[No public or meaningful name is available]

Endpoint summary

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Tissues received an additional overnight incubation, but were then used before expiry. SD of negative control viabilities was above acceptance criterion, but as the test item was unequivocally non-irritant, it was unnecessary to repeat the test.

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Not mentioned

Source strain:

other: Human

Vehicle:

not specified

Details on test system:

2 mL of maintenance medium, warmed to approximately 37 °C, was pipetted into the second column of 3 wells of the 12-well plate.
Triplicate tissues were treated with the test item for an exposure period of 15 minutes. The test item was applied topically to the corresponding tissues ensuring uniform covering. 5 µL of sterile distilled water was topically applied to the epidermal surface in order to improve contact between the test item and the epidermis. Approximately 10 mg (26.3 mg/cm2) of the test item was then applied to the epidermal surface. Triplicate tissues treated with 10 µL of DPBS served as the negative controls and triplicate tissues treated with 10 µL of SDS 5% w/v served as the positive controls. To ensure satisfactory contact with the positive control item the SDS solution was spread over the entire surface of the epidermis using a pipette tip (taking particular care to cover the center). After a 7-Minute contact time the SDS solution was re-spread with a pipette tip to maintain the distribution of the SDS for the remainder of the contact period (re-spreading is not required for the negative control or test item). The plates were kept in the biological safety cabinet at room temperature for 15 minutes.
At the end of the exposure period, each tissue was removed from the well using forceps and rinsed using a wash bottle containing DPBS with Ca++ and Mg++. Rinsing was achieved by filling and emptying each tissue insert for approximately 40 seconds using a constant soft stream of DPBS to gently remove any residual test item. The rinsed tissues were transferred to the second column of 3 wells containing 2 mL of maintenance medium in each well. The rinsed tissues were incubated at 37 °C, 5% CO2 in air for 42 hours. Following the 42-Hour post-exposure incubation period each 12-well plate was placed onto a plate shaker for 15 minutes to homogenize the released mediators in the maintenance medium. 1.6 mL of the maintenance medium from beneath each tissue was transferred to pre-labeled micro tubes and stored in a freezer at -14 to -30 ºC for possible inflammatory mediator determination.
2 mL of a 0.3 mg/mL MTT solution, freshly prepared in assay medium, was pipetted into the third column of 3 wells of the 12-well plates. The tissues were transferred to the MTT filled wells, being careful to remove any excess maintenance medium from the bottom of the tissue insert by blotting on absorbent paper. The tissues were incubated for 3 hours at 37 °C, 5% CO2 in air. At the end of the 3-Hour incubation period each tissue was placed onto absorbent paper to dry. A total biopsy of the epidermis was made using the EPISKINTM biopsy punch.
The epidermis was carefully separated from the collagen matrix using forceps and both parts (epidermis and collagen matrix) placed into labeled 1.5 mL micro tubes containing 500 µL of acidified isopropanol, ensuring that both the epidermis and collagen matrix were fully immersed. Each tube was plugged to prevent evaporation and mixed thoroughly on a vortex mixer. The tubes were refrigerated at 1 to 10 °C until Day 6 of the experiment, allowing the extraction of formazan crystals out of the MTT-loaded tissues.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

Approximately 10 mg (26.3 mg/cm2) of the test item was applied to the epidermal surface.

Duration of treatment / exposure:

15 minutes

Duration of post-treatment incubation (if applicable):

42 hour

Number of replicates:

Triplicate

Irritation / corrosion parameter:

% tissue viability

Value:

ca. 96.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Interpretation of results:

GHS criteria not met

Conclusions:

The test item was non-irritant in in vitro skin irritation testing.

Executive summary:

Skin irritation of the text item was evaluated according to OECD Guideline 439 (In vitro skin irritation). The test item (10 mg) was applied topically to reconstructed human epidermis model (EPISKIN) in a cell culture dish. After the 15-minute exposure period, the cells were incubated an additional 42 hours. After the post-exposure incubation period, the cells were analyzed for cell viability (cytotoxicity), a predictor for rabbit skin irritation. Exposure to the test item resulted in 97% cell viability, indicating that the test item is a non-irritant. This results in the test material being unclassified according to GHS.

  

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

other information

Study period:

Study initiation date: 05 December 2012; Experimental completion date: 31 December 2012

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Irritation parameter:

overall irritation score

Basis:

other: Draize Score

Time point:

24 h

Score:

16

Max. score:

110

Interpretation of results:

GHS criteria not met

Conclusions:

Notified Substance is expected to be a non-irritant to the eye.

Executive summary:

The Notified Substance is a product consisting of a mixture of octadecane-1,2-diol (CAS 20294-76-2) (35 -39%), hexadecane-1,2-diol (CAS 6920-24-7) (38 -42%), and polyether impurities (approximately 17 -25%). The eye irritation data gap can be filled by a read-across assessment using data available from structural analogues to predict toxicological outcomes of the Notified Substance. Using SUBSTANCE A (CAS No. 1384165-12-1) for the read-across approach, the Notified Substance is predicted to be non-irritating to the eye. Therefore, the Notified Substance is not classified for eye irritation.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

other information

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Interpretation of results:

GHS criteria not met

Conclusions:

The read-across assessment with structurally analogs concludes that the Notified Substance is non-irritating to the eye and thus not classified by EU CLP and UN GHS.

Executive summary:

The Notified Substance is a product consisting of a mixture of octadecane-1,2-diol (CAS 20294-76-2) (35 -39%), hexadecane-1,2-diol (CAS 6920-24-7) (38 -42%), and polyether impurities (approximately 17 -25%). The eye irritation data gap can be filled by read-across assessment using data available from structural analogues to predict toxicological outcomes of the Notified Substance. Using long-chain alcohols (i.e., C6-C22 primary aliphatic alcohols) with a focus on C16-C18 alcohol (CAS 67762-27-0), for the read-across approach, the Notified Substance is predicted to be non-irritating to the eye. Therefore, the Notified Substance is not classified for eye irritation. 

Reference 3

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Study initiation date: 05 December 2012; Experimental completion date: 31 December 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP 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
- Source: Male New Zealand White albino rabbits were received from Covance Research Products, Inc., Denver, PA
- Age at study initiation: The selected animals were approximately 7 months old.
- Weight at study initiation: 3118.1 g to 4061.0 g
- Fasting period before study: No fasting period.
- Housing: all animals were housed individually in clean, stainless steel cages. Enrichment devices were provided to all animals as appropriate throughout the study for environmental enrichment. One leaf of fresh kale was offered to each rabbit every Monday, Wednesday, and Friday beginning after the day of receipt.
- Diet (e.g. ad libitum): Basal diet (certified feed) provided at approximately 150 g/day was provided ad libitum
- Water (e.g. ad libitum): municipal water, delivered by an automatic watering system, was provided ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Set to maintain conditions of 19 ± 3°C
- Humidity (%): Set to maintain conditions of 50 ± 20%
- Air changes (per hr): 10 fresh air changes per hour
- Photoperiod (hrs dark / hrs light): Fluorescent lighting provided illumination for a 12-hour light (0600 hours to 1800 hours)/12-hour dark photoperiod.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100-mg dose of the test substance was instilled into the lower conjunctival sac of the right eye.

Duration of treatment / exposure:

Single application.

Observation period (in vivo):

14 days.

Number of animals or in vitro replicates:

3.

Details on study design:

Method of test substance administration:
The test substance was placed directly into the cupped lower conjunctival sac of each rabbit’s right (test) eye. The eyelid was held closed for approximately 1 second after instillation and then released. The left eye of each animal was manipulated in an identical manner to simulate the dosing of the right eye.

Initially, a single animal was dosed to evaluate the ocular irritative potential of the test substance. This single rabbit received a single, unwashed exposure. No severe ocular damage was observed in this animal. Based on these results, and to complete the study, 2 additional rabbits each received a single, unwashed exposure of the test substance.

Parameters evaluated:
Mortality:
The rabbits were observed twice daily, once in the morning and once in the afternoon, for mortality and moribundity.

Ocular observations:
Both eyes of all rabbits were examined for ocular abnormalities prior to initiation of dosing. The pre-initiation examination included the use of sodium fluorescein and a direct ophthalmoscope for detection of corneal abnormalities. Only rabbits with no pre-existing ocular abnormalities were assigned to study.

Both eyes of all rabbits were examined macroscopically for ocular irritation in accordance with the method of Draize scoring criteria for ocular reactions, at approximately 1, 24, 48, and 72 hours after dosing and on study days 4, 7, 10, and 14, if irritation persisted. A direct ophthalmoscope was used during these observations to examine the corneal tissue. In addition, both eyes were further examined at 24 hours and at all subsequent observations with sodium fluorescein.

Body Weights:
Body weights were obtained and recorded on study day 0 (initiation) and at each rabbit’s termination from the study.

Termination:
After study termination, the rabbits were euthanized by intravenous injection of sodium pentobarbital and discarded.

Irritation parameter:

overall irritation score

Basis:

other: Draize Score

Time point:

other: 24 hour

Score:

16

Max. score:

110

Irritant / corrosive response data:

Positive corneal irritation (grade 1) and iridial irritation (grade 1) were noted for 1 male (No. 9853), and positive conjunctival irritation (grade 2) was noted for the remaining 2 males (Nos. 9850 and 9851). Positive corneal and iridial irritation subsided by 24 hours post-instillation. Positive scores for conjunctival irritation subsided by 72 hours post-instillation. All ocular irritation subsided by study day 14.

Individual and average ocular irritation scores for treated eyes are presented in Table 1, and individual animal results (other findings), including sodium fluorescein examination results, are presented in Table 2 (see attached background material for Tables 1 and 2).

The left (control) eyes were free of evidence of ocular irritation and other findings for the duration of the study.

The Draize scale for scoring ocular irritation and method of score calculation are presented in attached background material.

Other effects:

Mortality:
There were no deaths during the study.

Body Weights:
There were no remarkable body weight changes noted during the study.

Ocular Irritation Results Summary:

Number of Animals with Positive Effect/Number of Animals Treated

Group	Cornea	Iris	Conjunctiva	Total	24 Hour Draize
100 mg/right eye, unwashed	1/3	1/3	2/3	3/3	16/110

 

Conclusions:

The 24-hour post-instillation Draize score for SP4834 was 16/110. All ocular irritation subsided by study day 14.

Executive summary:

Objective:

The objective of this study was to determine the primary ocular irritative potential of the test substance in albino rabbits.

Test Guidelines:

The protocol was designed to be in general compliance with the OECD Guidelines for Testing of Chemicals, Section 405 (2002).

Study Design:

Initially, a single animal was dosed to evaluate the ocular irritative potential of the test substance. This single rabbit received a single, unwashed exposure. No severe ocular damage was observed in this animal. Based on these results, and to complete the study, 2 additional rabbits received single, unwashed exposures of the test substance. Each 100-mg dose of the test substance was instilled into the lower conjunctival sac of the right eye. The eyelid was held closed for approximately 1 second and released. The left eye of each animal was manipulated in an identical manner to simulate the dosing of the right eye and served as a contralateral control.

Results:

There were no deaths or remarkable body weight changes during the study.

Positive corneal irritation (grade 1) and iridial irritation (grade 1) were noted for 1 animal and conjunctival irritation (grade 2) was noted for the remaining 2 animals. Positive corneal and iridial irritation subsided by 24 hours post-instillation. Positive conjunctival irritation subsided by 72 hours post-instillation. All ocular irritation subsided by study day 14.

Conclusion:

The 24-hour post-instillation Draize score for SP4834 was 16/110. All ocular irritation subsided by study day 14. The following table presents the individual ocular irritation scores at 24, 48, and 72 hours post-instillation:

Animal	Tissue	24 Hours	48 Hours	72 Hours
9853	Cornea (O-A)	0 0	0 0	0 0
	Iris	0	0	0
	Conjunctive  (R-C-D)	1 1 0	1 0 0	0 0 0
9850	Cornea (O-A)	0 0	0 0	0 0
	Iris	0	0	0
	Conjunctive  (R-C-D)	2 1 0	1 1 0	1 0 0
9851	Cornea (O-A)	0 0	0 0	0 0
	Iris	0	0	0
	Conjunctive  (R-C-D)	2 1 0	2 1 0	1 0 0

 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Justification for classification or non-classification

The test item was determined to be non-irritating to skin by an in vitro skin test and non-irritating to the eye by a read-across approach. The results are considered scientifically valid to support non-classification. Therefore, this material is not classified as irritating to skin or eye.