Sucrose and glycerol, reaction products with C12-18, C18unsatd. fatty acids

Endpoint summary

Description of key information

Skin irritation / corrosion
Sucroglyceride C12 -18, C18unsatd. is not irritating to the skin following in vitro application to RHE, the relative absorbance being 87,5 % (classification is required at 50% or below).
Sucroglyceride C12 -18, C18unsatd. 10% a.i. is not irritating in vivo following dermal treatment of rabbits.  
The read-across substances Sucroglyceride C12 10% a.i., Stearic acid, esters with methyl α-D-glucoside and Isostearic acid, esters with methyl α-D-glucoside are not irritating in vivo following dermal treatment of rabbits. 
Eye irritation
Sucroglyceride C12 -18, C18unsatd. 100 % and 25% a.i. are not irritating in vivo following ocular treatment of rabbits. 
Sucroglyceride C12 -18, C18unsatd. is not corrosive or severely irritating to the eye following in vitro application to bovina corneae, the measured score being 13,94 (classification is required at 55,1 or above).
The read-across substances Stearic acid, esters with methyl α-D-glucoside and Isostearic acid, esters with methyl α-D-glucoside are not irritating in vivo following ocular treatment of rabbits. 
General evaluation
All studies are GLP compliant guideline studies, the key studies have a Klimisch score 1 while the Klimisch scores of the supporting studies were changed from 1 to 2 to reflect the fact that they were conducted on read-across substances. 
Overall,  Sucroglyceride C12 -18, C18unsatd. is not considered a dermal or an ocular irritant. 

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011-09-21 to 2011-09-26

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: Human skin model: EpiSkin™

Strain:

other: EpiSkin™ Kit Lot No.: 11-EKIN-034

Type of coverage:

other: not applicable in this in vitro test system

Preparation of test site:

other: not applicable in this in vitro test system

Vehicle:

water

Controls:

other: refer to "details on study design"

Amount / concentration applied:

TEST MATERIAL
- Amount applied to each EpiSkin™ insert of the treatment group: 10 mg of test substance

VEHICLE
- Amount applied to each EpiSkin™ insert of the treamtent group: 15 μL of deionised water

Duration of treatment / exposure:

15 ± 1 min

Observation period:

refer to "details on study design"

Number of animals:

refer to "details on study design"

Details on study design:

TEST MODEL
For the test an EpiSkin™ Reconstituted Human Epidermis model was used. The model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis. It consists of organised basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiSkin™ tissues are cultured on specially prepared cell culture inserts.

PREPARATION OF TEST SYSTEM
EpiSkin™ tissues were shipped with ice packs on medium-supplemented agarose gels in a 12-well plate, they were delivered one day prior to the experiment, transferred to 12-well plates with maintenance medium and prewarmed for 23 hours.

TREATMENT
Three sets of triplicate EpiSkin™ tissues contained in 12-well plates were treated respectively with the negative and positive controls (10 μL were dosed per tissue) and with test substance. Due to its waxy consistency, firstly 10 mg of test substance were weighed on top of the stamp of a syringe, wetted with 15 μL of deionised water, and then evenly applied to each of triplicate tissues used for treatment. Incubation took place for 15 ± 1 min at 37 ± 1.5 °C and 5 ± 0.5 % CO2.
At the end of the treatment period the tissue inserts were immediately removed from the 12-well plate and gently rinsed with PBS to remove any
residual test substance. The rinsed inserts were placed in the plates with 2 mL maintenance medium. The tissues were incubated for about 42 hours at 37 ± 1.5 °C and 5 ± 0.5 % CO2.

MTT ASSAY FOR CELL VIABILITY
After 42 hours of incubation the tissues were transferred for the MTT cell viability assay to 12-well plates for a 3 hour incubation period (37 ± 1.5 °C and 5 ± 0.5 % CO2). Then, and following removal of MTT and rinsing, the tissue sampels were immersed into vials containing 0.5 mL extractant solution (isopropanol / 2 N HCl 49:1 (v/v)). Then the tissue samples were completely covered by isopropanol, sealed to inhibit isopropanol evaporation and the formazan salt was extracted for approximately 69 hours in the refrigerator.
Per each tissue sample 2 x 200 μL aliquots of the formazan blue solution were transferred into a 96-well flat bottom microtiter plate. OD was read in a microplate reader (Versamax® Molecular Devices, 85737 Ismaning, Germany) with 570 ± 1 nm filter. Mean values were calculated from the 2 wells per tissue sample.

CONTROL OF MTT REDUCING ACTIVITY OF TEST ITEM
MTT reducing capability of the test substance was tested: ~25 mg of the test item were added to 1 mL of MTT-containing medium and the mixture was incubated in the dark at room temperature for 60 minutes. Untreated MTT medium was used as control.

Irritation / corrosion parameter:

other: other: tissue viability: MTT reduction measured as Formazan salt formation

Remarks on result:

other:

Remarks:

Basis: other: mean of all replicates. Time point: 15 min. Reversibility: other: not applicable. Remarks: relative absorbance at 570 nm for test substance: 87.5 % (threshold for irritancy: ≤ 50 %),. (migrated information)

Irritant / corrosive response data:

Treatment of three EpiSkin™ tissue inserts with the test substance resulted in a mean relative absorbance value of the test substance at 570 nm, corresponding to the cell viability, of 87.5 % as compared to 100.0 % mean relative absorbance for the negative control and 33.0 % for the positive control. The threshold value for irritancy is ≤ 50 %, consequently the test substance was non irritant to skin in this test system.

Results after treatment with Sucroglyceride C12-18, C18unsatd. and controls

Dose group	Treatment Interval	Absorbance 570 nm Tissue 1	Absorbance 570 nm Tissue 2	Absorbance 570 nm Tissue 3	Mean Absorbance of 3Tissues	Relative Absorbance [%] Tissue 1, 2 + 3**	Standard Deviation[%]	Rel. Absorbance [% of Negative Control]***
Negative Control	15 min	0.801	0.814	0.801	0.805	99.5 101.1 99.5	0.9	100.0
Positive Control	15 min	0.269	0.273	0.255	0.266	33.4 33.8 31.7	1.1	33.0
Test Substance	15 min	0.697	0.698	0.719	0.705	86.6 86.6 89.3	1.6	87.5

* Mean of two replicate wells after blank correction

 

 ** relative absorbance per tissue [rounded values]:

 

100 x (absorbance_tissue)

__________________________________________________________

 (mean absorbance_{negative control})

 

 *** relative absorbance per treatment group [rounded values]:

 

100 x (mean absorbance_{test substance})

___________________________________________________________

 (mean absorbance_{negative control})

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: expert judgment

Conclusions:

Treatment of RHE (Reconstituted Human Epidermis) with Sucroglyceride C12-18, C18unsatd. resulted in a mean relative absorbance of 87.5% as compared to 100.0% mean relative absorbance for the negative control. According to the evaluation scheme used in the test system, a substance is considered irritant at a mean realtive absorbance ≤ 50%. Thus, the test substance Sucroglyceride C12-18, C18unsatd. was graded as not irritating to the skin in this in vitro assay.

Executive summary:

In an in vitro Human Skin Model Test, performed according to the OECD Test Guideline 439, the potential to cause irritation upon first contact with the skin was assessed by application of the test substance to RHE (Reconstituted Human Epidermis). 10 mg of Sucroglyceride C12 -18, C18unsatd. were applied together with 15 μL of water to an area of approximately 0.38 cm² of RHE and incubated for 15 min.

For evaluation of the potential of the test substance to cause skin irritation, the samples of treated reconstituted skin were then exposed to MTT for 3 h. The potential to reduce MTT and form a blue Formazan salt was assessed by extraction of Formazan over 69 h and subsequent determination of the absorbance at 570 nm.

Treatment with a 5% solution of SLS (Sodium lauryl sulphate) in deionised water, used as positive control, resulted in a mean relative absorbance of 33.0%, thus showing the normal functionality of the test system.

Treatment with the test substance resulted in a mean relative absorbance of 87.5% as compared to 100.0% mean relative absorbance for the negative control. According to the evaluation scheme used in the test system, a substance is considered irritant at a mean realtive absorbance ≤ 50 %. Thus, the test item Sucroglyceride C12 -18, C18unsatd. was graded as not irritating to the skin in this in vitro assay.

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:

07-11-2011 to 22-11-2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.16 - 2.67 kg
- Age: 12-20 weeks
- Housing: individual, suspended cages
- Diet: ad libitum, 2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK
- Water: ad libitum
- Acclimation period: 5 days at least

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23
- Humidity (%): 30-70
- Air changes (per hr): at least 15
- - Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

other: control: left untreated eye of each test animal

Amount / concentration applied:

0.1 mL of the test material (approximately 99 mg) was placed into the conjunctival sac of the right eye while the left eye served as control.

Duration of treatment / exposure:

exposure to test material not limited, not rinsed after application

Observation period (in vivo):

assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment

Number of animals or in vitro replicates:

2

Details on study design:

REMOVAL OF TEST SUBSTANCE:
no

SCORING SYSTEM:
according to Draize (from Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.48 to 49).

TOOL USED TO ASSESS SCORE:
examination of the eye was facilitated by the use ofthe light source from a standard ophthalmoscope

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2

Time point:

other: 1 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 24 h

Irritation parameter:

cornea opacity score

Basis:

animal: #1, #2

Time point:

other: 24 h, 48 h, 72 h

Score:

0

Max. score:

4

Reversibility:

other: n.a.

Irritation parameter:

iris score

Basis:

animal: #1, #2

Time point:

other: 1 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 24 h

Irritation parameter:

iris score

Basis:

animal: #1, #2

Time point:

other: 24 h, 48 h, 72 h

Score:

0

Max. score:

2

Reversibility:

other: n.a.

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2

Time point:

other: 1 h, 24 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2

Time point:

other: 48 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

animal: #1, #2

Time point:

other: 72 h

Score:

0

Max. score:

3

Reversibility:

other: n.a.

Irritation parameter:

chemosis score

Basis:

animal: #1, #2

Time point:

other: 1 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

chemosis score

Basis:

animal: #1, #2

Time point:

other: 24 h, 48 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

chemosis score

Basis:

animal: #1, #2

Time point:

other: 72 h

Score:

0

Max. score:

4

Reversibility:

other: n.a.

Irritation parameter:

other: discharge score

Basis:

animal: #1, #2

Time point:

other: 1 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

other: discharge score

Basis:

animal: #1, #2

Time point:

other: 24 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

other: discharge score

Basis:

animal: #1, #2

Time point:

other: 48 h, 72 h

Score:

0

Max. score:

3

Reversibility:

other: n.a.

Irritant / corrosive response data:

The test substance produced scattered or diffuse corneal opacity and iridial inflammation in both treated eyes one hour after treatment. Moderate conjunctival irritation was noted in both treated eyes one and 24 hours after treatment with minimal conjunctival irritation noted at the 48 hour observation. Both treated eyes appeared normal at the 72 hour observation.

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: CLP, EU GHS (Regulation (EC) No 1272/2008) and DSD (Directive 67/548/EEC)

Conclusions:

Based on the results of this study, the test substance was determined to be non-irritant to the eye. According to the criteria of OECD GHS, Regluation (EC) 1272/2008 and of DSD (Directive 67/548/EEC) no classification and labelling is required.

Executive summary:

In a primary eye irritation study performed according to the testing guideline OECD 405 0.1 mL (99 mg) of  Sucroglyceride C12-18, C18unsatd. (92.66% a.i.) was instilled undiluted into the conjunctival sac of one eye of 2 young adult New Zealand White rabbits. Eyes were not washed. Animals then were observed for 72 hours. Irritation was scored by the method of Draize.

Scattered or diffuse corneal opacity and iridial inflammation in both treated eyes one hour after treatment. Moderate conjunctival irritation was noted in both treated eyes one and 24 hours after treatment with minimal conjunctival irritation noted at the 48 hour observation. All observed effects were completely reversible within 72 hours and both treated eyes appeared normal at the 72 hour observation.

In this study, Sucroglyceride C12-18, C18unsatd. (93.66% a.i) is not an eye irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Assessment of skin irritation / corrosion

 

Key study on the registration substance Sucroglyceride C12-18, C18unsatd. (Evonik, 2011)

For the examination of skin irritation, Sucroglyceride C12-18, C18unsatd. was tested in vitro on Reconstructed Human Epidermis (RHE). The test substance is a waxy solid. Purity is 100% or 91-94%, respectively, by subtraction of the water content.

In an in vitro Human Skin Model Test, performed according to the OECD Test Guideline 439, the potential to cause irritation upon first contact with the skin was assessed by application of the test substance to RHE. 10 mg of the Sucroglyceride C12 -18, C18unsatd. were applied together with 15 μL of water to an area of approximately 0.38 cm² of RHE and incubated for 15 min.

For evaluation of the potential of the test substance to cause skin irritation, the samples of treated reconstituted skin were then exposed to MTT for 3 h. The potential to reduce MTT and form a blue Formazan salt was assessed by extraction of Formazan over 69 h and subsequent determination of its absorbance at 570 nm.

The positive control, a 5 % solution of SLS (Sodium lauryl sulphate) in water, resulted in a mean relative absorbance of 33.0%, the negative control value was 100%. Treatment with the test substance resulted in a mean relative absorbance of 87.5%.

In this test system, the threshold for considering a substance as irritant is a mean relative absorbance ≤ 50%. Thus, Sucroglyceride C12 -18, C18unsatd. is graded as not irritant to the skin.

 

Key study on the registration substance Sucroglyceride C12-18, C18unsatd. 10% a.i. (Evonik, 1985)

In an in vivo study, a solution of Sucroglyceride C12-18, C18unsatd. 10% a.i. was tested. The test substance is a white liquid. Purity is 10% a.i.

A primary dermal irritation study was performed according to Federal Register 1973, Volume 38, No. 187, 1500.41. The study design is in principle comparable to a guideline study (OECD 404) with some deviations. Treatment was on intact and abraded skin, exposure time was 24 h, readings were performed at 24 and 72 h.

0.5 mL of Sucroglyceride C12-18, C18unsatd. 10% a.i. were administered under occlusive dressing to intact and abraded skin segments, to 2.5 x 2.5 cm of the body surface area, of 6 young adult New Zealand White rabbits. The duration of exposure was 24 h, animals then were observed for 72 hours and skin reactions were scored by the method of Draize.

On intact skin, edema scores for all animals were 0 at the 24 and 72 h readings while the erythema score was 0 for one animal and 1 for five animals at the 24 h reading and 0 for all animals at 72 h.

On the abraded skin, edema scores were 0 for five animals and 1 for one animal at the 24 h reading and 0 for all animals after 72 h, while the erythema score was 1 for four animals and 2 for two animals at the 24 h reading and 0 for all animals at 72 h.

Thus, all dermal reactions were completely reversible after 72 h and in no case the edema or erythema scores obtained for Sucroglyceride C12-18, C18unsatd. 10% a.i. reached the threshold which would require classification. Sucroglyceride C12-18, C18unsatd. 10% a.i. is graded as not irritant to the skin.

 

Supporting study on the read-across substance Sucroglyceride C12 10% a.i. (Evonik, 1987)

The read-across substance Sucroglyceride C12 10% a.i., a structural analogue of the registration substance, was tested in vivo for its dermal irritation potential. The test substance is a white liquid, purity is 10% a.i.

A primary dermal irritation study was performed according to Federal Register 1973, Volume 38, No. 187, 1500.41. The study design is in principle comparable to a guideline study (OECD 404) with some deviations. Treatment was on intact and abraded skin, exposure time was 24 h, readings were performed at 24 and 72 h.

The study design and the preparation of the test substance was identical to the key study reported above.

On intact skin, edema scores were 0 for four animals and 1 for two animals at the 24 h reading and 0 for all animals after 72 h, while the erythema score was 1 for five animals and 2 for one animal at the 24 h reading and 0 for all animals at 72 h.

On the abraded skin, edema scores were 0 for five animals and 1 for one animal at the 24 h reading and 0 for all animals after 72 h, while the erythema score was 1 for five animals and 2 for one animal at the 24 h reading and 0 for all animals at 72 h.

Thus, all dermal reactions were completely reversible after 72 h and in no case the edema or erythema scores obtained for Sucroglyceride C12 10% a.i. reached the threshold which would require classification. Sucroglyceride C12 10% a.i. is graded as not irritant to the skin.

 

Supporting study on the read-across substance Stearic acid, esters with methyl α-D-glucoside (Evonik, 1990)

The read-across substance Stearic acid, esters with methyl α-D-glucoside, a structural analogue of the registration substance, was tested in vivo for its dermal irritation potential. The test substance is a solid. Purity is 100%.

In a primary dermal irritation study according to the OECD Guideline 404 (Acute Dermal Irritation / Corrosion), three New Zealand White rabbits were dermally exposed to 0.5 mL of Stearic acid, esters with methyl α-D-glucoside. The test substance was applied for 4 hours to intact shaved body surface areas of approximately 6 cm² under semi-occlusive dressing. Animals then were observed for 72 hours. Irritation was scored by the method described in OECD Guideline 404.

Edema scores and erythema scores were 0 for all animals after 30-60 min and after 24, 48 and 72 hours.

Thus, Stearic acid, esters with methyl α-D-glucoside is graded as not irritant to the skin.

 

Supporting study on the read-across substance Isostearic acid, esters with methyl α-D-glucoside (Evonik, 2009)

The read-across substance Isostearic acid, esters with methyl α-D-glucoside, a structural analogue of the registration substance, was tested in vivo for its dermal irritation potential. The test substance is a yellow paste. Purity is 100%.

In a primary dermal irritation study according to the OECD Guideline 404 (Acute Dermal Irritation / Corrosion), three young adult male New Zealand White rabbits were dermally exposed to 0.5 g of Isostearic acid, esters with methylα-D-glucoside as supplied by the sponsor for 4 hours, under semi-occlusive dressing. The animals then were observed for 14 days. Irritation was scored by the method described in the OECD Guideline 404.

Very slight erythema in the treated skin areas of all three animals was observed after 60 minutes, which had resolved within 24 h after exposure in two animals and within 7 days after exposure in one animal. Scaliness in the treated skin area of one animal was noted at 72 hours and 7 days post-treatment and had resolved within 14 days.

Edema scores were 0 for all animals after 30-60 min and after 24, 48 and 72 h.

Erythema scores were 0 for two animals and 1 for one animal after 24 h and 0 for all animals after 48 and 72 h.

Thus, all dermal reactions were completely reversible after 72 h and in no case the edema or erythema scores obtained for Isostearic acid, esters with methyl α-D-glucoside reached the threshold which would require classification. Isostearic acid, esters with methyl α-D-glucoside is graded as not irritant to the skin.

 

General evaluation of skin irritation / corrosion

Pure Sucroglyceride C12-18, C18unsatd. is not irritant in an in vitro assay and a solution containing 10% a.i. of the same substance is not irritant following 24 h exposure to intact and abraded skin. The overall evaluation of these data allow to conclude that Sucroglyceride C12-18, C18unsatd. is not a skin irritant.

 

For dermal irritation / corrosion, the registration substance Sucroglyceride C12-18, C18unsatd. and the three read-across substances Sucroglyceride C12, Stearic acid, esters with methyl α-D-glucoside and Isostearic acid, esters with methyl α-D-glucoside exhibit a comparable profile and are characterised by experimental results which lead to the same overall evaluation with regard to classification and labelling.

 

For the registration substance Sucroglyceride C12-18, C18unsatd. and for three read-across substances, the dermal irritation potential is very low or absent after a single dermal administration.

 

Based on the experimental evidence provided for skin irritation / corrosion, Sucroglyceride C12-18, C18unsatd. does not need to be classified for skin irritation according to CLP, EU GHS (Regulation (EC) No 1272/2008) and according to DSD (Directive 67/548/EEC) and labelling is not required.

 

 

Assessment of eye irritation

 

Key study on the registration substance Sucroglyceride C12-18, C18unsatd. (Evonik, 2012)

In order to assess its potential for eye irritation, Sucroglyceride C12-18, C18unsatd. was tested in an in vivo study. The test substance is a white waxy solid. Purity is 100% or 92.66%, respectively, by subtraction of the water content.

A primary eye irritation study was performed according to the OECD Guideline 405 (Acute Eye Irritation / Corrosion). In this study, eye irritation was tested on only two rabbits rather than three since the eye irritation scores of the two animals were identical. In the absence of differences in the irritation scores of the two rabbits, and for animal welfare reasons it is the default of the testing facility to avoid the use of a third animal. In fact, on the basis of the results obtained, it is not to be expected that treating a third rabbit would have modified the study outcome and the indications for classification and labelling.

0.1 mL (99 mg) of Sucroglyceride C12-18, C18unsatd. was instilled undiluted into the conjunctival sac of one eye of 2 young adult New Zealand White rabbits. Eyes were not washed. Animals were then observed for 72 hours. Irritation was scored by the method of Draize.

Scattered or diffuse corneal opacity and iridial inflammation was observed in both treated eyes 1 h after treatment. Moderate conjunctival irritation was noted in both treated eyes a 1 and 24 h after treatment with minimal conjunctival irritation noted at the 48 h observation.

For both animals the cornea score was 1 after 1 h and 0 at 24-48-72 h, the iris score was 1 after 1 h and 0 at 24-48-72 h, the conjunctivae score was 2 at 1-24 h, 1 at 48 h and 0 at 72 h, the chemosis score was 2 at 1 h, 1 at 24-48 h and 0 at 72 h, the discharge score was 2 at 1 h, 1 at 24 h and 0 at 48-72 h.

Thus, all observed ocular effects were completely reversible within 72 h. Both treated eyes appeared normal and in no case the Draize scores obtained for Sucroglyceride C12-18, C18unsatd. reached the threshold which would require classification. Sucroglyceride C12-18, C18unsatd. is graded as not irritant to the eye.

 

Key study on the registration substance Sucroglyceride C12-18, C18unsatd. 25% a.i. (Evonik, 1985)

In an in vivo study, a solution of Sucroglyceride C12-18, C18unsatd. 25% a.i. was tested. The test substance is a white viscous liquid. Purity is 25% a.i.

An ocular irritation study was performed according to Federal Register 1973, Volume 38, No. 187, 1500.42. The study design is in principle comparable to a guideline study (OECD 405). The eye irritation potential of the test substance was assessed after a single application to rabbit eye for 24 hours.The animals were observed for evidence of irritation at 24-48-72 h post application. Reactions were scored according to the scale of Draize.

Sucroglyceride C12-18, C18unsatd. (25% a.i.) undiluted was instilled into the conjunctival sac of one eye of 6 young adult New Zealand White rabbits. Eyes were not washed. Animals then were observed for 72 hours.

Minimal irritation observed in one animal at 24 hours reading (conjunctivae redness score 1) was completely reversible within 48 hours.

The cornea score was 0 for all animals at 24-48-72 h, the iris score was 0 for all animals at 24-48-72 h, conjunctivae score was 0 for five animals and 1 for one animal after 24 h and 0 for all animals at 48-72 h, chemosis score was 0 for all animals at 24-48-72 h.

Thus, the minor ocular effects observed were completely reversible within 48 h. In no case the Draize scores obtained for Sucroglyceride C12-18, C18unsatd. 25% a.i. reached the threshold which would require classification. Sucroglyceride C12-18, C18unsatd. 25% a.i. is graded as not irritant to the eye.

 

Key study on the registration substance Sucroglyceride C12-18, C18unsatd. (Evonik, 2011)

For the examination of eye irritation in vitro, Sucroglyceride C12-18, C18unsatd. was tested in the BCOP Assay. The test substance is a waxy solid. Purity is 100% or 91-94%, respectively, by subtraction of the water content.

In an in vitro Bovine Cornea Opacity and Permeability (BCOP) Assay, performed according to the OECD Test Guideline 437, the potential to cause irritation upon first contact with the eye was assessed by application of the test substance to three freshly isolated bovine corneae.0.75 mL of a 20 % (w/v) solution of test substance Sucroglyceride C12 -18, C18unsatd. in saline, i.e. 0.9% (w/v) NaCl in deionised water, were applied top each cornea and incubated for 240 min at 32 ± 1 °C.

After a first opacity measurement of the fresh bovine corneae (t₀), for evaluation of the potential of the test substance to cause corrosion / severe irritation to the eye, the 20% (w/v) suspension of the test item in saline, the positive, and the negative controls were applied to corneae and incubated for 240 minutes at 32 ± 1 °C. After the incubation phase the test item, the positive, and the negative controls were each rinsed from the corneae and opacity was measured again (t₂₄₀).

After the opacity measurements permeability of the corneae was determined by measuring spectrophotometrically the transfer of fluorescein after incubation in a horizontal position for 90 minutes at 32 ± 1 °C.

With the negative control (saline) neither an increase of opacity nor permeability of the corneae could be observed.

The positive control (10% (w/v) Benzalkonium chloride in saline) showed clear opacity of the corneae corresponding to a classification as corrosive / severe irritant to the eye (GHS / CLP Cat 1).

Treatment of bovine corneae with Sucroglyceride C12-18, C18unsatd. resulted in a mean in vitro irritation score of 13.94 as compared to 1.80 of the negative control and 195.88 of the positive control. According to the evaluation scheme used in the test system, a substance is considered corrosive / severe irritant to the eye at an in vitro irritation score ≥ 55.1. Thus, the test substance Sucroglyceride C12-18, C18unsatd. was graded not corrosive / severe irritant to the eye in this in vitro assay.

 

Supporting study on the read-across substance Stearic acid, esters with methyl α-D-glucoside (Evonik, 1990)

The read-across substance Stearic acid, esters with methyl α-D-glucoside, a structural analogue of the registration substance, was tested in vivo for its ocular irritation potential. The test substance is a solid. Purity is 100%.

A primary eye irritation study was performed according to the OECD Guideline 405 (Acute Eye Irritation / Corrosion). In this study, eye irritation was tested on three rabbits.

0.1 g of Stearic acid, esters with methylα-D-glucoside was instilled undiluted into the conjunctival sac of one eye of three New Zealand White rabbits. The untreated eye served as control. The eyes were not washed. Animals were then observed for 72 hours. Irritation was scored as stipulated by OECD Guideline 405.

No iridial irritation or corneal opacity was observed. An irritation of the conjunctivae, which consisted of redness and chemosis was observed in one eye of each rabbit within the first hour. The irritation was fully reversible within 24 hours.

The cornea score was 0 for all animals at 1-24-48-72 h, the iris score was 0 for all animals at 1-24-48-72 h, conjunctivae score was 0 for all animals at 24-48-72 h, chemosis score was 0 for all animals at 24-48-72 h.

Thus, the minor ocular effects observed were completely reversible within 24 h. In no case the Draize scores obtained for Stearic acid, esters with methyl α-D-glucoside reached the threshold which would require classification. Stearic acid, esters with methyl α-D-glucoside is graded as not irritant to the eye.

 

Supporting study on the read-across substance Isostearic acid, esters with methyl α-D-glucoside (Evonik, 2009)

The read-across substance Isostearic acid, esters with methyl α-D-glucoside, a structural analogue of the registration substance, was tested in vivo for its ocular irritation potential. The test substance is a yellow paste. Purity is 100%.

In a primary eye irritation study according to OECD Guideline 405 0.1 mL of Isostearic acid, esters with methyl α-D-glucoside, was instilled into the conjunctival sac of one eye, each of three New Zealand White rabbits. The eyes were not washed. Animals were then observed for 72 hours. Irritation was scored according to OECD Guideline 405.

After 1 h, instillation of the test substance resulted in irritation of the conjunctivae, which consisted of redness (score 1). The irritation completely resolved within 24 hours.

The cornea score was 0 for all animals at 1-24-48-72 h, the iris score was 0 for all animals at 1-24-48-72 h, conjunctivae score was 1 for all animals at 1 h and 0 for all animals at 24-48-72 h, chemosis score was 0 for all animals at 1-24-48-72 h, discharge score was 0 for all animals at 1-24-48-72 h.

Thus, the minor ocular effects observed were completely reversible within 24 h. In no case the Draize scores obtained for Isostearic acid, esters with methyl α-D-glucoside reached the threshold which would require classification. Isostearic acid, esters with methyl α-D-glucoside is graded as not irritant to the eye.

 

General evaluation of eye irritation

Sucroglyceride C12-18, C18unsatd. is not irritant to the eye in two in vivo assays, performed on pure substance and on 25% a.i. Furthermore, in an in vitro BCOP assay, a solution containing 20% a.i. of the same substance is not corrosive or severely irritant following 4 h exposure to freshly isolated bovine corneae. The overall evaluation of these data allow to conclude that Sucroglyceride C12-18, C18unsatd. is not an eye irritant.

 

For ocular irritation, the registration substance Sucroglyceride C12-18, C18unsatd. and the two read-across substances, Stearic acid, esters with methyl α-D-glucoside and Isostearic acid, esters with methyl α-D-glucoside exhibit a comparable profile and are characterised by experimental results which lead to the same overall evaluation with regard to classification and labelling.

 

For the registration substance Sucroglyceride C12-18, C18unsatd. and for two read-across substances, the ocular irritation potential is extremely low or absent after a single ocular administration.

 

Based on the experimental evidence provided for ocular irritation, Sucroglyceride C12-18, C18unsatd. does not need to be classified for eye irritation according to CLP, EU GHS (Regulation (EC) No 1272/2008) and according to DSD (Directive 67/548/EEC) and labelling is not required.

Justification for selection of skin irritation / corrosion endpoint:
Only the in vitro study was performed on concentrated Sucroglyceride C12-18, C18unsatd.
In the second key study, an vivo test, Sucroglyceride C12-18, C18unsatd. (10% a.i.) was used.

Justification for selection of eye irritation endpoint:
Only this in vivo study was performed on concentrated Sucroglyceride C12-18, C18unsatd.
In a second in vivo study Sucroglyceride C12-18, C18unsatd. (25% a.i.) was used. An in vitro study was performed on concentrated Sucroglyceride C12-18, C18unsatd. Further two studies report read-across data from supporting substancees (structural analogues).

Justification for classification or non-classification

With respect to skin irritation / corrosion the in vitro and in vivo results for the test substance and the in vivo read-across data from three studies with a chemically closely related substances show that edema and erythema scores are very low or absent and that there is no skin irritation potential. Consequently, unambiguous data are available to support that, according to the CLP Regulation (EC) No 1272/2008 and according to DSD (67/548/EEC), there is no need for classification of Sucroglyceride C12 -18, C18unsatd. for skin irritation / corrosion and no labelling is required.

Furthermore, with respect to eye irritation the in vitro and in vivo results for the test substance and the in vivo read-across data from two studies with chemically closely related substances show that scores for cornea, iris, conjunctivae, chemosis and discharge are very low or absent and that there is no eye irritation potential.

Consequently, unambiguous data are available to support that, according to the CLP Regulation (EC) No 1272/2008 and according to DSD (67/548/EEC), there is no need for the classification of Sucroglyceride C12 -18, C18unsatd. for eye irritation and no labelling is required.