Fatty acids, C18-unsaturated, reaction products with amines, polyethylenepoly-, (tetraethylenepentamine, pentaethylenehexamine, hexaethyleneheptamine fractions)

Endpoint summary

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

Study period:

1 Sep 2000 - 25 Sep 2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to OECD Guideline 406 and in compliance with GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

The study was performed before the application of the Reach guidance.

Species:

guinea pig

Strain:

Hartley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
Hartley Cd: (HA) BR, Caesarian obtained, Barrier sustained - Virus Antibody Free (COBS - VAF®)
- Source: Charles River France, 76410 Saint-Aubin-lès-Elbeuf, France.
- Age at study initiation: 1-3 months
- Weight at study initiation: 379 ± 13 g for the males and 374 ± 13 g for the females.
- Housing: individually in polycarbonate cages with stainless steel lid (48 cm x 27 cm x 20 cm) equipped with a polypropylene bottle. Dust-free sawdust was provided as litter (SICSA, 94142 Alfortville, France). Sawdust is analysed by the supplier for composition and contaminant levels.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2°C
- Humidity (%): 30 to 70%
- Air changes (per hr): 12 cycles/hour of filtered, non-recycled air.
- Photoperiod (hrs dark / hrs light): 12 h/12 h

IN-LIFE DATES: From: 1 Sep 2000 To: 25 Sep 2000

Route:

intradermal and epicutaneous

Vehicle:

physiological saline

Concentration / amount:

The dosage form preparation which could pass through a needle and into the dermis had a maximum concentration of 10% (w/w).
Intradermal induction: 0.1%
epicutaneous induction: 5%
epicutaneous challenge: 1%

Route:

epicutaneous, occlusive

Vehicle:

physiological saline

Concentration / amount:

The dosage form preparation which could pass through a needle and into the dermis had a maximum concentration of 10% (w/w).
Intradermal induction: 0.1%
epicutaneous induction: 5%
epicutaneous challenge: 1%

No. of animals per dose:

Control: 5 animals/sex
Treated group: 10 animals /sex

Details on study design:

RANGE FINDING TESTS:
Series of test substance concentrations were tested by intra dermal (0.1, 1, 5, 10% with and without FCA) and epicutaneous route (0.1, 1, 5, 10, 50 100%)
Intradermal: Concentrations of 1% and higher were corrosive. 0.1% resulted to irritation visible during all reported observation day from 24h to 6 days. This concentration was selected for the main study.
Epicutaneous: 5% was the lowest concentration leading to irritation (grade 2: moderate and confluent erythema). This concentration was used for epidermal induction. 1% was without signs of irritation, and was select for challenge concentration.

MAIN STUDY
A. INDUCTION EXPOSURE
-Day 1
The scapular region was clipped and three pairs of intradermal injections (0.1 mL/site) were made in this area as follows:
A) A 1:1 w/w mixture of FCA (Sigma, France) with 0.9% NaCl.
B) The test substance at a 50% concentration in 0.9% NaCl.
C) A 1:1 w/w mixture of the undiluted test substance and FCA.
Note: One of each pair was on each side of the midline and from cranial A) to caudal C).
Controls were treated similarly, but vehicle instead of testsubstance was used.

Day 7
The scapular area between the injection sites was clipped .

Day 8
a pad of filter paper (approximately 8 cm2) was fully-loaded with the test substance at the concentration of 5% (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
Day 21: Clipped and shaved each flank.
Day 22: the animals of treated and control groups received an application of the test substance and vehicle. The filter paper of a chamber (Finn Chamber®) was fully-loaded with the test substance at the concentration of 1% (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.
Twenty-four and 48 hours after removal of the dressing of the challenge application, both flanks of the treated and control animals were observed in order to evaluate cutaneous reactions, according to the following scale:
• 0 - no visible change
• 1 - discrete or patchy erythema
• 2 - moderate and confluent erythema
• 3 - intense erythema
Any observed oedema and other lesions were noted..

In life examinations:
At least one a day for clinical signs and mortality
Body weights: Prior to start and at termination of the study.

Positive control substance(s):

yes

Positive control results:

The sensitivity of the experimental technique is regularly assessed using a known moderate sensitizer, MERCAPTOBENZOTHIAZOLE. In a recent study performed under CIT experimental conditions, the strain of guinea pigs used showed a satisfactory sensitization response in 70% animals.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

1% w/w

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

Two animals skin injured by dressing, making scoring difficult/impossible; one of these animals also showed crusts.

Remarks on result:

other: see Remark

Remarks:

Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 1% w/w. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Two animals skin injured by dressing, making scoring difficult/impossible; one of these animals also showed crusts..

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

5% w/w

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

Two animals skin injured by dressing, making scoring difficult/impossible.

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 5% w/w. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Two animals skin injured by dressing, making scoring difficult/impossible..

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

5%

No. with + reactions:

19

Total no. in group:

19

Clinical observations:

One animal died study day 11 (before challenge). Dryness of skin and/or crusts observed in most animals.

Remarks on result:

other: see Remark

Remarks:

Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 5%. No with. + reactions: 19.0. Total no. in groups: 19.0. Clinical observations: One animal died study day 11 (before challenge). Dryness of skin and/or crusts observed in most animals..

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

5% w/w

No. with + reactions:

19

Total no. in group:

19

Clinical observations:

Dryness of skin and/or crusts observed in all animals.

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 5% w/w. No with. + reactions: 19.0. Total no. in groups: 19.0. Clinical observations: Dryness of skin and/or crusts observed in all animals..

Interpretation of results:

sensitising

Remarks:

Migrated information

Conclusions:

Reactions were observed in all animals of treatment group and none in the control. Cutaneous effects were attributed to delayed contact hypersenisitivity.

Executive summary:

The potential of the test substance BASE 136 (batch No. AB77l) to induce delayed contact hypersensitivity was evaluated in guinea pigs according to the maximization method of Magnusson and Kligman and to OECD (No. 406, 17th July 1992) and EC (96/S4/EEC, B.6, 30 July 1996) guidelines.

The study was conducted in compliance with the principles of Good Laboratory Practice Regulations.

 

 

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% with 0.9% NaCl (both groups),

- test substance at the chosen concentration in the chosen vehicle (treated group) or vehicle alone (control group),

- test substance at the chosen concentration in a mixture FCA/0.9% NaCl 50/50 (treated group) or vehicle at the concentration of SO% in a mixture FCAl/0.9% NaCl 50/50 (control group).

 

On day 8, the test substance (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 the treated and control groups were challenged by a cutaneous application of the test substance to the right flank. The left flank served as control and received the vehicle only. Test substance 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 substance concentrations were as follows:

 

Induction (treated group)

-       intradermal injections (day 1): BASE 136 at the concentration of 0.1% (w/w) in sterile isotonic saline solution (0.9% NaCl),

-       topical application (day 8): BASE 136 at the concentration of S% (w/w) in sterile isotonic saline solution (0.9% NaCl).

 

Challenge (all groups)

-       topical application (day 22): BASE 136 at the concentration of 1% (w/w) in sterile isotonic saline solution (0.9% NaCl).

 

At the end of the study, animals were killed without examination of internal organs.

Skin samples were taken from the challenge application sites of all the animals.

No histological examination was performed.

 

Results

No clinical signs and no deaths related to treatment were noted during the study.

 

These results on Fatty acids C16-18, C18 unsat reaction products with tetraethylenepentamine are fully valid for Tall oil, reaction products with tetraethylene-pentamine (Amidoamine/Imidazoline) as both substances have the same fatty acid distribution and both used TEPA as starting material.

After the challenge application, no relevant cutaneous reactions were observed in the animals of the control group.

In the treated group, a discrete or moderate erythema was noted in all animals. An oedema was recorded in 11/19 animals. Dryness of the skin and/or crusts were observed all animals. The observed cutaneous were attributed to delayed contact hypersensitivity.

 

Conclusion

Under our experimental conditions and according to the maximization method of Magnusson and Kligman, the test substance BASE 136 (batch No. AB771) induces delayed contact hypersensitivity in 19/19 (100%) guinea pigs.

According to the classification criteria laid down in Directive 93/21/EEC (27th April 1993) adapting to technical progress for the eighteenth time Council Directive 67/548/EEC, the test substance should be considered as a skin sensitizer.

These results lead to classification according to CLP (ATP 2): Skin sensitiser Cat.1A (GPMT = 30% positive at = 0.1% i.d. induction)

As explained in the category justification, for cross-reading in general use is made with data of same or lower EA-length where available, and that of Tall oil + DETA representing the worst case. This dossier is for the substance "Fatty acids C18 unsat, reaction products with Triethylenetetramine, tetraethylenepentamine, Pentaethylenehexamine and polyethylene amines" (or TO + TETA, TEPA, PEHA, PolyEA). As for the substance itself no toxicological information is available, cross-reading has been applied to TO + TEPA.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

As explained in the category justification, for cross-reading in general use is made with data of same or lower EA-length where available, and that of Tall oil + DETA representing the worst case. This dossier is for the substance Fatty acids C18 unsat, reaction products with polyethylene amines. As for the substance itself no toxicological information is available, cross-reading has been applied to TO + TEPA.

Fatty acids C16-18, C18 unsat reaction products with tetraethylenepentamine was found to be sensitising to skin in a GPMT test.

All substances within the AAI group show the same reactive groups, show similar composition of amide, imidazoline, and some dimer structures of both, with the length of original EA amines used for production as biggest difference. Inherent reactivity and toxicity is not expected to differ much between these substances, aspects which determine sensitization.

All skin sensitisation studies performed on AAI substances all indicate them to be sensitising.

The following information is available on sensitising properties of AAI substances:

    TO + DETA                            Sensitising: 40% positive reaction in GPMT, i.d. induction 0.1%

    TO + DETA                            Sensitising: LLNA: EC3: 0.3%.

    C16-18, C18 unsat + TEPA   Sensitising: 100% positive reaction in GPMT, i.d. induction 0.1%

    TO + Poly(Amide)                  Sensitising: 100% positive reaction in GPMT, i.d. induction 0.2%

 

Based on the available data, all substances in the AAI group should be considered sensitising to skin.

Classification according to CLP (ATP 2) skin sensitiser: Cat.1A (LLNA: EC 3 = 2%; GPMT = 30% positive at = 0.1% i.d. induction or = 60% positive at >0.1% i.d. induction)

Migrated from Short description of key information:
Fatty acids C16-18, C18 unsat reaction products with tetraethylenepentamine was found to be sensitising to skin in a GPMT test.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information:

There is no information on respiratory sensitisation, but AAI products are sensitising to skin.

The likelihood for exposure via inhalation and thus becoming sensitised to AAI, is however very low, based on the high boiling point (> 300 °C) and very low vapour pressure (0.00017 mPa at 25°C for DETA based AAI).

Migrated from Short description of key information:
No data.

Justification for classification or non-classification

Various studies with different AAI indicate that these substances can cause dermal sensitisation and should be classified as dermal sensitiser Cat.1A for CLP (Criteria ATP 2, Commission Regulation 286/2011 of 10 March 2011).

However, the actual risk of sensitisation is probably low, as AAI are corrosive to skin and consequently exposure will be low due to necessary protective measures to limit dermal exposure.

Also the likelihood for exposure via inhalation and thus becoming sensitised to AAI, is very low, based on the high boiling point (> 300 °C) and very low vapour pressure (0.00017 mPa at 25°C for DETA based AAI).