Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Only unsufficient data with Amines, polyethylenepoly-, tetraethylenepentamine fraction regarding repeated dose toxicity by oral route are available. A 90-day study according to OECD 408 inlcuding a developmental/reproduction screening is ongoing. As soon as the data is available the dossier will be updated.

Therefore, studies with the structural analoge substance TETA-2HCl (CAS 38260 -01 -4) are used and considered reliable to evaluate the repeated dose toxicity of Amines, polyethylenepoly-, tetraethylenepentamine fraction.

LOAEL (TETA-2HCl, rat, oral, subchronic) = 50 mg/kg bw/day

Reliable data regarding repeated dose toxicity by dermal route is available:

NOAEL (OECD 410, rabbit, dermal, subacute) systemic >= 200 mg/kg bw/day, local = 1.25 mg/cm²

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Remarks:
Summary of available data used for the endpoint assessment of the target substance
Adequacy of study:
key study
Justification for type of information:
refer to analogue justification provided in IUCLID section 13
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Dose descriptor:
NOAEL
Remarks:
rat
Effect level:
50 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: irreversible toxic changes in the lung
Remarks on result:
other: CAS 38260-01-4, Yanagisawa, 1998
Dose descriptor:
LOAEL
Remarks:
rat
Effect level:
50 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: irreversible toxic changes in the lung
Remarks on result:
other: CAS 38260-01-4, Yanagisawa, 1998
Dose descriptor:
NOAEL
Remarks:
dog
Effect level:
50 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
neuropathology
Remarks on result:
other: CAS 38260-01-4, Maemura, 1998
Dose descriptor:
NOAEL
Remarks:
mice
Effect level:
600 ppm
Based on:
test mat.
Remarks:
corresponds to 92 mg/kg bw
Sex:
male/female
Basis for effect level:
other: see remarks
Remarks on result:
other: CAS 38260-01-4, Greenman, 1996
Dose descriptor:
NOAEL
Remarks:
rats
Effect level:
>= 3 000 ppm
Based on:
test mat.
Remarks:
3000 ppm corresponds to 270 mg/kg bw
Sex:
male
Basis for effect level:
other: No test substance-related effects observed.
Remarks on result:
other: CAS 38260-01-4, Greenman, 1996
Dose descriptor:
NOAEL
Remarks:
rats
Effect level:
600 ppm
Based on:
test mat.
Remarks:
600 ppm corresponds to 60 mg/kg bw/day
Sex:
female
Basis for effect level:
other: uterine
Remarks on result:
other: CAS 38260-01-4, Greenman, 1996
Critical effects observed:
yes
Lowest effective dose / conc.:
50 mg/kg bw/day (actual dose received)
System:
respiratory system: lower respiratory tract
Organ:
lungs
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
not specified
Conclusions:
A 90-day study is availabe with the source substance triethylenetetramine dihydrochloride in mice and rats. Due to inflammation of the lung interstitium, hematopoietic cell proliferation of the spleen, liver periportal fatty infiltration, kidney weight reduction, reduced renal cytoplasmic vacuolization, and body weight gain reduction the NOAEL for mice was set to 92 mg/kg bw/day. In male rats no substance-related effects were observed and in female rats changes in the uterus were observed resulting in NOAEL (male) of 270 mg/kg bw/day and NOAEL (female) = 60 mg/kg bw/day. Sub-chronic treatment (26-weeks) of rats with the source substance triethylenetetramine dihydrochloride was associated with death and irreversible toxic changes in the lung. A dosage of 50 mg/kg bw/day was considered to be the NOAEL for females and for males the NOAEL was considered to be less than 50 mg/kg/day. Sub-chronic treatment (26-weeks) of dogs with the source substance triethylenetetramine dihydrochloride was associated with musculoskeletal effects. A dosage of 50 mg/kg bw/day was considered to be the NOAEL for female and male dogs. As explained in the analogue justification, it is considered that the target and the source substances are unlikely to lead to differences in repeated dose toxicity.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LOAEL
50 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
The available information comprises adequate, reliable and consistent studies, from a reference substances with similar structure and intrinsic properties. Read-across is justified based on similarities in PC/ECO/TOX properties. The selected study is thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.
System:
respiratory system: lower respiratory tract
Organ:
lungs

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

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
April-May 1986
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Purity of a.i. has been indicated; complete composition of the test substance is not available (available in departmental study file).
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 410 (Repeated Dose Dermal Toxicity: 21/28-Day Study)
Version / remarks:
1981
Deviations:
yes
Remarks:
20 instead of 21 days of exposure, occlusive dressing
GLP compliance:
yes
Limit test:
no
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Ray Nichols Rabbitry, Lumberton, TX, USA
- Age at study initiation: ca. 5 months
- Weight at study initiation: ca. 3.1 - 4.0 kg
- Fasting period before study: not applicable
- Housing: individually in stainless steel cages
- Diet: restricted to 4 oz per animal per day
- Water: ad libitum
- Acclimation period: at least 14 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 72 ± 5 degr F (22 ± 1.5 degr C)
- Humidity (%): 50
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12/12


Type of coverage:
occlusive
Vehicle:
water
Details on exposure:
TEST SITE
- Area of exposure: 10 x 15 cm
- % coverage: 100
- Type of wrap if used: an occlusive bandage of absorbent gauze and non-absorbent cotton; the bandage was held in place using a lycra/spandex jacket
- Time intervals for shavings or clipplings: periodically as required

REMOVAL OF TEST SUBSTANCE
- Washing (if done): not reported
- Time after start of exposure: ca. 6 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): a 10% solution was applied with the volume adjusted to provide the required dose
- Concentration (if solution): 10%
- Constant volume or concentration used: yes, constant concentration

VEHICLE
- Justification for use and choice of vehicle (if other than water): water
- Amount(s) applied (volume or weight with unit): 1 mL/kg BW

USE OF RESTRAINERS FOR PREVENTING INGESTION: no (but animals wore jackets)
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Test material concentration (100 mg/g) was measured 3 times during the study in triplo. % of target concentrations were 87.0, 95.3 and 90.9%, respectively. Stability was measured 2, 4 and 7 days after preparation. % of Day 0 was, 92.1, 59.4 and 59.9%, respectively.
Duration of treatment / exposure:
20 days
Frequency of treatment:
5 days/week, ca. 6h/day
Dose / conc.:
50 mg/kg bw/day (nominal)
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
200 mg/kg bw/day (nominal)
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on a probe study using 2 female rabbits per group, at levels of 0, 50, 100 or 200 mg/kg bw/day for 4 days
- Rationale for animal assignment (if not random): computer-generated tables of random numbers
- Rationale for selecting satellite groups: not used
- Post-exposure recovery period in satellite groups: not used
- Section schedule rationale (if not random): no info
Positive control:
not used
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily on working days (one additional routine monitoring on these days and on weekend and holidays for dead animals and check for water and food availability)

DETAILED CLINICAL OBSERVATIONS: No

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: daily upon removal when bandage and jacket were removed.

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION: Since the animals were maintained on a restricted diet and normally ate all their ration, measurement of food consumption was not doen. Qualitative changes in consumption were monitored by a periodic visual check.

FOOD EFFICIENCY: No

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: not applicable

HAEMATOLOGY: Yes
- Time schedule for collection of blood: immediately prior to necropsy
- Anaesthetic used for blood collection: Yes (no info which was used)
- Animals fasted: No
- How many animals: all
- Parameters examined: RBC, WPC, platelet count, haemoglobin, packed cell volume

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: immediately prior to necropsy
- Animals fasted: No
- How many animals: all
- Parameters examined: blood urea nitrogen, ALP, ALAT, glucose, total protein, albumin, Cl, Na, K, globulin (calculated)

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (including eyes)
HISTOPATHOLOGY: Yes (complete set) of control and high dose animals; skin (application site and a non-treated section), liver kidneys and gross pathologic lesions were aslo examined from low and mid dose animals
Organ weights: brain, liver, kidneys, adrenals, ovaries, testes
Other examinations:
None
Statistics:
Yes, for body weights, organ weights, clinical chemistry, haematology.
Clinical signs:
no effects observed
Dermal irritation:
effects observed, treatment-related
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
effects observed, treatment-related
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Other effects:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY: one control female was removed from the study on day 14 because of a broken back.
There were no signs other than local dermal effects. These were dose-related and consisted of red dermal test sites on days 1-2. By study Day 6, animals tretaed with 200 mg/kg bw showed very red and slightly swollen test sites; on Day 16 the skin of some of these females was severely irritated with some crusting and bleeding. Other animals of the 200 mg/kg bw group and those of the 100 mg/kg group had irritated skin that did not progress further.

BODY WEIGHT AND WEIGHT GAIN: no effects

FOOD CONSUMPTION: no effects

FOOD EFFICIENCY: not measured

WATER CONSUMPTION: not measured

OPHTHALMOSCOPIC EXAMINATION: not applicable

HAEMATOLOGY: no effects

CLINICAL CHEMISTRY: no effects

URINALYSIS: not measured

NEUROBEHAVIOUR: not measured

ORGAN WEIGHTS: no effects

GROSS PATHOLOGY: confined to the skin of the 100 and 200 mg/kg groups, characterised by multifocal areas of epidermal and dermal necrosis often covered with crusts comprised of dry serum and necrotic debris.

HISTOPATHOLOGY: NON-NEOPLASTIC
Only dose-related changes in the skin consisting of: multifocal necrosis of the epidermis with extension into the dermis, acanthotic and hyperkeratotic areas, underlying dermis had a subacute inflammatory response consisting of lymphocytes, macrophages and heterophils. Dermal arterioles contained marginated leukocytes, hair folicles showed keratinization. Grading of the skin lesions reflected the extent if the lesion and the degree of functional damage.

HISTOPATHOLOGY: NEOPLASTIC (not applicable)

HISTORICAL CONTROL DATA (not applicable)

OTHER FINDINGS: none
Dose descriptor:
NOAEL
Remarks:
local
Effect level:
1.25 mg/cm² per day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
dermal irritation
Dose descriptor:
NOEL
Remarks:
systemic
Effect level:
>= 200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical signs
gross pathology
haematology
mortality
Critical effects observed:
no

The NOAEL for the local effects is 50 mg/kg bw/day. Taking into account a mean body weight of this dose group of 3745 g and an exposed are of 150 cm² 50 mg/kg bw/day corresponds to about 1.25 mg/cm²/day.

Conclusions:
At 100 and 200 mg/kg bw/day, the only lesions noted were skin lesions; the NOEL was 50 mg/kg bw/day.
Executive summary:

Groups of 5 male and 5 female rabbits were treated on their skin with 50, 100 or 200 mg Amines, polyethylenepoly-, tetraethylenepentamine fraction per kg/bw ca. 6 h/day day, 5 days/week for a period of 31 days (under occlusive conditions). These levels were based on a 4 -day RF study using the same levels using 2 females per group. Controls were treated with the vehicle, distilled water. No changes were observed in body weights, food intake, haematology, clinical chemistry, and organ weights. The only lesions observed were local skin lesions; the degree of irritation was dose-related; effects in the 200 mg/kg group were generally more severe than in the 100 mg/kg group. There were no indications of systemic toxicity. Because no changes were seen in the 50 mg/kg group, the NOEL was 50 mg/kg bw/day.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
200 mg/kg bw/day
Study duration:
subacute
Species:
rabbit
Quality of whole database:
The key study is GLP compliant and comparable to guideline, with Klimisch score 2.

Repeated dose toxicity: dermal - local effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
April-May 1986
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
Purity of a.i. has been indicated; complete composition of the test substance is not available (available in departmental study file).
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 410 (Repeated Dose Dermal Toxicity: 21/28-Day Study)
Version / remarks:
1981
Deviations:
yes
Remarks:
20 instead of 21 days of exposure, occlusive dressing
GLP compliance:
yes
Limit test:
no
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Ray Nichols Rabbitry, Lumberton, TX, USA
- Age at study initiation: ca. 5 months
- Weight at study initiation: ca. 3.1 - 4.0 kg
- Fasting period before study: not applicable
- Housing: individually in stainless steel cages
- Diet: restricted to 4 oz per animal per day
- Water: ad libitum
- Acclimation period: at least 14 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 72 ± 5 degr F (22 ± 1.5 degr C)
- Humidity (%): 50
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12/12


Type of coverage:
occlusive
Vehicle:
water
Details on exposure:
TEST SITE
- Area of exposure: 10 x 15 cm
- % coverage: 100
- Type of wrap if used: an occlusive bandage of absorbent gauze and non-absorbent cotton; the bandage was held in place using a lycra/spandex jacket
- Time intervals for shavings or clipplings: periodically as required

REMOVAL OF TEST SUBSTANCE
- Washing (if done): not reported
- Time after start of exposure: ca. 6 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): a 10% solution was applied with the volume adjusted to provide the required dose
- Concentration (if solution): 10%
- Constant volume or concentration used: yes, constant concentration

VEHICLE
- Justification for use and choice of vehicle (if other than water): water
- Amount(s) applied (volume or weight with unit): 1 mL/kg BW

USE OF RESTRAINERS FOR PREVENTING INGESTION: no (but animals wore jackets)
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Test material concentration (100 mg/g) was measured 3 times during the study in triplo. % of target concentrations were 87.0, 95.3 and 90.9%, respectively. Stability was measured 2, 4 and 7 days after preparation. % of Day 0 was, 92.1, 59.4 and 59.9%, respectively.
Duration of treatment / exposure:
20 days
Frequency of treatment:
5 days/week, ca. 6h/day
Dose / conc.:
50 mg/kg bw/day (nominal)
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
200 mg/kg bw/day (nominal)
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on a probe study using 2 female rabbits per group, at levels of 0, 50, 100 or 200 mg/kg bw/day for 4 days
- Rationale for animal assignment (if not random): computer-generated tables of random numbers
- Rationale for selecting satellite groups: not used
- Post-exposure recovery period in satellite groups: not used
- Section schedule rationale (if not random): no info
Positive control:
not used
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily on working days (one additional routine monitoring on these days and on weekend and holidays for dead animals and check for water and food availability)

DETAILED CLINICAL OBSERVATIONS: No

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: daily upon removal when bandage and jacket were removed.

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION: Since the animals were maintained on a restricted diet and normally ate all their ration, measurement of food consumption was not doen. Qualitative changes in consumption were monitored by a periodic visual check.

FOOD EFFICIENCY: No

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: not applicable

HAEMATOLOGY: Yes
- Time schedule for collection of blood: immediately prior to necropsy
- Anaesthetic used for blood collection: Yes (no info which was used)
- Animals fasted: No
- How many animals: all
- Parameters examined: RBC, WPC, platelet count, haemoglobin, packed cell volume

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: immediately prior to necropsy
- Animals fasted: No
- How many animals: all
- Parameters examined: blood urea nitrogen, ALP, ALAT, glucose, total protein, albumin, Cl, Na, K, globulin (calculated)

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER:
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (including eyes)
HISTOPATHOLOGY: Yes (complete set) of control and high dose animals; skin (application site and a non-treated section), liver kidneys and gross pathologic lesions were aslo examined from low and mid dose animals
Organ weights: brain, liver, kidneys, adrenals, ovaries, testes
Other examinations:
None
Statistics:
Yes, for body weights, organ weights, clinical chemistry, haematology.
Clinical signs:
no effects observed
Dermal irritation:
effects observed, treatment-related
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
effects observed, treatment-related
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Other effects:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY: one control female was removed from the study on day 14 because of a broken back.
There were no signs other than local dermal effects. These were dose-related and consisted of red dermal test sites on days 1-2. By study Day 6, animals tretaed with 200 mg/kg bw showed very red and slightly swollen test sites; on Day 16 the skin of some of these females was severely irritated with some crusting and bleeding. Other animals of the 200 mg/kg bw group and those of the 100 mg/kg group had irritated skin that did not progress further.

BODY WEIGHT AND WEIGHT GAIN: no effects

FOOD CONSUMPTION: no effects

FOOD EFFICIENCY: not measured

WATER CONSUMPTION: not measured

OPHTHALMOSCOPIC EXAMINATION: not applicable

HAEMATOLOGY: no effects

CLINICAL CHEMISTRY: no effects

URINALYSIS: not measured

NEUROBEHAVIOUR: not measured

ORGAN WEIGHTS: no effects

GROSS PATHOLOGY: confined to the skin of the 100 and 200 mg/kg groups, characterised by multifocal areas of epidermal and dermal necrosis often covered with crusts comprised of dry serum and necrotic debris.

HISTOPATHOLOGY: NON-NEOPLASTIC
Only dose-related changes in the skin consisting of: multifocal necrosis of the epidermis with extension into the dermis, acanthotic and hyperkeratotic areas, underlying dermis had a subacute inflammatory response consisting of lymphocytes, macrophages and heterophils. Dermal arterioles contained marginated leukocytes, hair folicles showed keratinization. Grading of the skin lesions reflected the extent if the lesion and the degree of functional damage.

HISTOPATHOLOGY: NEOPLASTIC (not applicable)

HISTORICAL CONTROL DATA (not applicable)

OTHER FINDINGS: none
Dose descriptor:
NOAEL
Remarks:
local
Effect level:
1.25 mg/cm² per day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
dermal irritation
Dose descriptor:
NOEL
Remarks:
systemic
Effect level:
>= 200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical signs
gross pathology
haematology
mortality
Critical effects observed:
no

The NOAEL for the local effects is 50 mg/kg bw/day. Taking into account a mean body weight of this dose group of 3745 g and an exposed are of 150 cm² 50 mg/kg bw/day corresponds to about 1.25 mg/cm²/day.

Conclusions:
At 100 and 200 mg/kg bw/day, the only lesions noted were skin lesions; the NOEL was 50 mg/kg bw/day.
Executive summary:

Groups of 5 male and 5 female rabbits were treated on their skin with 50, 100 or 200 mg Amines, polyethylenepoly-, tetraethylenepentamine fraction per kg/bw ca. 6 h/day day, 5 days/week for a period of 31 days (under occlusive conditions). These levels were based on a 4 -day RF study using the same levels using 2 females per group. Controls were treated with the vehicle, distilled water. No changes were observed in body weights, food intake, haematology, clinical chemistry, and organ weights. The only lesions observed were local skin lesions; the degree of irritation was dose-related; effects in the 200 mg/kg group were generally more severe than in the 100 mg/kg group. There were no indications of systemic toxicity. Because no changes were seen in the 50 mg/kg group, the NOEL was 50 mg/kg bw/day.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
1.25 mg/cm²
Study duration:
subacute
Species:
rabbit
Quality of whole database:
The key study is GLP compliant and comparable to guideline, with Klimisch score 2.

Additional information

Oral:

There are only insufficient data available on repeated dose toxicity with Amines, polyethylenepoly-, tetraethylenepentamine fraction. In order to fulfil the standard information requirements set out in Annex VII-IX, 8.6., in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006, read-across from structurally related substances was conducted. In accordance with Article 13 (1) of Regulation (EC) No 1907/2006, "information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set out in Annex XI are met.” In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests, which includes the use of information from structurally related substances (grouping or read-across). Having regard to the general rules for grouping of substances and read-across approach laid down in Annex XI, Item 1.5, of Regulation (EC) No 1907/2006 whereby substances may be predicted as similar provided that their physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity.

Triethylenetetramine dihydrochloride (CAS 38260-01-4) and Amines, polyethylenepoly-, tetraethylenepentamine fraction (CAS 90640 -66 -7) are considered to be similar on the basis of the structural similar properties and/or activities. It has to be noted that target substance is a multiconstituent substance whereas the source substance is considered to be a monoconstiutent substance which is however not completely characterized with regards to the presence of impurities. Therefore, taking into account the latter mentioned deficiency in the read-across approach, the registrants agree to conduct further testing on the registration substance that is also in line with the request in the final decision received from ECHA on September 2nd 2019 (CCH-D-2114482142 -55 -01/F). Until the study data on the registration substance are available, the data on the source subtance TETA x 2 HCl will be used as interim approach to conduct quantitative risk assessment. A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).

 

Short-term toxicity

A key study similar to OECD 407 with the structural analogue substance TETA-2HCl is available (Yanagisawa, 1998). Triethylenetetramine dihydrochloride (TETA-2HCl), a copper chelating agent used to treat Wilson's disease, was administered orally to male and female F-344 rats for 4 or 8 weeks at dosages of 0, 100, 350 or 1200 mg/kg bw/day. Two males receiving 1200 mg/kg bw/day died during week 8 of treatment. In males receiving 1200 mg/kg bw/day during weeks 5 to 8 of treatment, body weight gain and food consumption were decreased and hunched posture and thin build were observed. During week 4 or 8 of treatment urinalysis revealed, for males receiving 100 mg/kg bw/day or animals receiving 350 mg/kg bw/day or more, increased electrolyte outputs possibly due to the hydrochloride nature of TETA-2HCl, with low plasma alkaline phosphatase activities evident in animals receiving 350 or 1200 mg/kg bw/day. After 4 and 8 weeks, and during 8 weeks of treatment, high lung weights and bronchiolar epithelium hypertrophy and broncho-alveolar pneumonia were recorded for animals receiving 1200 mg/kg bw/day, and submucosal acute inflammation within the glandular region of the stomach was recorded for males receiving 350 or 1200 mg/kg bw/day and in all treated female groups. Those effects lead to a systemic NOAEL of 350 mg/kg bw/day.

 

A 7-day repeated dose toxicity study with the target substance Amines, polyethylenepoly-, tetraethylenepentamine fraction is available which is used as supporting study (Myers, 1979). The test substance was added to the diet and fed to groups of 5 Wistar rats (30 days of age) per sex for 7 days. In the first part of the study 4 groups were used, these were dosed at 0, 0.50, 1.25 and 3.15 g/kg bw. Because no effects were observed on body weight gain, food intake and liver and kidneys weights, in the second part of the study, 2 groups were used, these were dosed at 0 and 5.00 g/kg bw. Dosages attained (calculated based on nominal levels and food intake) were: 0, 0, 0.42, 1.05, 2.80 and 3.99 g/kg bw for males, and 0, 0, 0.47, 1.26, 3.14 and 3.63 g/kg bw for females. Animals treated at the highest dose showed a decrease in food intake, body weight loss, and decreased absolute and relative liver weight and decreased relative kidney weight. The NOAELs, therefore, were, 2.80 and 3.14 g/kg bw for males and females, respectively.

 

Sub-chronic toxicity

A key study similar to OECD 408 with the structural analogue substance TETA.2HCl is available (Greenman, 1996). Mice and rats received TETA.2HCl in the drinking water at concentrations of 0, 120, 600, or 3000 ppm for up to 92 days; they were fed diets containing nutritionally adequate levels of copper. An additional control group of rats and mice received a Cu-deficient diet. This low copper diet resulted in Cu-deficiency symptoms, such as anemia, liver periportal cytomegaly, pancreatic atrophy and multifocal necrosis, spleen hematopoietic cell proliferation, and increased heart weight, together with undetectable levels of plasma copper in rats but not in mice. TETA.2HCl lowered plasma copper levels somewhat (at 600 and 3000 ppm) in rats, but did not induce the usual signs of copper deficiency. TETA.2HCl caused an increased frequency of uterine dilatation at 3000 ppm in rats fed the adequate copper diet but was not noted in females fed the Cu-deficient diet. This observation may reflect an altered endocrine milieu since uterine dilatation is a normal phenomenon of the estrous cycle and is known to be sensitive to changes in estrogen exposure. TETA.2HCl toxicity occurred only in mice in the highest dose group. Increased frequencies of inflammation of the lung interstitium and liver periportal fatty infiltration were seen in both sexes, and hematopoietic cell proliferation was seen in the spleen of males. Kidney and body weights were reduced in males as was the incidence of renal cytoplasmic vacuolization. There were no signs of copper deficiency in mice exposed to TETA.2HCl. Based on the effects in mice the NOAEL was 600 ppm or 92 mg/kg bw/day for males and 99 mg/kg bw/day for females. In rats the NOAEL was 3000 ppm or 270 mg/kg bw/day for males and 600 ppm or 60 mg/kg bw/day for females.

 

A further study similar to OECD 408 with the structural analogue substance TETA.2HCl is available (Yanagisawa, 1998). Triethylenetetramine dihydrochloride (TETA.2HCl), a copper chelating agent used to treat Wilson's disease, was administered orally to male and female F-344 rats for 26 weeks at dosages of 0, 50, 175 or 600 mg/kg bw/day. One male receiving 175 mg/kg bw/day and three males receiving 600 mg/kg bw/day died, showing lung changes. With regard to the lungs, histopathology revealed a dose-related incidence and severity of focal chronic interstitial pneumonitis accompanied by fibrosis of the alveolar walls in females receiving 175 mg/kg/day or more and all treated male groups. Apart from the histological changes found in the lung, all other changes were reversible. It was concluded that the NOAEL of TETA.2HCl in this 26 -week study was considered to be 50 mg/kg bw/day for females and less than 50 mg/kg bw/day for males; the latter, therefore, was considered a LOAEL.

The authors of the study assume that TETA.2HCl could be taken up into the bronchiolar epithelium and/or alveolar epithelium by an endogenous polyamine uptake process and therefore could accumulated in the specific tissue site of the lung. Polyamines are generally irritating agents for mucous membranes, upper respiratory tract and skin. In the 4 - or 8 -week study, the histopathological changes observed in the stomach (submucosal inflammation within the glandular region) indicates that TETA.2HCl was slightly irritating. Therefore, it is likely that the chronic interstitial pneumonitis was caused by the cytotoxic effect of TETA.2HCl which could have accumulated in the bronchiolar epithelial cells and the alveolar pneumocytes.

 

In addition a subchronic study in dogs similar to OECD 409 is available as supporting study (Maemura, 1998). Beagle dogs received TETA.2HCl orally at dosages of 50, 100 or 200 mg/kg bw/day for 26 weeks followed by a 13 -week reversibility phase. However, in view of the severe signs which resulted in the sacrifice for human reasons of two males and one female receiving 200 mg/kg bw/day during week 9 of treatment, surviving dogs of this group were only treated for 10 weeks. Signs before killing included marked underactivity, body tremors, abnormal gait, limited use of limbs and prone posture. The ante mortem neurological examination generally indicated depressed postural and flexor withdrawal reactions. The signs were rapidly reversible except in one female which was killed humanly on day 2 of the reversibility period. Abnormal "stiff legged" gait and underactivity were evident, from week 23 of treatment, in two males and one female receiving 100 mg/kg bw/day. In the absence of any macroscopic or histopathologic findings, even after the examination of additional samples of muscle and nerve, the exact nature of this condition could not be elucidated. In all treated groups low copper and zinc concentrations in the livers and high urinary copper and zinc concentrations were found. The NOAEL was considered to be 50 mg/kg bw/day.

 

Overall, the results from the 26-week key study in rats (Yanagisawa, 1998) and the 13 -week study in mice (Greenman, 1996) showed focal chronic interstitial pneumonitis. In the 13-week study in rats described by Greenman, 1996 female rats resulted effects in the uterine only. The 26-week study in dogs (Maemura, 1998) did not show lung toxicity but showed neurological effects instead. The NOAELs in these studies were in the same order of magnitude, i.e. between 50 and 99 mg/kg bw/day, although for male rats the level of 50 mg/kg bw/day should be considered a LOAEL.

 

The above study results with the structural analogue substance triethylenetetramine dihydrochloride (CAS 38260-01-4) indicate that the registration substance (CAS 90640-66-7) could also result in adverse effects after repeated oral exposure. To confirm whether the registration substance will cause adverse systemic effects after repeated oral exposure, further testing will be conducted on the registration substance itself. In the meanwhile, the LOAEL of 50 mg/kg bw/day is used for DNEL derivation until data on the registration substance are available.

 

Dermal:

One 28 -day dermal toxicity study in rabbits with Amines, polyethylenepoly-, tetraethylenepentamine fraction as test substance similar to OECD 410 and in compliance with GLP is available (Szabo, 1986).

Groups of 5 male and 5 female rabbits were treated on their skin with 50, 100 or 200 mg test substance per kg/bw/day ca. 6 h/day day, 5 days/week for a period of 31 days (under occlusive conditions). These levels were based on a 4 -day range-finding study using the same levels using 2 females per group. Controls were treated with the vehicle, distilled water. No changes were observed in body weights, food intake, haematology, clinical chemistry, and organ weights. The only lesions observed were local skin lesions; the degree of irritation was dose-related; effects in the 200 mg/kg bw/day group were generally more severe than in the 100 mg/kg bw/day group. There were no indications of systemic toxicity. As not systemic toxicity was observed the NOAEL for systemic toxicity was >= 200 mg/kg bw/day. Due to local effects on the skin the NOAEL for local effects was 50 mg/kg bw/day which corresponds to 1.25 mg/cm².

Justification for classification or non-classification

No reliable repeated dose toxicity data on the registration substance are available but further testing is planned. In the meanwhile, reliable repeated dose toxicity data from a structural analogue was read-across and inidicated adverse systemic effects after repeated oral exposure. However, due to deficiencies in the read-across approach the data on the source substance is only used as interim approach for quantitiative risk assessment. Therefore, a conclusion on the classification of the registration substance according to Rgulation (EC) No. 1272/2008 cannot be made and is thus deemed inconclusive until further repeated dose toxicity data on the registration substance becomes available.

Reliable data from the test substanc on repeated dose toxicity by dermal route indicate that the test substance does not meet the criteria for classification according to Regulation (EC) No. 1272/2008.