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Description of key information

Oral:
Subacute Toxicity Study 28days, oral, rat, OECD 422, up to 400mg/kg bw/day tested: LOAEL >= 50mg/kg bw/d (BASF SE, 2013, 85R0674/12X380). Specific target organ toxicity: heart (cardiomyopathy)
Dermal:
No data available
Inhalation:
No data available

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: oral
Remarks:
combined repeated dose and reproduction / developmental screening
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline and GLP compliant study
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
adopted 22. March, 1996
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EPA OPPTS 870.3650, July 2000
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories, B.V., Horst, Netherlands (RccHanTM:WIST(SPF)
- Age at study initiation: 11 weeks
- Weight at study initiation: weight range: male 330 to 383 g; female 217 to 255 g
- Housing: in groups of five in Makrolon type-4 cages with wire mesh tops up to the day of randomization, afterwards individually in Makrolon type-3 cages
- Diet: Pelleted standard Harlan Teklad 2018C rodent maintenance diet, ad libitum
- Water: Community tap-water, ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
End of April until start of May the temperature was occasionally reduced up to 17.9 °C maximally due to technical problems.
- Humidity (%): 30 - 70
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
The dose formulations were prepared weekly using the test item as supplied by the Sponsor. Amines, tri-C8-10-alkyl were be weighed into a glass beaker on a tared precision balance and the vehicle were added (w/v). Using an amagnetic stirrer, a homogeneous solution were prepared. Separate formulations were prepared for each concentration. Homogeneity of the test item in the vehicle was maintained during the daily administration period using a magnetic stirrer.

Dose formulations were stored at room temperature (15 - 25°C) in glass beakers. Based upon the results of stability analyses performed within the Harlan Laboratories study no. D71061 for at least 10 days at room temperature (15 - 25°C).

Animals were treated orallay by gavage. The applicated volume was 4ml/kg body weight diluted in corn oil. Dosages are 0, 50, 150 and 400 mg/kg bw/d.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
On the first dose formulation day samples from the control group as well as three samples (top, middle and bottom) of about 1 g of each concentration were taken prior to dosing for analysis of concentration and homogeneity. Samples of about 1 g of each test item concentration were taken from the middle to confirm the stability (8 and 10 days at room temperature 15-25°C). Towards the end of the study, samples were taken from the middle to confirm concentration. The aliquots for analysis of dose formulations were frozen (-20 ± 5 °C) and delivered on dry ice to Dr. B. Ludwig (Harlan Laboratories Ltd., Zelgliweg 1, 4452 Itingen / Switzerland) and stored there at -20 ± 5 °C until analysis.
The samples will be analyzed by HPLC-ELSD. The test item will be used as the analytical standard.
Duration of treatment / exposure:
males: approximately 6 weeks
females: approximately 6 weeks
Frequency of treatment:
daily, 7 days/week
Remarks:
Doses / Concentrations:
50, 150, 400 mg/kg bw
Basis:
actual ingested
No. of animals per sex per dose:
12
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Based on the results of a 14-day range-finding study (BASF project number 01R0674/12X381) the dose levels of 50, 150, and 400 mg/kg bw per day were selected for the present study.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once daily
Viability/Mortality: twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once weekly
Once prior to the first administration of the test item (day 6 of acclimatization) and weekly thereafter (in the gestation period on day 0, 6, 13 and 20 post coitum), detailed clinical observations were performed outside the home cage in a standard arena. Animals were observed for the following: changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies or bizarre behavior were also reported.

BODY WEIGHT: Yes
- Time schedule for examinations: daily, from treatment start to day of necropsy

FOOD CONSUMPTION:
- Food consumption for each animal determined: Males: Pre-pairing period days 1 - 4, 4 - 8, 8 - 11 and 11 – 14, after pairing period days 1 – 7 and 7-10. Females: Pre-pairing period days 1 - 4, 4 - 8, 8 - 11 and 11 - 14; gestation days 0 – 7, 7 14 and 14 – 21 and days 1 - 4 of the lactation. No food consumption was recorded during the pairing period.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: from males on day of necropsy and from females on day 5 post partum
- Anaesthetic used for blood collection: Yes (isofluorane)
- Animals fasted: Yes
- How many animals: 5 males and 5 females from each group
- Parameters checked: Complete Blood Cell Count: Erythrocyte count, Hemoglobin, Hematocrit, Mean corpuscular volume, Red cell volume distribution width, Mean corpuscular hemoglobin, Mean corpuscular hemoglobin concentration, Hemoglobin concentration distribution width, Leukocyte count (total), Differential leukocyte count: Platelet count & Reticulocytes; Coagulation: Prothrombin time (= Thromboplastin time), Activated partial Thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: from males of each group on day of necropsy and from females from each group on day 5 post partum
- Animals fasted: Yes
- How many animals: 5 males and 5 females from each group
- Parameters checked: Glucose, Urea, Creatinine, Bilirubin (total), Cholesterol (total), Triglycerides, Aspartate aminotransferase, Alanine aminotransferase, Alkaline phosphatase, Gamma-glutamyl-transferase, Bile acids, Sodium, Potassium, Chloride, Calcium, Phosphorus, Protein (total), Albumin, Globulin, Albumin/Globulin ratio

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: at one time during the study (males shortly before the scheduled sacrifice and in females of groups 1 and 2 on day 3 post partum and in females of groups 3 and 4 on day 23 or 24 post coitum)
- Dose groups that were examined: 5 males and 5 females randomly selected from each group
- Battery of functions tested:
• Cage-side observations: faeces-balls, urine and posture as well as resistance to removal.
• Hand-held observations: muscle tone, constitution, skin, pupil size, palpebral closure, lacrimation, salivation, reaction to handling and general abnormalities.
• Open field observations: level of ambulatory activity including rearing (one minute evaluation), unusual body movements (e.g. spasms, convulsions), gait evaluation, behavior, hair coat, respiration, quantity of faeces-balls and urine.
• Reflexes: blinking, palpebral closure, pinna reflex, extensor thrust response, paw pinch, responsiveness to sharp noise, righting reflex and hearing ability (Preyer’s reflex).
• Measurements / Counts: hind limb / fore limb grip strength, rectal temperature.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
At the scheduled sacrifice, the testes and epididymides of all parental males were weighed separately.
In addition, from 5 males and 5 females killed at the end of the study which were selected for hematology and clinical chemistry examination from each group, the following organs were trimmed from any adherent tissue, as appropriate, and their wet weight taken:
Adrenal glands (weighed as pairs), brain, heart, kidneys (weighed as pairs), uterus (including cervix), prostate, liver, thymus, spleen, thyroid (after fixation), ovaries (weighed as pairs), seminal vesicles (inclusive coagulating gland)

The following tissues from all parental males were preserved in neutral phosphate buffered 4% formaldehyde solution:
Prostate, Seminal vesicles with coagulating gland, Testes, Epididymides (males); Ovaries, Uterus (females);

In addition, from all males and females the following tissues were preserved in neutral phosphate buffered 4% formaldehyde solution:
Gross lesions, Brain, Spinal chord (cervical, thoracic, lumbar), Small and large intestines (incl. Peyer’spatches), Stomach (forestomach and glandular stomach), Liver, Kidneys, Adrenals, Spleen, Lymph nodes (axillary and mesenteric),Urinary bladder, Aorta, Eyes with optic nerve and harderian gland, Lacrimal gland, Larynx, Nasal caity, Esophagus, Heart, Thymus, Thyroids and parathyroids, Trachea and lungs (preserved by inflation with fixative and then immersion), Pituitary gland, Spleen, Peripheral nerve (sciatic), Bone marrow (femur), Femur with knee joint, Mammary gland (male and female), Pancreas, Salivary glands – mandibular, sublingual, Skeletal muscle, Sternum with bone marrow, Pharynx

All organ and tissue samples to be examined by the study pathologist were processed, embedded and cut at an approximate thickness of 2 - 4 micrometers and stained with hematoxylin and eosin. Additionally, the testis was stained by PAS-hematoxylin.

Testes, epididymides, prostate, seminal vesicles, ovaries, oviduct, vagina and uterus from all animals of the control and high-dose group were examined. The same applied to all occurring gross lesions. The remaining organs/tissues of 5 randomly selected males and females of the control and high-dose group, respectively, were examined histopathologically.

Special emphasis was made on the stages of spermatogenesis and histopathology of interstitial cell structure.

Histological examination of ovaries was carried out on the females that did not give birth.

A histopathology peer review was performed by W. Henderson from Harlan Laboratories Ltd. The phase report is included in Appendix VII on p.{ DA }.

ORGAN WEIGHTS:
At the scheduled sacrifice, the testes and epididymides of all parental males will be weighed separately.
In addition, from 5 males and 5 females killed at the end of the study which were selected for hematology and clinical chemistry examination from each group, the following organs will be trimmed from any adherent tissue, as appropriate, and their wet weight taken: Adrenal glands (weighed as pairs), Brain, Heart, Kidneys (weighed as pairs), Uterus (including cervix), Prostate, Liver, Thymus, Spleen, Thyroid (after fixation), Ovaries (weighed as pairs), Seminal vesicles (inclusive coagulating gland)
Statistics:
The following statistical methods were used to analyze food consumption, body and organ weights, clinical laboratory and reproduction data, locomotor activity and macroscopical findings:
• Means and standard deviations of various data were calculated.
• The Dunnett-test [see References (2)] (many to one t-test) based on a pooled variance estimate was applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.
• The Steel-test [see References (3)] (many-one rank test) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.
• Fisher's exact-test [see References (4)] was applied if the variables could be dichotomized without loss of information.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
no unplanned deaths; for more informations see details on results.
Mortality:
mortality observed, treatment-related
Description (incidence):
no unplanned deaths; for more informations see details on results.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
see details on results
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
see details on results
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
see details on results
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
see details on results
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
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
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
see details on results
Histopathological findings: neoplastic:
not specified
Details on results:
CLINICAL SIGNS AND MORTALITY
There were no unplanned deaths.
In two males at 400 mg/kg bw/day, slightly ruffled fur was noted during six consecutive days during pairing. This was accompanied by reddish nasal secretion in both animals for two consecutive days. One male at 50 mg/kg bw/day also showed one instance of reddish nasal secretion towards the middle of the after pairing. No further clinical signs were noted in males at these dose levels.
Due to the low number of animals affected, it could be deemed that this effect was not adverse although it cannot be excluded that it was a signs of discomfort associated with treatment with the test item.
Incidentally, in one male at 400 mg/kg bw/day a left eye slightly reduced in size was noted from the last two days of pre-pairing onwards.
No clinical signs were noted in males at 0 or 150 mg/kg bw/day.
At 400 mg/kg bw/day, one female showed bedding in mouth and salivation on days 5 to 7 of gestation, which was similar to the males and considered rather as sign of discomfort associated with treatment with the test item than an adverse effect of the test item.
One female at 150 mg/kg bw/day had vaginal bleeding on the last day of pairing and the first day of gestation period. This isolated finding and vaginal to this early time point was considered to be incidental.
In females, no clinical signs were noted at 0 and 50 mg/kg bw/day.

BODY WEIGHT AND WEIGHT GAIN
In males treated at 400 mg/kg bw/day, body weights and body weight gain were lower than controls during the major part of the study after onset of treatment at the beginning of the pairing period. This effect reached statistical significance on several occasions during the study. This was consistent with the statistically significant reduction in food consumption noted in males during pre-pairing.
The overall differences in mean body weight gain at the dose levels of 0, 50, 150 and 400 mg/kg bw/day were: +11%, +12%, +11% and +6% during the pre-pairing period, +10%, +10%, +11% and +8% during the pairing period and +1%, +2%, +3% and +3% during the after pairing period (percentages refer to the body weight gain within the period).
In females at 400 mg/kg bw/day, reduced body weight gain was noted from the beginning of pre-pairing onwards. This was similar to the effect in males at this dose level, although in females no statistically significant reduction in body weights was noted.
In females at 50 mg/kg bw/day, there was a tendency towards higher body weights when compared with controls however mean body weight gain was similar in the gestation period and reduced in the gestation period (statistically significant on single days). Higher body weights were considered to be not an effect of treatment with the test item, since the difference was already noted during acclimatization and the start of pre-pairing.
The overall differences in mean body weight gain at the dose levels of 0, 50, 150 and 400 mg/kg bw/day were +6%, +7%, +8% and +4% during the pre-pairing period, +59% and +52% during gestation and +3% and +3% during the lactation period (percentages refer to the body weight gain within the period) at a dose level of 0 and 50 mg/kg bw/day.

FOOD CONSUMPTION AND COMPOUND INTAKE
In males treated at 400 mg/kg bw/day, mean food consumption was statistically significantly lower than controls during the major part of pre-pairing (up to -17% between days 4 to 8). However, towards the end of pre-pairing and during the remainder of the study, no statistically significant effect on food consumption was noted in males at 400 mg/kg bw/day. Therefore, this transient reduction in food consumption was deemed not to be adverse. No effect of treatment was noted in males at 150 and 50 mg/kg bw/day.
At 400 mg/kg bw/day during pre-pairing, a statistically significant reduction in food consumption (up to 20.5% between days 4 to 11) was noted. Since the reduction was less marked towards the end of pre-pairing and in analogy with the findings in males it can be deemed that this effect would have been reversible and not adverse. No effect of treatment was noted in females at 150 and 50 mg/kg bw/day.

HAEMATOLOGY
In males at 400 mg/kg bw/day percentage and number of neutrophils were statistically significantly higher from controls and outside the range of the historical control data (28% versusu 12% in controls, 1.8-fold increase in number), with a concomitant decrease in the percentage of lymphocytes. This increase could be test item-related, since histopathologically inflammation in the heart and macrophages in the lung were found. Furthermore in males at 400 mg/kg bw/day, hemoglobin and hematokrit values were statistically significantly lower than controls and outside the range of the historical control data. However, these alterations were considered to be of no toxicological relevance, since the values were borderline to the historical control data, evident in one gender only and/or with less statistical power. Other statistically significantly differences (lower mean corpuscular hemoglobin and basophils levels) were in the range of the historical control data. For females, all values were in the range of the historical control data.

CLINICAL CHEMISTRY
Higher concentrations of aspartate aminotransferase (not statistically significantly different), of alanine aminotransferase (males only) and of alkaline phosphatase were noted in both genders at 400 mg/kg bw/day. In both genders, one male and one female, showed high activities. A pale discolored liver was noted in the male animal during necropsy. However, the increased liver values were considered to be test item-related. Other statistically significant differences (creatinine, potassium, chloride, protein and albumin) were in the range of the historical control data. One female at 400 mg/kg bw/day, showed unusually high values in several parameters.

ORGAN WEIGHTS
In males, statistically significantly reduced absolute testes and epididymides were noted at a dose level of 400 mg/kg bw/day. Calculated relative to body weights, the epididymides weights were also statistically significant reduced. The values for testes and epidymides were in the range of the historical control data (at the lower limit) but due to histopathological findings in testes and epididymides, these reductions were considered to be test item-related.
In females, no test item-related effects on organ weights were noted.

GROSS PATHOLOGY
At 150 and 400 mg/kg bw/d, testes were flaccid and/or reduced in size in single males, which was considered to be test item-related. One male at 400 mg/kg bw/d, had a pale discolored liver, which correlates to his high values of aspartate aminotransferase and alanine aminotransferase.
All other findings in males and females were typical of this strain and age of rat and were considered to be incidental.

HISTOPATHOLOGY: NON-NEOPLASTIC
At 50, 150 and 400 mg/kg bw/day in the heart, a minimal to moderate multifocal dose-dependent cardiomyopathy (myocardial necrosis and/or degeneration with mononuclear inflammatory infiltration, and often accompanied by fibrosis/fibroplasias) was observed in the majority male and female rats. The males showed higher incidence and severity than the females.
Minimal to moderate foamy macrophages accumulation was observed at 50, 150 and 400 mg/kg bw/day in the lungs. The distribution of the lesion was focal to multifocal, and was mainly in the alveolar spaces around bronchioles and/or in terminal bronchioles. The males appeared to be more affected in incidence and severity.
Minimal to moderate increase in the formation of the residual bodies of abnormal shape and size was observed at 150 and 400 mg/kg bw/day. In the epididymides, the exfoliated residual bodies (cell debri) were noted at 400 mg/kg bw/day.

OTHER FINDINGS
In males at 400 mg/kg bw/day, the incidental reduction in eye size already noted during daily clinical signs was recorded. Another male showed salivation, which was also observed in one female of this dose level during the daily clinical signs observation. No further findings were noted at functional observational battery test.
In females, statistically significantly lower locomotor activity was noted at 50 mg/kg bw/day. Although no statistics could be applied to results from females treated at 150 and 400 mg/kg bw/day, a similar lower locomotor activity towards the end of the measurement period was observed. However, since this difference was only due to low activity in individual animals rather than to a trend within the whole dose group and since the values were borderline to historical control data, this finding was deemed incidental and unrelated to treatment with the test item.








Dose descriptor:
LOAEL
Remarks:
systemic
Effect level:
<= 50 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Specific target organ toxicity: heart (cardiomyopathy)
Critical effects observed:
not specified
Conclusions:
Based on the myocardiopathy found, a LOAEL of 50 mg/kg bw/d was determined.
Executive summary:

A study investigating the toxicological effects of the test item Amines, tri-C8-10-alkyl to rats according to OECD guideline 422 resulting from repeated oral-gavage administration is performed. Amines, tri-C8-10-alkyl was administered in corn oil as vehicle at dosages of 50, 150 and 400 mg/kg body weight/day, and controls received the vehicle only. Amines, tri-C8-10-alkyl was administered to male rats for 43 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 postpartum.

All animals survived until scheduled necropsy. There were no test item related clinical signs in both genders at 50 mg/kg bw/day. At 150 mg/kg bw/day, no signs of discomfort were observed; food consumption and body weights were similar to controls. A reduction in food consumption and body weight was noted in males and females at 400 mg/kg bw/day when compared with controls. Furthermore, higher values of aspartate aminotransferase, alanine aminotransferase and of alkaline phosphatase were noted in both genders at 400 mg/kg bw/day, without a histopathological correlate in the liver.

At a dose level of 150 and 4000 mg/kg bw/day, no female gave birth. Females had only implantation sites (minority of females) or embryonic resorptions (majority of the females). At 50 mg/kg bw/day, a higher mean incidence of post-implantation loss was noted, which was not statistically significantly different compared to the control group but out of the range of historical control data. Due to the total post implantation loss at 150 and 400 mg/kg bw/day, it cannot be excluded, that this was a test-item-related effect.

In males, reduced testes and epididymides weights were noted at 400 mg/kg bw/day. Microscopically, minimal to moderate increase in the formation of the residual bodies of abnormal shape and size was observed in the testes at 150 and 400 mg/kg bw/day. In the

epididymides, the exfoliated residual bodies (cell debri) were noted at 400 mg/kg bw/day.

 

In all treatment groups (50, 150 and 400 mg/kg bw/day), cardiomyopathy was noted in the heart, and foamy macrophages accumulation was observed in the lungs. The males showed higher incidence and severity than the females, respectively.

 

Based on these results, no NOAEL (No Observed Adverse Effect Level) could be established for general toxicity and reproduction in this study. Nevertheless, according to the applicant a LOAEL of 50 mg/kg bw/d based on cardiomyopathy in males has to be established and used for calculation.

 

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

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

There are valid sub acute / chronic study data available to assess the repeated dose toxicity of Amines, tri-C8- 10-alkyl.

Studies according to OECD guidelines:

A study investigating the toxicological effects of the test item Amines, tri-C8-10-alkyl to rats according to OECD guideline 422 resulting from repeated oral-gavage administration is performed (BASF SE, 2013, 85R0674/12X380). Amines, tri-C8-10-alkyl was administered in corn oil as vehicle at dosages of 50, 150 and 400 mg/kg body weight/day, and controls received the vehicle only. Amines, tri-C8-10-alkyl was administered to male rats for 43 days and to female rats for 14 days prior to pairing, through the pairing and gestation periods until the F1 generation reached day 4 postpartum.

All animals survived until scheduled necropsy. There were no test item related clinical signs in both genders at 50 mg/kg bw/day. At 150 mg/kg bw/day, no signs of discomfort were observed; food consumption and body weights were similar to controls. A reduction in food consumption and body weight was noted in males and females at 400 mg/kg bw/day when compared with controls. Furthermore, higher values of aspartate aminotransferase, alanine aminotransferase and of alkaline phosphatase were noted in both genders at 400 mg/kg bw/day, without a histopathological correlate in the liver.

At a dose level of 150 and 400 mg/kg bw/day, no female gave birth. Females had only implantation sites (minority of females) or embryonic resorptions (majority of the females). At 50 mg/kg bw/day, a higher mean incidence of post-implantation loss was noted, which was not statistically significantly different compared to the control group but out of the range of historical control data. Due to the total post implantation loss at 150 and 400 mg/kg bw/day, it cannot be excluded, that this was a test-item-related effect.

In males, reduced testes and epididymides weights were noted at 400 mg/kg bw/day.Microscopically, minimal to moderate increase in the formation of the residual bodies ofabnormal shape and size was observed in the testes at 150 and 400 mg/kg bw/day. In the epididymides, the exfoliated residual bodies (cell debri) were noted at 400 mg/kg bw/day.

 In all treatment groups (50, 150 and 400 mg/kg bw/day), cardiomyopathy was noted in the heart, and foamy macrophages accumulation was observed in the lungs. The males showed higher incidence and severity than the females, respectively.

Based on these results, no NOAEL (No Observed Adverse Effect Level) could be established for general toxicity and reproduction in this study. Nevertheless, according to the applicant a LOAEL of 50 mg/kg bw/d based on cardiomyopathy in males is established and used for calculation. As the effects observed show a dose reponse it is assumed that the effects decrease to a normal range (compareable to the controls, based on the trend observed from 400 to 50 mg/kg bw/d) when lowering the dosage to 5 mg/kg bw. To adress this assumption in Risk assessment a factor for LOAEL to NOAEL conversion of 10 is added.

Furthermore these assumption is used for the classification of the substance.

 

Key study assignment:

As there is only one reliable and relevant study investigating the subacute effects of Amines, tri-C8-10-alkyl

this study is integrated as key study.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
only available reliable result

Justification for classification or non-classification

Based on the result presented above, no NOAEL could be established for general toxicity in this study.


The LOAEL was 50mg/kg bw/d, which was the lowest concentration tested and a dose response correlation was observed by the leading effects (cardiomyopathy). Due to the trend in dose response it is assumed that these effects observed may be decreased if the dose will be reduced to 5 mg/kg bw/d. Therefore this worst case value is used as threshold for classification.


 


Based on the classification criteria of Directive 67/548/EEC and Regulation (EC) No 1272/2008 the substance has therefore labeled as:


 


GHS: STOT RE 1, H372: Causes damage to organs (heart) through prolonged or repeated exposure if swallowed.


DSD: T, R48/25 Toxic: danger of serious damage to health by prolonged exposure if swallowed.