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Description of key information

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
September 24, 2013
Reliability:
1 (reliable without restriction)
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Remarks:
no major deviations only minor deviations that have no impact on results
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
- Diet: SDS/DIETEX STANRAB (P) SQC feed was available ad libitum.
- Water:Drinking water was available ad libitum in polycarbonate bottles with a stainless steel nipple. A specimen of water is sent approximately every 6 months to Laboratoire de la Touraine - ZA n°1 du Papillon - Rue de l0Aviation - 37210 Parçay-Meslay - France, for analysis. The criteria for acceptable levels of contaminants in the water supplied were within the limits of the analytical specifications.
-Housing: Observation was performed at the time of delivery of the animals and daily during the acclimatisation period. Animals were housed individually in cages of standard size. These cages were placed in an air-conditioned (15- 21°C) animal house kept at between 45 % and 65 % relative humidity (except during the cleaning slot) in which non-recycled filtered air is changed approximately 10-15 times per hour. These parameters are continuously recorded using a computerised acquisition system. The artificial day/night cycle was 12 hours light and 12 hours darkness with light on at 7.30 a.m.
- IN-LIFE DATES: Start of the study (signature of the study plan by the Study Director): 02 May 2013, Start of the experimental period: 06 May 2013, End of the experimental period: 16 May 2013.
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
0.5 mL of "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623 was applied to the skin of the right or the left flank region. The test item was applied to the selected area on the right or left flank of each animal. Adjacent surfaces of non-treated skin of each animal served as a control for the trial.
The test item was placed directly on the skin, over an area of approximately 6 cm² then covered with a non-irritating gauze square of the same surface area. This gauze square was protected by a pad consisting of a semi-occlusive micro-porous and non-allergenic dressing, held in place with a soft and aerated fastening system consisting of elastic, adhesive, hypoallergenic tape. The animals were fitted with this pad for the duration of the exposure period. At the end of the exposure period, the dressing was removed. Since the application of the test item induced a brown colouration, the application sites were rinsed using a waterlogged gauze and a control zone was also rinsed and served as control. The standard amounts applied were not adjusted to body weight.
Duration of treatment / exposure:
3 min, 1h, 4h on the skin for the first animal, 4h for the two others.
Observation period:
For the initial test in one animal, due to the colouration of the treatment sites, the application site was not examined immediately after the patch has been removed. One hour, 24 hours, 48 hours and 72 hours approximately after patch removal dermal reactions on the right or the left flank of each animal were graded and recorded.
Number of animals:
3 females
Details on study design:
The rabbit was chosen because of its acceptance as a predictor of irritant/corrosive effects of chemicals in man and the recognition by regulatory authorities that this species is suitable for skin irritation studies.
The number of animals per group is the minimum number enabling an accurate assessment of the studied toxicological effect.
Approximately 24 hours before the test, fur was removed by closely clipping the dorsal area of the trunk of the animals (at least 14 cm x 5 cm). Care was taken to avoid abrading the skin, and only animals with healthy, intact skin were used.
Three test patches were applied sequentially to separate skin sites on the animal. The first patch was removed after three minutes. Since no serious skin reaction was observed, a second patch was applied at a different site and removed after one hour. Since the observations at this stage indicated that exposure could humanely be allowed to extend to four hours, a third patch was applied and removed after four hours and any responses were graded.
Since no corrosive effect was observed in the initial test, the irritant or negative response was confirmed using two additional animals, each with one patch, for an exposure period of four hours.
For the initial test in one animal, due to the colouration of the treatment sites, the application site was not examined immediately after the patch has been removed. One hour, 24 hours, 48 hours and 72 hours approximately after patch removal dermal reactions on the right or the left flank of each animal were graded and recorded.
At the end of the study, euthanasia of animals was performed by an overdose of sodium pentobarbital injected by the intravenous route. No necropsy was performed at the end of the study.
Irritation parameter:
erythema score
Basis:
animal #3
Score:
ca. 0
Max. score:
0
Reversibility:
other: not applicable
Irritation parameter:
edema score
Basis:
animal #3
Score:
ca. 0
Max. score:
0
Reversibility:
other: not applicable
Irritant / corrosive response data:
The application of the test item induced a dark brown to black colouration of the application site due to the same colouration of the test item. The application site was rinsed using a waterlogged gauze.
There was no cutaneous lesion in any animal at all times of examination.
Mean Indices were 0 in 3 out of 3 animals, therefore Mean Index of erythema and oedema was 0.
Other effects:
There was no change in body weight gain between D1 and D4.

Acute skin lesions - Individual scores - Test item - Local observations

Clinical signs

(range of scores; normal score)

 

Time

Animal serial number

Erythema and eschar (0-4 ;0)

Timmediate

1

2

3

4

5

T 1H

NE

NE

NE

NE

NE

T 24H

0

0

0

0

0

T 48H

0

0

0

0

0

T 72H

0

0

0

0

0

Oedema (0-4 ;0)

T 1H

0

0

0

0

0

T 24H

0

0

0

0

0

T 48H

0

0

0

0

0

T 72H

0

0

0

0

0

NE: not evaluable.

 

Mean Index (M.I.)

Treatment

Animal number

Erythema

Oedema

Test item

130407- T 3MIN

0

0

Test item

130407- T 1H

0

0

Test item

130407- T 4H

0

0

Test item

130408 - T 4H

0

0

Test item

130409 - T 4H

0

0

 

Effect on body weight (individual values)

Treatment

Animal number

 

D1

D4

 

Test item

130407

1.898

2.040

 

 Test item

130408

2.397

2.380

 

 Test item

130409

2.120

2.360

 

Results expressed in kg

D: Day

 

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: OECD GHS
Conclusions:
Under the experimental conditions adopted, "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623 (Batch No. SMOU/13/06) was found to be non irritant for the skin of the rabbit.
Classification according to Council Directive No. 67/548/EEC and subsequent amendments and CLP Regulation No. 1272/2008:
Symbol: none is required.
Indication of danger: none is appropriate.
Risk phrase: none is applicable.
Executive summary:

The aim of the study was to determine any irritant property and/or degree of corrosion of "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623 (Batch No. SMOU/13/06) in the rabbit following a single semi-occluded application to intact skin in accordance with General Requirements of O.E.C.D Guideline No. 404 (April 24, 2002), method B4 of Commission Directive No. 2004/73/EC (April 29, 2004) adapting to technical progress for the 29th Council Directive No. 67/548/EEC and

subsequent amendments and of Council Regulation No. 440/2008 (30th May 2008) and Regulation No. 1907/2006 (REACH).

Three animals were used for the study.

0.5 mL of test item was applied on the skin.

Semi-occlusive dressings held the test item in place for 3 minutes, 1 hour and 4 hours on the skin of the first animal and for 4 hours for the two other animals.

Any cutaneous lesion was evaluated approximately one hour, 24, 48 and 72 hours after removal of the dressing.

Mean indices were calculated from results obtained from each rabbit at times 24, 48 and 72 hours.

The application of the test item induced a brown colouration of the application site. The application site was rinsed using a waterlogged gauze.

There was no cutaneous lesion in any animal at all times of examination.

Mean Index of erythema and oedema was 0.

There was no change in body weight gain.

Conclusion: Under the experimental conditions adopted, "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623 was found to be non irritant for the skin of the rabbit. Substance was not classificied according to Council Directive No. 67/548/EEC and subsequent amendments and CLP Regulation No. 1272/2008.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
September 24, 2013
Reliability:
1 (reliable without restriction)
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Remarks:
no major deviations only minor deviations that have no impact on results
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
- Diet: SDS/DIETEX STANRAB (P) SQC feed was available ad libitum.
- Water:Drinking water was available ad libitum in polycarbonate bottles with a stainless steel nipple. A specimen of water is sent approximately every 6 months to Laboratoire de la Touraine - ZA n°1 du Papillon - Rue de l0Aviation - 37210 Parçay-Meslay - France, for analysis. The criteria for acceptable levels of contaminants in the water supplied were within the limits of the analytical specifications.
-Housing: Observation was performed at the time of delivery of the animals and daily during the acclimatisation period. Animals were housed individually in cages of standard size. These cages were placed in an air-conditioned (15- 21°C) animal house kept at between 45 % and 65 % relative humidity (except during the cleaning slot) in which non-recycled filtered air is changed approximately 10-15 times per hour. These parameters are continuously recorded using a computerised acquisition system. The artificial day/night cycle was 12 hours light and 12 hours darkness with light on at 7.30 a.m.
- IN-LIFE DATES: Start of the study (signature of the study plan by the Study Director): 07 May 2013, Start of the experimental period: 13 May 2013 , End of the experimental period: 24 May 2013
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
The following procedures were applied to avoid or minimize pain and distress in ocular safety testing procedures.

• 45 minutes prior to test item application, buprenorphine 0.01 mg/kg was administered by subcutaneous injection to provide a therapeutic level of systemic analgesia.
• 5 minutes prior to test item application, one drop of a topical ocular anaesthetic (oxybuprocaine chloryde or equivalent) was applied to each eye in order to avoid possible interference with the study. The eye of each animal which was not treated with a test article, but treated with topical
anaesthetics, served as a control.

0.1 mL of "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623 was introduced into the conjunctival sac of one eye of each of the animals. The conjunctival sac was opened by gently pulling the lower lid away from the eyeball. Eyelids were held closed for approximately 1 second in order to
avoid any loss of "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623. The non-treated eye was used as control. Following treatment, animals were kept for approximately one hour in a restraint cage, in order to prevent rubbing of the eye.
Removal of test item: Since the test item was not removed from the eye by a natural mechanism, then after 1 hour the eye was rinsed with 20 mL of saline.
Exposure of one animal: Without information on the test item and/or the active ingredient, a single animal test was considered in order to anticipate marked effects.
Exposure of two additional animals: Since no corrosive effect was observed after instillation of test item in the first animal, the irritant or negative response was confirmed using up to two additional animals.
The standard amount applied was not adjusted to body weight.
Duration of treatment / exposure:
1 hour
Observation period (in vivo):
Test animals were routinely evaluated for the entire duration of the study for clinical signs of pain and/or distress at least twice daily.

Examination of the eye: Lesions of the conjunctiva, iris and cornea were monitored by direct examination. One hour, 24 hours, 48 hours and 72 hours approximately after the instillation of "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623, all possible conjunctival, iris or corneal lesions were evaluated on the left or right eye of each animal.

Ocular lesions were graded using the following scoring system in accordance with the OECD Guideline
No. 405
Number of animals or in vitro replicates:
3 females
Details on study design:
Three animals are required for the complete test. The rabbit was chosen because of its acceptance as a predictor of irritant/corrosive
effects of chemicals in man and the recognition by regulatory authorities that this species is suitable for eye irritation studies.

The day of instillation was taken as the first study day D1. Following a pretreatment with a systemic analgesic and induction of appropriate topical anesthesia, "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623 was instilled once only.
Justification of the number of animals: The number of animals is the minimum number enabling an accurate assessment of the studied toxicological effect.
Examination after fluorescein instillation: Confirmation of the absence of corneal damage was carried out at predose, 24 hours, 48 hours or
72 hours by examination after instillation of a 0.5% aqueous solution of sodium fluorescein and rinsing with sterile and pyrogen free isotonic saline.
Corneal lesions seen after the instillation of fluorescein were graded using the following evaluation scale described below.
Individual body weight was taken at the beginning and at the end of the experiment.
Euthanasia of animals was performed by an overdose of sodium pentobarbital injected by the intravenous route.
No necropsy was performed at the end of the study.
Calculation of ocular primary irritation indices: Mean indices were calculated from results obtained for each rabbit by direct examination at times
24 hours, 48 hours and 72 hours. The surface area of corneal opacity was not taken into account in the calculation of mean index. For each parameter, mean of indices for each rabbit for the three times was noted (Mean index per parameter, per animal for the three times = Mi). The maximum theoretical score is 4 (chemosis), 3 (redness), 2 (iris lesions) and 4 (corneal lesions).
Criteria of classification, choice of symbol, indication of danger and risk phrase: "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623 was classified irritant or non-irritant on the basis of results obtained, in accordance with Council Directive No. 67/548/EEC and subsequent amendments.


Irritation parameter:
conjunctivae score
Remarks:
redness of the conjunctivae
Basis:
animal #3
Score:
ca. 1
Max. score:
1
Reversibility:
not fully reversible within: 24h
Irritant / corrosive response data:
The application of the test item did not induce colouration of the application site and did not interfere with grading of any ocular lesion.
One hour after instillation, all animals had redness of the conjunctivae (score 1). There was no ocular lesion from 24 to 72 hours after instillation.
Mean index values were 0
Other effects:
There was no change in body weight gain between D1 and D4.

Acute eye lesions - Individual scores - test item - local observations

Clinical signs (range of scores; normal score)

Time

Animal

1

2

3

Chemosis (0-4; 0)

T 1H

0

0

0

 

T 24H

0

0

0

 

T 48H

0

0

0

 

T 72H

0

0

0

Lacrimation (0-3; 0)

T 1H

0

0

0

 

T 24H

0

0

0

 

T 48H

0

0

0

 

T 72H

0

0

0

Redness (0-3; 0)

T 1H

1

1

1

 

T 24H

0

0

0

 

T 48H

0

0

0

 

T 72H

0

0

0

Iris (0-2; 0)

T 1H

0

0

0

 

T 24H

0

0

0

 

T 48H

0

0

0

 

T 72H

0

0

0

Cornea opacity (0-4; 0)

T 1H

0

0

0

 

T 24H

0

0

0

 

T 48H

0

0

0

 

T 72H

0

0

0

Area of cornea involved (0-4; 0)

T 1H

0

0

0

 

T 24H

0

0

0

 

T 48H

0

0

0

 

T 72H

0

0

0

Mean Index (M.I.)

Treatment

Animal number

Chemosis

Redness

Iris

Cornea

Test item

130410

0

0

0

0

Test item

130411

0

0

0

0

Test item

130412

0

0

0

0

 

Body weight (individual values)

Treatment

Animal number

D1

D4

Test item

130410

2.351

2.415

 

130411

2.577

2.562

 

130412

2.508

2.593

Results expressed in kg

D: day

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the experimental conditions adopted, "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623 was found to be non irritant for the eye of the rabbit.
Classification according to Council Directive No. 67/548/EEC and subsequent amendments and CLP Regulation No. 1272/2008:
Symbol: none is required.
Indication of danger: none is appropriate.
Risk phrase: none is applicable.
Executive summary:

The aim of the study was to determine any irritant property and/or degree of corrosion of "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623 following a single ocular instillation in the rabbit in accordance with the General Requirements of OECD Guideline No. 405 (Oct 2012) and method B5 of Commission Directive No. 2004/73/EC (April 29, 2004) adapting to technical progress for the 29th time Council Directive No. 67/548/EEC and subsequent amendments and of Council Regulation No. 440/2008 (30th May 2008) and Regulation No. 1907/2006 (REACH).

Three animals were used for the study.

0.1 mL of test item was introduced into the conjunctival sac of the left or the right eye of each of the animals. The untreated eye served as a control. Any conjunctival, iris and corneal changes were evaluated approximately one hour, 24 hours, 48 hours, 72 hours after instillation. Mean indices were calculated from results obtained for each rabbit at times 24, 48 and 72 hours.

The application of the test item did not induce colouration of the application site and did not interfere with grading of any ocular lesion. One hour after instillation, all animals had redness of the conjunctivae (score 1). There was no ocular lesion from 24 to 72 hours after instillation. There was no ocular lesion from 24 to 72 hours after instillation. Mean index values were 0.

There was no change in body weight gain.

Under the experimental conditions adopted, "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623 was found to be non irritant for the eye of the rabbit. Substance was not classificied according to Council Directive No. 67/548/EEC and subsequent amendments and CLP Regulation No. 1272/2008.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Additional information

Justification for selection of skin irritation / corrosion endpoint:
key study V1

Justification for selection of eye irritation endpoint:
key study V1

Justification for classification or non-classification

Under the experimental conditions adopted (OECD 404), "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623 was found to be non irritant for the skin of the rabbit with a Mean Index of erythema and oedema was 0. Substance was not classified according to Council Directive No. 67/548/EEC and subsequent amendments and CLP Regulation No. 1272/2008.

Under the experimental conditions adopted (OECD 405), "hydrate de carbone et sucres, hexitol, anhydro" LAB 4623 was found to be non irritant for the eye of the rabbit. The application of the test item did not induce colouration of the application site and did not interfere with grading of any ocular lesion. One hour after instillation, all animals had redness of the conjunctivae (score 1). There was no ocular lesion from 24 to 72 hours after instillation. There was no ocular lesion from 24 to 72 hours after instillation. Mean index values were 0. Substance was not classified according to Council Directive No. 67/548/EEC and subsequent amendments and CLP Regulation No. 1272/2008.