Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Description of key information

The test substance is a skin sensitizer.

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
Study period:
10 September 2008 - 18 September 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)
Specific details on test material used for the study:
- Expiration date: Augustus 2009
- Storage conditions: 15-30°C
- Physical condition: Clear colorless liquid
Species:
mouse
Strain:
CBA:J
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Jackson Laboratories, Bar Harbor, ME 04609
- Age at study initiation: 10-12 weeks
- Weight at study initiation: 21-25 gram
- Housing: Animals were group housed 5 per cage
- Diet: Harlan Teklad Certified Rodent Chow 7012C, ad libitum
- Water: Tap water, ad libitum
- Acclimation period: six days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18.3 - 23.3
- Humidity (%): 43 - 81
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
other: 1:3 Diethyl phthalate/Ethanol
Concentration:
1, 2.5, 5, 10, 25% (v/v)
No. of animals per dose:
5
Details on study design:
In-Life Observations and Measurements
- Mortality/Morbidity: Daily on days 1 to 6
- Clinical Observations: Observations were performed prior to dose administration and following dose administration. Clinical observations were performed once daily on days 4-6. Particular attention was given to the application sites. Any significant alteration to the application sites, and the general appearance of the pinnae, including build up of test article, was recorded.
- Dermal irritation: Animals were examined daily for signs of erythema and edema. Irritation was scored an recorded using the Draize scoring system. Scoring was performed prior to dosing on days 1-3.
- Body weight: Animals were weighted on days 1 and 6.

Method of performance
- Mice were treated on the dorsal surface of both ears (25 µL/ear), once per day on days 1, 2, and 3. Approximately 24±2 hours between applications of test article was maintained. On day 6 the mice were injected i.v. with 20 µCi of 3H-thymidine in 250 µl of sterile saline. Five hours later the mice were euthanized by CO2 asphyxiation and the draining auricular lymph nodes were removed. At removal, the number of nodes collected per animal was recorded, and the nodes were examined for size/appearance and the data recorded. Any unexpected observations were noted in study records. The lymph nodes from each group were pooled and a single cell suspension was prepared. Cells were washed twice with phosphate buffered saline (PBS) and precipitated with 5% trichloroacetic acid (TCA) overnight at 2-8°C. The pellets were recovered by centrifugation and resuspended in 1 mL of TCA and transferred to a vial containing scintillation fluid. An additional 1 mL of TCA was used to rinse the tube, and it was also transferred to the scintillation fluid. Incorporation of 3H-thymidine was measured in a beta-scintillation counter. DPM for each group was determined Increases in 3H-thymidine incorporation relative to the vehicle-treated control was derived for each group and recorded as stimulation indices (SI).
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
SYSTAT version 9.01 F developed by SPSS, Inc was used for statistical analysis. Body weights on Days 1 and 6 and body weight changes were also evaluated using SYS TAT version 9.01. The evaluation of the equality of means for body weight data was made by a one-way analysis of variance using the F distribution to assess statistical significance. If statistically significant differences between the means are found, a Dunnett's test was used to determine the degree of significance from the control means.
Positive control results:
In September 2008, for HCA, SI of 5.5, 6.1, and 19.5 were determined at concentrations of 5, 15, and 35%, respectively.
Key result
Parameter:
EC3
Value:
4.2
Test group / Remarks:
5 animals per group
Parameter:
SI
Value:
1.2
Test group / Remarks:
1%
Parameter:
SI
Value:
1.9
Test group / Remarks:
2.5%
Parameter:
SI
Value:
3.5
Test group / Remarks:
5%
Parameter:
SI
Value:
3.6
Test group / Remarks:
10%
Parameter:
SI
Value:
12.8
Test group / Remarks:
25%
Cellular proliferation data / Observations:
- Mortality: There was no mortality and all animals appeared normal throughout the study.
- Clinical Observations: No erythema or edema was noted in any of the mice in the vehicle group or in those dosed with the test article at 1, 2.5, 5, 10 or 25% (v/v). Ears appeared wet in mice treated with 25% from days 2-4. There were no other findings.
- Body Weights: The mean body weights of the mice treated on days 1 and 6 showed no statistically significant differences. Therefore, administration of the test article did not appear to exert overt toxicity.
- Local Lymph Node Assay: At termination, the lymph nodes from the mice in the vehicle group and the test article treated animals at 1, 2 5, 5, or 10% (v/v) were normal in size and appearance with the exception of lymph nodes from mice treated with 25% were enlarged but appeared normal. Exposure to 1, 2.5, 5, 10 or 25% (v/v) resulted in stimulation indices of 1.2, 1.9, 3.5, 3.6, and 12.8, respectively. Since the data indicated a positive response the EC3 was calculated and determined to be 4.2%.
Interpretation of results:
Category 1B (indication of skin sensitising potential) based on GHS criteria
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

Animals

In a GLP-compliant local lymph node assay, performed in accordance with OECD Guideline 429, groups of 5 CBA/J female mice were treated on the dorsal surface of both ears once per day for 3 days with 1, 2.5, 5, 10 or 25% (v/v) of test substance or with the vehicle (EtOH/DEP). On Day 6, the mice were injected, i.v. with 20 µCi of 3H-thymidine in sterile saline. Five hours later, the mice were euthanized and the draining auricular lymph nodes were removed. The lymph node cells were pooled then precipitated with 5% trichloroacetic acid (TCA) with the pellets counted in a beta-scintillation counter to determine incorporation of the 3 H-thymidine. There was no mortality and all animals appeared normal throughout the study. No erythema or edema was noted in any of the mice in the vehicle group or in those dosed with the test article. Ears appeared wet in mice treated with 25% test substance from days 2-4. The mean body weights of the mice treated with the test substance on Days 1 and 6 showed no statistically significant differences. At termination, the lymph nodes from the mice in the vehicle group and the test article treated animals at 1, 2.5, 5, or 10% (v/v) were normal in size and appearance with the exception of lymph nodes from mice treated with 25% test substance which were enlarged but appeared normal. Exposure to 1, 2.5, 5, 10, or 25% (v/v) resulted in stimulation indices of 1.2, 1.9, 3.5, 3.6, and 12.8, respectively. Since the data indicated a positive response the EC3 was calculated and determined to be 4.2%.

Human

Twenty-five volunteers were expoded to 4% test substance in petrolatum. The test substance is applied under occlusive dressing to the same site (forearm or scapular back) for five 48-hour periods. Prior to each exposure, the site was pre-treated with 2.5% aqueous SLS under occlusion for 24 hours. Following a 10-day rest period, a challenge patch of the test substance is applied to a different site for 48 hours under occlusion. The challenge site is pre-treated for 1 hour with 5% to 10% aqueous SLS under occlusion. Observations are made immediately after removal of the challenge patch and 24 hours thereafter. No effects were observed.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

The test substance has to be classified as Skin sensitisation 1B (H317) according to Regulation (EC) No 1272/2008.