Fatty acids C18 unsat, reaction products with triethylenetetramine

Skin sensitisation

Administrative data

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.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

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. QA statement)

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Study available before Reach guidance

Test material

Test material information

Constituent 1

Reference substance name:

1H-Imidazole-1-ethanamine, 4,5-dihydro-, 2-nortall-oil alkyl derivs.

EC Number:

270-500-3

EC Name:

1H-Imidazole-1-ethanamine, 4,5-dihydro-, 2-nortall-oil alkyl derivs.

Cas Number:

68442-97-7

Constituent 2

Reference substance name:

Tall oil diethylenetriamine imidazoline

IUPAC Name:

Tall oil diethylenetriamine imidazoline

Details on test material:

- Name of test material (as cited in study report): Tall oil diethylenetriamine imidazoline
- Substance type: Clear slightly viscous amber liquid (determined at NOTOX)
- Physical state: Liquid
- Analytical purity: 98%
- Impurities (identity and concentrations): Free diethylenetriamine (3.6%) + Free fatty accid (2.0%)
- Composition of test material, percentage of components: C-chain distribution: C16 (<=10%) + C18 (>=85%) + C20-24 (<=5%)
- Lot/batch No.: S000922
- Expiration date of the lot/batch:02 July 2017
- Stability under test conditions:
StabilityTall oil diethylenetriamine imidazoline under storage conditions = Stable
Stability Tall oil diethylenetriamine imidazoline in vehicle (Corn oil) = Not indicated
- Storage condition of test material: At room temperature in the dark under nitrogen

In vivo test system

Test animals

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

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

corn oil

Induction 2

Concentration / amount:

intradermal inductie: 0.1%
epidermal inductie: 0.5%
Challange: 0 and 0.5%

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

corn oil

Challenge 2

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

Results and discussion

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.

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

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.

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

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.

Results 3

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.

Results 4

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.

Applicant's summary and conclusion

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.