2-(2-aminoethylamino)ethanol

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline Study OECD 429 and GLP compliance

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

Animals:
- Origin: Harlan Winkelmann GmbH, Gartenstr. 27, 33178 Borchen, Germany
- Sex: Female
- Identification of the animals: The single housed animals were identified by cage cards
- Age at the beginning of the study: ca. 7 weeks
- Body weight range at day 0: 17.1 g – 20.7 g
- Reasons for the selection: The test method was developed in mice.

Housing conditions:
- Air conditions: The animals were housed in fully air-conditioned rooms in which a central air-conditioning system ensured a temperature in the range of 20 - 24 °C and a relative humidity in the range of 30 - 70 %.

- Illumination period: 12 h light (6.00 a.m. - 6.00 p.m.) , 12 h darkness (6.00 p.m. - 6.00 a.m.)
- Type of cage: Makrolon type I
- No. of animals per cage: 1
- Type of diet: Kliba-Labordiät (Maus / Ratte Haltung “GLP”), Provimi Kliba SA, Kaiseraugst, Basel, Switzerland, ad libitum
- Watering: Tap water ad libitum
- Bedding: Granulat Typ ¾ (staubfrei); SSNIFF
- Acclimatization period: 8 days before the first test substance application

Vehicle:

other: acetone

Concentration:

1) 3 %
2) 10 %
3) 30 %
4) vehicle acetone

No. of animals per dose:

6

Details on study design:

The study comprised three treatment groups and a vehicle control group. Each group consisted of 6 mice. A check for dead or moribund animals was made twice each workday and once on Saturdays, Sundays and on public holidays.
No detailed clinical examination of the individual animals was performed but any obvious signs of systemic toxicity and/or local inflammation at the application sites were noted in the raw data.

- Body weights of the individual animals were determined on study day 0 prior to the first application and on day 5 prior to the sacrifice of the animals
- Test substance application: Epicutaneous application is simulating dermal contact with the compound which is possible to occur under practical use conditions.
- Application volume: 25 μL per ear
- Site of application: Dorsal part of both ears
- Frequency of application: 3 consecutive applications (day 0 – day 2) to the same application site
- On study day five (about 66 to 72 hours after the last application of test substance to the ears) the mice were injected intravenously (i.v.) with 20 μCi of 3H-thymidine in 250 μl of sterile saline into a tail vein.

- The animals were sacrificed on study day 5 about 5 hours after 3H-thymidine injection by cervical dislocation.

- Immediately after the death of each animal ear thickness was measured with a suitable gauge (Oditest®-device (measurement steps 0.01 mm), Kroeplin, Germany). Thereafter a circular piece of tissue (diameter 0.8 cm) was punched out of the apical part of each ear of all animals. The weight of the pooled punches was determined for each animal. These measurements serve for detecting a potential inflammatory ear swelling.

- Immediately after removal of the ear punches the left and right auricular lymph nodes were dissected. The weight of the pooled lymph nodes from both sides was determined for each animal.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

Mean values and standard deviations of the measured parameters were calculated for the test and control groups from the individual values. The stimulation indices of cell count, 3H-thymidine incorporation, lymph node weight, ear weight and ear thickness measurements were calculated as the ratio of the test group mean values for these parameters divided by those of the vehicle control group. Further statistical analyses ( Cell counts, 3H-thymidine incorporation, lymph node weight, ear weights as well as ear thickness) were performed with the WILCOXON-Test.

Positive control results:

The Murine Local Lymph Node Assay is able to show the skin sensitizing effect of Alpha-Hexylcinnamaldehyde, techn. 85 % under the test conditions chosen. The threshold concentration for sensitization induction was between 5 % and 10 % under the test conditions chosen. The EC 1.5 for cell count and the EC 3 for 3H-thymidine incorporation was calculated by linear regression from the results of these concentrations to be 6.9 % and 10.5 % , respectively.

Parameter:

SI

Remarks on result:

other: control (vehicle) 1.0, 3 % AEEA 2.0, 10 % AEEA 1.72, 30 % AEEA 6.6 EC 3: 15.2 %

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: Means: control (vehicle) 444.9, 3 % AEEA 891.1, 10 % AEEA 766.1, 30 % AEEA 2938.3

- Cell counts, 3H-thymidine incorporation and lymph node weights

When applied as 30 % preparation in acetone, the test substance induced a statistically significant and biologically relevant (increase to 1.5 fold or above of control value = stimulation index (SI) 1.5) increase in cellularity of the auricular lymph nodes. Concomitantly, the increase of 3H-thymidine incorporation into the cells was statistically significant and biologically relevant (increase above the cut off stimulation index of 3). The 10 % and 3 % test substance preparation caused small but statistically significant increases in cellularity of the auricular lymph nodes and 3H-thymidine incorporation into the lymph node cells, which failed to reach the cut off stimulation indices and thus lie below the threshold of immunologic relevance. The lymph node weights were statistically significantly increased in all substance treated groups.

- Ear weights and ear thickness

Statistically significant increases in ear weights and ear thickness were observed in mice treated with the 30 % test substance preparation, which was accompanied by incrustation of the ear skin observed before the third application and on the day of lymph node removal. The 10% test substance preparation caused a statistically significant increase in ear weights, but not in ear thickness. The magnitude of ear skin irritation observed in the test group treated with a 30 % preparation might be considered relevant for evaluating the sensitizing potential of the test substance. As the stimulation indices at this concentration, especially that for cellularity, are well beyond the cut off criteria, an immunologic background of the lymph node response cannot be excluded.

Interpretation of results:

sensitising

Remarks:

Migrated information

Executive summary:

In a dermal sensitisation study with AEEA (BASF AG, Experimental Toxicology and Ecology, 2006) groups of 6 female CBA/Ca mice each were treated with 3 %, 10 % and 30 % w/w preparations of the test substance in acetone or with the vehicle alone using the Local Lymph Node Assay.AEEA shows a skin sensitizing effect under the test conditions chosen. The threshold concentration for sensitization induction was between 10 % and 30 %. The EC 3 for ³H-thymidine incorporation was calculated by linear regression from the results of these concentrations to be 15.2 %.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

In a dermal sensitisation study with AEEA (BASF AG, Experimental Toxicology and Ecology, 2006) groups of 6 female CBA/Ca mice each were treated with 3 %, 10 % and 30 % w/w preparations of the test substance in acetone or with the vehicle alone using the Local Lymph Node Assay.

AEEA shows a skin sensitizing effect under the test conditions chosen. The threshold concentration for sensitization induction was between 10 % and 30 %. The EC 3 for ³H-thymidine incorporation was calculated by linear regression from the results of these concentrations to be 15.2 %.

 

In a dermal sensitisation study with AEEA in acetone/olive oil 4:1 (AOO) Balb/c mice were tested using the Local Lymph Node Assay (Dearman, 2001). The test concentrations were 2.5, 5.0, 10 and 20 %. The stimulation index for AEEA was > 3, with a maximal value of nearly 15 achieved at the maximum applied concentration of 20 %. The EC 3 concentration was calculated to be 5.3 %. AEEA was a sensitizer in this assay.

 

In a dermal sensitisation study (Leung, 1997) with AEEA in deionized distilled water young adultpigs (10/sex) were tested using the method of Magnusson and Kligman (Induction 5 % intracutaneous, Induction 50 % epicutaneous, challenge 25 % epicutaneous) according to OECD Guideline 406. In this study 40 % (8/20) of the animals treated with AEEA showed a skin sensitization response (score≥1). AEEA was rated as a moderate dermal sensitiser.

Migrated from Short description of key information:
In three sensitisation studies with AEEA both LLNAs and the guinea pig maximisation test were positive.

Justification for selection of skin sensitisation endpoint:
The key study (BASF AG, 2006) was selected.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available (further information necessary)

Additional information:

Whitin the LLNA test published by Dearman (2001) decribed above the ability of AEEA to induce cytokines was tested.

According to the authors, the positive response within the LLNA test and the significant expression of the type 1 cytokine Interferon-gamma within the Cytokine Fingerprinting Assay indicate that AEEA possesses contact allergic potential, but lacks significant respiratory sensitizing potential.

Migrated from Short description of key information:
There are no human data available indicative of a respiratory sensitisation. Cytokine profiling performed in a LLNA study using mice indicated that AEEA lacks a significant respiratory sensitisation potential.

Justification for classification or non-classification

Based on the results of the sensitisation studies, AEEA is subject to classification and labelling as follows:

R43 (may cause sensitisation by skin contact) according to Directive 67/548/EEC and Skin sensitizer Cat 1B / H317 (may cause an allergic skin reaction) according to Regulation 1272/2008/EC.