L-gluco-Octitol, 1,5-anhydro-6,8-dideoxy-, (7.xi.)-

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 November - 30 December 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP study conducted in compliance with OECD Guideline 406 with minor deviation: temperature in the animal room was slightly outside the target range

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

yes

Remarks:

temperature in the animal room was slightly outside the target range

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material (as cited in study report): Mexoryl SBF
- Physical state: Thick brownish paste (very sticky)
- Analytical purity: 92 %
- Lot/batch No.: 003D001
- Date of receipt: 18 November 2005
- Expiration date of the lot/batch: October 2006
- Storage condition of test material: Stored at +4 °C and protected from light

In vivo test system

Test animals

Species:

guinea pig

Strain:

Hartley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories France, L’Arbresle, France
- Age at study initiation: 1-2 months
- Weight at study initiation: Control group (males: 353 ± 5 g; females: 341 ± 14 g); treated group (males: 359 ± 9 g; females: 347 ± 16 g)
- Housing: Animals were housed individually in polycarbonate cages with stainless steel lid.
- Diet: 106 pelleted diet (SAFE, Epinay-sur-Orge, France), ad libitum
- Water: Drinking water filtered by a FG Millipore membrane (0.22 µm), ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 30-70 %
- Air changes: Approximately 12 cycles/h of filtered, non-recycled air
- Photoperiod: 12 h dark / 12 h light

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

other: intradermal and cutaneous

Vehicle:

other: 0.9 % NaCl for intradermal injections and purified water for topical applications

Induction 2

Concentration / amount:

Preliminary test:
- Intradermal route: 5 and 10 % w/w
- Cutaneous route (induction phase): 10 and 25 % w/w
- Cutaneous route (challenge phase): 10 and 25 % w/w

Main test:
Induction phase: Intradermal route - 10 % w/w; Cutaneous route - 25 % w/w
Challenge phase: Cutaneous route - 25 % w/w

Challenge

Challenge 1

Route:

other: cutaneous, occlusive

Vehicle:

other: 0.9 % NaCl for intradermal injections and purified water for topical applications

Challenge 2

Concentration / amount:

Preliminary test:
- Intradermal route: 5 and 10 % w/w
- Cutaneous route (induction phase): 10 and 25 % w/w
- Cutaneous route (challenge phase): 10 and 25 % w/w

Main test:
Induction phase: Intradermal route - 10 % w/w; Cutaneous route - 25 % w/w
Challenge phase: Cutaneous route - 25 % w/w

No. of animals per dose:

- Preliminary test: 1/sex/dose
- Main test: 5 and 10/sex/dose for control and treatment groups, respectively

Details on study design:

PRELIMINARY TEST:
- Intradermal route: Guinea pigs (1/sex) received intradermal injections (0.1 mL) of test item at concentrations of 5 and 10 % w/w and evaluated for local reactions at 24 and 48 h and 6 days after injections.
- Topical induction exposure: One female and one male guinea pig was applied topically with filter paper (approximately 8 cm2) saturated with 10 and 25 % w/w test item, respectively via occlusive patch for 48 h and evaluated for cutaneous reactions at 24 and 48 h after removal of the dressings.
- Topical challenge exposure: Two guinea pigs (male and female) were applied topically with filter paper saturated with 10 and 25 % w/w test item via occlusive patch for 24 h and evaluated for cutaneous reactions at 24 and 48 h after removal of the dressings.

MAIN STUDY
A. INDUCTION EXPOSURE: INTRADERMAL
- No. of exposures: One
- Test groups: : On Day 1, three injections of 0.1 mL of FCA (50 % v/v in 0.9 % NaCl), test item at 10 % w/w in 0.9 % NaCl and test item at 10 % w/w in the mixture of FCA/0.9 % NaCl (50/50) were made in to each side of anterior, middle and posterior sites of interscapular region, respectively.
- Control group: On Day 1, three injections of 0.1 mL of FCA (50 % v/v in 0.9 % NaCl), 0.9 % NaCl and vehicle at 50 % (w/v) in mixture of FCA/0.9 % NaCl (50/50) were made in to each side of anterior, middle and posterior sites of interscapular region, respectively.
- Site: Interscapular region
- Duration: Days 1-7

B. INDUCTION EXPOSURE: TOPICAL
- No. of exposures: One
- Exposure period: 48 h
- Test groups: Filter paper patch saturated with test item at the concentration of 25 % w/w, topically applied on Day 8 via occlusive patch
- Control group: Filter paper patch saturated with vehicle alone topically applied on Day 8 via occlusive patch
- Site: Interscapular region
- Frequency of applications: Single application
- Duration: Days 8-21

C. CHALLENGE EXPOSURE: TOPICAL
- No. of exposures: One
- Day of challenge: Day 22
- Exposure period: 24 h
- Test groups: Filter paper patch saturated with test item at the concentration of 25 % w/w, topically applied on Day 22 via occlusive patch
- Control group: Filter paper patch saturated with vehicle alone topically applied on Day 22 via occlusive patch
- Site: Test item and vehicle was applied to the skin of the posterior right and left flank, respectively
- Evaluation (h after removal of challenge patch): 24 and 48 h

OTHER:
- Clinical examinations: Animals were observed at least once a day during the study to check for clinical signs and mortality.
- Body weight: Animals were weighed individually on the day of allocation into the groups, on Days 1 and 25 of the study.
- Pathology: At the end of the study, all animals were killed by carbon dioxide asphyxiation. No necropsy was performed and no skin samples were taken.

Challenge controls:

A filter paper patch saturated with vehicle only was applied to the posterior left flank on Day 22 via occlusive patch.

Positive control substance(s):

yes

Remarks:

Mercaptobenzothiazole (Study no. 30329 RDG – September 2005)

Study design: in vivo (LLNA)

Concentration:

Not applicable

No. of animals per dose:

Not applicable

Details on study design:

Not applicable

Statistics:

Not applicable

Results and discussion

Positive control results:

Mercaptobenzothiazole induced positive skin sensitization reactions in 100 % (10/10) guinea pigs.

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

25 % w/w

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

none

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25 % w/w. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: none.

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

25 % w/w

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

none

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25 % w/w. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: none.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: Not applicable

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: Not applicable

Any other information on results incl. tables

Clinical examinations:

- Marked local reactions (but not necrosis) at the intradermal injection sites were noted in a few animals of both control and treated groups, between Days 11 and 18.

- No systemic clinical signs and no deaths were observed during the study.

Body weight:

- Reduced body weight gain was observed in 1/10 males and 1/10 females during the study period. The body weight change of the other treated animals was similar to that of controls.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under these test conditions, Mexoryl SBF is not classified as sensitizing to the skin according to the Annex VI to the Directive 67/548/EEC and the CLP Regulation (EC) N° (1272-2008).

Executive summary:

In a Magnusson & Kligman maximisation study (GPMT) performed according to OECD Guideline 406 and in compliance with GLP, groups of 20 Hartley Crl: (HA) BR guinea pigs (10/sex) were induced with three pairs of intradermal injections of 0.1 mL of FCA (50 % v/v in 0.9 % NaCl), test item at 10 % w/w in 0.9 % NaCl and test item at 10 % w/w in mixture of FCA/0.9 % NaCl (50/50) on Day 1 on three different sites on each side of interscapular region. Control group of 10 animals (5/sex) was intradermally induced with FCA (50 % v/v in 0.9 % NaCl), 0.9 % NaCl and vehicle at 50 % w/v in a mixture of FCA/0.9 % NaCl (50/50). After one week the same area was topically induced with test item at 25 % w/w via occluded filter paper patch for 48 h for the treated group. Control group was patched with filter paper saturated with vehicle alone. After 2 weeks of rest period, a challenge filter paper patch of vehicle and test item at 25 % w/w was applied to posterior left and right flank of all animals, respectively. The test concentrations for the main study were determined from a sighting study using two animals.

No deaths and no systemic clinical signs were noted during the study. No cutaneous reactions were noted at the challenge sites of the test or control group animals at the 24 or 48 h observations. Mexoryl SBF produced a 0 % (0/20) sensitisation rate and was considered to be a non-sensitizer to guinea pig skin. Historical data on positive control (mercaptobenzothiazole) exhibited evidence of sensitisation.

Under these test conditions, Mexoryl SBF is not classified as sensitizing to the skin according to the Annex VI to the Directive 67/548/EEC and the CLP Regulation (EC) N° (1272-2008).