Reaction mass of Amines, N-tallow alkyltrimethylenedi-, mono(2-ethylhexanoates), Amines, N-tallow alkyltrimethylenedi-, acetates and n-tallow-1,3-diaminopropane ditallate

Endpoint summary

Description of key information

The starting material for this reaction mass is n-tallow alkyltrimethyenediamine CAS No 61791-55-7, which was registered for REACH as N-C16-18-alkyl(evennumbered) C18 unsaturated propane-1, 3-diamine(CAS No 1219010-04-4), that is reacted with tall oil fatty acids (CAS No 61790-12-3), 2-ethylhexanoic acid (CAS No 149-57-5) and acetic acid (CAS No 64-18-7). The n-tallow alkyltrimethyenediamine is ca. 52.2% of the reaction mixture with the 2-ethylhexanoic acid at 10.6% compared to the tall oil fatty acids at 32.2% and the acetic acid 4.6%.  The acetic acid, tall oil fatty acids and the 2-ethyhexanoic acid, have no potential for skin sensitisation. Therefore any skin sensitisation potential would have to come from the main (52.2%) component in the reaction mass, the N-C16-18-alkyl(evennumbered) C18 unsaturated propane-1, 3-diamine. Therefore the assessment for skin sensitisation potential for the reaction mass can be based on this substance. There is no data on this substance as it is corrosive to skin 1B and therefore it is not ethical to tests in animals for skin sensitisation potential. However there is animal data in the form of a Guinea Pig maximisation test (M&K) on the acetate version, Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates which is less irritant. As the acetate part of the substance will not affect the skin sensitisation this is considered to be a suitable source substance for read across.

Profiling and QSARs indicate a low risk for sensitisation, and due to use in industrial and professional setting only, with the application of adequate PPE related to the severe corrosive properties of the diamines, exposures are limited. There are no reports on incidents of sensitisation to diamines available.

There is a Guinea Pig Maximisation study with CAS No 1313206-64-2, Amines, N- C16-18 and C18 unsaturated alkyl) trimethylenedi-,diacetates which is representative for this reaction mass.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Assay performed on a mixture containing 30-40% of Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates, 25-35% of butylglycol and water until complement to 100%. As the existing data on butylglycol indicate no sensitization potential and a very slight irritating potential, the presence of this solvent does not compromise the interpretation of the results.

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

yes

Remarks:

On day 1, the third pair of intradermal injections was performed with the test item at the concentration of 50% (v/W) in a mixture FCA/0.9% NaCl (50/50) (control group).

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

yes

Remarks:

See above

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Data performed before Reach guidance.

Species:

guinea pig

Strain:

Hartley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Saint-Aubin-lès-Elbeuf, France.
- Age at study initiation: 1-2 months old
- Weight at study initiation: 335 ± 9 g for the males and 344 ± 11 g for the females.
- Housing: individually in polycarbonate cages
- Diet (e.g. ad libitum): free access to 106 pelleted diet (UAR, Villemoisson, Epinay-sur-Orge, France).
- Water (e.g. ad libitum): Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum.
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
· temperature: 22 ± 2°C
· relative humidity: 30 to 70%
· light/dark cycle: 12 h/12 h
· ventilation: approximately 12 cycles/hour of filtered, non-recycled air.

IN-LIFE DATES: From: July 30, 2003 To: September 5, 2003

Route:

intradermal and epicutaneous

Vehicle:

other: 0.9% NaCl

Concentration / amount:

Induction (treated group):
• intradermal injections (day 1): test item at the concentration of 0.1% (w/w) in 0.9% NaCl,
• topical application (day 8): test item at the concentration of 25% (w/w) in 0.9% NaCl.

First challenge (all groups): topical application (day 22): test item at the concentration of 10% (w/w) in 0.9% NaCl.

Route:

epicutaneous, open

Vehicle:

other: 0.9% NaCl

Concentration / amount:

Induction (treated group):
• intradermal injections (day 1): test item at the concentration of 0.1% (w/w) in 0.9% NaCl,
• topical application (day 8): test item at the concentration of 25% (w/w) in 0.9% NaCl.

First challenge (all groups): topical application (day 22): test item at the concentration of 10% (w/w) in 0.9% NaCl.

No. of animals per dose:

- four males and four females for the preliminary test,
- in the main test : control group = 5 animals/sex , treated group = 10 animals/sex/dose

Details on study design:

RANGE FINDING TESTS:
A preliminary test was conducted in order to determine the concentrations to be tested in the main study.
By intradermal route (tested concentrations: 25%, 10%, 5%, 1% and 0.1% (w/w)):
• 24 hours before treatment, the dorsal region of the animals was clipped,
• intradermal injections of the dosage form preparations (0.1 mL) were performed in the interscapular region,
• cutaneous reactions were evaluated approximately 24, 48 hours and 6 days after the injections.
By cutaneous route : Under the conditions of the induction phase (tested concentrations: 100%, 50% (w/w), 25% (w/w) and 10% (w/w)):
• 24 hours before treatment, both flanks of the animals were clipped and shaved,
• the filter paper of a chamber (Finn Chamber®) was fully-loaded with a dosage form preparation. The chamber was then applied to the clipped area of the skin. The chamber was held in place by means of an occlusive dressing for 48 hours,
• cutaneous reactions were evaluated 24 and 48 hours after removal of the dressings.


Under the conditions of the challenge phase (tested concentrations: 100%, 50% (w/w), 25% (w/w) and 10% (w/w)):
• 24 hours before treatment, both flanks of the animals were clipped and shaved,
• the filter paper of a chamber (Finn Chamber®) was fully-loaded with a dosage form preparation. The chamber was then applied to the clipped area of the skin (one concentration per flank). The chamber was held in place by means of an occlusive dressing for 24 hours,
• cutaneous reactions were evaluated 24 and 48 hours after removal of the dressings.

In order to respect the criteria for the selection of concentrations (the concentration should be well-tolerated systemically and locally, intradermal injections should cause moderate irritant effect but no necrosis or ulceration of the skin), concentration chosen for the main study was 0.1% (w/w).
In order to respect the criteria for the selection of concentrations (the concentrations should be well-tolerated systemically and locally, cutaneous application for the induction should cause at most weak or moderate skin reactions or be the maximal practicable concentration, cutaneous application for the challenge phase should be the highest concentration which does not cause irritant effect), concentration chosen for the topical application of the induction phase (day 8)was 25% (W/W) and for the challenge application (day 22) 10% (w/w).

MAIN STUDY
A. INDUCTION EXPOSURE (intradermal injection and topical administration)
On day 1, six intradermal injections were made deep into the dermis of a 4 cm x 2 cm clipped interscapular area, using a needle (diameter: 0.50 x 16 mm) mounted on a 1 mL plastic syringe (0.01 mL graduations). 2 injections per site.
As the test item was shown to be irritant during the preliminary test, a topical application with sodium lauryl sulfate was not necessary on day 7.
On day 8, a pad of filter paper (approximately 8 cm2) was fully-loaded with the test item at the concentration of 25% (w/w) and was then applied to the interscapular region of the animals of the treated group.
The animals of the control group received an application of the vehicle alone under the same experimental conditions.
The pad was held in place for 48 hours by means of an adhesive hypoallergenic dressing and an adhesive anallergenic waterproof plaster.
B. CHALLENGE EXPOSURE
First challenge application
On day 22, the animals of treated and control groups received an application of the test item and vehicle. The filter paper of a chamber was fully-loaded with the test item at the concentration of 10% (w/w) and was then applied to a clipped area of the skin of the posterior right flank of all animals.
The vehicle was applied under the same experimental conditions to the skin of the posterior left flank.
The chambers were held in contact with the skin for 24 hours by means of an adhesive anallergenic waterproof plaster.

Challenge controls:

no

Positive control substance(s):

yes

Remarks:

MERCAPTOBENZOTHIAZOLE. In a recent study performed under CIT experimental conditions, the strain of guinea pigs used showed a satisfactory sensitization response in 100% animals.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

0%

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

10%

No. with + reactions:

2

Total no. in group:

10

Clinical observations:

discrete erythema

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 10%. No with. + reactions: 2.0. Total no. in groups: 10.0. Clinical observations: discrete erythema.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

0%

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

10%

No. with + reactions:

4

Total no. in group:

10

Clinical observations:

discrete to moderate erythema and/or dryness of skin

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 10%. No with. + reactions: 4.0. Total no. in groups: 10.0. Clinical observations: discrete to moderate erythema and/or dryness of skin.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

0%

No. with + reactions:

0

Total no. in group:

19

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 19.0.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

10%

No. with + reactions:

9

Total no. in group:

19

Clinical observations:

discrete to moderate erythema

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 10%. No with. + reactions: 9.0. Total no. in groups: 19.0. Clinical observations: discrete to moderate erythema.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

0%

No. with + reactions:

0

Total no. in group:

19

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 19.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

10%

No. with + reactions:

12

Total no. in group:

19

Clinical observations:

discrete to moderate erythema and/or dryness of skin

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10%. No with. + reactions: 12.0. Total no. in groups: 19.0. Clinical observations: discrete to moderate erythema and/or dryness of skin.

No systemic clinical signs and no deaths related to treatment were noted during the study. The body weight gain of the treated animals was similar to that of controls. Marked local reactions at the intradermal injection sites were noted in a few animals of the treated group, between days 10 and 14. Therefore, one of these animals (Male No. 156) was sacrificed on day 14 for ethical reasons.

A discrete erythema (grade 1) was noted in 2/10 and 1/10 animals of the control group at the 24 and 48 -hour readings, respectively. Dryness of the skin, which masked the evaluation of the erythema in one animal, was also observed in 4/10 animals at the 48 -hour reading. In the treated group, at the 24 -hour reading, a discrete or moderate erythema (grade 1 or 2) was recorded in 7/19 and 2/19 animals, respectively. At the 48 -hour reading, a discrete or moderate erythema (grade 1 or 2) was seen in 8/19 and 2/19 animals, respectively. Dryness of the skin was also noted in 8/19 animals at the 48 -hour reading.

The persistent cutaneous reactions observed in 3/19 animals of the treated group, which were of higher severity than those recorded in the animals of the control group, were attributed to possible delayed contact hypersensitivity. According to the classification criteria laid down in Regulation EC n°1272/2008 (CLP), the test item should not be considered as a skin sensitizer.

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

According to the classification criteria laid down in Regulation EC n°1272/2008 (CLP), the test item, Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates should not be considered as a skin sensitizer.

Executive summary:

The potential of the test item, Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates to induce delayed contact hypersensitivity was evaluated in guinea pigs according to the maximization method of Magnusson and Kligman and to OECD 406 guideline. The assay was performed on a mixture containing 30-40% of Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates, 25-35% of butylglycol and water until complement to 100%. As the existing data on butylglycol indicate no sensitization potential and a very slight irritating potential, the presence of this solvent does not compromise the interpretation of the results.

Thirty guinea pigs were allocated to two groups: a control group of five males and five females and a treated group of ten males and ten females. On day 1, three pairs of intradermal injections were performed in the interscapular region of all animals: Freund's complete adjuvant (FCA) diluted at 50% (v/v) with 0.9% NaCl (both groups), test item at the chosen concentration in the chosen vehicle (treated group) or vehicle alone (control group), and test item at the chosen concentration in a mixture FCA/0.9% NaCl (50/50, v/v) (treated group) or vehicle at the concentration of 50% (w/v) in a mixture FCA/0.9% NaCl (50/50, v/v) (control group). On day 8, the test item (treated group) or the vehicle (control group) was applied topically to the same test site, which was then covered by an occlusive dressing for 48 hours. On day 22, all animals of both groups were challenged by a cutaneous application of the test item to the right flank. The left flank served as control and received the vehicle only. The test item and vehicle were maintained under an occlusive dressing for 24 hours. Skin reactions were evaluated approximately 24 and 48 hours after removal of the dressing.

Test item concentrations were as follows: Induction (treated group) = intradermal injections (day 1): test item at the concentration of 0.1% (w/w) in 0.9% NaCl, + topical application (day 8): test item at the concentration of 25% (w/w) in 0.9% NaCl. Challenge (all groups) : topical application (day 22): test item at the concentration of 10% (w/w) in 0.9% NaCl. At the end of the study, animals were killed without examination of internal organs. No skin samples were taken from the challenge application sites of all the animals.

No systemic clinical signs and no deaths related to treatment were noted during the study. The body weight gain of the treated animals was similar to that of controls. Marked local reactions at the intradermal injection sites were noted in a few animals of the treated group, between days 10 and 14. Therefore, one of these animals was sacrificed on day 14 for ethical reasons.

A discrete erythema (grade 1) was noted in 2/10 and 1/10 animals of the control group at the 24 and 48 -hour readings, respectively. Dryness of the skin, which masked the evaluation of the erythema in one animal, was also observed in 4/10 animals at the 48 -hour reading. In the treated group, at the 24 -hour reading, a discrete or moderate erythema (grade 1 or 2) was recorded in 7/19 and 2/19 animals, respectively. At the 48 -hour reading, a discrete or moderate erythema (grade 1 or 2) was seen in 8/19 and 2/19 animals, respectively. Dryness of the skin was also noted in 8/19 animals at the 48 -hour reading.

The persistent cutaneous reactions observed in 3/19 (16%) animals of the treated group, which were of higher severity than those recorded in the animals of the control group, were attributed to possible delayed contact hypersensitivity.

According to the classification criteria laid down in Regulation EC n°1272/2008 (CLP), the test item,Amines, N-(C16-18 and C18 -unsatd. alkyl)trimethylenedi-, diacetates should not be considered as a skin sensitizer.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

The profiling of alkyl-diamines (QSAR Toolbox v.4.1) indicates that no alerts are found for protein binding, thiol reactivity is not expected, and that the structure is not represented among the categories of high, moderate or low reactivity in DPRA (direct peptide reactivity assays) for either cysteine or lysine depletion. Additionally, the molecular structure of the diamines does not contain toxicophores indicating a concern for sensitization, and also read across to data available on structurally related branched triamine (Dodecyl dipropylene triamine) and primary amines in general do not indicate a concern.

(The automated workflow in QSAR Toolbox for sensitisation results to a positive prediction, which is based on cross-reading to a single positive GPMT result from decanol, and a negative LLNA for 1,14-Tetradecanediol. As both substances are not even amine structures, this prediction is not considered valid)

Information from QSARs:

- VEGA (Skin Sensitisation model (CAESAR) version 2.1.6): Predicts sensitizer, but with low validity: it indicates that the substance is out of model Applicability Domain and that there are no similar compounds in the training set. (All listed positive structures are not amines)

- DEREK (Derek Nexus: 3.0.1, Nexus: 1.5.0): predicts that skin sensitisation is plausible. This is based on observation of sensitisation to diamine as diaminoethane, ethyleneamines and 3-aminopropyldimethylamine. These are not considered to be predictive to the cationic surfactant diamines.

- TOPKAT (Accelrys ADMET Toxicity Prediction (Extensible)) predicts non-sensitizer, with highest reliability for longer (C18) chain length.

 

There are no reports on incidences of sensitisation from industrial production and use of the substance.

 

There is a Guinea Pig Maximisation study with CAS No 1313206-64-2,Amines, N-C16-18 and C18 unsaturated alkyl) trimethylenedi-,diacetates which is representative for this reaction mass. Also read-across to data derived from animal testing available for the structurally related primary amines do not indicate a concern.

(For information on the applicability of the read-across from various diamines, see document "Category polyamines - 20170518.pdf" added to IUCLID Ch. 13.)

 

As indicated under skin corrosion/irritation, the skin reactions have a large inflammatory component. These inflammatory reactions lead to local lymph node growth at extreme low concentrations, suggesting of a very potent sensitisation reaction, although the few available data involving guinea pig studies (on other cationic surfactants do not show increased reactions upon challenge. Finally, also for polyamine products used as biocide, there are no specific concerns for sensitisation following handling or use.

 

Discussion: dermal

There is no sensitisation data available for Diamine C16-18, C18-unsaturated (“tallow-diamine”). her is however data for a Guinea Pig Maximisation study with CAS No 1313206-64-2,Amines, N-C16-18 and C18 unsaturated alkyl) trimethylenedi-,diacetates which is representative for this reaction mass.

Available studies indicate that the reaction mass is corrosive to skin cat 1C. .There are no consumer exposures, only industrial/professional use under circumstances involving the use of PPM following the classification as corrosive cat. 1C.

 

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information:

Diamine C16-18, C18-unsaturated (“tallow-diamine”) is a paste with mp of 37°C and has a vapour pressure of less than 0.0015 Pa at 20°C. Its use is limited to industrial and professional users and does not involve the forming of aerosols, particles or droplets of an inhalable size. So exposure to humans via the inhalation route will be unlikely to occur.

Additionally, information from profiling for expected protein interaction and QSARs for sensitisation result to a low concern.

Justification for classification or non-classification

Based on limited exposures by dermal route (substance is severely corrosive) or by inhalation (very low vapour pressure) and a low concern based on profiling for expected protein interaction and QSARs for sensitisation, sensitisation is not expected to be an issue.However, as a firm conclusion from a study with this compound is lacking, no definite conclusion might be drawn for classification purposes.