acetalization product between glucose and C16-18(even numbered)- alcohol

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007-08-22 to 2007-09-13

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

The test was performed in 2007, whilst the LLNA method (OECD 442 B) was adopted in 2010 , also the (OECD tests n°442 C , D) was adopted in 2015.

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): LCE07050
- Physical state: solid, powder
- Analytical purity: 100%
- Composition of test material, percentage of components: 67762-27-0, 54549-27-8, 27836-65-3
- Lot/batch No.: T70615
- Expiration date of the lot/batch: 07 February 2009 (re-test)
- Storage condition of test material: at the room temperature

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

16 female albino guinea pigs of Dunkin-Hartley strain, supplied by CHARLES RIVER (F-69592 L’ARBRESLE).
The animals were housed either in groups of 2 or 3 in polycarbonate containers, the flooring of which was covered with dust-free cuttings and the top fitted a stainless steel lid with a feeding device and drinking device of 500 ml.
The environmental parameters of the main test were :
- Temperature : between 19°C and 23°C
- Relative humidity : between 38%
The drinking water (tap water from public distribution system) and food were supplied freely.
Microbiological verification and chemical analysis were conducted every six months by the Institut Européen de l’Environnement de Bordeaux (I.E.E.B.).

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

other: olive oil for the induction and liquid paraffin for the challenge

Induction 2

Concentration / amount:

(1) Preliminary study
-determination by intrademal injection of the Maximal Non Necrotizing Concentration (MNNC): 0.39% ; 0.78% ; 1.56% ; 3.125% ; 6.25% ; 25% in olive oil.
-determination by topical application of the Pre-Maximal Non Irritant Concentration (Pre-MNIC): 6.25 ; 12.5 ; 25 and 50% in liquid paraffin.
-determination by topical application of the Maximal Non Irritant Concentration (MNIC): 6.25 ; 12.5; 25 and 50% in liquid paraffin
(2) Main study
intradermal induction with the product at 12.5% in olive oil
topical induction with the product at 50% in liquid paraffin
challenge with the product at 3.125% and 6.25% in liquid paraffin

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

other: olive oil for the induction and liquid paraffin for the challenge

Challenge 2

Concentration / amount:

(1) Preliminary study
-determination by intrademal injection of the Maximal Non Necrotizing Concentration (MNNC): 0.39% ; 0.78% ; 1.56% ; 3.125% ; 6.25% ; 25% in olive oil.
-determination by topical application of the Pre-Maximal Non Irritant Concentration (Pre-MNIC): 6.25 ; 12.5 ; 25 and 50% in liquid paraffin.
-determination by topical application of the Maximal Non Irritant Concentration (MNIC): 6.25 ; 12.5; 25 and 50% in liquid paraffin
(2) Main study
intradermal induction with the product at 12.5% in olive oil
topical induction with the product at 50% in liquid paraffin
challenge with the product at 3.125% and 6.25% in liquid paraffin

No. of animals per dose:

5 animals for the preliminary assay
11 animals per dose
5 negative controls

Details on study design:

(1) Preliminary study
- for the determination of the MNNC, two animals received increasing concentration of the substance by intradermal injection on both sides of the spine. A macroscopic evaluation of the cutaneous reactions was conducted 24 hours later.
- for the determination of the pre-MNIC, the product was applied on the dorso-lumbar zone of 2 animals shorn beforhand, with occlusive dressing for 24 hours. A macroscopic evaluation of the cutaneous reactions was conducted 24 hours after removal of the dressing.
- for the determination of the MNIC, 3 animals were treated according to the same treatment as negative controls for the induction phase (ie 2 intradermic injection (ID) of Freund's Complete Adjuvant (FCA) diluted at 50% in isotonic sodium chloride + 2 ID of olive oil + 2 ID a mixture with equal volumes v/v FCA at 50% and isotonic sodium chloride.
A macroscopic evaluation of the cutaneous reactions was conducted 24 hours after removal of the occlusive dressing.
(2) Main study
A- Induction
1st Intradermal Induction:
Day 0
After shearing the scapular zone, three (3) pairs of intradermal injections (ID) of 0.1 ml were performed on the scapular zone in such a way as an injection on each pair is placed to either side of the spine as follows :
GROUP 1 (Negative control):
• 2 ID: Freund’s Complete Adjuvant diluted at 50 % in isotonic sodium chloride.
• 2 ID: olive oil
• 2 ID: a mixture with equal volumes v/v :
- Freund’s Complete Adjuvant at 50% and isotonic sodium chloride,
GROUP 2 (Treated):
• 2 ID: Freund’s Complete Adjuvant diluted by 50 % in isotonic sodium chloride,
• 2 ID: test item at 12.5%,
• 2 ID a test mixture in equal volumes v/v :
- Freund’s Complete Adjuvant at 50% and the test item at 25%.
2nd Topical Induction:
Day 6
The scapular zone of all the animals in each group, shorn beforehand, was brushed with a solution of sodium lauryl sulfate at 10% in thick vaseline, in
order to create a local irritation.
Day 7
A topical application under occlusive dressing for 48 hours was performed on the injection sites of each animal.
GROUP 1 (Negative control): 0.5 ml of liquid paraffin
GROUP 2 (treated): 0.5 ml of the test item at 50%
B - Rest phase
The animals of both groups were left for 10 days.
C - Challenge phase
Day 21
The experimental procedure of this phase was identical for both groups GROUP 1 (Negative control) and GROUP 2 (Treated) submitted to this experimentation: on the previously shorn dorso-lumbar zone, an application on either side of the spine, under occlusive dressing, was performed during 24 hours:
- 1 sample cup containing the test item at 6.25% (MNIC) and at 3.125% (1/2 MNIC).

Challenge controls:

Negative controls : 5 animals

1st Intradermal Induction:
Day 0
After shearing the scapular zone, three (3) pairs of intradermal injections (ID) of 0.1 ml were performed on the scapular zone in such a way as an injection on each pair is placed to either side of the spine as follows :

GROUP 1 (Negative control):
• 2 ID: Freund’s Complete Adjuvant diluted at 50 % in isotonic sodium chloride.
• 2 ID: olive oil
• 2 ID: a mixture with equal volumes v/v :
- Freund’s Complete Adjuvant at 50% and isotonic sodium chloride,


2nd Topical Induction:
Day 6
The scapular zone of all the animals in each group, shorn beforehand, was brushed with a solution of sodium lauryl sulfate at 10% in thick vaseline, in
order to create a local irritation.
Day 7
A topical application under occlusive dressing for 48 hours was performed on the injection sites of each animal.
GROUP 1 (Negative control): 0.5 ml of liquid paraffin

Rest phase
The animals of control groups were left for 10 days.

Challenge phase
Day 21
The experimental procedure of this phase was identical for both groups GROUP 1 (Negative control) and GROUP 2 (Treated) submitted to this experimentation: on the previously shorn dorso-lumbar zone, an application on either side of the spine, under occlusive dressing, was performed during 24 hours:
- 1 sample cup containing the test item at 6.25% (MNIC) and at 3.125% (1/2 MNIC).

Positive control substance(s):

yes

Remarks:

alpha-hexylcinnamaldehyde

Results and discussion

Positive control results:

positve control at 50% : between 70% and 100% of animals sensitized after 24h
positve control at 50% : between 70% and 90% of animals sensitized after 48h
positve control at 25% : between 50% and 100% of animals sensitized 24h
positve control at 25% : between 30% and 90% of animals sensitized 48h

In vivo (non-LLNA)

Results

Results 1

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

6.25%

No. with + reactions:

0

Total no. in group:

11

Clinical observations:

none

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 6.25%. No with. + reactions: 0.0. Total no. in groups: 11.0. Clinical observations: none.

Results 2

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

3.125%

No. with + reactions:

0

Total no. in group:

11

Clinical observations:

none

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 3.125%. No with. + reactions: 0.0. Total no. in groups: 11.0. Clinical observations: none.

Results 3

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

6.25%

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 6.25%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

Results 4

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

3.125%

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 3.125%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

Results 5

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

50%

No. with + reactions:

10

Total no. in group:

10

Results 6

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Dose level:

25 %

No. with + reactions:

10

Total no. in group:

10

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

In conclusion, in view of these results, under these experimental conditions, the test item LCE07050
need not to be classified, in accordance with the criteria for classification, packaging and labelling of
dangerous substances and preparations of the E.E.C. Directives 67/548, 2001/59 and 99/45. No
symbol and risk phrase are required.

Executive summary:

The aim of the study was to evaluate the possible allergenic activity of the test item LCE07050 after intradermic injection and topical administration in guinea pigs. After induction (intradermic injection at 12.5 % and topical application at 50 %) of 11 Guinea Pigs of treated group with the test item LCE07050 and a 10-day rest phase, the challenge phase, under occlusive dressing for 24 hours, consisted to a single topical application of the test item diluted at 6.25% and at 3.125% in liquid paraffin. The experimental protocol was established according the O.E.C.D. guideline n°406 dated July 17th, 1992 and the method B.6 of the E.E.C. n°96/54 dated July 30th, 1996. No macroscopic cutaneous reactions attributable to allergy was recorded during the examination following the removal of the occlusive dressing (challenge phase) from the animals of the treated group with the test item. No cutaneous intolerance reaction was recorded in animals from the negative control group. In conclusion, in view of these results, under these experimental conditions, the test item LCE07050 need not to be classified, in accordance with the criteria for classification, packaging and labelling of dangerous substances and preparations of the E.E.C. Directives 67/548, 2001/59 and 99/45. No symbol and risk phrase are required.