Registration Dossier

Administrative data

Description of key information

Skin Irritation/Corrosion: Not corrosive but mildly irritating to skin (not classified under CLP or GHS); OECD 404: S. Yang., 2018

Eye Irritation/Corrosion: Not corrosive but mildly irritating to the eye (not classified under CLP or GHS); OECD 405: S. Yang., 2018)

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:
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.
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
2015
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
Species:
rabbit
Strain:
New Zealand White
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Controls:
other: Untreated site
Amount / concentration applied:
0.5 g
Duration of treatment / exposure:
4 hours
Observation period:
1, 24, 48 and 72 hours plus additional observation until day 4
Number of animals:
3
Details on study design:
In the initial test, two sites of one rabbit’s clipped back along the midline were selected as application sites. One site was designated as test substance site. The other site was served as control site. 0.5 g of test substance was placed on the test substance site, moistened with water for injection and held in contact with the skin by a semi-occlusive dressing for 4 hours. No severe irritant effect was observed in the initial test, thus a confirmatory test was conducted using two additional animals. The observation of application sites for skin response was conducted at 1, 24, 48 and 72 hours after patch removal and the skin response scored according to the ‘Evaluation of skin reactions (Method of Draize: 1959)1)’. The degree of skin irritation was classified according to the Draize’s method).In both the initial and confirmatory tests, skin irritation was observed at 72 hours after patch removal. Therefore, additional dermal observations were conducted until Day 4.
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
mean
Time point:
other: 24 hr and 4 days
Score:
ca. 0.8
Max. score:
4
Reversibility:
other: irritation
Remarks on result:
probability of weak irritation
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 0.8
Max. score:
4
Reversibility:
other: irritant
Remarks on result:
probability of weak irritation
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 0.8
Max. score:
4
Reversibility:
other: irritant
Remarks on result:
probability of weak irritation
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
ca. 0.8
Max. score:
4
Reversibility:
other: irriatnt
Remarks on result:
probability of weak irritation
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
ca. 0.8
Max. score:
4
Reversibility:
other: irritant
Remarks on result:
probability of weak irritation
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
ca. 0.8
Max. score:
4
Reversibility:
other: irritant
Remarks on result:
probability of weak irritation
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
ca. 0.8
Max. score:
4
Reversibility:
other: irritant
Remarks on result:
probability of weak irritation
Irritant / corrosive response data:
The substance is not corrosive but slightly irritating

Table 3. Irritation scores

Applied area

Group

Animal ID

Score of erythema and eschar/Score of edema Intervals (hour)

Score of erythema and eschar/Score of edema Intervals

IndividualP.I.I.a)

P.I.I.b)

Classification of skin irritation

1

24

48

72

4

100% test Item

Initial

1101

0/0

1/0

1/0

1/0

0/0

0.8

0.8

Slightly irritant

Confirmatory

1102

0/0

1/0

1/0

1/0

0/0

0.8

1103

0/0

1/0

1/0

1/0

0/0

0.8

Control (only patch)

Initial

1101

0/0

0/0

0/0

0/0

0/0

0/0

0.0

Non irritant

Confirmatory

1102

0/0

0/0

0/0

0/0

0/0

0/0

1103

0/0

0/0

0/0

0/0

0/0

0/0

hour: Observation after patch removal

a: Individual P.I.I.(Primary skin irritation index) was determined by dividing the total scores of reactions for erythema and eschar/edema formation observed at 1, 24, 48 and 72 hours after patch removal by 4.

b: P.I.I. was determined by dividing the totaled individual P.I.I. by 3 animals to obtain the average

Table 4. Body weights

Applied area

Group

Animal ID

Body weight (kg/day)

Gain (kg)

1

4

 

G1

Initial

1101

2.05

2.16

0.11

Confirmatory

1102

2.06

2.18

0.12

1103

2.19

2.30

0.11

Mean

2.10

2.21

0.11

SD

0.08

0.08

0.01

N

3

3

3

Interpretation of results:
GHS criteria not met
Conclusions:
Under the condition of the study, the primary irritation index (PII) of the test item was 0.8, following exposure to clipped rabbit skin under semi-occlusive condition. Therefore the test item does 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.
Executive summary:

OECD 404 (2018): The irritant or corrosive effects of the test item, was assessed following application to the clipped skin of white New Zealand rabbit. 0.5 g of test item was placed on the test substance site, moistened with water for injection and held in contact with the skin by a semi-occlusive dressing for 4 hours. No severe irritant effect was observed in the initial test, thus a confirmatory test was conducted using two additional animals. The observation of application sites for skin response was conducted at 1, 24, 48 and 72 hours after patch removal and the skin response scored according to the ‘Evaluation of skin reactions (Method of Draize: 1959)’. The degree of skin irritation was classified according to the Draize’s method. In both the initial and confirmatory tests, skin irritation was observed at 72 hours after patch removal. Therefore, additional dermal observations were conducted until Day 4.

Animals were observed for mortality, toxicological and pharmacological effects at least once daily. Body weights were recorded pre-test and at termination.

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


The test item sites did not reveal any sign of adverse skin reactions such as erythema and edema in any of three animals at one hour after patch removal. Erythema of score 1 was observed in three animals from 24 hours to 72 hours after patch. Skin reactions were not observed on Day 4.  In both the initial and confirmatory tests, the control sites did not reveal any sign of adverse skin reactions during the observation period. 


The individual primary skin irritation index (Individual P.I.I.) of the test substance was ‘0.8’ for all animals. Therefore, the P.I.I. of the test substance was ‘0.8’. The degree of irritation of the test substance was classified as ‘Slightly irritant’ according to the Draize’s method. 


However, the test item does not meet the criteria for classification under the Globally Harrmonised Classification System or the Regulation (EC) No. 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.

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:
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.
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
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
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.
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.

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

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.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

OECD 404 (2018): The irritant or corrosive effects of the test item was assessed following application to the clipped skin of white New Zealand rabbit. 0.5 g of test item was placed on the test substance site, moistened with water for injection and held in contact with the skin by a semi-occlusive dressing for 4 hours. No severe irritant effect was observed in the initial test, thus a confirmatory test was conducted using two additional animals. The observation of application sites for skin response was conducted at 1, 24, 48 and 72 hours after patch removal and the skin response scored according to the ‘Evaluation of skin reactions (Method of Draize: 1959)1)’. The degree of skin irritation was classified according to the Draize’s method(1). In both the initial and confirmatory tests, skin irritation was observed at 72 hours after patch removal. Therefore, additional dermal observations were conducted until Day 4.

Animals were observed for mortality, toxicological and pharmacological effects at least once daily. Body weights were recorded pre-test and at termination.

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


The test item sites did not reveal any sign of adverse skin reactions such as erythema and edema in any of three animals at one hour after patch removal. An erythema of score of 1 was observed in three animals from 24 hours to 72 hours after patch removal. Skin reactions were not observed on Day 4.  In both the initial and confirmatory tests, the control sites did not reveal any sign of adverse skin reactions during the observation period. 


The individual primary skin irritation index (Individual P.I.I.) of the test substance was ‘0.8’ for all animals. Therefore, the P.I.I. of the test substance was ‘0.8’. The degree of irritation of the test substance was classified as ‘Slightly irritant’ according to the Draize’s method. 


Under the condition of the study, it was concluded that the test item was non-corrosive but caused slightly irritation to the rabbits’ skin. However, the test item did not meet the criteria for classification according to the Globally Harmonized Classification System or Regulation (EC) No. 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixture.

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.

However, the test item did not meet the criteria for classification according to the Globally Harmonized Classification System or Regulation (EC) No. 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixture.

Justification for classification or non-classification

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.