N,N'-bis{N-[N-(4-chlorophenyl)carbamimidoyl]carbamimidoyl}hexane-1,6-bis(aminium) diundec-10-enoate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August 11- September 26, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

N,N'-bis{N-[N-(4-chlorophenyl)carbamimidoyl]carbamimidoyl}hexane-1,6-bis(aminium) diundec-10-enoate

EC Number:

813-944-0

Cas Number:

1884575-91-0

Molecular formula:

C44H70Cl2N10O4

IUPAC Name:

N,N'-bis{N-[N-(4-chlorophenyl)carbamimidoyl]carbamimidoyl}hexane-1,6-bis(aminium) diundec-10-enoate

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

OTHER SPECIFICS:SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 001/CHUA/01 16
- Expiration date of the lot/batch: December, 2017
- Purity test date: January, 2016

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room Temperature
- Stability under test conditions: Two years

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source:
- Age at study initiation:
- Weight at study initiation:
- Housing:
- Diet (e.g. ad libitum):
- Water (e.g. ad libitum):
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
- Humidity (%):
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light):

IN-LIFE DATES: From: To:

Test system

Vehicle:

water

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):

VEHICLE
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:

Duration of treatment / exposure:

21 days

Observation period (in vivo):

The eyes were examined at approximately 1, 24, 48, 72 hours, days 7, 14 and 21

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done):
- Time after start of exposure:

SCORING SYSTEM:

TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein

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:

4

Reversibility:

not fully reversible within: 21 days

Results 2

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 21 days

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 4

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 14 days

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

not fully reversible within: 21 days

Results 6

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

not fully reversible within: 21 days

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 14 days

Results 9

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

not fully reversible within: 21 days

Results 10

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

not fully reversible within: 21 days

Results 11

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritant / corrosive response data:

No corrosive effects were observed on the cornea of the treated rabbits.

Other effects:

The test item did not result in any staining of the treated eye of the rabbits. No abnormalities were detected internally in any of the treated animal during necropsy at terminal sacrifice.
Moderate corneal opacity and slightly conjunctivae edematous was present in left eye of Animal No. 01. Marked corneal opacity, conjunctiva edema slightly present in left eye of Animal No. 02 and 03 were observed externally during necropsy at terminal sacrifice.

Any other information on results incl. tables

EYE IRRITATION SCORES - MEAN VALVES AFTER 24, 48 AND 72 HOURS (TREATED - LEFT EYE) FOR INDIVIDUAL ANIMAL

Animal number	Sex	Eye reaction
		Corneal opacity	Iris	Conjunctivae	Chemosis
01	Male	1.00	1.00	1.67	3.00
02		1.67	1.33	1.33	3.00
03		1.67	1.33	1.33	3.00

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

Since the treated animals exhibited redness of conjunctivae ≤ 1.67, corneal opacity ≤ 1.67, chemosis ≤ 3.00, lesions in iris ≤ 1.33 and the lesions in the eye were not reversed at day 21 observation in 3 out of 3 treated animals, hence the outcome of result qualify for category 1 of the classification criteria, hence CHUA is classified as per Harmonised Integrated Classification System as "irreversible effects" to rabbit eyes.

Executive summary:

The acute eye irritation/corrosion study of CHUBA in Rabbits was investigated according to OECD test guideline 405. The test item, 0.1g was placed in the conjunctiva! sac of the left eye of a single male rabbit. Eye reactions were observed up to 72 hours approximately post treatment, the test was completed using the remaining two male rabbits to confirm the findings of the initial test. The scoring of eye reactions was performed at 1, 24, 48, 72 hour, day 7, 14 and 21 for all the treated animals post test item instillation. The mean score was calculated across 3 scoring intervals (24, 48 and 72 hour post treatment) for each animal for corneal opacity, iris, conjunctival redness and chemosis. The individual mean score of opacity, iris, conjunctivae and chemosis for Animal No. 01 was 1.00, 1.00, 1.67 and 3.00, for Animal No. 02 and 03 were 1.67, 1.33, 1.33 and 3.00.

In Animal No. 01, instillation of test item in rabbit left eye caused redness of conjunctivae with some blood vessels definitely hyperaemic at 1 hour observation post test item instillation. Redness of conjunctivae with diffuse, crimson color; individual vessels not easily discernible was observed at 24 and 48 hour observation. Animal continued to exhibit conjunctival redness, the severity was slightly reduced to conjunctivae with some blood vessels definitely hyperaemic at 72 hour and the lesion fully reversed and appeared normal on day 7. Some swelling above normal (includes nictitating membranes) of chemosis was observed at 1 hour observation post test item instillation. Chemosis with obvious swelling with partial eversion of lids was observed at 24 hour observation. Swelling, with lids about half closed was observed at 48 hour and swelling, with lids about more than half closed was observed at 72 hour observation. The severity of Chemosis was slightly reduced to some swelling above normal (includes nictitating membranes) on day 7 and appeared normal on day 14 of observation. No opacity was observed at 1 hour and 24 hour observation whereas diffuse areas of opacity involving one quarter but not zero and details of iris clearly visible was observed at 48 hour and easily discernible translucent area of opacity with greater than one quarter, but less than half was observed at 72 hour observation. The severity increased to opaque cornea and iris not discernible through the opacity greater than three quarters, upto whole area on was observed on day 7, 14 and 21 observation. No lesion in iris was observed at 1, 24, hour and iris reactive to light (a sluggish reaction is considered to be an effect) was observed at 48 hour observation. At 72 hour the severity increased to no reaction to light and continued to exhibit on days 7, 14 and 21 observation.

In Animal No. 02 and 03, instillation of test item in rabbit left eye caused redness of conjunctivae with some blood vessels definitely hyperaemic at 1 hour observation post test item instillation. Redness of conjunctivae with diffuse, crimson color; individual vessels not easily discernible was observed at 24 and the severity was slightly reduced to conjunctivae with some blood vessels definitely hyperaemic at 48 and 72 hour observation. This lesion fully reversed and appeared normal on day 7. Some swelling above normal (includes nictitating membranes) of chemosis was observed at 1 hour observation post test item instillation. Chemosis with obvious swelling with partial eversion of lids was observed at 24 hour observation. Swelling, with lids about half closed was observed at 48 hour and swelling, with lids about more than half closed was observed at 72 hour observation. The severity of Chemosis was reduced to some swelling above normal (includes nictitating membranes) on day 7 and appeared normal on day 14 of observation. No opacity was observed at 1 hour observation whereas diffuse areas of opacity involving one quarter but not zero and details of iris clearly visible was observed at 24 hour and easily discernible translucent area of opacity with greater than one quarter, but less than half was observed at 48 and 72 hour observation. The severity of lesion increased to opaque cornea and iris not discernible through the opacity greater than three quarters, upto whole area was observed on day 7, 14 and 21 observation. No lesion in iris was observed at 1, 24, hour and no reaction to light was observed at 48 and 72 hour observation. This lesion was continued to exhibit on days 7, 14 and 21 observation. After 21 days of observation all the treated animals (Animal No. 01, 02 and 03) were sent for terminal sacrifice.

Based on the above findings, in 3 out of 3 animals, the eye lesions persisted till day 21 observation. The experiment was terminated on day 21 considering the test item had "irreversible effects" on the eye. No clinical signs were observed in any of the test animals throughout the acclimatization period and during experimental period. The body weights of all the animals were considered to be within the normal range of variability commonly observed for this species, strain and age. No abnormalities were detected internally in any of the treated animal during necropsy at terminal sacrifice. Moderate corneal opacity and slightly conjunctivae edematous was present in left eye of Animal No. 01. Marked corneal opacity, conjunctiva edema slightly present in left eye of Animal No. 02 and 03 were observed externally during necropsy at terminal sacrifice.