[No public or meaningful name is available]

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

January 15, 2019 to April 28, 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Ministry of Environmental Protection of the People's Republic of China State Department of environmental protection: The Guideline for Testing of Chemicals (HJ/T 153-2004)-405.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Sulfonic Acids, C18-alkane hydroxyl and C18-alkene, potassium salts

Cas Number:

2210238-56-3

IUPAC Name:

Sulfonic Acids, C18-alkane hydroxyl and C18-alkene, potassium salts

Test material form:

solid

Remarks:

Color: White to pale yellow This sample was obtained from the original liquid form by removing water.

Specific details on test material used for the study:

-Storage conditions
Store container tightly closed in well-ventilated place. Store in corrosive resistant container with a resistant inliner. Store locked up. Don't store with acids.

-Test item stability under storage conditions
Stable in container after opening. Stable in water and in light. Stability in soil is not applicable.

Test animals / tissue source

Species:

rabbit

Strain:

other: Japanese white large-eared rabbits

Details on test animals or tissues and environmental conditions:

Sex: Female, nulliparous and non-pregnant
Age: 84 - 140 d (at receipt)
Body Weight Range: 2.00 - 2.05 kg (at receipt)
Acclimatization: The animal was introduced for health examination within 24 hours, and the auricle number and cage card were used for unique identification within 24 hours, and the animal was weighed. After the introduction of single cage feeding, the animals were transferred from the stock animals to the experiment for 32 d of adaptation time, and the animals were observed every day during the adaptation period.


Husbandry: Animals are housed in building C room C127 at the start of study. Animals are individually raised in suspended, stainless wire cages (L38 cm x W50 cm x H35 cm) on cage racks (L200 cm x W65 cm x H 170 cm) during the acclimation and observation period. Animal use license No.: SYXK (Liao) 2015-0006.
Conditions: The ranges for room temperature and relative humidity are 18.04 - 23.00℃ and 40.00 - 67.40%, respectively. The animals are provided with a light cycle of 12 h light and 12 h dark.
Diet and Water: Animals are provided with pellet breeding rabbit diet by Liaoning Changsheng Biotechnology Co., Ltd. (Lot number 18112751, 19022261), License number: SCXK(Liao) 2015-0003. All the nutrition components and contaminants are within the permitted limits described in the national standard (GB 14924.3-2010 and GB/T 14924-2-2001). Results of diet analyses are supplied by the supplier.
Drinking water are provided ad libitum, which is analyzed once at least annually. All parameters are within the permitted limits described in the national drinking water standard (GB 5749-2006).
The diet and water are considered not to contain any contaminants that could affect the purpose or integrity of the study. Animals eat and drink freely during the experiment.


Animal use comply with national animal welfare laws and regulations (instructive notions with respect to caring for laboratory animals) (2006, PRC Ministry of Science). The animal care and use activities required for conduct of this study are reviewed and approved by the test facility Animal Care and Use Committee (IACUC). All survival animals were euthanized under CO2 after completing test and their corpse treatment were delivered to entrusted specialized agencies.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g

Duration of treatment / exposure:

The eyes were not washed within 24 hours after exposure.

Observation period (in vivo):

The treated eyes of each animal were examined 1 h, 24 h, 48 h, 72 h, 4-21 d once daily after treatment.

Number of animals or in vitro replicates:

3

Details on study design:

Testing was initially conducted in one animal.


In the initial test, no severe irritation or corrosiveness was observed in the eyes of the subjects
with in 48 hours of exposure, and two additional animals were used for verification.


Approximately 24 hours prior to administration, both eyes of each animal will be examined with binocular loupe and 2% aqueous of sodium fluorescein (vehicle: physiological saline) in a binocular loupe. One drop of 2% aqueous of sodium fluorescein (vehicle: physiological saline) will be placed in the conjunctivae sac of both eyes, and then eyes will be washed by using physiological saline after 15 seconds. Corneas of both eyes will be examined in the binocular loupe to see whether there is fluorescein detention in the cornea or not. No eye abnormalities were noted for all treated eye prior to test.


The rabbit was held firmly but gently until quiet. 0.1 g of the grind test chemical was placed in the conjunctival sac of right eye of each animal after gently pulling the lower lid away from the eyeball. The lids were gently held together for approximately one second in order to prevent loss of the material. The left eye remained untreated and served as a control.


-Clinical observations
Immediately after administration of the test chemical, the initial reaction was observed and recorded. The test animal was observed twice daily from first to third day, the 2 observation intervals were not less than 6 h, and once daily on forth day for clinical signs. Any adverse systemic effects were also recorded .
-Eye Examination
The treated eyes of each animal were examined by using a binocular loupe at approximately 1 h, 24 h, 48 h, 72 h, 4-21 d once daily after treatment. At each observation point discharge and the area of cornea opacity was recorded, but not scored. Any other lesions in the eye were also reported.
-Fluorescent Examination
About 24 hours after exposure, fluorescein was detected in the eyes of animals, and sodium fluorescein retention was observed in the right eye. Fluorescein examination was performed at 48 h, 72 h, 4 d-7 d, 14 d, 18 d, 21 d.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 14 d

Results 2

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.7

Max. score:

1

Reversibility:

fully reversible within: 72 h

Results 4

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.7

Max. score:

2

Reversibility:

fully reversible within: 7 d

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 18 d

Results 6

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48 h

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 4 d

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 6 d

Results 9

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 12 d

Results 10

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 11

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 4 d

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.7

Max. score:

2

Reversibility:

fully reversible within: 5 d

Irritant / corrosive response data:

The individual score of animal eye reaction is shown in Table 2 and the average score of Animal Eye reaction is shown in Table 3 of Attachments.

Other effects:

All animals showed expected gain in body weights during the study.
The individual weight and weight changes of animals are shown in Table 1.

Any other information on results incl. tables

Table 1. Animal body weight individual value

Animal number	Dose (g/animal)	Individual body weight (kg)			Body weight gain (kg)
		Receiving day	Administration day	Last day
2100	0.1	2.00	2.70	3.20	0.50
2101	0.1	2.00	2.50	3.05	0.55
2102	0.1	2.05	2.80	3.35	0.55

Applicant's summary and conclusion

Interpretation of results:

Category 2A (irritating to eyes) based on GHS criteria

Conclusions:

The mean score of corneal/iris/conjunctival redness/conjunctival chemosis in the right eye of all animals at 24 h, 48 h and 72 h after exposure were 1.0/0/0.7/1.7, 1.0/0.3/1.0/2.0, 1.0/0/1.0/ 1.7, respectively. According to the GHS criteria for eye irritation and corrosivity, the sample is a class 2A ocular irritant.

Executive summary:

<Introduction> This experiment is in accordance with the requirements of "Chemical testing guidelines" (HJ/T 153-2004) of the Ministry of Environmental Protection of China and on the method of the testing of Chemicals-Health effect volume (the 2nd edition, Beijing, China Environment Press, 20 l 3). The acute eye i rritat ion/corrosion test of C18K was carried out in Japanese white rabbits.

<Method> Three rabbits were used for the study. Each animal was administered 0.1 g of the test chemical into right eye. Untreated left eye served as the control. Eye reactions were observed immediately, and at 1 h, 24 h, 48 h, 72 h, 4-21 d after administration and mean scores were calculated at 24 h, 48 h and 72 h after administration.

<Results> No. 2100 animal showed eye-closing at immediately observation after administration. Eye-closing, conjunctival edema observed on day 0. Eye-closing, conjunctival edema, secretions and maculae cornea were observed on day 1 and lasted until day 2. On the 3rd day, semi-closed eyes, conjunctival edema, secretions and maculae cornea were observed. On the 4th day, semi-closed eyes, conjunctiva) edema and maculae cornea were observed. On the 5th day, conjunctival edema and maculae cornea were observed and lasted until day 6. On the 7th day, maculae cornea were observed and lasted until day 13. No anomalies on the 14th day and lasted until day 21.

No. 2101 animal showed eye-closing at immediately observation after administration. Eye­ closing, conjunctival edema observed on day 0. Eye-closing, conjunctival edema, secretions and maculae cornea were observed on day 1 and lasted until day 2. On the 3rd day, semi­ closed eyes, conjunctival edema, secretions and maculae cornea were observed and lasted until day 4. On the 5th day, conjunctival edema and maculae cornea were observed. On the 6th day, maculae cornea were observed and lasted until day 17. No anomalies on the 18th day and lasted until day 21.

No. 2102 animal showed eye-closing at immediately observation after administration. Eye­ closing, conjunctival edema observed on day 0. Eye-closing, conjunctival edema, secretions and maculae cornea were observed on day 1. On the 2nd day , semi-closed eyes, conjunctival edema, secretions and maculae cornea were observed. On the 3th day , conjunctival edema and maculae cornea were observed and lasted until day 4. On the 5th day, maculae cornea were observed and lasted until day 11. No anomalies on the 12th day and lasted until day 21. No. 2100. 2101 animals were tested for fluorescein on 24 h, 48 h, 72 h, 4 d-7 d. 14 d. 18 d and 21 d after exposure, all had sodium fluorescein retention. For No. 2102 animal was tested for fluorescein on 24 h, 48 h, 72 h, 4 d-7 d, 14 d and 18 d after exposure, all had sodium fluorescein retention and the retention of sodium fluorescein disappeared on the 21st day.

 For number 2100, 2101 and 2102 animals, the mean scores of corneal/iris/conjunctival redness/conjunctival chemosis at 24 h, 48 h, 72 h were 1.0/0/0.7/1.7, 1.0/0.3/1.0/2.0, 1.0/0/1.0/1.7, respectively.

All animals showed expected gain in body weights during the study.

<Conclusion> The mean score of corneal/iris/conjunctival redness/conjun ctival chemosis in the right eye of all animals at 24 h, 48 h and 72 h after exposure were 1.0/0/0.7/1.7, 1.0/0.3/0/2.0, 1.0/0/1.0/1.7, respectively. According to the GHS criteria for eye irritation and corrosivity, the sample is a class 2A ocular irritant.