Registration Dossier

Administrative data

Description of key information

No data on sensibilisation is available for "Fatty acids C18 unsat, reaction products with triethylenetetramine" (or TO + TETA). Read-across is performed with "Fatty acids C18 unsat, reaction products with diethylenetetramine" (or TO + DETA).

There is no information on respiratory sensitisation, but AAI-DETA is sensitising by dermal route.

AAI-DETA is a viscous liquid with a boiling point over 300°C, and has a vp of 1.7 x 10-7 Pa at 25°C. Use of the substance is limited to industrial and professional users and does not result in aerosols, particles or droplets of an inhalable size. So exposure to humans via the inhalation route will be unlikely to occur.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09 March 2010 - 16 April 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study has been performed according to OECD and/or EC guidelines and according to GLP principles.
Qualifier:
according to
Guideline:
other: OECD Guideline 406 (Skin Sensitisation) EU Method B.6 (Skin Sensitisation) EPA OPPTS 870.2600 (Skin Sensitisation) Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000, including the most recent
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Study available before Reach guidance
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Kisslegg, Germany.
- Age at study initiation: approx. 6 weeks old
- Weight at study initiation: no data
- Housing: Group housing of maximally 5 animals per labeled cage (74 cm x 54 cm x 25 cm height) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France).
- Diet (e.g. ad libitum):Complete breeding diet for guinea pigs (SSNIFF® MS-Z, V2273; SSNIFF® Spezialdiäten GmbH, Soest, Germany). Hay (TecniLab-B MI BV, Someren, The Netherlands) was provided at least twice a week.
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: The acclimatization period was at least 5 days before the start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.5 – 20.7ºC
- Humidity (%): 23 - 95%)
- Air changes (per hr): approximately 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours darkness per day.

IN-LIFE DATES: From: To: 09 March 2010 - 16 April 2010
Route:
intradermal and epicutaneous
Vehicle:
corn oil
Concentration / amount:

intradermal inductie: 0.1%
epidermal inductie: 0.5%
Challange: 0 and 0.5%
Route:
epicutaneous, occlusive
Vehicle:
corn oil
Concentration / amount:

intradermal inductie: 0.1%
epidermal inductie: 0.5%
Challange: 0 and 0.5%
No. of animals per dose:
Experimental group: 10 females.
Control group: 5 females.
Details on study design:
RANGE FINDING TESTS:
Series of test substance concentrations were tested (0.5, 1, 2 and 5%. )
The test system and procedures were identical to those used during the main study, unless otherwise specified. The six animals selected were between 4 and 9 weeks old. No body weights were determined.


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 Freunds' Complete Adjuvant (Difco, Detroit, U.S.A.) with water for injection (Fresenius AG, Bad Homburg, Germany).
B) The test substance at a 0.1% concentration.
C) A 1:1 w/w mixture of the test substance, at twice the concentration used in (B) and Freunds' Complete Adjuvant.
Note: One of each pair was on each side of the midline and from cranial A) to caudal C).

Day 3
The dermal reactions caused by the intradermal injections were assessed for irritation.


Day 8
The scapular area between the injection sites was clipped and subsequently treated with 0.5 mL of a 0.5% test substance concentration using a Metalline patch (2x3 cm) mounted on Medical tape, which was held in place with Micropore tape and subsequently Coban elastic bandage.

The dressing was removed after 48 hours exposure, the skin cleaned of residual test substance using water and the dermal reactions caused by the epidermal exposure were assessed for irritation.


B. CHALLENGE EXPOSURE
-Day 21
One flank of all animals was clipped and treated by epidermal application of a 0.5% test substance concentration and the vehicle (0.1 mL each), using Patch Test Plasters (Curatest®, Lohmann, Almere, The Netherlands). The patches were held in place with Micropore tape and subsequently Coban elastic bandage.

The dressing was removed after 24 hours exposure and the skin cleaned of residual test substance and vehicle using water. The treated sites were assessed for challenge reactions 24 and 48 hours after removal of the dressing.

After termination, animals were sacrificed by intra peritoneal injection of pentobarbital (Euthesate®; Sanofi Sante B.V., Maassluis, The Netherlands).

In life examinations:

Mortality/Viability: Twice daily

Toxicity: At least once daily.

Body weights: Prior to start and at termination of the study.

Irritation:
Skin reactions were graded according to the following numerical scoring systems. Furthermore, a description of all other (local) effects was recorded.
To facilitate scoring, the epidermally treated skin-areas were clipped at least 3 hours before the 48-hour reading of these areas in the preliminary irritation study and challenge phase.

Positive control substance(s):
yes
Remarks:
Alpha-Hexylcinnamicaldehyde
Positive control results:
Reliability test:

Was performed not more than 6 months previously. Similar procedures were used in the reliability test and in this study.

The skin reactions observed in seven experimental animals in response to the 20% test substance concentration in the challenge phase were considered indicative of sensitisation, based on the absence of any response in the control animals. These results lead to a sensitisation rate of 70 per cent to the 20% concentration.

From these results, it was concluded that the female guinea pig of the Dunkin Hartley strain is an appropriate animal model for the performance of studies designed to evaluate the sensitizing potential of a substance in a Maximization type of test.


Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
0.5%
No. with + reactions:
4
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.5%. No with. + reactions: 4.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
0.5%
No. with + reactions:
1
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.5%. No with. + reactions: 1.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
20%
No. with + reactions:
7
Total no. in group:
10
Remarks on result:
other:
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control group. Dose level: 20%. No with. + reactions: 7.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other:
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: vehicle control group. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 5.0.
Interpretation of results:
sensitising
Remarks:
Migrated information
Conclusions:
The results indicate a sensitization rate of 40 per cent.
Executive summary:

Assessment of Contact Hypersensitivity to Tall oil diethylenetriamine imidazoline in the Albino Guinea Pig( Maximization Test).

The study was carried out based on the guidelines described in:

OECD No. 406 (1992) "Skin Sensitization"

EC No 440/2008; B6: "Skin Sensitization: Guinea-Pig Maximization Test (GPMT)"

EPA OPPTS 870.2600 (2003) “Skin Sensitization”

JMAFF Guidelines (2000) including the most recent partial revisions.

The study was based on the method described by Magnusson and Kligman, "Allergic Contact Dermatitis in the Guinea Pig - Identification of Contact Allergens" (1970).

 

Test substance concentrations selected for the main study were based on the results of a preliminary study.

 

In the main study, ten experimental animals were intradermally injected with a 0.1% concentration and epidermally exposed to a 0.5% concentration. Five control animals were similarly treated, but with vehicle alone (corn oil).

 

Two weeks after the epidermal application all animals were challenged with a 0.5% test substance concentration and the vehicle.

 

Skin reactions of grade 1 were observed in four experimental animals in response to the 0.5% test substance concentration. No skin reactions were evident in the control animals.

 

The skin reactions observed in response to a 0.5% test substance concentration in 4 (of the ten) experimental animals in the challenge phase were considered indicative of sensitization, based on the absence of any response in the control animals. These results indicate a sensitization rate of 40 per cent.

The 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" (or TO + TETA). As for the substance itself no toxicological information is available, cross-reading has been applied to TO + DETA.

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20 Feb 2007 - 28 Feb 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study has been performed according to OECD and/or EC guidelines and according to GLP principles.
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: approximately 9 weeks
- Weight at study initiation: mean body weight ± standard deviation of 20.9 ± 1.0 g.
- Housing: individually in disposable crystal polystyrene cages (22.00 cm x 8.50 cm x 8.00 cm). Each cage contained autoclaved sawdust (SICSA, Alfortville, France)
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 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: 20 Feb 2007 To: 28 Feb 2007
Vehicle:
acetone/olive oil (4:1 v/v)
Concentration:
0, 0.05, 0.1, 0.25 and 1 % (v/v)
No. of animals per dose:
4 females/dose
Details on study design:
RANGE FINDING TESTS:
- Irritation: Erythema and crusts were noted on ears of all animals given the concentrations = 2.5%. A significant increase in ear thickness was recorded at concentrations = 2.5%, showing the irritant potential of the test item at these concentrations. The highest concentration retained for the main test was therefore 1%.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method:
- Criteria used to consider a positive response:

TREATMENT PREPARATION AND ADMINISTRATION:
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
SI = dpm of treated group / dpm of control group
skin sensitizer when the SI for a dose group is = 3.
EC3 derived by linear interpolation of points on the dose-response curve, immediately above and below the 3-fold threshold.
Positive control results:
HCA 25%, SI = 14.38. The study was therefore considered valid.
Parameter:
SI
Value:
>= 1 - <= 18.65
Test group / Remarks:
overall
Parameter:
EC3
Value:
0.3
Test group / Remarks:
Overall
Parameter:
SI
Value:
1
Test group / Remarks:
0.05%
Parameter:
SI
Value:
1.88
Test group / Remarks:
0.1%
Parameter:
SI
Value:
1.67
Test group / Remarks:
0.25%
Parameter:
SI
Value:
5.91
Test group / Remarks:
0.5%
Parameter:
SI
Value:
18.95
Test group / Remarks:
1%

No mortality and no clinical signs were observed during the study.

The body weight change of treated animals was similar to that of control animals.

Local irritation:

A dryness of the skin of the ears was noted on day 6 in animals given the concentrations of 0.5 and 1%.

Increases in ear thickness were noted at concentrations of 0.5% (19.19% increase) and 1% (30.10% increase).

Proliferation assay:

The quantity of cells obtained in each group was satisfactory and the cellularity correlated with incorporation of 3H-TdR. The cell viability was higher than 70% in each group.

In the positive control group given HCA at the concentration of 25%, a moderate increase in cellularity and a stimulation index exceeding the threshold value of 3 (SI = 14.38) were noted. The study was therefore considered valid.

A dose-related increase in the SI was recorded and the threshold positive value of 3 was exceeded at 0.5%.

In the absence of an excessive local irritation at the concentration of 0.5% (increase in ear thickness < 30%), the significant lymphoproliferative response observed was attributed to delayed contact hypersensitivity.

The EC3 value for the test item IMIDAZOLINE 4900 is equal to 0.3%.

Study results

Group

Treatment concentration

Cell count

viable %

amount of cells
(x 10^6)

Cellularity index

Number of nodes per group

dpm per group

dpm per node

Stimulation Index (SI)

Increase in ear thickness (% between day 1 and day 6)

Irritation level

EC3 value (%)

viable

dead

1

Vehicle

42

8

84.00

4.20

 

8

375.97

47.00

 

1.98

 

 

2

Test Item 0.05%

46

9

83.64

4.60

1.10

8

375.95

47.12

1.00

0.98

I

 

3

Test Item 0.1%

66

10

86.84

6.60

1.57

8

707.84

88.48

1.88

1.00

I

0.3

4

Test Item 0.25%

67

8

89.33

6.70

1.60

8

629.60

78.70

1.67

8.16

I

5

Test Item 0.5%

154

40

79.38

15.40

3.67

8

2221.13

277.64

5.91

19.19

II

 

6

Test Item
1%

258

52

83.23

25.80

6.14

8

7012.20

876.53

18.65

30.10

III

 

7

HCA
0.05%

250

32

88.65

25.00

5.95

8

5406.58

675.82

14.38

 

 

 

Interpretation of results:
sensitising
Remarks:
Migrated information
Conclusions:
Test item should be considered as a strong sensitizer.
Executive summary:

The aim of this study was to evaluate the potential of the test item IMIDAZOLINE 4900 (batch No. pilote du 27/07/06) to induce delayed contact hypersensitivity using the murine Local Lymph Node Assay (LLNA). Evaluation of local irritation was also carried out in parallel.

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

 

Methods

A preliminary test was first performed in order to define the concentrations of test item to be used in the main test.

In the main test, twenty-eight female CBA/J mice were allocated to seven groups:

- five treated groups of four animals receiving the test item IMIDAZOLINE 4900 at the concentration of 0.05, 0.1, 0.25, 0.5 or 1%,

- one negative control group of four animals receiving the vehicle (mixture acetone/olive oil (4/1, v/v)),

- one positive control group of four animals receiving the reference item,a-hexylcinnamaldehyde (HCA), a moderate sensitizer, at the concentration of 25%.

During the induction phase, the test item, vehicle or reference item was applied over the ears (25 µL per ear) for 3 consecutive days (days 1, 2 and 3). After 2 days of resting, the proliferation of lymphocytes in the lymph node draining the application site was measured by incorporation of tritiated methyl thymidine (day 6). The obtained values were used to calculate stimulation indices (SI).

 

The irritant potential of the test item was assessed in parallel by measurement of ear thickness on days 1, 2, 3 and 6.

 

Results

The test item was not soluble in any of the five recommended vehicles, consequently, an homogenous suspension, obtained in acetone/olive oil (4/1, v/v), was used for the treatment.

The maximum practicable concentration in this vehicle was 50%. Consequently, the concentrations selected for the first preliminary test were 10, 25, 50 and 100%.

Since the test item was irritant at all these concentrations, a complementary preliminary test was undertaken at lower concentrations: 0.5, 1, 2.5 and 5%.

Since the test item was irritant in the complementary preliminary test at the concentrations of 2.5 and 5%, the highest concentration retained for the main test was 1%.

 

Systemic clinical signs and mortality

No mortality and no clinical signs were observed during the study.

 

Local irritation

A dryness of the skin of the ears was noted on day 6 in animals given the concentrations of 0.5 and 1%.

Increases in ear thickness were noted at concentrations of 0.5% (19.19% increase) and 1% (30.10% increase).

Proliferation assay

A significant lymphoproliferation was noted in the positive control group given HCA, the study was therefore considered as valid.

A dose-related increase in the SI was recorded and the threshold positive value of 3 was exceeded at 0.5%.

The results are presented in the following table:

Treatment

Concentration (%)

Irritation level

Stimulation Index (SI)

Test item

0.05

non-irritant

1

Test item

0.1

non-irritant

1.88

Test item

0.25

non-irritant

1.67

Test item

0.5

slightly-irritant

5.91

Test item

1

irritant

18.65

HCA

25

-

14.38

   

In the absence of an excessive local irritation at 0.5% (increase in ear thickness < 30%), the significant lymphoproliferative response observed at this concentration was attributed to delayed contact hypersensitivity.

The EC3value for the test item IMIDAZOLINE 4900 is equal to 0.3%.

 

Conclusion

Under our experimental conditions, the test item IMIDAZOLINE 4900 (batch No. pilote du 27/07/06) induced delayed contact hypersensitivity in the murine Local Lymph Node Assay.

According to the EC3 value obtained in this experiment, the test item IMIDAZOLINE 4900 should be considered as a strong sensitizer.

The results lead to classification according to CLP (ATP 2) skin sensitiser Cat.1A (EC 3 = 2%)

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 triethylenetetramine" (or TO + TETA). As for the substance itself no toxicological information is available, cross-reading has been applied to TO + DETA.

Tall oil + DETA was found to be sensitising to skin in a GPMT and in a LLNA.

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)

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available
Additional information:

There is no information on respiratory sensitisation, but Tall oil + DETA is sensitising by dermal route.

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).

Justification for classification or non-classification

Various studies with different AAI indicate that these substances can cause dermal sensitisation. Tall oil + DETA was found to be sensitising to skin in a GPMT, and in a LLNA, resulting to an EC3 of 0.3% indicating that it should be considered as a strong sensitizer and should be classified as dermal sensitiser Cat.1A for CLP (Criteria ATP 2, Commission Regulation 286/2011 of 10 March 2011).

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).


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