Bis[2-(2-ethoxyethoxy)ethyl] cyclohexane-1,4-dicarboxylate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November 19, 2007 - January 11, 2008

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was performed similar to OECD guidelines, except that a 50% dilution was used, and according to GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: Guide to quasi-drug and cosmetic regulations in Japan (February, 2006).

Deviations:

no

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Bis[2-(2-ethoxyethoxy)ethyl] cyclohexane-1,4-dicarboxylate

EC Number:

618-829-7

Cas Number:

922165-31-9

Molecular formula:

C20H36O8

IUPAC Name:

Bis[2-(2-ethoxyethoxy)ethyl] cyclohexane-1,4-dicarboxylate

Details on test material:

- Name of test material (as cited in study report): CH-CA
- Description: Transparent liquid
- Stability under test conditions: Unknown
- Storage condition of test material: Stored at room temperature in a sealed container

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: ORIENTBIO INC., Korea
- Age at study initiation: 17 weeks old.
- Weight at study initiation: 2.40~2.65 kg.
- Housing: Individually housed in labeled stainless cages with an automatic flushing system, 380Wx490Dx350H (mm).
- Diet (e.g. ad libitum): Purina experimental diet for rabbit 5302 (lot no.: KSN:11:09:MKS, KSN:11:26:KMS, Supplier: Agribrands Purina Korea Inc.) was available during the study period ad libitum.
- Water (e.g. ad libitum): Purified water that was filtered and irradiated by ultraviolet rays from municipal tap water was provided ad libitum.
- Acclimatization period: 8 days.

Health conditions were examined and released for study use by a staff veterinarian.

The diet was analyzed periodically by manufacturer to quantify nutrient contents and contaminants. There were no known contaminants in the diet at levels that would be expected to interfere with or affect the results of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.9ºC -20.9ºC
- Humidity (%): 48.7 - 63.8%
Cages, fodder tub and water bottles were washed using MICRO QUAT (component: Alkyldimethylbenzyl ammonium chloride, diluted one-four hundredth, ECOLAB., Inc., Korea) solution and cleased with municipal tap water.
- Air changes (per hr): 10-15 (all fresh air method)
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours darkness per day

Test system

Vehicle:

other: 1,3-Butylene glycol (1,3-BG)

Controls:

other: One eye of each animal remained untreated and served as the r eference control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): The test substance in a concentration of 50% was chosen in compliance with the Sponsor.

VEHICLE
- Lot/batch no.: 02262CC (Sigma Chemical Co., Ltd.

Duration of treatment / exposure:

Single instillation on Day 1.

Observation period (in vivo):

The eyes of each animal were examined approximately 1, 24, 48, 72 and 96 hours after instillation of the test substance. The test was terminated since an eye irritation was not observed at 96 hours after exposure.

Number of animals or in vitro replicates:

3 males

Details on study design:

REMOVAL OF TEST SUBSTANCE
-Washing (if done): No

OBSERVATIONS
- Clinical signs: During the observation period, all animals were observed for clinical signs daily.
- Body Weight: Individual body weights were measured on the first day (Day 0) and the last day (Day 4) during the observation period using the balance.
- Irritation:
The response scores for the cornea, iris and conjunctiva were recorded at 1, 24, 48, 72 and 96 hours after instillation of the test substance in accordance with Draize's method. The test was terminated since an eye irritation was not observed at 96 hours after exposure. At each observation, the highest scores given were recorded:

CORNEAL IRRITATION
Opacity: degree of density (area most dense taken for reading)
0: No ulceration or opacity (may include slight dulling of normal luster)
1: Scattered or diffuse areas of opacity, details of iris clearly visible
2: Easily discernible translucent area, details of iris slightly obscured
3: Nacreous area, no details of iris visible, size of pupil barely discernible
4: Opaque cornea, iris not discernible through the opacity

With additionally for area of cornea:
0: No ulceration or opacity
1: One quarter or less but not zero
2: Greater than one quarter, but less than half
3: Greater than half, but less than three quarters
4: Greater than three quarters, up to whole area

IRIS
0: Normal
1: Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia, or injection, any of these or combination thereof, iris still reacting to light (sluggish reaction is positive)
2: No reaction to light, hemorrhage, gross destruction (any or all of these)

CONJUNCTIVAL IRRITATION
Redness (refers to palpebrae and sclera, excluding cornea and iris):
0: Blood vessels normal
1: Some blood vessels definitely hyperaemic (injected)
2: Diffuse, crimson color, individual vessels not easily discernible
3: Diffuse beefy red

Chemosis (refers to lids and/or nictitating membranes):
0: No swelling
1: Any swelling above normal (includes nictitating membranes)
2: Obvious swelling with partial eversion of lids
3: Swelling with lids about half closed
4: Swelling with lids more than half closed

Discharge:
0: No discharge (may include small amounts observed in inner canthus of normal animals)
1: Any amount different from normal and/or lacrimation
2: Discharge with moistening of the lids and hairs just adjacent to lids
3: Discharge with moistening of the lids and hairs (considerable area around the eye)

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

mean

Remarks:

animal #1, #2 and #3

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

4

Results 2

Irritation parameter:

iris score

Basis:

mean

Remarks:

animal #1, #2 and #3

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

2

Results 3

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Remarks:

animal #1, #2 and #3

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

3

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

animal #1, #2 and #3

Time point:

other: 24, 48 and 72 hrs.

Score:

0

Max. score:

4

Irritant / corrosive response data:

At 1, 24, 48, 72 and 96 hours reading after exposure of 50% test substance, adverse effects on the conjunctiva, cornea and iris were not observed in all animals.

Other effects:

During the observation period, no abnormal clinical signs were observed in all animals.

During the observation period, normal body weight development was seen in all animals; mean body weight gains were recorded to be 0.06 kg.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information 50% solution Criteria used for interpretation of results: EU

Conclusions:

In an eye irritation test performed according to OECD 405 guideline and GLP principles, the test substance (50% CH-CA) was not irritating to eyes of rabbits. No conclusions can be drawn on 100% test substance.