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Administrative data

Description of key information

90 d NOAEL = 300 mg/kg bw/d (subchronic (90 day) repeated dose toxicity study, OECD TG 408, rat m/f, oral: gavage), RL1; GLP

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:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1990-03-19 to 1990-07-26
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Qualifier:
according to guideline
Guideline:
EU Method B.26 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Rodents)
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
The test item was suspended in the vehicle to the appropriate dose levels and was administered orally by gavage, at a constant volume of 10 mL/kg bw/day
The amount of the test item was adjusted to each animal’s actual body weight weekly.
The test item-vehicle mixture was freshly prepared every day, in consideration of test substance density.

VEHICLE
- distilled water
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
95 or 96 days exposure period, treatment 5 days/week (monday to friday), in total 68 or 69 applications (depending on section date)
Recovery period: Additional animals in exposure groups 1 and 4 (control and high dose group) were leaved untreated for 35 days after finalisation of the exposure to determine the reversibility of possible compound related findings.
Frequency of treatment:
once daily (5 days per week (monday to friday))
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10 males and 10 females per group
recovery groups: in exposure groups 1 (control) and 4 (high dose group) additional 5 males and 5 females, each
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: testing up to and including the guideline limit dose of 1000 mg/kg bw/day
- Rationale for animal assignment (if not random): animal assignment by random number
- Rationale for selecting satellite groups: determination of the reveribility of possible compound related findings
- Post-exposure recovery period in satellite groups: 35 day recovery period; 5 males and 5 females per group for control and high-dose group.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily,

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: twice daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION : Yes
Food consumption per cage on a weekly basis.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION : yes
Drinking water consumption per cage on a weekly basis

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: on day before section
- Dose groups that were examined: group 1 and 4
The eyes of all animals were examined with a slit lamp Topcon SL-S D, Fa. Bon, D-Wahlstedt, Germany. Prior to examination mydriasis was produced
by instillation of Mydriaticum Roche R (Hoffmann-La Roche, Grenzach-Wyhlen)

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 6 weeks of treatment and at study termination
- Anaesthetic used for blood collection: no data
- Animals fasted: no data
- How many animals: all
- Parameters examined: erythrocytes, haematocrit value, mean corpuscular volume (MCV), haemoglobin content, leucocytes, platelets, differential blood count (neutrophilic stab granulocytes, neutrophilic segmented granulocytes, lymphocytes, eosinophiles, monocytes, basophiles, myelocytes, juveniles, other)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after 6 weeks of treatment and at study termination
- Animals fasted: no data
- How many animals: all
- Parameters examined in serum:
-- kidney parameter: urea, sodium, potassium, creatinin
-- adrenal gland parameter: glucose, sodium, potassium
-- mineral metabolism and parathyroid parameter: calcium, alkaline phosphatase
-- liver parameter: aspartate aminotransferase, alanine aminotransferase, gamma glutamyl transferase, bilirubin
-- electrolyte and water metabolism: sodium, potassium, chloride, protein total
-- lipid metabolism: cholesterol

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
All animals were sacrificed by ether overdose, exsanguinated, thereafter subcutis, abdomen, thoracic and cranial cavity were dissected, organs were
inspected macroscopically in situ and after removal, weighed and fixed in 4% neutral formaline; eye balls were fixed by method of Davidson.
Relative and absolute organ weights of brain, testicles, heart, liver, spleen, adrenal glands, kidneys and thymus were recorded.

Animals of recovery group were pathologically examined in the same way after a 35 day treatment–free interval.

HISTOPATHOLOGY: Yes
- The following organs of 9 male and 10 female animals of groups 1 and 4 (control and high dose) were examined histologically after haematoxylin-
eosin staining: aorta thoracica, eyes, large intestine, small intestine, glandular stomach, cerebrum, urinary bladder, skin, heart, testicles, cerebellum,
liver, trachea, lung, lymph nodes (maxillar and mesenteric), spleen, epididymis, adrenal glands, nerve peripheric, kidneys, ovary, pancreas, prostate, spinal cord (vertebral body), seminal gland, thyroid, salivary gland, oesophagus, skeletal muscles, thymus, uterus, forestomach, tongue.
- In dose groups 2 (low dose, 10 males and 10 females) and in dose group 3 (mid dose, 9 males and 10 females) as well as in the both recovery groups (control and high dose, 5 males and 5 females, each) the following organs were examined histologically: forestomach, liver and urinary bladder (possible target organs)
Statistics:
The following statistical methods were used:
- t-test (body weight, biochemical and hematological parameter)
- Steel-test (organ weights)
Clinical signs:
no effects observed
Description (incidence and severity):
Substance related effects were not observed. The results of the patho-histological examination of animal 34 revealed no substance related effects.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
All animals survived the treatment with the test compound, except animal 34 (male, group 4, 1000 mg/kg bw/day).
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
The total body weight gain in all test groups was comparable to the control, with the exception of animal 34, which lost weight after 11 weeks of treatment.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
The mean food consumption in the male and female treatment groups showed some dose independent variations, which were considered not to be treatment related.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
effects observed, non-treatment-related
Description (incidence and severity):
The mean water consumption in the male and female treatment groups showed some dose independent variations, which were considered not to be treatment related.
Ophthalmological findings:
no effects observed
Description (incidence and severity):
The examination of the eyes with a slit lamp microscope showed no compound related effects.
Haematological findings:
no effects observed
Description (incidence and severity):
The haematological examinations revealed a decrease of the MCV for the female group 4. As the value lies in the normal range for animals of this strain, this observation was considered not to be substance related. In all other groups the parameters were comparable to the control.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
The biochemical examinations revealed some compound and dose independent variations which were within historical controls and therefore considered not to be relevant. The significant increase of the ALT-values (alanine-aminotransferase) in the male and female animals of the group 4 are considered to be possibly indicative for substance related liver effects.
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
The relative organ weights of the heart showed some slight variations which were in the normal range for animals of this strain and were therefore considered not to be treatment related. Absolute organ weights were comparable to controls.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
The macroscopical examination of the organs displayed in all groups some findings like hydrometra and hydronephrosis. Enlargement of the mandibulary lymph nodes in all groups including controls and recovery groups were considered to be related to the bacterial infection (Tyzzer´s Disease).
The observed edema of the forestomach mucosa in the highest dose group (2/9 males and 3/10 females) were considered to be a reaction due to the irritating properties of the test compound.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
- The microscopical examination revealed substance-related effects in the highest dose group. These effects were located in the forestomach and the urinary bladder.
--The mucosa of the forestomach of 2/9 male and 3/10 female animals of group 4 showed effects indicating local irritation. These effects were not seen in the animals of the group 2 and 3 as well as in the animals of the recovery group 4.
-- In 6/9 male animals of the treatment group 4 the urinary bladder showed increased desquamation and localized regressive changes of the epithelium. Kidney, kidney pelvis and urether were free of such symptoms and therefore the effects on the urinary bladder epithelium are considered to be a result of local physiological accumulation of the test compound or possible metabolites. The urinary bladders of the animals of the recovery group 4 and the treatment group 2 and 3 were free of such effects.

A bacterial infection (Tyzzer´s Disease induced by Clostridium piliforme) occurred in all groups, including control and recovery group. The following effect were considered to be related to this infection:

- In the male and female animals of all groups the livers showed effects which were considered to be a result of this bacteriosis.
- The macroscopical and the microscopical findings (germinal hyperplasia) at the mandibulary lymph node can be interpreted as a consequence of this bacteriosis.

- Histopathological findings in reproductive organs: The following reproductive organs were histopathologically examined: testes, epididymis, prostata, seminal vesicle, ovary and uterus. No abnormalities were detected.

- Reversibility of effects: The treatment related histopatholgical alterations of the forestomach and the urinary bladder seen at termination of treatment in study group 4 (high dose) were completely reversible in the high dose recovery group .
Histopathological findings: neoplastic:
not examined
Key result
Dose descriptor:
NOEL
Effect level:
300 mg/kg bw/day (actual dose received)
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Critical effects observed:
yes
Lowest effective dose / conc.:
1 000 mg/kg bw/day (actual dose received)
System:
urinary
Organ:
bladder
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
yes
Critical effects observed:
yes
Lowest effective dose / conc.:
1 000 mg/kg bw/day (actual dose received)
System:
gastrointestinal tract
Organ:
other: forestomach
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
no

The interpretation of this study is somewhat hampered due the occurrence of a bacterial infection (Tyzzer´s Disease induced by Clostridium piliforme) in all dose groups. Macroscopical or microscopical findings in the mandibular lymph node and the liver in all dose groups including the control were judged by the study authors as directly related to this infection. Bacterial foci were verified in the liver.

Apart from these effects, animals of the high dose groups displayed potentially substance related increases of the activity of the alanine transaminase (ALT) in blood serum, signs of forestomach irritation and regressive epithelial changes in the urine bladder. However, an interaction with the bacterial infection cannot be entirely excluded.

The significant increase of the ALT-values in the male and female animals of the high dose group was considered to be indicative for a substance related liver effect by the study author. However, a confounding effect by the infection with Clostridium piliforme is likely. Beside the intestines and the myocardium, the liver is a common target organ of an infection by Clostridium piliforme (Tyzzer´s Disease). Typical degenerative histopathological findings associated with the infection were seen in all dose groups including the control. Therefore, an induction of the enzyme alanine aminotransferase as a marker of liver damage in addition to these histopathological alterations is likely.

However, the histopathological liver findings and thus the induction of an enzyme commonly associated with pathological liver alterations most likely primarily induced by the infection with Clostridium piliforme might have been intensified by the substance application.

The observed edema of the forestomach mucosa in 2/9 male and 3/10 female animals of the highest dose group were considered to be a reaction due to the irritating properties of the test substance.

35 days after termination of the treatment, the animals of the high dose recovery group were free of any forestomach findings. Forestomach findings related to an irritant activity of the test item are common findings in rat gavage studies. The stomach of rats is anatomically different from the human stomach. The rat stomach consists of two anatomically distinct parts of approximately equal size: a non-glandular forestomach and a glandular stomach. The forestomach is connected to the oesophagus at the gastro-oesophageal junction, and is clearly separated from the glandular stomach by a distinct border called the limiting ridge. The forestomach is not present in humans. The main function of the rat forestomach is storage and trituration of ingested food prior to digestion in the glandular stomach. The forestomach is a holding compartment and due to this function a long exposure time to orally - especially by gavage as bolus dose - administered doses in this organ occurs. Thus, an irritant test item can easily take effect at the forestomach squamous epithel. The forstomach injury due to irritant test items is usually a subject to reversibility. Also in this study, a complete reversibility of the forestomach effects has been seen in the recovery group left 35 days untreated after termination of the substance application. A functional equivalent to the rat forestomach is missing in humans. A histological similar in humans is the oesophagus, because the epithelium of the human oesophagus is morphologically of the same type as the forestomach epithelium of rats. However, tissue exposure in the human oesophagus is likely to be minimal compared to tissue exposure in the rat forestomach since the epithelial contact is much shorter in the human oesophagus compared to the rat forestomach and it seems very unlikely that exposure to concentrations far below those having an irritating potential is hazardous to man. Therefore, forestomach findings in rat studies resulting from irritation associated with high-concentration gavage dosing should not form the basis for determining systemic toxicity or quantitative systemic toxicity potency estimates. They are attributable to the specific anatomy of the test species and the non-physiological bolus application by gavage and therefore judged as not relevant in view of a potential serious health risk for humans.

In 6/9 male animals of the high dose group the urinary bladder showed increased desquamation and localized regressive changes of the epithelium. The effects on the urinary bladder epithelium were considered by the study author to be a result of local physiological accumulation of the test compound or possible metabolites. A complete reversibility of the urinary bladder effects has been seen in the high dose recovery group left 35 days untreated after termination of the substance application. Due to this complete reversibility, the effects on the urinary bladder epithelium seen only in the high dose male animals can be judged as transient and do not pose a severe adverse health risk relevant to humans.

 

The histological examination including the reproductive organs (testes, epididymis, prostata, seminal vesicle and uterus) did not reveal any treatment related abnormalities.

On the basis of this study, a NOEL of 300 mg/kg bw/day can be derived.

Conclusions:
The interpretation of this study is somewhat hampered due the occurrence of a bacterial infection (Tyzzer's Disease) in all dose groups. Some of the macroscopical or microscopical findings were directly related to this infection by the study investigators. Apart from these effects, animals of the high dose groups displayed potentially substance related increases of blood liver enzyme (ALT), signs of gastric (forestomach) irritation and
regressive epithelial changes in the urine bladder. The histopathological effects were completely reversible within a recovery period of 35 days.
The NOEL for the partially unsaturated TEA-Esterquat (90 % a.i.) was determined to be 300 mg/kg bw/day.
Executive summary:

In a subchronic toxicity study comparable to OECD guideline 408 (1981) the partially unsaturated TEA-Esterquat was administered to 10 Sprague-Dawley rats/sex/dose by gavage at dose levels of 0, 100, 300 and 1000 mg/kg bw/day. The animals were treated 5 days/week and received in total 68 or 69 applications (depending on section date). Study performed according to the OECD guideline 408 of 1981, which allowed for exposure during working days, only, and stalling exposure at weekends. A satellite group of 5 males and 5 female for the control and high dose group was included to assess the reversibility of effects after a 35 day recovery period.

In the experimental animals no clinical effects and no impairment of body weight gain, food and water consumption, hematological parameters, organ or body weights related to the test substance administration were seen. The examination of the eyes with a slit lamp microscope showed no test substance related effects. The interpretation of this study is somewhat hampered due the occurrence of a bacterial infection (Tyzzer´s Disease induced by Clostridium piliforme) in all dose groups, including control and recovery group. Macroscopic or microscopic findings in the mandibular lymph node and the liver in all dose groups including the control were judged as directly related to this infection. Bacterial foci were detected in the liver.

Apart from these effects, animals of the high dose groups displayed potentially substance related increases of the activity of the alanine transaminase (ALT) in blood serum, signs of forestomach irritation and regressive epithelial changes in the urine bladder. However, an interaction with the bacterial infection cannot be entirely excluded. The significant increase of the ALT-values in the male and female animals of the high dose group was considered by the study author to be indicative for substance related liver effects. However, a confounding effect by the infection with Clostridium piliforme is likely. Beside the intestines and the myocardium, the liver is a common target organ of an infection by Clostridium piliforme (Tyzzer´s Disease). Typical degenerative histopathologic findings associated with the infection were seen in all dose groups and the control.

An induction of the enzyme alanine aminotransferase as a marker of liver damage in addition to these histopathologic alterations was evident. The histopathologic liver findings as well as the induction of the enzyme ALT - commonly associated with pathological liver alterations – are most likely due to the infection with Clostridium piliforme might have been intensified by the substance application. The observed edema of the forestomach mucosa in 2/9 male and 3/10 female animals of the highest dose group were considered to be a reaction due to irritating properties of the test substance observed in some (but not all) skin and eye irritation studies. The animals of the high dose recovery group were free of any forestomach findings, 35 days after termination of the treatment. Forestomach findings related to an irritant activity of the test item are common findings in rat gavage studies. They are attributable to the specific anatomy of the test species and the non-physiological bolus application by gavage and therefore judged as not relevant in view of a potential serious health risk for humans.

In 6/9 male animals of the high dose group the urinary bladder showed increased desquamation and localized regressive changes of the epithelium. The effects on the urinary bladder epithelium were considered by the study author to be a result of local physiological accumulation of the test compound or possible metabolites. A complete reversibility of the urinary bladder effects has been seen in the high dose recovery 35 days group. Therefore, the effects on the urinary bladder epithelium seen only in the high dose male animals can be judged as transient and do not pose a severe adverse health risk relevant to humans.

The histological examination of the reproductive organs (testes, epididymis, prostata, seminal vesicle, ovary and uterus) did not reveal any treatment related abnormalities.

On the basis of this study, a NOEL of 300 mg/kg bw/day for the partially unsaturated TEA-Esterquat (90 % a.i.) can be derived.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
300 mg/kg bw/day
Study duration:
subchronic
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

In a subchronic toxicity study according to OECD guideline 407 (1995) and EU method B7 (1996) the test substance partially unsaturated TEA-Esterquat, was administered to 5 CD rats/sex/dose by gavage at dose levels of 0,100, 300 and 1000 mg/kg bw/day for 28 days. A satellite group of 5 males and 5 female for the control and high dose group was included to assess the reversibility of any effects after a 2-week recovery period.


No mortality and no influence on behavior, external appearance, body weight, food and drinking water consumption, the eyes or optic region, the haematological and clinical-biochemical parameter and the relative or absolute organ weights at any of the tested dose levels was noted. No test item-related changes were revealed during neuropharmacological functional observations in any of the dosed groups.


Macroscopic post mortem examination and histopathology including the reproductive organs (epididymis, ovary, prostate, testicle and uterus) revealed no test item related changes in treated animals. There is no evidence for a specific target organ toxicity in this study.


Under test conditions, the NOAEL was above 1000 mg/kg bw /day in this 4- week subchronic toxicity study.


The analytical verification of dosing solutions demonstrated agreement of actual initial concentrations with nominal test concentrations.


 


In a subchronic toxicity study comparable to OECD guideline 408 (1981) partially unsaturated TEA-Esterquat was administered to 10 Sprague-Dawley rats/sex/dose by gavage at dose levels of 0, 100, 300 and 1000 mg/kg bw/day. The animals were treated 5 days/week and received in total 68 or 69 applications (depending on section date). Study performed according to the OECD guideline 408 of 1981, which allowed for exposure during working days, only, and stalling exposure at weekends. A satellite group of 5 males and 5 female for the control and high dose group was included to assess the reversibility of effects after a 35 day recovery period.


In the experimental animals no clinical effects and no impairment of body weight gain, food and water consumption, hematological parameters, organ or body weights related to the test substance administration were seen. The examination of the eyes with a slit lamp microscope showed no test substance related effects. The interpretation of this study is somewhat hampered due the occurrence of a bacterial infection (Tyzzer´s Disease induced by Clostridium piliforme) in all dose groups, including control and recovery group. Macroscopic or microscopic findings in the mandibular lymph node and the liver in all dose groups including the control were judged as directly related to this infection. Bacterial foci were detected in the liver.


Apart from these effects, animals of the high dose groups displayed potentially substance related increases of the activity of the alanine transaminase (ALT) in blood serum, signs of forestomach irritation and regressive epithelial changes in the urine bladder. However, an interaction with the bacterial infection cannot be entirely excluded. The significant increase of the ALT-values in the male and female animals of the high dose group was considered by the study author to be indicative for substance related liver effects. However, a confounding effect by the infection with Clostridium piliforme is likely. Beside the intestines and the myocardium, the liver is a common target organ of an infection by Clostridium piliforme (Tyzzer´s Disease). Typical degenerative histopathologic findings associated with the infection were seen in all dose groups and the control.


An induction of the enzyme alanine aminotransferase as a marker of liver damage in addition to these histopathologic alterations was evident. The histopathologic liver findings as well as the induction of the enzyme ALT - commonly associated with pathological liver alterations – are most likely due to the infection with Clostridium piliforme might have been intensified by the substance application. The observed edema of the forestomach mucosa in 2/9 male and 3/10 female animals of the highest dose group were considered to be a reaction due to irritating properties of the test substance observed in some (but not all) skin and eye irritation studies. The animals of the high dose recovery group were free of any forestomach findings, 35 days after termination of the treatment. Forestomach findings related to an irritant activity of the test item are common findings in rat gavage studies. They are attributable to the specific anatomy of the test species and the non-physiological bolus application by gavage and therefore judged as not relevant in view of a potential serious health risk for humans.


In 6/9 male animals of the high dose group the urinary bladder showed increased desquamation and localized regressive changes of the epithelium. The effects on the urinary bladder epithelium were considered by the study author to be a result of local physiological accumulation of the test compound or possible metabolites. A complete reversibility of the urinary bladder effects has been seen in the high dose recovery 35 days group. Therefore, the effects on the urinary bladder epithelium seen only in the high dose male animals can be judged as transient and do not pose a severe adverse health risk relevant to humans.


The histological examination of the reproductive organs (testes, epididymis, prostata, seminal vesicle, ovary and uterus) did not reveal any treatment related abnormalities.


On the basis of this study, a NOEL of 300 mg/kg bw/day for partially unsaturated TEA-Esterquat (90 % a.i.) can be derived.


 


There are no data gaps for the endpoint repeated dose toxicity. No human data are available. However, there is no reason to believe that these results from rat and rabbits would not be applicable to humans.


 


Similar results were obtained with the source substance MDEA-Esterquat C16-18 and C18 unsatd.: one 28 day repeated dose study comparable to OECD Guideline 407 and one sub-chronic toxicity study comparable to OECD Guideline 408 are available for the oral route of administration. The no effect level (NOEL) was determined to be 500 mg/kg/day (highest dose tested) for both studies. These data are included into the dossier to demonstrate, that both substances have a similar toxicological profile.

Justification for classification or non-classification

Based on the available data, the substance does not need to be classified for repeated dose toxicity according to regulation (EC) 1272/2008. Thus, no labelling is required.