Registration Dossier

Administrative data

Description of key information

Based on histopathological changes seen in all treated groups (LD, MD, HD) in the combined repeated dose toxicity and reproduction/ developmental toxicity screening test, no NOAEL (No Observed Adverse Effect Level) could be established for general toxicity. The LD of 100 mg/kg/day could be considered the LOAEL in this study.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: oral, other
Remarks:
combined repeated dose and reproduction / developmental screening
Type of information:
experimental study
Adequacy of study:
key study
Study period:
July 6 - August 26, 2010
Reliability:
1 (reliable without restriction)
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Qualifier:
according to guideline
Guideline:
other: OPPTS 870.3650: Combined repeated dose Toxicity study with the Reproduction/ developmental Toxicity screening Test
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Previous names of the test substance: MK 195KSF; Naphthenic acids, reaction products with diethylenetriamine.
Batch No.: MK195K/BM/198/067/A/001
Physical state at RT: Hazy solid
Colour: orange/brown
Active Components (%): 100
Storage Conditions: at room temperature
Species:
rat
Strain:
Wistar
Details on species / strain selection:
Young healthy male and nulliparous, non pregnant female rats [strain: Wistar Crl:WI] (Full-Barrier), were used in this study. The animals were derived from a controlled full barrier maintained breeding system (SPF) (Source: Charles River, 97633 Sulzfeld, Germany).
At the beginning of the study, the age of the animals was 8-9 weeks.
The range of the body weight was:
Females: 154 to 183 g, (mean: 167.93 g, ± 20%= 33.59 g)
Males: 213 to 233 g, (mean: 222.30 g, ± 20%= 44.46 g)
Sex:
male/female
Details on test animals or test system and environmental conditions:
The animals were randomly assigned (using Microsoft Excel template) to the dose/control groups, each animal was assigned a unique identification number and caged individually. The animals were acclimatised for at least five days before the first dose administration.

After an adequate acclimatisation period (at least five days) the animals were barrier maintained (full barrier) in air-conditioned rooms under the following conditions:
- 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)
- Housed individually in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding
- Certificates of food, water and bedding are filed at the testing laboratory.
Route of administration:
oral: gavage
Details on route of administration:
The test item was administered by gavage using a gavaging cannula. The maximum dose volume administered was 5 mL / kg body weight.
Vehicle:
corn oil
Details on oral exposure:
The test item was dissolved/ suspended in corn oil (Batch No.: 11KBC6753; Storage conditions: at room temperature).

The vehicle was chosen as suggested by the test item‟s solubility.
Before dose formulation preparation test item was heated up to 80º C. The test item was weighed and the required volume of vehicle was added for better and speedy dissolution.
The test item formulation was prepared freshly on each administration day before the administration procedure.
Based on the available information from other toxicity studies following doses were selected:
Control: 0 mg/kg bw
LD: 100 mg/kg bw
MD: 300 mg/kg bw
HD: 750 mg/kg bw
The highest dose level is chosen with the aim of inducing toxic effects but not death or severe suffering. Thereafter, a descending sequence of dose levels is selected with a view to demonstrate any dosage related response and no-observed-adverse effects at the lowest dose level (NOAEL).
The animals in the control group were handled in an identical manner to the test group subjects and received the vehicle in the volume same as treated groups.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Each dosing concentration was analysed for nominal concentration.
Homogeneity of the test item in the vehicle was analysed for the low and high dose concentrations.
Samples for the nominal concentration verification were taken in study week 1 (first week of pre mating period), 3 (first week of mating), 5 (gestation) and 7 (gestation/lactation).
Samples for homogeneity were taken from the top, middle and bottom of the high dose and low dose preparation in study week 1 and 5.
All formulation samples were analysed on the same day of collection.
The results of the concentration measurements revealed recoveries between 85.3 % and 98.5% in HD group, between 86.4 % and 97.2 % in MD group and between 87.6 % and 95.5 % in LD group. Homogeneous distribution of the test substance in the preparation was approved.
Results are given as concentration in mg/mL and as recovery referring to nominal concentration.
Duration of treatment / exposure:
The test substance was administered daily during 14 days pre mating and 14 days mating in both male and in female, during gestation period and up to post natal day 3 in females. Males were dosed for 28-30 days.
Frequency of treatment:
daily; The animals were dosed with the test item on 7 days per week basis.
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
750 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
80 animals (10 females and 10 males/group) were included in the study.
Control animals:
yes, concurrent vehicle
Details on study design:
Four groups comprising 10 adult males and 10 non pregnant nulliparous female rats (Wistar Crl:WI) were dosed daily by oral gavage with 100, 300 and 750 mg/kgbody weight per day.
The test item was formulated in corn oil with administration volume of 5 mL/kg body weight.
Control animals were handled identically as treated groups and received corn oil in similar volume as treated groups.
The test item formulation was prepared freshly and administered daily during 14 days pre mating and 14 days mating period in both male and in female, during gestation period and up to post natal day 3 in females. Males were dosed for 28-30 days. Dose volumes were adjusted weekly based on the body weight measurement.
After 14 days of treatment to both male and female, animals were paired (1:1) for maximum 14 days.
The subsequent morning onwards the vaginal smears of females were checked to confirm the evidence of mating.
After the confirmation of the mating, females were separated. Each litter was examined as soon as possible after delivery of the dam to establish the number and sex of pups, stillbirths, live births, runts and the presence of gross abnormalities. Live pups were counted, sexed and litters weighed within 24 hours of parturition and on day 4 post-partum.
Males and females were sacrificed on treatment day 29-31, post natal day 4 respectively and subjected to necropsy. Non Pregnant females were sacrificed on their respective day 26 after the evidence of mating.
Animals were examined for the daily clinical signs, mortality, weekly detailed clinical observations, pre treatment and at termination for functional observations. Body weight and food consumption was measured weekly except food consumption was not measured during the mating period (female) and mating/post mating period (male). Haematological and clinical biochemistry evaluations were performed on blood samples collected at terminal sacrifice from five males and five randomly selected females from each group. Urinalysis was performed on samples collected at terminal sacrifice from five randomly selected males from each group.
Observations and examinations performed and frequency:
Animals were examined for the daily clinical signs, mortality, weekly detailed clinical observations, pre treatment and at termination for functional observations.
Body weight and food consumption was measured weekly except food consumption was not measured during the mating period (female) and mating/post matingperiod (male).
Haematological and clinical biochemistry evaluations were performed on blood samples collected at terminal sacrifice from five males and five randomly selected females from each group.
Urinalysis was performed on samples collected at terminal sacrifice from five randomly selected males from each group.
Sacrifice and pathology:
Males and females were sacrificed on treatment day 29-31, post natal day 4 respectively and subjected to necropsy. Non Pregnant females were sacrificed on their respective day 26 after the evidence of mating.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Test item related clinical findings namely piloerection, vocalization, pushing the bedding and salivation were recorded in most animals of treatment groups during the study. Occasionally findings like reduced spontaneous activity, weight loss, dyspnoea, diarrhoea, nasal discharge (bloody or plain) and dehydration were also observed.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
During the study twelve rats were found dead or sacrificed moribund. Out of these, 1 male from control, 1 male and 1 female from LD group and 3 males/ 6 females of HD group died or sacrficed.
The death of three females (animal no.s 34, 35, 39) could not be determined due to advanced autolysis and as a reason no pathological evaluation could be performed.
The histopathological evaluation has revealed that the intercurrent deaths observed might possibly be due to gavaging error.
In addition in most of the HD intercurrent dead animals similar findings in the mesenteric lymph node, small intestine and stomach were found corresponding to those seen in terminal kill animals of this group which may have accounted for the degraded health status observed before death in these animals.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
In males, the statistical analysis revealed significantly lower body weight development during premating period in HD group when compared with the controls, which further recovered during mating/post mating period.
Significantly low weight development was also observed on first week of mating/post mating period in MD group, which further recovered during second week of mating/post mating period. Overall from premating day 1 to mating/post mating day 14, the body weight development was significantly lower in MD and HD group.
In females, the statistical analysis revealed significantly lower body weight development during second week of premating period in HD group. Significant low weight development was also recorded during GD 0-7 (MD and HD groups), GD 7-14 (HD group), GD 14-20 (MD and HD groups), GD 0-20 (MD and HD groups). There was also a consistent dose response pattern observed for all treatment groups when compared to the corresponding controls.
The low body weight development in MD or HD groups might be considered due to treatment.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
In males and females, statistical analysis reavelaed significantly lower food consumption during premating period in HD group when compared to the corresponding controls.
In females, significantly lower food consumption was also recorded during GD 0-7 and 14-20 in MD and HD groups when compared to corresponding controls. During GD 7-14 the measurement of food consumption was inadvertently not measured. Hence, there is no data available.
The food intake and weight development are correlated to each other. Hence, the lower food intake in MD or HD group is considered to be due to treatment related.
Food efficiency:
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
Statistically significant deviation was for few hematological parameters MD or HD groups when compared to corresponding control values and also some individual values were below or above the historical control range, which are presented as follows.
WBC: In male, 1 individual value in LD group was below the historical control range. In females, 2 individual values in Control and 2 individual values in LD group were below the historical control range. Statistical significance was observed for male HD group and female MD and HD group compared to corresponding controls.
RBC: In males, all individual and mean values were within the historical control range. In females, 1 individual value each in control, LD and MD were below thehistorical control range.
HGB: In males, all individual and mean values were within the historical control range. In females, 1 individual value each in control and LD, 2 individual values each in MD and HD groups were below the historical control range.
MCV: In males, all individual and mean values were within the historical control range. In females, 1 individual value each in Control, LD and MD groups were above the historical control range. Statistical significance was observed for female HD group when compared to the control.
MCH: All individual values in male and female control and treatment groups were with historical control range. The statistical evaluation revealed significantly low value in female HD group compared to control.
Platelet: All individual values in male and female control and treatment groups were with historical control range. The statistical evaluation revealed significantly high value in male HD and female MD group compared to corresponding control.
aPTT: In males, 1 individual value in control was above the historical control range, while 1 individual values each in LD and MD groups were below the historical control range. In females, 1 individual value each in LD, MD and HD groups were below the biological range. There was no statistical deviation observed between the treatment and control groups.
PT: The statistical evaluation revealed significantly high value in male and female HD group.
Neutrophils: In males, 2 individual values and a mean value in MD group, 3 individual values and a mean value in HD group were above the historical control range. In females, 1 individual value in LD group, 3 individual values and mean value in MD group and all individual values and a mean values in HD group were above the historical control range. The statisatical significance was observed for both male and female MD and HD groups when compared to corresponding controls.
Lymphocytes: In males, 2 individual values and a mean value in MD group, 3 individual values and mean value in HD group were below the historical control range. Statistical significance was observed for male and female MD and HD groups when compared to the controls.
Monocytes: All individual values in male and female control and treatment groups were with historical control range. Statistically significant high values were observed in male MD and HD groups and female HD group compared to corresponding controls.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Statistically significant deviation was for few clinical biochemistry parameters in treatment groups when compared to corresponding control values and also some individual values were below or above the historical control range, which are presented as follows.
GLU: In males, 1 individual value in control, 4 individual values and a mean value in LD group, 2 individual values in MD group were above the historical control range. In females, 1 individual value in MD group was above the historical control range. There was no statistical deviation observed between treatment and control groups. The deviation from the normal historical range could be due to differential feeding as the animals were not starved overnight.
CHOL: In males, all individual and mean values in control, LD (except 1 individual value) and HD groups, 3 individual values and a mean value in MD group were below the historical control range. In females, all individual and mean values in control, MD and HD, 3 individual values and a mean value in LD were below the historical control range. But there was no statistical deviation observed between the treatment and control groups.
TP: In males, all in individual and mean values of treatment and control groups were below the historical control range. In females, 2 individual values in control, all individual values and a mean value in LD and MD groups, 2 individual values and a mean value in HD group were below the biological range. There was no statistical deviation observed between the treatment and control groups.
ALBB: in males, 2 individual values in HD group were below the historical control range. In females, all individual and mean values were with the historical control range. The statistical deviation was observed for females LD, MD and HD groups compared to control.
SGPT: In males and females, all individual and mean values were within the historical control range. The statistical evaluation revealed significantly high value in male HD group compared to controls.
Na: In males and females, all individual and mean values were within the historical control range. The statistical evaluation revealed significantly low value in female HD group compared to controls.
Urinalysis findings:
no effects observed
Description (incidence and severity):
The urinalysis revealed no significant differences in any of the treatment groups compared to corresponding control.
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
No relevant differences were observed concerning functional and behavioural examination in males and females.
No abnormalities were recorded concerning posture, gait, palpebral closure, lacrimation and arousal, except for piloerection and respiration observed in MD or HD group animal during the study.
No convulsions, tremors, stereotypy or bizarre behaviour were observed for the animals of any groups.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
In males, statistically significant low mean values were observed for absolute weight of heart (MD and HD groups), thymus (HD group) and seminal vesicle + coagulating gland (HD group) when compared with the corresponding controls.
The relative weight of liver indicated significantly higher values for MD and HD groups compared to control. The relative organ weight was significantly deviated for spleen (HD group), testes (MD group), thymus (MD group), Seminal vesicles + coagulating gland (HD group). The statistically significant deviation observed for absolute weight of heart (MD and HD group) and thymus (HD group) did not support the findings as the relative organ weight of the same organs did not deviate from the control group.
In females, statistically significant lower values were observed for absolute weight of heart (LD and HD groups) and uterus with cervix (HD group). The relative organ weight was significantly higher for spleen (MD group), liver (MD and HD groups), low for heart (LD and HD groups) and uterus with cervix (HD group).
The deviation observed in relative organ weight of liver, spleen and thymus may be due to histopathological organ changes considered secondary to a general inflammatory state caused by test item effects in treated animals, or to stress. The deviation observed for the relative organ weight of reproductive organs (seminal vesicles+coagulating gland and uterus with cervix) could be due to histopathological changes considered not to be directly related to test item, but attributed to general stress situation.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
In males, the macroscopic findings observed were as follows:
Control - small testes (1/10), small epididymides (1/10), discoloured axillary lymph node (1/10), discoloured thymus (1/10), small spleen (1/10), large and discoloured lung (1/10);
LD- discoloure lung (1/10), enlarged mesenteric lymph node (1/10);
MD - discoloured axillary lymph node (1/10), discoloured thymus (1/10), discoloured,enlarged and foam filled lung (1/10), lung with small black spots (1/10);
HD- small testes (2/10), small epididymides (1/10), small spleen (2/10), enlarged lung (2/10), discoloured lung (2/10), small Seminal vesicle+coagulating gland (2/10), discoloured liver (2/10), discoloured pancrea (1/10), gas filled stomach and intestine (3/10), small thymus (3/10), bloody/foamy nasal discharge (2/10), bloody lung (1/10).
In females, the macroscopic findings observed were as follows:
Control - cyst on ovary (1/10);
LD - cyst on ovary (1/10), discoloured intestine (1/10), discoloured lung (1/10), enlarged and foam filled lung (1/10);
MD- bloody lung (1/10);
HD- bloody lung (2/7), gas filled stomach and intestine (3/7), small spleen (1/7), small liver (1/7), small thymus (2/7), dilated intestine (1/7), enlarged mesenteric lymph node (2/7), discoloured Kidney (2/7).
Neuropathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
At terminal sacrifice, histopathological changes considered to be directly test item-related and toxicologically relevant were seen in the small intestine (jejunum and ileum), mesenteric lymph node, spleen, lung and adrenal gland.
In HD group, minor aggregations of foamy histiocytes were seen in the villi of small intestine. In all treatment groups, minimal to marked histiocytic infiltrates were observed in the mesenteric lymph node (draining lymph node of the small intestine). The effect on mesenteric lymph node was strongly dose-related and in many cases associated with foci of necrosis and/or abscessation as well as in some cases also with other secondary changes. Histiocytic infiltrates were also noted in the spleen or lung of isolated animals of MD or HD groups. In the female MD and HD group, there were also higher grades of alveolar macrophage aggregations in the lung. In the inner cortex of the adrenal gland, minor vacuolation was noted in isolated animals and/or minor mononuclear cell infiltrates were seen in a proportion of animals of MD or HD groups and in a single female of LD group.
Histopathological lesions of the tracheal epithelium (diffuse epithelial alteration) and gastric nonglandular mucosa of stomach (extensive or diffuse epithelial hyperplasia) were seen in occasional decedents and/or terminal kill animals, predominantly of the high dose group, and were considered to be likely due to a local irritant effect of the test item. Further histopathological organ changes in the spleen, lung, liver, bone marrow and thymus were considered secondary to a general inflammatory state caused by test item effects in treated animals, or to stress.
Among the decedents, two HD group females did not show physiological estrous cycle, and in two decedent HD group males, there was minor testicular degeneration and/or decreased secretion in the sexual glands. As no effects on reproductive organs were noted on terminal sacrifice animals, these minor changes were not considered to be directly test item-related, but might rather be a consequence of a general stress situation in the animals concerned.
Histopathological findings: neoplastic:
no effects observed
Dose descriptor:
NOAEL
Effect level:
< 100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
histopathology: non-neoplastic
Remarks on result:
other:
Remarks:
At dosages of 100, 300 and 750 mg/kg body weight revealed test item-related findings in males and females of all treatment group. No NOAEL could be established for general toxicity.
Key result
Dose descriptor:
LOAEL
Effect level:
<= 100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
histopathology: non-neoplastic
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
100 mg/kg bw/day (actual dose received)
System:
other: gastrointestinal tract; endocrine system; respiratory system: lower respiratory tract
Organ:
adrenal glands
ileum
jejunum
lungs
mesenteric lymph node
spleen
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
not specified
Conclusions:
The repeated dose administration of the substance in corn oil to the male (28-30 days) and female (maximum 54 days) rats at dosages of 100, 300 and 750 mg/kg body weight revealed test item-related findings in males and females of all treatment group.
Based on the data generated from this combined repeated dose toxicity and reproduction/ developmental toxicity screening test with Fatty acids C10-20, reaction product with diethylenetriamine, no NOAEL (No Observed Adverse Effect Level) could be established for general toxicity.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LOAEL
100 mg/kg bw/day
Study duration:
subacute
Species:
rat
Organ:
adrenal glands
ileum
jejunum
lungs
mesenteric lymph node
spleen

Additional information

A GLP experimental study has been performed to cover systemic toxicity effects after repeated administration by oral route (dietary) for 28 -days in rodents combined with reproductive end-points (OECD 422 study). No studies were performed for dermal and/or inhalation treatment following the discussed waiving criteria.

 

Mortality

During the study twelve rats were found dead or sacrificed moribund. Out of these, 1 male from control, 1 male and 1 female from LD group and 3 males/ 6 females of HD group died. The death of animals were histopathologically termed possibly due to misgavaging except for three dead females (animal no.s 34, 35, 39) in HD group, whose death could not be determined due to advanced autolysis.

 

Clinical findings

Test item related clinical findings viz., piloerection, vocalization, pushing the bedding and salivation were recorded in most animals of treatment groups

during the study. Occasionally findings like reduced spontaneous activity, weight loss, dyspnoea, diarrhoea, epistaxis, nasal discharge, dehydration,

etc were also observed.

 

Body Weight

In males, the statistical analysis revealed significantly low body weight development during pre-mating period in HD group and during first week of mating/post mating period in MD group, which further recovered with the progress of the study. But, overall the body weight development was significantly lower in MD and HD group. In females, significantly low body weight development were observed during second week of pre-mating period in HD group and during GD 0-7 (HD group), GD 14-20 (MD group), GD 14-20 (HD group). There was also a consistent dose response pattern observed for all treatment groups when compared to the corresponding controls.

 

Food consumption

In males and females, statistical analysis revealed significantly low food consumption during premating period in HD group when compared to the corresponding controls. In females, significantly low food consumption was also recorded during GD 0-7 and 14-20 in MD and HD groups when compared to corresponding controls.

 

Functional Examination

No relevant differences were observed concerning functional and behavioural examination in males and females.

No abnormalities were recorded concerning posture, gait, palpebral closure, lacrimation, arousal and vocalization, expect for piloerection which was observed in MD or HD group animal during the study.

No convulsions, tremors, stereotypy or bizarre behaviour were observed for the animals of any groups.

For supported and unassisted rears no abnormalities were detected. Responses to reflex testing were normal in all groups.

 

Blood analysis

Statistically significant deviation was observed for few hematological parameters viz., WBC, MCV, MCH, platelet, PT, Neutrophil, lymphocytes and monocytes in MD or HD groups when compared to corresponding control values.

Statistically significant deviation was obsereved for few clinical biochemistry parameters viz., ALBB, SGPT and Na in LD or MD or HD groups when compared to corresponding control values.

 

Urinalysis

The urinalysis revealed no significant differences in any of the treatment groups compared to corresponding control.

 

Macroscopic findings

In males, the macroscopic findings observed were as follows, Control- small testes (1/10), small epididymides (1/10), discoloured axillary lymph node (1/10), discoloured thymus (1/10), small spleen (1/10), large and discoloured lung (1/10); LD- discoloured lung (1/10), enlarged mesenteric lymph node (1/10); MD- discoloured axillary lymph node (1/10), discoloured thymus (1/10), lung with small black spots (1/10); HD- small testes (2/10), small epididymides (1/10), small spleen (2/10), enlarged lung (2/10), discoloured lung (2/10), small Seminal vesicle+coagulating gland (2/10), discoloured liver (2/10), discoloured pancrea (1/10), gas filled stomach and intestine (2/10), small thymus (2/10), foamy nasal discharge (1/10).

In females, control- cyst on ovary (1/10); LD- cyst on ovary (1/10), discoloured intestine (1/10); MD- bloody lung (1/10); HD- bloody lung (2/7), gas filled stomach and intestine (3/7), small spleen (1/7), small liver (1/7), small thymus (1/7), dilated intestine (1/7), enlarged mesenteric lymph node (2/7), discoloured Kidney (2/7).

 

Organ weight

In males, statistically significant low mean values were observed for absolute weight of heart (MD and HD groups), thymus (HD group) and seminal vesicle + coagulating gland (HD group) when compared with the corresponding controls. But, the relative weight of liver indicated significantly high values for MD and HD groups compared to control. The relative organ weight was significantly deviated for spleen (HD group), testes (MD group), thymus (MD group), Seminal vesicles + coagulating gland (HD group).

In females, statistically significant low values were observed for absolute weight of heart (LD and HD groups) and uterus with cervix (HD group); significantly high value for spleen (MD group). The relativeorgan weight was significantly higher for liver (HD group), low for heart (LD and HD groups) and uterus with cervix (HD group).

 

Microscopic findings (Histopathology)

At terminal sacrifice, histopathological changes considered to be directly test item-related and toxicologically relevant were seen in the small intestine (jejunum and ileum), mesenteric lymph node, spleen, lung and adrenal gland in LD or MD or HD group animals. The histopathological organ changes observed in the spleen, lung, liver, bone marrow and thymus were considered secondary to a general inflammatory state caused by test item effects in treated animals, or to stress.

 

Conclusion

Based on observed effect seen in all treated groups (LD, MD, HD) no NOAEL (No Observed Adverse Effect Level) could be established for general toxicity. The LD of 100 mg/kg/day could be considered the LOAEL in this study.

The 90-day study by oral route was not deemed necessary as the 28-day study showed a sufficient level of toxicity and leads to classify the substance as STOT RE 2, H373. Furthermore the DNEL calculated is considered enough low to cover also the possible effects that could arise in the 90-day study.

Justification for classification or non-classification

Based on observed effects and LOAEL value, the substance is classified as STOT RE 2, H373, according to CLP criteria.