Fatty acids, C10-20-neo, potassium salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 September 2012 to 23 October 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

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:

Naphthenic acids, potassium salts

EC Number:

266-120-2

EC Name:

Naphthenic acids, potassium salts

Cas Number:

66072-08-0

IUPAC Name:

266-120-2

Test material form:

other: liquid (unspecified)

Details on test material:

- Appearance: dark brown liquid
- Storage conditions: room temperature in the dark
- pH of test material: 10.3

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Strain: Hsdlf:NZW.
- Age at study initiation: 12 to 20 weeks.
- Weight at study initiation: 2.29 or 2.40 kg.
- Housing: The animals were individually housed in suspended cages.
- Diet (e.g. ad libitum): ad libitum.
- Water (e.g. ad libitum): Free access to mains drinking water.
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23 °C
- Humidity (%): 30 to 70 %
- Air changes (per hr): At least 15 per hour.
- Photoperiod (hrs dark / hrs light): The lighting was controlled by a time switch to give twelve hours continuous light (0600 to 1800) and twelve hours darkness.

IN-LIFE DATES: From: 17 September 2012 To: 23 October 2012

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: The left eye remained untreated and was used for control purposes.

Amount / concentration applied:

0.1 mL

Duration of treatment / exposure:

The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test material, and then released.

Observation period (in vivo):

21 days

Number of animals or in vitro replicates:

2
After consideration of the ocular responses produced in the first animal, a second animal was treated.
In order to minimise pain on the application of the test material to the second animal treated, one drop of local anaesthetic (Tetracaine hydrochloride 0.5 %) was instilled into both eyes 1 to 2 minutes prior to treatment.

Details on study design:

SCORING SYSTEM: Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the Draize numerical evaluation.
Additional observations were made on Days 7, 14 and 21 to assess the reversibility of the ocular effects. Individual bodyweights were recorded on Day 0 (the day of dosing) and at the end of the observation period. Any clinical signs of toxicity, if present, were also recorded.

Draize Scale for Scoring Ocular Irritation

1. CONJUNCTIVAE

Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
Vessels normal............................................................................................................................................0
Vessels definitely injected above normal................................................................................................1
More diffuse, deeper crimson red, individual vessels not easily discernible....................................2
Diffuse beefy red.........................................................................................................................................3

Chemosis
No swelling..................................................................................................................................................0
Any swelling above normal (includes nictitating membrane).............................................................1
Obvious swelling with partial eversion of lids.......................................................................................2
Swelling with lids about half closed.........................................................................................................3
Swelling with lids half closed to completely closed...............................................................................4

Discharge
No discharge...............................................................................................................................................0
Any amount different from normal (does not include small amounts observed in inner
canthus of normal animals)......................................................................................................................1
Discharge with moistening of the lids and hairs just adjacent to lids...............................................2
Discharge with moistening of the lids and hairs a considerable area around the eye...................3

2. IRIS

Values
Normal.........................................................................................................................................................0
Folds above normal, congestion, swelling, circumcorneal injection (any or all
of these or combination of any thereof) iris still reacting to light
(sluggish reaction is positive).................................................................................................................1
No reaction to light, haemorrhage, gross destruction (any or all of these)....................................2

3. CORNEA

Degree of Opacity (most dense area used)
No opacity...................................................................................................................................................0
Scattered or diffuse areas, details of iris clearly visible......................................................................1
Easily discernible translucent areas, details of iris slightly obscured..............................................2
Opalescent areas, no details of iris visible, size of pupil barely discernible....................................3
Opaque, iris not discernible through the opacity................................................................................4

Area of Cornea Involved
One quarter (or less) but not zero..........................................................................................................1
Greater than one quarter but less than half...........................................................................................2
Greater than half but less than three quarters......................................................................................3
Greater than three quarters, up to whole area.......................................................................................4

TOOL USED TO ASSESS SCORE: Any additional ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 2 animals

Time point:

other: mean score at 24, 48 and 72 hours

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7 days

Results 2

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 2 animals

Time point:

other: mean score at 24, 48 and 72 hours

Score:

0.83

Max. score:

1

Reversibility:

fully reversible within: 7 days

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

of 2 animals

Time point:

other: mean score at 24, 48 and 72 hours

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 14 days

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 2 animals

Time point:

other: mean score at 24, 48 and 72 hours

Score:

1.83

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 5

Irritation parameter:

other: discharge

Basis:

mean

Remarks:

of 2 animals

Time point:

other: mean score at 24, 48 and 72 hours

Score:

1.83

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Individual scores for ocular irritation are given in Table 1.
Scattered or diffuse corneal opacity was noted in both treated eyes one hour after treatment and at the 24, 48 and 72 hour observations. Vascularisation, with a localised ingrowth of vessels for approximately 4 mm, was also noted in one treated eye at the 14 day observation. No corneal effects were noted in this treated eye at the 21 day observation.
Iridial inflammation was noted in all treated eyes one hour after treatment, at the 24 and 48 hour observations and in one treated eye at the 72 hour observation.
Moderate conjunctival irritation was noted in both treated eyes one hour after treatment and at the 24, 48 and 72 hour observations with minimal conjunctival irritation at the 7 day observation.
Small areas of petechial haemorrhage on the nictitating membrane were noted in one treated eye at the 7 day observation.

One treated eye appeared normal at the 14 day observation and the other treated eye appeared normal at the 21 day observation.

Other effects:

BODYWEIGHT
Both animals showed the expected gain in bodyweight during the study.

Any other information on results incl. tables

Table 1 Individual Scores for Ocular Irritation

Rabbit Number and Sex	72424 Male							72568 Male
	IPR = 3							IPR not recorded*
Time After Treatment	1 hour	24 hours	48 hours	72 hours	7 days	14 days	21 days	1 hour	24 hours	48 hours	72 hours	7 days	14 days
Cornea Degree Area	1 2	1 1	1 2	1 2	0 0	0† 0	0 0	1 2	1 2	1 2	1 1	0 0	0 0
Iris	1	1	1	0	0	0	0	1	1	1	1	0	0
Redness	2	2	2	2	1	0	0	2	2	2	2	1‡	0
Chemosis	2	2	2	2	1	0	0	3	2	2	1	1	0
Discharge	3	2	3	2	0	0	0	2	2	1	1	0	0

IPR = Initial pain reaction; a score of 3 indicates moderate initial pain. The rabbit holds the eye shut and puts pressure on lids, may rub eye with paw.

*Due to technician error, the initial pain reaction was not recorded. One drop of localised anaesthetic was instilled into both eyes 1 to 2 minutes before treatment.

†Vascularisation, with a localised ingrowth of vessels for approximately 4 mm.

‡Small areas of petechial haemorrhage on the nictitating membrane.

Applicant's summary and conclusion

Interpretation of results:

other: Category 2

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study, the test material causes irritation to the eye and requires classification as Category 2 in accordance with EU criteria.

Executive summary:

The irritancy potential of the test material was assessed in vivo in accordance with the standardised guidelines OECD 405 and EU Method B.5.

A single application of the test material was sequentially administered to the right eye of two New Zealand White rabbits. The treated eyes were not irrigated; the left eye remained untreated and served as the control. The animals were observed for 21 days.

Administration of the test material produced scattered or diffuse corneal opacity (vascularisation was noted in one eye at the 14 day observation), iridial inflammation and moderate conjunctival irritation (petechial haemorrhage on the nictitating membrane were noted in one eye at the 7 day observation). One treated eye appeared normal at the 14 day observation; the other treated eye appeared normal at the 21 day observation.

Under the conditions of this study, the test material causes irritation to the eye and requires classification as Category 2 in accordance with EU criteria.