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Description of key information

In a Buehler test according to OECD Guideline 406, the test substance was observed not to be sensitising to the skin (Lemoncelli, 2012).

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:
2012-06-19 to 2012-07-19
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
Well documented GLP study performed according to OECD Guideline 406 with some minor deviations (temperature and relative humidity levels) that no impact the outcome of the study.
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
Temperature and relative humidity ranges were broader than indicated by the OECD Guideline, however there was no impact on study outcome; timing of scoring, however no impact on the study
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
The Buehler test method is the non-adjuvant test given preference over other non-adjuvant methods in the OECD 406 guideline.
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 1J513
- Composition of test material, percentage of components: 0.08 wt % water

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature: 15-30°C

OTHER SPECIFICS:
- Name of test material (as cited in study report): JEFFCAT DMDEE
- Substance type: Colorless to yellow liquid
- Physical state: Liquid
Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Elm Hill Breeding Laboratories
- Age at study initiation: 6 weeks (dose range); 7 weeks at start of dosing (day 1) of the main study
- Weight at study initiation: 343-373 grams (dose range); 336-458 grams at the outset (day 1) of the main study
- Housing: Animals were individually housed upon receipt. No other species were kept in the same room.
- Diet (e.g. ad libitum): ad libitium
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Study animals were acclimated to their housing for a minimum of 5 and 7 days respectively for the dose range and main assay prior to their first day of dosing.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16 to 26°C
- Humidity (%): 25 to 97%
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

Route:
epicutaneous, occlusive
Vehicle:
other: induction: ethanol; challenge: acetone
Concentration / amount:
Dose-range-finding study (primary irritation screen): four concentrations were dosed on four separate sites: 10%, 25%, 50% or 100%
Main assay: Based on the results of the dose-range-finding study, an induction dose and challenge dose of 100% (test item as received) was chosen for this study.
Route:
epicutaneous, occlusive
Vehicle:
other: induction: ethanol; challenge: acetone
Concentration / amount:
Dose-range-finding study (primary irritation screen): four concentrations were dosed on four separate sites: 10%, 25%, 50% or 100%
Main assay: Based on the results of the dose-range-finding study, an induction dose and challenge dose of 100% (test item as received) was chosen for this study.
No. of animals per dose:
Total number:
Dose range primary irritation: 4 (2 male/2 female)
Test article group: 20 (10 male/10 female)
Vehicle control group: 10 (5 male/5 female)
Positive control group: 6 (3 male/3 female)
Details on study design:
RANGE FINDING TESTS: Initially, a dose range was performed in four naive animals with the test article JEFFCAT DMDEE at 10%, 25%, 50% and 100% (as received). Based on these results, an induction dose and challenge dose of 100% (as received) was chosen for this study.

MAIN STUDY
A. INDUCTION EXPOSURE
For the induction phase of this study, twenty guinea pigs (10/sex) in the test article group were induced with three six-hour occluded dermal applications of JEFFCAT DMDEE (as received). A vehicle group of ten animals (5/sex) was induced in the same manner with 0.9% sodium chloride (saline). A positive control group of six animals (3/sex) was induced in the same manner as above with a known dermal sensitizer: 0.3 % 1-chloro-2,4-dinitrobenzene (DNCB) in 80% ethanol.

B. CHALLENGE EXPOSURE
Fourteen days after the last induction, all animals were dermally challenged with occluded applications at naive test sites. Animals in the test article and vehicle control group were challenged with JEFFCAT DMDEE (as received) and 0.9% sodium chloride (saline), respectively. One the day following the challenge, animals were depilated and approximately three hours later were scored for dermal irritation (24 hour). Scoring was repeated approximately 24 hours later for the 48 hour score.
Challenge controls:
Animals in the positive control group were challenged with 1-chloro-2,4-dinitrobenzene (DNCB) at 0.2% in acetone.
Positive control substance(s):
yes
Remarks:
1-chloro-2,4-dinitrobenzene
Positive control results:
The positive control group, induced and challenged with DNCB, exhibited the anticipated positive responses at challenge, indicating that the methods employed in this study were valid.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
Jeffcat DMDEE at 100%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No signs of systemic toxicity and all animals gained weight during the main study
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: Jeffcat DMDEE at 100%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No signs of systemic toxicity and all animals gained weight during the main study.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
Saline
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No signs of systemic toxicity and all animals gained weight during the main study
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: Saline. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No signs of systemic toxicity and all animals gained weight during the main study.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
Jeffcat DMDEE at 100%
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
No signs of systemic toxicity and all animals gained weight during the main study
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: Jeffcat DMDEE at 100%. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: No signs of systemic toxicity and all animals gained weight during the main study.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
Saline
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No signs of systemic toxicity and all animals gained weight during the main study
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: Saline. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No signs of systemic toxicity and all animals gained weight during the main study.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.2% DNCB in Acetone
No. with + reactions:
6
Total no. in group:
6
Clinical observations:
1/6 with score 1, 5/6 with score 2; No signs of systemic toxicity and all animals gained weight during the main study
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: 0.2% DNCB in Acetone. No with. + reactions: 6.0. Total no. in groups: 6.0. Clinical observations: 1/6 with score 1, 5/6 with score 2; No signs of systemic toxicity and all animals gained weight during the main study.
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
0.2% DNCB in acetone
No. with + reactions:
6
Total no. in group:
6
Clinical observations:
1/6 with score 1, 5/6 with score 2; No signs of systemic toxicity and all animals gained weight during the main study
Remarks on result:
other: see Remark
Remarks:
Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 0.2% DNCB in acetone. No with. + reactions: 6.0. Total no. in groups: 6.0. Clinical observations: 1/6 with score 1, 5/6 with score 2; No signs of systemic toxicity and all animals gained weight during the main study.

Clinical observations:

There were no signs of systemic toxicity and all animals gained weight during the main study.

Challenge phase:

The positive control group, induced and challenged with DNCB, exhibited the anticipated positive responses at challenge, indicating that the methods employed in this study were valid.

All dermal irritation scores at the 24- and 48 -hour observations were zero for the vehicle control animals challenged with the test article and the vehicle (saline).

A single score of ± was recorded in the test article group at the test article site at 24 hours. All dermal irritation scores at the 48 -hour observations were zero for the test article group challenged with the test article and the vehicle (saline).

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study, induction with the test substance as received did not elicit a delayed contact hypersensitivity response in guinea pigs when challenged as received.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

Lemoncelli (2012) investigated the sensitising properties of the test substance in a buehler test with male and female Hartley guinea pigs. The study examined the skin sensitising effect on guinea pigs, using 4 animals in the dose range finding test, 20 animals in the test article group, 10 animals in the vehicle control group and 6 animals in the positive control group. Under the conditions of this study, induction with the substance as received did not elicit a delayed contact hypersensitivity response in guinea pigs when challenged as received.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Based on the study results and the criteria of the CLP Regulation, the test substance should not be classified as skin sensitising substance.