Registration Dossier

Administrative data

Description of key information

The substance is severey irritating to the skin and causes irreversible effects on the eye.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
08 June 2016 - 19 July 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
2015
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Version / remarks:
May 2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Version / remarks:
August 1998
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147.
Version / remarks:
November 2000, including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
date certificate 3 November 2015
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
- Source: Charles River France, L’Arbresle Cedex, France.
- Age at study initiation: between 12 and 13 weeks old.
- Weight at study initiation: between 2.351 and 3.211 kg.
- Housing: Animals were housed individually in cages with perforated floors and shelters.
- Diet: Pelleted diet for rabbits (Global Diet 2030 from Harlan Teklad, Italy) approximately 100 grams per day. Hay and wooden sticks were abailable during the study period.
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS (set to maintain)
- Temperature (°C): 18 – 24
- Humidity (%): 40 - 70
- Air changes (per hr): approx 10
- Photoperiod (hrs dark / hrs light): 12/12

Deviations from the maximum level of daily mean relative humidity occurred. These deviations were mostly transient and not adversely impact the outcome of the study.

IN-LIFE DATES: From: 08 June 2016 to 19 July 2016
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
other: Adjacent areas of the untreated skin of each animal served as controls.
Amount / concentration applied:
TEST MATERIAL
- Amount applied: 0.5 mL
Duration of treatment / exposure:
One hour and 4 hours in sentinel animal and 4 hours in two other animals.
Observation period:
14 days.
Number of animals:
3 males.
Details on study design:
STUDY DESIGN
The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel) with a stepwise exposure regime. The two other animals were treated with a single patch applied for 4 hours 18 days later, after considering the degree of skin irritation observed in the first animal.

TEST SUBSTANCE PREPARATION
The test item was applied undiluted as delivered by the Sponsor.

TEST SITE
Approximately 24 hours before treatment, the dorsal fur was clipped with electric clippers, exposing an area of approximately 150 square centimeters (10x15 cm). To facilitate scoring, treated skin areas were re-clipped at least 3 hours before the observations.

The test substance was applied to two separate skin sites of one flank, using a patch of 2x3 cm.

REMOVAL OF TEST SUBSTANCE
One hour or 4 hours after the application, the dressing was removed and the skin cleaned of residual test substance using tap water, watery ethanol (50% v/v) and watery acetone (50% v/v).

OBSERVATIONS
- Mortality/Viability: Twice daily.
- Toxicity: At least once daily.
- Body Weight: Day of treatment (prior to application) and on the day of the final observation.
- Necropsy: No necropsy for gross macroscopic examination was performed according to study plan.
- Irritation:
In the initially treated animal, the skin reactions of all visible treated sites were assessed immediately after removal of a dressing and approximately 1, 24, 48, 72 hours after the removal of the last dressing and test item. After the 4 hours exposure in two further animals, the skin reactions were assessed approximately 1, 24, 48, 72 hours after the removal of the dressing and test item. For the duration of the skin reactions, further observations were made 7 and 14 days after exposure. The irritation scores and a description of all other (local) effects were recorded.

SCORING SYSTEM:
The irritation was assessed according to the numerical scoring system according to OECD 404.
Irritation parameter:
erythema score
Basis:
animal #1
Remarks:
(mean)
Time point:
24/48/72 h
Score:
3
Max. score:
4
Reversibility:
not fully reversible within: 14 days
Remarks:
; Reduced flexibility, scaliness, bald skin
Remarks on result:
other:
Irritation parameter:
erythema score
Basis:
animal #2
Remarks:
(mean)
Time point:
24/48/72 h
Score:
1.3
Max. score:
4
Reversibility:
not fully reversible within: 14 days
Remarks:
; Bald skin
Remarks on result:
other:
Irritation parameter:
erythema score
Basis:
animal #3
Remarks:
(mean)
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
edema score
Basis:
animal #1
Remarks:
(mean)
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 14 days
Remarks:
; Very slight oedema
Irritation parameter:
edema score
Basis:
animal #2
Remarks:
(mean)
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
edema score
Basis:
animal #3
Remarks:
(mean)
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritant / corrosive response data:
There was no evidence of a corrosive effect on the skin.
Exposure to 0.5 mL of di-C16-C18 (evennumbered) alkyl tripropylenetetramine for 3 minutes, 1 hour or 4 hours in the sentinel animal, all resulted in severe erythema (max. grade 4) and slight oedema (max. grade 2) in the treated skin areas. Four hour exposures to 0.5 mL of di-C16-C18 (evennumbered) alkyl tripropylenetetramine in further two animals resulted in well defined erythema (max. grade 2) and very slight (grade 1) or slight (grade 2) oedema. Highest grades of erythema and oedema were observed at 72 hours and 7 days after application.
Moreover, reduced flexibility of the skin, scabs, scaliness, fissuring of the skin and/or scattered erythema were noted from 72 hours after application onwards in two animals. This had completely resolved at 14 days after application in one of these animals but did not resolve in the other animal. The third animal showed reduced flexibility of the skin from 24 hours after application onwards and scaliness at 7 days after application which had resolved within 14 days after application. The above mentioned effects resulted in bald skin in two animals at 14 days after application.
Other effects:
Remnants of the test item were present on the skin between Days 1 and 8 for all animals. No signs of systemic toxicity were observed in the animals during the test period and no mortality occurred.
Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
In a skin irritation study with rabbits, performed according to OECD/EC test guidelines, di-C16-C18 (evennumbered) was classified as skin irritant (Category 2) based on the skin reactions observed.
Executive summary:

A primary skin irritation/corrosion study with di-C16-C18 (evennumbered) alkyl tripropylenetetramine in the rabbit was performed according to OECD/EC guidelines and in compliance with GLP principles. Initially, one rabbit was exposed to three samples of 0.5 mL of di-C16-C18 (evennumbered) alkyl tripropylenetetramine applied to separate skin-sites on intact, clipped skin using a semiocclusive dressing. The exposure periods were 3 minutes, 1 hour and 4 hours, respectively. Skin reactions were assessed at least once daily on Days 1-4 after treatment and 7 and 14 days after exposure. Based on the absence of very severe skin reactions, two further animals were exposed for 4 hours to 0.5 mL di-C16-C18 (evennumbered) alkyl tripropylenetetramine at a later stage.

Skin reactions were assessed 1, 24, 48 and 72 hours and 7 and 14 days after exposure. The exposures in the sentinel animal (3 minutes, 1 hour oand 4 hours) all resulted in severe erythema (max. grade 4) and slight oedema (max. grade 2) in the treated skin areas. Four hour exposures to 0.5 mL of di-C16-C18 (evennumbered) alkyl tripropylenetetramine in further two animals resulted in well defined erythema (max. grade 2) and very slight (grade 1) or slight (grade 2) oedema. Highest grades of erythema and oedema were observed at 72 hours and 7 days after application. Moreover, reduced flexibility of the skin, scabs, scaliness, fissuring of the skin and/or scattered erythema were noted from 72 hours after application onwards in two animals. The third animal showed reduced flexibility of the skin from 24 hours after application onwards and scaliness at 7 days after application. The skin reactions had completely resolved within 14 days after exposure in one animal, whereas the other two animals still showed bald skin respectively reduced flexibility, scaliness and bald skin after 14 days.

Based the skin reactions of the first animal and on the persistence of the skin reactions:

- according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments), di-C16-C18 (evennumbered) alkyl tripropylenetetramine should be classified as: skin irritant (Category 2).

- according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments), di-C16-C18 (evennumbered) alkyl tripropylenetetramine should be classified as Irritant (Category 2) and labeled as H315: Causes skin irritation.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
October 03, 2016 - October 04, 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
2012
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
May 2008, including most recent amendments.
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Version / remarks:
August 1998.
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147.
Version / remarks:
November 2000; including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
pH: 8.5-10.5 at concentration of 1%
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
- Source: Charles River France, L’Arbresle, France.
- Age at study initiation: Animal used withihn the study was 13 weeks old .
- Weight at study initiation: 2636 gram.
- Housing: Individually housed in cages with perforated floors and shelter.
- Diet: Approximately 100 g/day pelleted diet for rabbits (Global Diet 2030 Harlan Teklad, Italy). Hay and wooden sticks were available during the study period.
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS set to maintain
- Temperature (°C): 40 – 70
- Humidity (%): 18 - 24
- Air changes (per hr): approx 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 03 October 2016 to 04 October 2016
Vehicle:
unchanged (no vehicle)
Controls:
other: One eye of each animal remained untreated and served as the reference control.
Amount / concentration applied:
TEST MATERIAL
- Amount applied: 0.1 mL
Duration of treatment / exposure:
Single instillation on Day 1.
Observation period (in vivo):
The eyes of the animal were examined approximately 5 minutes and 1 hour after instillation of the test item.
An extra observation was done at 6 minutes after instillation of the test item.
Number of animals or in vitro replicates:
One male animal
Details on study design:
STUDY DESIGN
The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). No other animals were treated, after considering the degree of eye irritation observed in the first animal.

TREATMENT
The animal was treated by instillation of 0.1 mL of the test item, in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test item. The other eye remained untreated and served as the reference control.

REMOVAL OF TEST SUBSTANCE
-Washing (if done): No

OBSERVATIONS
- Mortality/Viability: Twice daily.
- Toxicity: At least once daily.
- Body Weight: Day of treatment (prior to instillation).
- Necropsy: No necropsy was performed on the animal sacrificed for severe irritation/corrosion of the eye.
- Irritation:
The eyes of the animal were examined approximately 5 minutes and 1 hour after instillation of the test item.

SCORING SYSTEM
The irritation scores and a description of all other (local) effects were recorded.The irritation was assessed according to OECD 405.

TOOL USED TO ASSESS SCORE: ophthalmic examination lamp
Irritation parameter:
cornea opacity score
Remarks:
(opacity)
Basis:
animal #1
Time point:
other: 6 minutes
Score:
2
Max. score:
4
Irritation parameter:
cornea opacity score
Remarks:
(opacity)
Basis:
animal #1
Time point:
other: 1 hour
Score:
4
Max. score:
4
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: 6 minutes
Score:
1
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: 1 hour
Max. score:
2
Remarks on result:
not determinable
Remarks:
due to the severe corneal damage
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal #1
Time point:
other: 6 minutes
Score:
2
Max. score:
3
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
animal #1
Time point:
other: 1 hour
Score:
2
Max. score:
3
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: 6 minutes
Score:
2
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: 1 hour
Score:
4
Max. score:
4
Irritation parameter:
conjunctivae score
Remarks:
(Discharge)
Basis:
animal #1
Time point:
other: 6 minutes
Score:
3
Max. score:
3
Irritation parameter:
conjunctivae score
Remarks:
(Discharge)
Basis:
animal #1
Time point:
other: 1 hour
Score:
3
Max. score:
3
Irritant / corrosive response data:
Instillation of test item into one eye of one rabbit resulted in severe effects on the eye and signs of pain and stress (restless behaviour, closing of the eye). Due to these immediate severe effects an extra observation was done at 6 minutes after instillation. At one hour after instillation the effects worsened and blood in the lacrimal fluid was seen. The iris could not be scored at this point due to the severe corneal damage.
The animal was sacrificed for humane reasons approximately 1 hour after application of the test item. The other animals assigned to the study were not treated.
Other effects:
Remnants of the test item were present in the eye shortly after application but not at one hour after application.
Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
In an eye irritation study with one rabbit, performed according to OECD/EC test guidelines, severe effects were observed including opacity of the cornea (grade 4/4) and chemosis of the conjunctivae (grade 4/4). Based on the results di-C16-C18 (evennumbered) alkyl tripropylenetetramine should be classified as Irreversible effects on the eye (Category 1) and labeled as H318: Causes serious eye damage.
Executive summary:

An acute eye irritation/corrosion study was performed with di-C16-C18 (evennumbered) alkyl tripropylenetetramine in the rabbit according to OECD/EC guidelines and in compliance with GLP principles. Instillation of approximately 0.1 mL of di-C16-C18 (evennumbered) alkyl tripropylenetetramine into one eye of one rabbit resulted in severe effects on the eye and signs of pain and stress (restless behaviour, closing of the eye). At 6 minutes after installation opacity of the cornea (grade 2/4) and injury of the iris (grade 1/2) were observed. The injury of the conjunctivae included redness (grade 2/3), chemosis (grade 2/4) and discharge (grade 3/3). At one hour after installation the effects worsened into opacity of the cornea (grade 4/4) and chemosis of the conjunctivae (grade 4/4). Blood was seen in the lacrimal fluid and the iris could not be scored due to the severe corneal damage. Remnants of the test item were present in the eye shortly after application but not at one hour after application. The animal was sacrificed for humane reasons approximately 1 hour after application of the test item. The other animals assigned to the study were not treated.

Based on these severe effects:

- according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments), di-C16-C18 (evennumbered) alkyl tripropylenetetramine should be classified as : having irreversible effects on the eyes (Category 1).

- according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments), di-C16-C18 (evennumbered) alkyl tripropylenetetramine should be classified as Irreversible effects on the eye (Category 1) and labeled as H318: Causes serious eye damage.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Di-C16-C18 (evennumbered) alkyl tripropylenetetramine was evaluated in an in vitro dermal corrosion studies using human reconstructed epidermis (OECD 431). Cell viability of 69% after 3 minutes 84% after 1 hour, indicate that the substance is non-corrosive. However, validation of this test system has shown it to be unsuitable for the evaluation for dermal corrosive effects of fatty amines, as so far all tested fatty amines have been falsely predicted to be non-corrosive (Houthoff et al., 2015, TiV 29(6),1263–1267).

 

As the results from in vitro tests cannot be trusted, the substance was subsequently tested in an in vivo skin irritation/corrosion study according to OED 404. Exposure to 0.5 mL of di-C16-C18 (evennumbered) alkyl tripropylenetetramine for 3 minutes, 1 hour or 4 hours in the sentinel animal, all resulted in severe erythema (max. grade 4) and slight oedema (max. grade 2) in the treated skin areas. Four hour exposures to 0.5 mL of di-C16-C18 (evennumbered) alkyl tripropylenetetramine in further two animals resulted in well-defined erythema (max. grade 2) and very slight (grade 1) or slight (grade 2) oedema. Highest grades of erythema and oedema were observed at 72 hours and 7 days after application. Moreover, reduced flexibility of the skin, scabs, scaliness, fissuring of the skin and/or scattered erythema were noted from 72 hours after application onwards in two animals. This had completely resolved at 14 days after application in one of these animals but did not resolve in the other animal. The third animal showed reduced flexibility of the skin from 24 hours after application onwards and scaliness at 7 days after application which had resolved within 14 days after application. The above mentioned effects resulted in bald skin in two animals at 14 days after application.

The skin reactions had completely resolved within 14 days after exposure in one animal, whereas the other two animals still showed bald skin respectively reduced flexibility, scaliness and bald skin after 14 days. There was however no evidence of a corrosive effect on the skin, and based on the GHS criteria, this substance is considered to be a skin irritant.

 

Di-C16-C18 (evennumbered) alkyl tripropylenetetramine further evaluated in vitro for eye irritation potential in the Isolated Chicken Eye (ICE) test. Three main parameters were measured to disclose possible adverse eye effects: corneal thickness (expressed as corneal swelling), corneal opacity and fluorescein retention of damaged epithelial cells. In addition, histopathology of the corneas was performed.

The test substance caused corneal effects consisting of slight corneal swelling (mean of 8%), moderate-to-severe opacity (mean score of 2.5) and moderate or moderate-to-severe fluorescein retention (mean score of 2.2). Microscopic examination of the corneas revealed very slight erosion with or without very slight vacuolation of the epithelium. According to the AISE histopathology criteria, upgrading to category 1 on basis of the histopathology of the corneas is not required. The toxicological relevance of the observed adhesive properties of the test substance was not taken into account in the assessment of the irritation classifications.

Applying the classification criteria of the ICE, the results point at a classification as eye irritant.

 

As the results from the in vitro ICE study are not decisive for classification purposes, a further in vivo eye irritation study in rabbit (OECD 405) was performed.

Instillation of approximately 0.1 mL of di-C16-C18 (evennumbered) alkyl tripropylenetetramine into one eye of one rabbit resulted in severe effects on the eye and signs of pain and stress (restless behaviour, closing of the eye). Due to these immediate severe effects an extra observation was done at 6 minutes after instillation. The effects consisted of opacity of the cornea (grade 2/4), injury of the iris (grade 1/2) and effects on the conjunctivae. The injury of the conjunctivae included redness (grade 2/3), chemosis (grade 2/4) and discharge (grade 3/3). Further, the animal showed a tilted head, a sign of pain and stress.

At one hour after instillation the effects worsened into opacity of the cornea (grade 4/4) and chemosis of the conjunctivae (grade 4/4). Moreover, blood in the lacrimal fluid was seen. The iris could not be scored at this point due to the severe corneal damage.

Remnants of the test item were present in the eye shortly after application but not at one hour after application. The animal was sacrificed for humane reasons approximately 1 hour after application of the test item. The other animals assigned to the study were not treated.

Justification for classification or non-classification

Studies on di-C16-C18 (evennumbered) alkyl tripropylenetetramine have shown it to be irritating to the skin and damaging to the eyes leading to the classification

- Irritating to the skin (Category 2 - H315: Causes skin irritation) and

- Irreversible effects on the eye/serious damage to eyes (Category 1 - H318 Causes serious eye damage.)

 

There is no information is available following exposure via inhalation. However, with a vapour pressure of 1.3 x 10-6 Pa at 20°C, the potential for inhalation of vapours is limited. Also the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route will be unlikely to occur. Consequently, despite the irritant nature of the substance, respiratory irritation is not expected, and classification STOT-SE Cat.3 for respiratory irritation is not indicated.