Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 27 MAR 2015 to 29 MAY 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP and OECD test guideline-compliant study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Remarks:
Homogenous Liquid
Specific details on test material used for the study:
Trade name: CYPHOS IL 101 Phosphonium Salt
Physical state: liquid
Composition of test material, percentage of components: Trihexyl(tetradecyl)phosphonium chloride (258864-54-9), > 95%
Lot/batch No.: WEC031280
Expiration date of the lot/batch: >1 year when stored at room temperature and protected from direct contact with water (hydrophobic)

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Covance Research Products, Inc., Denver, PA
- Age at study initiation: 18 week old
- Weight at study initiation: 2.7 kg
- Housing: individually housed in a suspented cage
- Diet: Fresh PMI Rabbit Chow (Diet#5321) was provided daily
- Water: available ad libitum
- Acclimation period: At 5 five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): temperature controlled
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Test animal was instilled with 0.1 mL 

VEHICLE: none (undiluted)
Duration of treatment / exposure:
Single application, not washed
Observation period (in vivo):
60 min, 24, 48 and 72 hours post dose
Number of animals or in vitro replicates:
1 male
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): not washed

SCORING SYSTEM: Occular reactions were graded according to the numerical Draize technique

TOOL USED TO ASSESS SCORE: Sodium fluorescein dye procedure were used at the 24-, 48-, 72-hour observation intervals

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
other: 1 Hour
Score:
3
Max. score:
4
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: 1 Hour
Score:
1
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: 1 Hour
Score:
2
Max. score:
3
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: 1 Hour
Score:
3
Max. score:
4
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Reversibility:
fully reversible within: 48h
Remarks on result:
other: Although the score for 48 and 72 hours is 0, pale areas were noted
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
4
Max. score:
4
Remarks on result:
other: No expectation of reversibility after 72-hour post dose
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Remarks on result:
not determinable
Remarks:
Due to chemosis
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Remarks on result:
not determinable
Remarks:
Due to chemosis
Irritant / corrosive response data:
See Table 1
The study was terminated at 72 hours since there was no improvement in chemosis of the treated eye. Reversibility of the occular reactions was not expected.

After 72 hours of exposure, severe ocular lesions were observed, mainly on conjunctiva, with swelling and abundant lachrymal discharge.
Due to chemosis, corneal opacity and iritis could not be determined at the 24-, 48-, 72-hour time points.

The animal lost weight. The animal had diminished food consumption at 72 hours post-dose.

Any other information on results incl. tables

Table 1: Occular findings at each observation time

Tissue Reading 1 Hour 24 Hour 48 Hour 72 Hour
Cornea Opacity Max. Score: 4 3 ? ? ?
Area Max. Score: 4 2 ? ? ?
Iris Max. Score: 2 1 ? ? ?
Conjunctiva Redness Max. Score: 3 2 3 0 p  0 p 
Chemosis Max. Score: 4 3 4 4 4
Discharge Max. Score: 3 3 3 3 3

Details of eye irritation scores (according to Draize)

The control eye appeared normal at all observation periods

? = unable to determine due to chemosis

p = pale areas

Table 2: Systemic Observations

Time periods
Day 1 A/A
Day 2 A/A
Day 3 1/1

A/A = Appeared normal (a.m./p.m. observations)

1/1 = diminished food consumption (a.m./p.m. observations)

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
Trihexyltetradecylphosphonium chloride is considered to cause irreversible tissue damage to the eye. Conjunctival chemosis was still observed at 72-hour post dose without expectation of reversibility
Executive summary:

Trihexyltetradecylphosphonium chloride, has been tested for acute eye irritation on 1 male New Zealand White rabbit, in accordance with OECD test guideline 405, and in compliance with Good Laboratory Practice.

The test article was applied as such in a single conjunctival dose of 0.1 mL in the right eye, the untreated eye being used as a control. Ocular examinations, grading cornea, iris and conjunctiva (redness, chemosis and discharge) reactions, were performed at 1, 24, 48, 72 hours after instillation of the test article.

Within the first hour post dose, the test article induced conjunctival abnormalities including hyperhaemia grade 2 (diffuse, crimson color, individual vessels not easily discernible), edema grade 3 and conjunctivae discharge (grade 3). Corneal opacity and injection of the iris (grade 3 and 1, respectively) were also observed.

At the observations carried out 24, 48 and 72 hours after instillation, hyperhaemia chemosis of grade 4 and discharge of grade 3 were observed. Due to chemosis, corneal opacity and iritis could not be determined at the 24-, 48-, 72-hour time points.

The study was terminated at 72 hours since there was no improvement in chemosis of the treated eye. Reversibility of the ocular reactions was not expected.

 

Based on these results, Trihexyltetradecylphosphonium chloride was considered to cause irreversible tissue damage to the eye.