Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP - Guideline study. According to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Groupe interministeriel des produits chimiques GIPC - DGCIS - SI BP 80001 - 67 rue Barbes - 94201 Ivry-sur-Seine, France
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): anhydro-D-glucitol trioleate
- Physical state: amber liquid
- Batch No.: 444978
- Storage condition of test material: at room temperature in darkness

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (F-69592 L'Arbresle)
- Age at study initiation: 4 weeks
- Weight at study initiation: 230-290 g
- Housing: individually or 2 animals in polycarbonate containers
- Diet (ad libitum): SDS, FD1
- Water (ad libitum): tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: olive oil (intradermal injections), liquid paraffin (topical applications)
Concentration / amount:
Intradermal induction: 2%
Epicutaneous induction: 100%
Challenge: 50 and 100%
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: olive oil (intradermal injections), liquid paraffin (topical applications)
Concentration / amount:
Intradermal induction: 2%
Epicutaneous induction: 100%
Challenge: 50 and 100%
No. of animals per dose:
5 (negative control)
10 (test groups)
Details on study design:
RANGE FINDING TESTS:
Three preliminary studies were performed.
For determination of the maximal non necrotising concentration after intradermal injection of the test substance, two animals received a volume of 0.1 mL of the test item on both sides of the spine, at concentrations of 10, 20, 50 and 100% in olive oil, respectively. A macroscopic evaluation of the cutaneous reactions was conducted 24 hours after the injections. Due to necrosis observed at all concentrations, the same animals received a volume of 0.1 mL of the test item on both sides of the spine, at 3 concentrations: diluted at 1, 2 and 5% in olive oil, respectively. A macroscopic evaluation of the cutaneous reactions was conducted 24 hours after the injections. No necrosis was observed at the concentration of 2 %.
To determine the pre-maximal non-irritant concentration after topical application, the test item was applied on the dorso-lumbar zone of two guinea pigs shorn beforehand, with occlusive dressing for 24 hours, at 4 different concentrations: 40, 60, 80 and 100% in liquid paraffin. A macroscopic evaluation of the cutaneous reactions was conducted 24 hours after removal of the dressing. No macroscopic findings were noted on the treated areas up to 100% test substance concentration.
To determine the maximal non-irritant concentration after topical application, three guinea pigs were treated according to the same treatment as animals from the negative control for the intradermal and epicutaneous induction phase in the main test (i.e. olive oil and liquid paraffin). During the challenge phase, the animals were treated with the test item placed onto the selected treatment sites and covered with an occlusive dressing for a period of 24 hours at 4 different concentrations: 100, 80, 60 and 40% in liquid paraffin. A macroscopic evaluation of the cutaneous reactions was conducted 24 and 48 hours after removal of the occlusive dressing and rinsed with distilled water. As no macroscopic findings were observed on the treated areas, concentrations of 50 and 100% test substance concentration for chosen for the challenge phase.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single injection (intradermal) and 48 h (epicutaneous)
- Negative control group:
Intradermal (3 pairs of injections, each 0.1 mL):
Injection 1: 1:1 mixture (v/v) FCA/isotonic sodium chloride
Injection 2: olive oil
Injection 3: 1:1 mixture (v/v) FCA/olive oil
- Test group:
Intradermal (3 pairs of injections):
Injection 1: 1:1 mixture (v/v) FCA/isotonic sodium chloride
Injection 2: 2 % test substance in olive oil
Injection 3: FCA at 50% and the test item at 4 % in olive oil

Epicutaneous:
- Test group: 0.5 mL of the test item at 100%
- Negative control: 0.5 mL of liquid paraffin

- Site: scapular zone (intradermal + epicutaneous)
- Frequency of applications: single
- Duration: Days 0-8 (On day 6, one day prior to epicutaneous induction, the shorn skin of all animals in each group was brushed with a solution of sodium lauryl sulphate at 10% in thick vaseline, in order to create a local irritation).

B. CHALLENGE EXPOSURE
- No. of exposures: 1 (challenge)
- Day(s) of challenge: 20 (challenge)
- Exposure period: 24 h
- Test groups: 50 and 100 % test substance
- Control group: 50 and 100 % test substance
- Site: shorn dorso-lumbar zone, on either side of the spine
- Concentrations: 50 and 100 %
- Evaluation (hr after challenge): 48 and 72 h

C. RECHALLENGE EXPOSURE
- No. of exposures: 1 (rechallenge)
- Day(s) of challenge: 28 (rechallenge)
- Exposure period: 24 h
- Test groups: 25 and 50 % test substance
- Control group: 25 and 50 % test substance
- Site: shorn dorso-lumbar zone, on either side of the spine
- Concentrations: 25 and 50 %
- Evaluation (hr after challenge): 48 and 72 h
Challenge controls:
The control group is actually a challenge control.
Positive control substance(s):
yes
Remarks:
alpha-Hexylcinnamaldehyde (CAS No 101-86-0, routinely evaluated every 6 month at challenge concentrations of 6.25 and 12.5%, thus meeting the reliabilty criteria)

Results and discussion

Positive control results:
Alpha-Hexylcinnamaldehyde (at 6.25 and 12.5% challenge concentrations) induced sensitisation in up to 100% of the treated animals.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
50% (at challenge)
No. with + reactions:
5
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 50% (at challenge). No with. + reactions: 5.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
2% (induction), 50% (at challenge)
No. with + reactions:
6
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 2% (induction), 50% (at challenge). No with. + reactions: 6.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
100% (at challenge)
No. with + reactions:
5
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 100% (at challenge). No with. + reactions: 5.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
2% (induction), 100% (at challenge)
No. with + reactions:
9
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 2% (induction), 100% (at challenge). No with. + reactions: 9.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
50% (at challenge)
No. with + reactions:
3
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50% (at challenge). No with. + reactions: 3.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
2% (induction), 50% (at challenge)
No. with + reactions:
4
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 2% (induction), 50% (at challenge). No with. + reactions: 4.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
100% (at challenge)
No. with + reactions:
4
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 100% (at challenge). No with. + reactions: 4.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
2% (induction), 100% (at challenge)
No. with + reactions:
5
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 2% (induction), 100% (at challenge). No with. + reactions: 5.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
25% (at challenge)
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 25% (at challenge). No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
2% (induction), 25% (at challenge)
No. with + reactions:
1
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 2% (induction), 25% (at challenge). No with. + reactions: 1.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
50% (at challenge)
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: 50% (at challenge). No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
2% (induction), 50% (at challenge)
No. with + reactions:
2
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 2% (induction), 50% (at challenge). No with. + reactions: 2.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
25% (at challenge)
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 25% (at challenge). No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
2% (induction), 25% (at challenge)
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 2% (induction), 25% (at challenge). No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
50% (at challenge)
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 50% (at challenge). No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
2% (induction), 50% (at challenge)
No. with + reactions:
1
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 2% (induction), 50% (at challenge). No with. + reactions: 1.0. Total no. in groups: 10.0.

Any other information on results incl. tables

During the induction phase, no cutaneous reaction was observed. Dryness was recorded in one animal of the control group (1/5) and in nine animals of the treatment group (9/10), 24 hours after the epicutaneous induction.

No abnormality was recorded in the body weight gain of both groups. No mortality was registered during the main test.

In the treatment group (treatment dose of 100%), slight to moderate erythema was noted in 90% (9/10), in 50% (5/10) and in 10% (1/10) of the animals, 24, 48 and 72 hours, respectively, after the challenge phase, on the treated area. Slight oedema was noted in 20% (2/10) of the animals, only 24 hours after the challenge phase, on the treated area.

In the control group (associated with the treatment dose of 100%), slight to moderate erythema was noted in 100% (5/5,) in 80% (4/5) and in 60% (3/5) of the animals, 24, 48 and72 hours, respectively, after the challenge phase, on the area challenged with the test substance at 100%. Slight oedema was noted in 80% (4/5) of the animals, only 24 hours after the challenge phase, on the treated skin.

In the treatment group (treatment dose of 50%), slight to moderate erythema was noted in 60% (6/10), in 40% (4/10) and in 10% (1/10) of the animals, 24, 48 and 72 hours, respectively, after the challenge phase, on the treated area. Slight oedema was noted in 10% (1/10) of the animals, only 24 hours after the challenge phase, on the treated area.

In the control group (associated with the treatment dose of 50%), slight to moderate erythema was noted in 100% (5/5), in 60% (3/5) and in 20% (1/5) of the animals, 24, 48 and 72 hours, respectively, after the challenge phase, on the area cheallenged with the test item at 50%. Slight oedema was noted in 20% (2/5) of the animals, only 24 hours after the challenge phase, on the treated area.

As irritation was observed in the control group animals, only reactions in the test group animals that exceeded the most severe reactions seen in the control group animals were attributed to skin sensitisation. Therefore, no sensitisation reaction was noted in animals from the treated group with the test item at 100 and 50%. To clarify these results, a rechallenge phase was performed with the test item diluted at 50 and 25% after a rest phase of 6 days.

In the treatment group (treatment dose of 50%), slight erythema was noted in 20% (2/10) and in 10% (1/10) of the animals from the treated group, 24 and 48 hours after the rechallenge phase, on the treated area. No skin reaction was noted at the reading time 72 hours. No cutaneous intolerance reaction was recorded in animals from the negative control group after the rechallenge phase, on the treated area, with the test item at 50%.

In the treatment group (treatment dose of 25%), slight erythema was noted in 10% (1/10) of the animals from the treated group, only 24 hours after the rechallenge phase, on the treated area. No skin reaction was noted at the reading time 72 hours. No cutaneous intolerance reaction was recorded in animals from the negative control group after the rechallenge phase, on the treated area, with the test item at 25%.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
CLP: not classified
DSD: not classified