Phosphoric acid, C4 and C12-14(even-numbered) Alkyl Esters, Ammonium Salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 October 1987 to 02 November 1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1987

Report date:

1987

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

only one animal exposed to test material without washing of the eye

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Phosphoric acid, mono-C4 and C12-14(even numbered) alkyl esters, Ammonium salts

Molecular formula:

Not applicable (UVCB substance)

IUPAC Name:

Phosphoric acid, mono-C4 and C12-14(even numbered) alkyl esters, Ammonium salts

Constituent 2

Reference substance name:

Phosphoric acid, di-C4 and C12-14(even numbered) alkyl esters, Ammonium salts

Molecular formula:

Not applicable (UVCB substance)

IUPAC Name:

Phosphoric acid, di-C4 and C12-14(even numbered) alkyl esters, Ammonium salts

Specific details on test material used for the study:

- Physical form: White to light tan paste
- Purity: 80 % active ingredient
- Lot: 751

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Remarks:

male

Details on test animals or tissues and environmental conditions:

ANIMAL HUSBANDRY
- Young adult male and female New Zealand White rabbits were received from Hare Marland, Hewitt, New Jersey.
- Rabbits were housed singly in suspended, stainless steel, wire-mesh cages.
- Each rabbit was assigned a unique identification number which was recorded on a card affixed to the cage.
- Purina Certified Rabbit Chow #5322 and water were available ad libitum except during the 24-hour period when exposure to test material took place.
- Rabbits were weighed and observed for general health for approximately two weeks.
- Animal rooms were maintained on a timer-controlled 12-hour light and 12-hour dark cycle.
- Target environmental conditions of the animal rooms were 20 ± 2 °C with relative humidity of 50 ± 10 %. Any excursions outside these ranges were of small magnitude and/or brief duration and did not adversely affect the validity of the study.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated left eye served as the control in both animals

Amount / concentration applied:

0.01 g

Duration of treatment / exposure:

20 seconds in one animal (test item removed from eye by rinsing) and 7 days in the second animal

Observation period (in vivo):

3 days in the animal where test item removal took place and 7 days in the second animal.

Number of animals or in vitro replicates:

Two

Details on study design:

PROTOCOL
- On the day of the study initiation, the eyes of two male New Zealand White rabbits were examined using fluorescein dye.
- Animals showing pre-existing corneal or conjunctival injury or irritation were not used in the study.
- Rabbits weighed 2578 g and 2767 g on the day of treatment.
- An aliquot (0.01 g) of test material was introduced into the lower conjunctival sac of the right eye in both rabbits. The left eyes served as controls.
- The treated and control eyes of one animal remained unwashed.
- Approximately 20 seconds after the test item was administered, both eyes of the second animal were washed with lukewarm tap water for one minute.
- Rabbits were examined for evidence of eye irritation after approximately 1 hour and 4 hours then 1, 2 and 3 days after treatment.
- The rabbit that had been exposed to test material without rinsing of the eyes was also examined 7 days after treatment.
- At each observation the treated eyes were examined using illumination and magnification. The treated eyes were scored for ocular reactions at each examination using the Draize scale (see Table I, attached). The untreated eye of each animal was also examined and used for comparison.
- Any unusual effects such as pannus, blistering of the conjunctiva, ulceration or other effects indicative of corrosive action were also noted.
- Fluorescein dye was used to evaluate corneal ulceration and irritation starting at the 1-day observation and at each subsequent observation. A sketch was drawn to indicate areas of stain retention.
- Biomicroscopic examinations fpr corneal injury were conducted at the 1-day observation and at each subsequent observation. Treated eyes were scored according to the system presented in Table II (attached).

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

unwashed rabbit eye

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other:

Remarks:

not applicable

Remarks on result:

other: Animal 21953

Remarks:

fluorescein examinations negative for corneal injury at all intervals

Results 2

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

unwashed rabbit eye

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other:

Remarks:

not applicable

Remarks on result:

other: Animal 21953

Results 3

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Remarks:

unwashed rabbit eye

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 d

Remarks on result:

other: Animal 21953

Results 4

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

unwashed rabbit eye

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other:

Remarks:

not applicable

Remarks on result:

other: Animal 21953

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

washed rabbit eye

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other:

Remarks:

not applicable

Remarks on result:

other: Animal 21954

Remarks:

fluorescein examinations negative for corneal injury at all intervals

Results 6

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

washed rabbit eye

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other:

Remarks:

not applicable

Remarks on result:

other: Animal 21954

Results 7

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Remarks:

washed rabbit eye

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 3 d

Remarks on result:

other: Animal 21954

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

washed rabbit eye

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other:

Remarks:

not applicable

Remarks on result:

other: Animal 21954

Irritant / corrosive response data:

- Individual eye irritation scores are given in Tables III and IV (attached).
- The test item produced slight corneal opacity, moderate iritis and mild conjunctival redness in both treated eyes.
- Test material also produced moderate chemosis and copious blood-tinged discharge in the unwashed eye (confirmed with Hemastix reagent strips).
- Slight chemosis was also observed in the washed eye.
- Fluorescein and biomicroscopic examinations revealed no corneal damage throughout the study.
- All occular irritation had resolved in the washed eye by day 3 and in the unwashed eye by day 7.

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Conclusions:

The test item caused reversible effects on the cornea, iris and conjunctivae in one washed and one unwashed rabbit eye and was considered to be a moderate eye irritant under the conditions of the study. However, the reversibility of the effects was not confirmed by exposing a third animal to test material for 7 days without rinsing of the treated eye, and these data cannot be used to classify for eye irritation under the terms of Regulation (EC) No 1272/2008 in the absence of supporting information.

Executive summary:

The test material was evaluated for acute eye irritation in two male rabbits. An aliquot (0.01 g) of test material was introduced into the lower conjunctival sac of the right eye in both rabbits. The left eyes served as controls. The treated and control eyes of one animal remained unwashed. Approximately 20 seconds after the test item was administered, both eyes of the second animal were washed with lukewarm tap water for one minute. Rabbits were examined for evidence of eye irritation after approximately 1 hour and 4 hours then 1, 2 and 3 days after treatment. The rabbit that had been exposed to test material without rinsing of the eyes was also examined 7 days after treatment. Treated eyes were examined using illumination, fluorescein plus biomicroscopic examination and the effects were scored. Slight corneal opacity, moderate iritis and mild conjunctival redness were seen in both the washed and unwashed eye. Moderate chemosis and copious blood-tinged discharge were observed in the unwashed eye. Slight chemosis was observed in the washed eye. Fluorescein and biomicroscopic examinations revealed no damage to the cornea throughout the study. All ocular irritation had resolved by day 3 in the washed eye and by day 7 in the unwashed eye. The test item was considered to be a moderate eye irritant under the conditions of the study. However, the reversibility of the effects was not confirmed by exposing a third animal to test material for 7 days without rinsing of the treated eye, and these data cannot be used to classify for eye irritation under the terms of Regulation (EC) No 1272/2008 in the absence of supporting information.