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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23 April - 14 June 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
The study was conducted under GLP in accordance with the international guideline.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
2017
Deviations:
no
Qualifier:
according to
Guideline:
other: “Regulation on Test Methods for Chemical Substances” Notification No. 2017-4, National Institute of Environmental Research, Republic of Korea
Version / remarks:
2017
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Storage condition of test material: At room temperature

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: ORIENTBIO INC.,Republic of Korea
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 10 - 11 weeks
- Weight at study initiation: 1.76 - 2.12 kg
- Fasting period before study: N/A
- Housing: Animals were individually housed in Stainless wire mesh cage, . 450W×600D×360H (mm), One animal/cage (during the quarantine–acclimation and observation periods).
- Diet (e.g. ad libitum): Purina experimental diet for rabbit 38302AF (lot no. KSN:18/04/07:LNS, KSN:18/04/21:LNS) placed in feeders and provided ad libitum. 
- Water (e.g. ad libitum): Public tap water in Cheongju-si was filtered and irradiated by ultraviolet light and provided ad libitum.
-Acclimation: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18.0−24.0°C
- Humidity (%): 30.0−70.0%
- Air changes (per hr): 10−15 clean, fresh, filtered air changes per hour
- Photoperiod (hrs dark / hrs light): 12 : 12
IN-LIFE DATES: 30 April - 14 June 2018

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: Untreated Left Eye
Amount / concentration applied:
0.1g
Duration of treatment / exposure:
One second
Observation period (in vivo):
1, 24, 48 and 72 hours
Duration of post- treatment incubation (in vitro):
n/a
Number of animals or in vitro replicates:
n/a
Details on study design:
All preparations were conducted immediately prior to use. 0.1 g of test substance was weighed (ENTRIS423i-1S, Sartorius, Germany).
Sixty minutes prior to test substance application, buprenorphine at 0.01 mg/kg was administered by subcutaneous injection (SC) to provide a therapeutic level of systemic analgesia.
Five minutes prior to test substance application, one drop of 0.5% proparacaine hydrochloride was applied to the right and left eyes.
In both the initial and confirmatory tests, irritation effects were shown from one hour after test substance application. Therefore, buprenorphine 0.01 mg/kg and meloxicam 0.5 mg/kg were administered by SC at eight hours after test substance application to provide a continued therapeutic level of systemic analgesia. Twelve hours later, buprenorphine 0.01 mg/kg SC was administered. In both the initial and confirmatory tests, no irritation effect was observed at 24 hours after test substance application; therefore, no systemic analgesics were administered.
0.1 g of test substance was instilled into the conjunctival sac of the right eye of animal using a spatula after gently pulling over the lower eyelid away from the eyeball. The eyelid was gently held together for approximately one second in order to prevent the loss of the test substance. The left eye was untreated and served as control.
No severe irritation was observed in the initial test. Therefore, a confirmatory test was conducted using two additional rabbits.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no opacity
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no effect on iris
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no conjunctivitis
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no chemosis
Remarks on result:
no indication of irritation
Irritation parameter:
maximum mean total score (MMTS)
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no irritation
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no opacity
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no effect on iris
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no conjunctivitis
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no chemosis
Remarks on result:
no indication of irritation
Irritation parameter:
maximum mean total score (MMTS)
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no irritation
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no effect on iris
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no opacity
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no conjunctivitis
Remarks on result:
no indication of irritation
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no chemosis
Remarks on result:
no indication of irritation
Irritation parameter:
maximum mean total score (MMTS)
Basis:
animal #3
Time point:
24/48/72 h
Score:
ca. 0
Reversibility:
other: no irritation
Remarks on result:
no indication of irritation
Irritation parameter:
maximum mean total score (MMTS)
Basis:
animal #1
Time point:
other: 1 hour
Score:
9
Reversibility:
fully reversible within: 24 hours
Remarks on result:
probability of mild irritation
Irritation parameter:
maximum mean total score (MMTS)
Basis:
animal #2
Time point:
other: 1 hour
Score:
9
Reversibility:
fully reversible within: 24 hours
Remarks on result:
probability of mild irritation
Irritation parameter:
maximum mean total score (MMTS)
Basis:
animal #3
Time point:
other: 1 hour
Score:
9
Reversibility:
fully reversible within: 24 hours
Remarks on result:
probability of mild irritation
Irritant / corrosive response data:
The test item was not corrosive but it was considered to be ‘Minimally irritating’ to rabbit eyes under the conditions of this study, due to effects seen one hour after application. In both the initial and confirmatory tests, congestion of iris of score 1, redness of the conjunctivae of score 1 and chemosis of the conjunctivae of score 1 were observed in three animals at one hour after application. The mean total score (MTS) after one hour was 9.0.

Any other information on results incl. tables

Table 3. Body Weights

Test Group

Phase

Animal ID

Body weight (kg/day)

Gain (kg)

1

3

 

G1

Initial

1101

2.06

2.17

0.11

Confirmatory

1102

2.12

2.25

0.13

1103

2.06

2.15

0.09

Mean

2.08

2.19

0.11

SD

0.03

0.05

0.02

N

3

3

3

Table 4. Summary of Eye Irritation Scores

Test group (Conc.)

Phase

No. of animal

Mean total score (MTS)

MMTS

Classification of Eye Irritation

1 hour

24hour

48 hour

72 hour

G1 (100%)

Initial

1

9.0

0.0

0.0

0.0

9.0

Minimally irritating

Confirmatory

2

MMTS:Maximum mean total score

hour: Observation after application

Conc.: Concentration

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the condition of the study the test item did not produce any evidence of ocular corrosivity but mild irritation was observed. However, the test item is not classified according to the Globally Harmonized Classification System and does not meet the criteria for classification under the Regulation (EC) No. 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.
Executive summary:

OECD 405 - 2018: Three healthy male New Zealand White rabbits free from evidence of ocular irritation and corneal abnormalities, were placed on the study. An analgesic, Buprenorphine, at 0.1 mg/kg and meloxicam 0.5 mg/kg were administered subcutaneously eight hour prior to instillation of the test item. In the initial test, 0.1 g of the test substance was instilled to the conjunctivae sac of the right eye of one animal. No severe irritation effect was observed in the initial test; thus, a confirmatory test was conducted using two additional animals in the same manner as the initial test. Eye irritation was scored according to the method of Draize at 1, 24, 48 and 72 hours after test substance application. The degree of eye irritation was classified by the method of Kay and Calandra. The control eyes were observed at the same time periods. Sodium fluorescein dye procedures were used at the 24-hour observation interval. Animals were evaluated for mortality, toxicity and pharmacological effects twice daily. Body weights were recorded pre-test and at termination.

No adverse signs were observed in any animal throughout the course of the study. No abnormal signs of pain or distress were observed in three animals during the observation period. All animals exhibited normal body weight gain. The mean body weight gain throughout the observation period was 0.11 kg.

In both the initial and confirmatory tests, a congestion of iris score of 1, redness of the conjunctivae score of 1 and chemosis of the conjunctivae score of 1 were observed in three animals at one hour after application. The mean total score (MTS) was ‘9.0’.

No reaction on the conjunctivae, cornea and iris was observed in any of three animals at 24, 48, 72 hours. MTS was ‘0.0’, respectively. Also, fluorescein sodium dye spots were examined at 24 hours. As a result, fluorescein sodium dye spots were not observed in the corneas of three animals.

No reaction on the conjunctivae, cornea or iris was observed in the control eye during the observation period.  The maximum mean total score (MMTS) was ‘9.0’ at one hour with full reversiblity within 24 hours post exposure.

The MMTS of the test substance was ‘9.0’. Therefore, the test substance was classified as ‘Minimally irritating’ according to the provisional evaluation of eye irritation (Method of Kay and Calandra). The final evaluation of irritation to an eye was classified as ‘Minimally irritating’, the same as previous evaluation as the MTS was ‘0’ at 48 hours according to the same method.

Under the conditions of the study the test item did not produce any evidence of ocular corrosivity but mild irritation was observed. However, the test item did not meet the criteria for classification according to the Globally Harmonized Classification System. or the criteria for classification under the Regulation (EC) No. 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.