Tall Oil Fatty Acids, compounds with 2-(2-aminoethoxy) ethanol

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 May 2015 to 22 February 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Tall Oil Fatty Acids, compounds with 2-(2-aminoethoxy) ethanol

Molecular formula:

Too complex

IUPAC Name:

Tall Oil Fatty Acids, compounds with 2-(2-aminoethoxy) ethanol

Test material form:

liquid

Details on test material:

- Appearance/ Physical State: light brown slightly viscous liquid
- Storage conditions: room temperature, in the dark

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

ANIMALS AND ANIMAL HUSBANDRY
- Three New Zealand White rabbits were supplied by Envigo RMS (UK) Limited, Leicestershire, UK.
- At the start of the study the animals weighed 2.79 kg to 3.04 kg and were 12 to 20 weeks old.
- After an acclimatisation period of at least 5 days each animal was given a number unique within the study, which was written with a black indelible marker-pen on the inner surface of the ear and on the cage label.
- Animals were individually housed in suspended metal cages.
- Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study.
- The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
- Temperature and relative humidity were set to achieve limits of 17 °C to 23 °C and 30 % to 70 % respectively. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study.
- The rate of air exchange was at least 15 changes per hour.
- Lighting was controlled by a time switch to give 12 hours continuous light (06:00 to 18:00) and 12 hours darkness.
- Animals were provided with environmental enrichment items, which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.1 mL

Duration of treatment / exposure:

Single application

Observation period (in vivo):

14 days

Number of animals or in vitro replicates:

Three

Details on study design:

TEST ITEM FORMULATION
- The absorption of the test material was not determined.

MEASUREMENT OF pH
- The pH of the test item was determined prior to commencement of the study and found to be 9.31 (undiluted as supplied), 8.88 (90 % v/v aqueous preparation of test item) and 8.51 (90 % v/v aqueous preparation of test item after 10 minutes).

PROCEDURE
- Immediately before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard opthalmoscope.
- Only animals free of ocular damage were used.
- Initially, a single rabbit was treated. A subcutaneous injection of buprenorphine (0.01 mg/kg) was administered 60 minutes prior to test item application to provide a therapeutic level of systemic analgesia. Five minutes prior to test item application, pre-dose anaesthesia of ocular anaesthetic was applied to each eye as two drops of 0.5 % tetracaine hydrochloride).
- A volume of 0.1 mL of the test material was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower lids were held together for about one second immediately after treatment, to prevent loss of the test material, and then released.
- The left eye remained untreated and was used for control purposes.
- Immediately after administration of the test material, an assessment of the initial pain reaction was made according to the six point scale shown in Appendix 1 (attached).
- Eight hours after test item application, a subcutaneous injection of post-dose analgesia (buprenorphine 0.01 mg/kg and meloxicam 0.5 mg/kg) was administered to provide a continued therapeutic level of systemic analgesia.
- The treated animal was checked for signs of pain and suffering approximately 12 hours later. No further analgesia was required.
- After consideration of the ocular responses produced in the first treated animal, two additional animals were similarly treated.
- Assessment of ocular damage was made approximately 1 hour and 24, 48 and 72 hours following treatment according to the numerical evaluation (Draize scale) given in Appendix 3 (attached).
- Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard opthalmoscope.
- Additional observations were made on Days 7 and 14 to assess the reversibility of the ocular effects.
- In order to confirm the absence or presence of corneal opacity, examination under ultra violet light following treatment with sodium fluorescein BP was performed in one treated eye at the 24, 48 and 72-hour plus the 7-day observations.
- Examination under ultra violet light following treatment with sodium fluorescein BP was performed in one other treated eye at the 72-hour, 7-day and 14-day observations.
- The cornea, conjunctivae and iris were also examined for lesions.
- The control eyes were similarly treated with sodium fluorescein BP.
- Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

INTERPRETATION OF RESULTS
- Mean scores following grading at 24, 48 and 72 hours after instillation of the test item were calculated for corneal opacity, iridial inflammation, conjunctival redness and chemosis (see Table 2, attached).
- Interpretation according to a modified version of the system described by Kay and Calandra (1962) is presented in Appendices 2 and 4 (attached).
- If evidence of irreversible ocular damage was noted, the test item was considered to be corrosive to the eye.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

mean score

Time point:

other: 24, 48 and 72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: 75268 (male)

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

mean score

Time point:

other: 24, 48 and 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible

Remarks:

14 d

Remarks on result:

other: 75257 (male)

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

mean score

Time point:

other: 24, 48 and 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: 75264 (male)

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

mean score

Time point:

other: 24, 48 and 72 h

Score:

0.33

Max. score:

2

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: 75268 (male)

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

mean score

Time point:

other: 24, 48 and 72 h

Score:

0.67

Max. score:

2

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: 75257 (male)

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

mean score

Time point:

other: 24, 48 and 72 h

Score:

0.67

Max. score:

2

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: 75264 (male)

Results 7

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Remarks:

mean score

Time point:

other: 24, 48 and 72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 7 d

Remarks on result:

other: 75268 (male)

Results 8

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Remarks:

mean score

Time point:

other: 24, 48 and 72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 14 d

Remarks on result:

other: 75257 (male)

Results 9

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Remarks:

mean score

Time point:

other: 24, 48 and 72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 d

Remarks on result:

other: 75264 (male)

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

mean score

Time point:

other: 24, 48 and 72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 7 d

Remarks on result:

other: 75268 (male)

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

mean score

Time point:

other: 24, 48 and 72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 d

Remarks on result:

other: 75257 (male)

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Remarks:

mean score

Time point:

other: 24, 48 and 72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 14 d

Remarks on result:

other: 75264 (male)

Irritant / corrosive response data:

OCULAR REACTIONS
- Individual scores and individual total scores for ocular irritation are given in Table 1 (attached).
- Individual and mean scores required for labelling regulations are presented in Table 2 (attached).
- Diffuse corneal opacity was noted in all treated eyes at the 24-hour observations, in two treated eyes at the 48 and 72-hour observations and persisted in one treated eye at the 7-day observation.
- Iridial inflammation was noted in all treated eyes at one and 24 hours after treatment and persisted in two treated eyes at the 48-hour observation.
- Moderate conjunctival irritation was noted in all treated eyes at one and 24 hours after treatment. Moderate conjunctival irritation was noted in two treated eyes with minimal conjunctival irritation noted in one treated eye at the 48 and 72-hour observations. Minimal conjunctival irritation was noted in two treated eyes at the 7-day observation.
- One treated eye appeared normal at the 7-day observation and two treated eyes appeared normal at the 14-day observation.

Other effects:

BODY WEIGHT
- Individual body weights and body weight changes are given in Table 3 (attached).
- All animals showed expected gain in body weight during the study.

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

The eye irritancy potential of the test item was assessed in accordance with OECD Guideline 405. The test item produced individual mean scores of 0.3, 1.0 and 1.0 for corneal opacity; 0.3, 0.7 and 0.7 for iritis; 2.0 for conjunctival redness (all three animals) and 1.3, 2.0 and 1.7 for conjunctival chemosis. Since the mean scores calculated from observations at 24, 48 and 72 hours were ≥ 1 in two out of three animals for corneal opacity and ≥ 2.0 in all three animals for conjunctival redness, classification as irritating to eyes (category 2) is required in accordance with Regulation (EC) No 1272/2008.

Executive summary:

GUIDELINE

The study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The method was designed to meet the requirements of OECD Guidelines for the Testing of Chemicals No 405 "Acute Eye Irritation/Corrosion" (adopted 02 October 2012) and Method B.5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No. 440/2008.

RESULTS

A single application of the test item to the non-irrigated eye of three rabbits produced diffuse corneal opacity, iridial inflammation and moderate conjunctival irritation. One treated eye appeared normal at the 7 -day observation and two treated eyes appeared normal at the 14 -day observation.

CONCLUSION

The test item produced individual mean scores of 0.3, 1.0 and 1.0 for corneal opacity; 0.3, 0.7 and 0.7 for iritis; 2.0 for conjunctival redness (all three animals) and 1.3, 2.0 and 1.7 for conjunctival chemosis. Since the mean scores calculated from observations at 24, 48 and 72 hours were ≥ 1 in two out of three animals for corneal opacity and ≥ 2.0 in all three animals for conjunctival redness, classification as irritating to eyes (category 2) is required in accordance with Regulation (EC) No 1272/2008.