following macroscopic findings were noted in animal no. 29 that was euthanized in a bad health condition: The thoracic cavity was filled with white fluid and the heart and thymus were abnormally dark.
At scheduled necropsy at the end of the treatment period the pericard of female animals 30 (LD group), 32 (MD group) and 40 (HD group) was filled with a yellow mucoid fluid. The reason for these specific findings observed in 3/30 dosed animals or 3/15 male dose animals is unclear. In absence of histopathological findings of the heart of HD animals and any other signs of toxicity in this study, this is not considered adverse in the context of this study. However, a relation to Sa 190 cannot be excluded.

A red thymus, axillary and mandibular lymph nodes was seen in animals no. 22, 24 (controls), 28 (LD group), 32 (MD group) and 38 (HD group). A red thymus was seen in animal no. 39 (HD group) and the thymus of LD group female no. 30 had an abnormal gelatinous consistency. Mesenteric lymph nodes were red in control female no. 24 and LD group female no. 28. As also control animals were affected these findings were not assumed to be caused by Sa 190.
Besides, single findings that are considered incidental were: a fluid-filled thoracic cavity of male animal no. 14 of the MD group and a red left caudal lung lobe and fluid-filled uterus of animal no. 26 of the LD group.

Organ Weight
Absolute kidney weight was slightly higher in male – but not female animals – independent of dose level in LD, MD and HD groups when compared to the control group (17 %, 13 % and 19 % above controls, respectively). Statistical significance was reached in LD and HD groups. Also when related to body weight was kidney weight statistically significantly lower in the HD group, when compared to controls. A cause of the test item cannot be excluded. However, in absence of histopathological findings this is not considered adverse.
A tendency towards a higher liver weight was observed in male animals of the LD, MD and HD groups (8 %, 10 % and 11 % above controls, respectively). In contrast, in female animals lower absolute liver weight was found in the LD and HD groups (11 % and 17 % below controls, respectively). The difference was statistically significant at the HD level. Due to the slightness and in absence of histopathological findings this is not considered adverse.

Histopathology
There were no histological lesions that distinguished controls from test item-treated animals.
Female no. 29 of group 2 was sacrificed during the course of the study. Pericardial inflammation, pleural inflammation and mediastinal inflammation at the thymus indicated gavage error.
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:
behaviour (functional findings)
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
gross pathology
haematology
histopathology: neoplastic
histopathology: non-neoplastic
mortality
ophthalmological examination
organ weights and organ / body weight ratios
urinalysis
Key result
Critical effects observed:
no

Dose Formulation Analysis

Concentration analysis of formulation samples was determined at three concentrations, 25 mg/mL, 75 mg/mL and 250 mg/mL in study weeks 1 and 3. The mean recoveries observed for the LD dose group were 88.5 % and 104.6 % of the nominal value, 94.8 % and 98.5 % for the MD dose group and 101.0 % and 106.4 % of the nominal value for HD dose group. The mean recoveries observed in the low dose (LD), medium dose (MD) and high dose (HD) groups were 96.6 %, 96.7 %, and 103.7 % of the nominal concentration, respectively.

Nominal concentrations were confirmed for all dose groups, as measured concentrations were within acceptance criterion of 10 %.

Conclusions:
On the basis of this 28-Day Repeated Dose Oral Toxicity study with Sa 190 in male and female Wistar rats with dose levels of 100, 300, and 1000 mg/kg body weight day, the following conclusions can be made:
Under the conditions of this study, the test item Sa 190 did not cause indicators of toxicity. The NOAEL may be established at 1000 mg/kg bw.
Executive summary:

The aim of this study was to assess the possible health hazards which could arise from repeated exposure of Sa 190 via oral administration to rats over a period of 28 days.

The test item was administered daily in graduated doses to 3 groups of test animals, one dose level per group for a treatment period of 28 days. Animals of an additional control group were handled identically as the dose groups but received corn oil, the vehicle used in this study. The 4 groups comprised of 5 male and 5 female Wistar rats.

During the period of administration, the animals were observed precisely each day for signs of toxicity. Animals that died were examined macroscopically and at the conclusion of the test, surviving animals were sacrificed and observed macroscopically.

Body weight and food consumption were measured weekly.

At the conclusion of the treatment period, all animals were sacrificed and subjected to necropsy. The wet weight of a subset of tissues was taken and a set of organs/tissues was preserved.

A full histopathological evaluation of the tissues was performed on high dose and control animals.

The following doses were evaluated:

Control:                           0       mg/kg body weight

Low Dose:                    100     mg/kg body weight

Medium Dose:              300    mg/kg body weight

High Dose:                  1000   mg/kg body weight

The test item was emulsified in corn oil and administered daily during a 28-day treatment period to male and female animals. After stability testing (Eurofins Munich Study No. 178290) test item formulations were used within 10 days after preparation. Dosing volumes were adjusted individually based on weekly body weight measurements.

Summary Results

A single female of the LD group was euthanized in a bad health condition in this study. Pericardial inflammation, pleural inflammation and mediastinal inflammation at the thymus indicated gavage error. Besides, no mortality occurred during the treatment period of this study. There were no clinical signs of systemic toxicity in this study. Transiently occurring moving the bedding and slight salivation were considered to be slight local reactions of Sa 190 almost exclusively at 1000 mg/kg bw/d. No relevant effects of Sa 190 were observed in any of the parameters of the functional observation battery. Body weights developed normally in all groups and no effect on food consumption was seen. There were no biologically relevant differences in body temperature between the groups. Sa 190 had no effect on haematological parameters, coagulation parameters and clinical biochemistry parameters determined at the end of the treatment period. Urinary parameters were also not affected by Sa 190.

At necropsy a pericard filled with a yellow mucoid fluid was found in single female animals of the LD, MD and HD group, respectively. In absence of histopathological findings of the heart of HD animals and any other signs of toxicity in this study, this is not considered adverse in the context of this study. However, a relation to Sa 190 cannot be excluded. Besides, non-test item related findings were noted at necropsy.

Independently of dose kidney and liver weight was slightly higher in male– but not female animals.

Histopathologically, there were no lesions that distinguished controls from test item-treated animals.

Gavaging error was confirmed histopathologically as cause of the only mortality in this study.

Conclusion

On the basis of this 28-Day Repeated Dose Oral Toxicity study with Sa 190 in male and female Wistar rats with dose levels of 100, 300, and 1000 mg/kgbody weight day, the following conclusions can be made:

Under the conditions of this study, the test item Sa 190 did not cause indicators of toxicity. The NOAEL may be established at 1000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 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: 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

Justification for classification or non-classification

Since no substance related adverse effects were observed at the highest dosage group, the substance does not need to be classified for repeated dose toxicity according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.