Fatty acids, C16-18 and C18-unsatd., compds. with triethanolamine

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study is conducted according to GLP and standard methods, therefore it is considered to be relevant, adequate and reliable.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

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:

Fatty acids, C16-18 and C18-unsatd., compds. with triethanolamine

EC Number:

270-279-3

EC Name:

Fatty acids, C16-18 and C18-unsatd., compds. with triethanolamine

Cas Number:

68424-19-1

Molecular formula:

Not applicable as the substance is a UVCB

IUPAC Name:

Fatty acids, C16-18 (even numbered) and C18-unsatd., compds. with triethanolamine.

Test material form:

semi-solid (amorphous): gel

Remarks:

paste

Details on test material:

- Name of test material (as cited in study report): Fatty acids, C16-18 and C18-unsatd., compds. with triethanolamine
- Physical state: Viscous, translucent yellowish waxy paste
- Analytical purity: >95%
- Impurities (identity and concentrations): See confidential details
- Composition of test material, percentage of components: See confidential details
- Purity test date: 01/2013
- Lot/batch No.: DDR02/91
- Expiration date of the lot/batch: October 2014
- Storage condition of test material: At +10° to +25°C
- Other: Manufacturer/Supplier: OLEON NV, Assenedestraat 2, 9940 Ertvelde, Belgium

Test animals / tissue source

Species:

rabbit

Strain:

Himalayan

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: LPT Laboratory of Pharmacology and Toxicology GmbH & Co. KG, branch Löhndorf, 24601 Löhndorf/Post Wankendorf, Germany
- Age at dosing: Approx. 8 - 9 months
- Weight at dosing:
Animal no. 1: 3.3 kg
Animal no. 2: 2.9 kg
Animal no. 3: 3.5 kg
- Fasting period before study:
- Housing: For 8 hours following test item application, the animals are kept singly in restrainers which allow free movement of the head but prevent a complete body turn, wiping of the eyes by the paws and exclude irritation of the eye by excrements and urine.
During the acclimatisation period and after the 8-hour period in restrainers, the animals are kept singly in cages with dimensions of 380 mm x 425 mm x 600 mm (manufacturer: Dipl.Ing. W. EHRET GmbH, 16352 Schönwalde, Germany).
- Diet (e.g. ad libitum): Ssniff® K-H (ssniff Spezialdiäten GmbH, 59494 Soest, Germany) served as food. The food was available ad libitum.
- Water (e.g. ad libitum): Tap water was offered ad libitum.
- Acclimation period: At least 20 adaptation days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.9°C – 21.7°C (20°C ± 3°C (maximum range))
- Humidity (%): 39.7 – 84.1%. A malfunctioning of the air conditioning system, caused by servicing, resulted in relative humidity values that exceeded the maximum range of 30% - 70% on five test days.
- Air changes (per hr): Not provided
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: May 24, 2013 To: June 04, 2013

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: the untreated eye served as control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL of the test item were administered into one eye each of three animals.

VEHICLE: No vehicle

Duration of treatment / exposure:

Single instillation into the conjunctival sac

Observation period (in vivo):

8 days

Number of animals or in vitro replicates:

3: Initial test 1; Confirmatory test 2

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: No

SCORING SYSTEM:
Cornea
Opacity: degree of density ( area most densetaken for reading)
No ulceration or opacity: 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible: 1
Easily discernible translucent area, details of iris slightly obscured: 2
Nacreous area, no details of iris visible, size of pupil barely discernible: 3
Opaque cornea, iris not discernible through the opacity : 4
Iris
Normal: 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia, or injection, iris reacting to light (a sluggish reaction is considered to be an effect): 1
Haemorrhage, gross destruction, or no reaction to light: 2
Conjunctivae
Redness: (refers to palpebral and bulbar conjunctivae, excluding cornea and iris)
Normal: 0
Some blood vessels hyperaemic (injected): 1
Diffuse, crimson colour, individual vessels not easily discernible: 2
Diffuse beefy red: 3
Chemosis
Swelling (refers to lids and/or nictitating membranes)
Normal: 0
Some swelling above normal: 1
Obvious swelling, with partial eversion of lids: 2
Swelling with lids about half closed: 3
Swelling, with lids more than half closed: 4

Any further lesions are listed.

Fluorescein-Test:
DEGREE OF STAINING
0: no staining
1: scattered or diffuse colouration, details of iris clearly visible
2: easily discernible translucent area, details of iris slightly obscured
3: opalescent areas, details of iris not discernible, extent of pupil difficultly
Determinable
4: opaque cornea, iris not discernible through the opacity
INVOLVED AREA OF CORNEA
0: none
1: up to 1/4 of the surface
2: 1/4 to 1/2 of the surface
3: 1/2 to 3/4 of the surface
4: 3/4 to whole surface.

TOOL USED TO ASSESS SCORE: hand-slit lamp / fluorescein
The eyes were examined ophthalmoscopically with a slit lamp prior to the administration and 1, 24, 48, 72 hours and 4 and 5 days after the administration (animal nos. 1 and 3) and prior to the administration and 1, 24, 48, 72 hours and 4 to 8 days after the administration (animal no. 2). The eye reactions were observed and registered.
24 hours after administration, fluorescein was applied to the eyes before being examined to aid evaluation of the cornea for possible lesions. The fluorescein test was repeated on each day of observation.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24, 48 and 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 8 days (animal no.2)

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24, 48 and 72 h

Score:

0

Max. score:

2

Remarks on result:

other: The irises were not affected by instillation of the test item.

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 24, 48 and 72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 8 days (animal no.2)

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24, 48 and 72 h

Score:

0.78

Max. score:

4

Reversibility:

fully reversible within: 8 days (animal no. 2)

Irritant / corrosive response data:

Cornea opacity (grade 1) was observed in all animals 60 minutes to 4 days, in animal no. 2 until 7 days after instillation.
Conjunctival redness (grade 1) was observed in all animals 60 minutes to 4 days, in animal no. 2 until 7 days after instillation.
Chemosis was observed in all animals:
- animal no. 1: 60 minutes (grade 2) and 24 to 72 hours (grade 1) after instillation;
- animal no. 2: 60 minutes (grade 2) and 24 hours to 4 days (grade 1) after instillation;
- animal no. 3: 60 minutes (grade 2) and 24 hours (grade 1) after instillation.
The fluorescein tests performed on each day of observation revealed corneal staining (grade 1) in all animals up to 3/4 of the surface.
All findings observed had disappeared in animal nos. 1 and 3 until 5 days, in animals no. 2 until 8 days after instillation.
The irises were not affected by instillation of the test item.
There were no systemic intolerance reactions concerning behaviour, body weight and food consumption.

Any other information on results incl. tables

Table 1. Examination of the treated eye

 

Time after administration	Cornea	Iris	Conjunctivae		Fluorescein Test
	Opacity		Redness*	Chemosis**	Grade	Area
Animal no.: 1/2/3
Right eye: 0.1 mL Test item/animal
24 hours	1/1/1	0/0/0	1/1/1	1/1/1	1/1/1	3/3/3
48 hours	1/1/1	0/0/0	1/1/1	1/1/0	1/1/1	3/3/3
72 hours	1/1/1	0/0/0	1/1/1	1/1/0	1/1/1	3/3/3
24-72 hours	1/1/1	0/0/0	1/1/1	1/1/0.33	1/1/1	3/3/3
Mean 24-72 h	1	0	1	0.78	1	3

* refers to palpebral and bulbar conjunctivae; excluding cornea and iris

** swelling: refers to lids and/or nictitating membrane

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

According to the EC-Commission directive 67 /548/EEC and its subsequent amendments on the approximation of the laws, regulations and administrative provision relating to the classification, packaging and labelling of dangerous substances and the results obtained under the present test conditions, Fatty acids, C16-18 and C18-unsatd., compds. with triethanolamine is non- irritating to eyes, hence, no labelling is required.
According to the EC Regulation 1272/2008 and subsequent regulations, the test item required labelling with 'Warning' and 'H319: Causes serious eye irritation'. According to the classification of the Globally Harmonized Classification System (GHS) the test item should be assigned to the hazard category 2A and labelled with an exclamation mark, 'Warning' and 'Causes serious eye irritation'.

Executive summary:

Fatty acids, C16-18 and C18-unsatd., compds. with triethanolamine was tested in an acute eye irritation/corrosion test in rabbits according to EC method B.5. (Regulation (EC) No. 440/2008) and OECD guideline 405 (2002). A single instillation of 0.1 mLper animal into the conjunctival sac of the right eye of three rabbits caused the following changes:
Cornea opacity (grade 1) was observed in all animals 60 minutes to 4 days, in animal no. 2 until 7 days after instillation.

Conjunctival redness (grade 1) was observed in all animals 60 minutes to 4 days, in animal no. 2 until 7 days after instillation.

Chemosis was observed in all animals:

- animal no. 1: 60 minutes (grade 2) and 24 to 72 hours (grade 1) after instillation;

- animal no. 2: 60 minutes (grade 2) and 24 hours to 4 days (grade 1) after instillation;

- animal no. 3: 60 minutes (grade 2) and 24 hours (grade 1) after instillation.

Mean 24 -72h scores for cornea, iris, conjuctiva and chemosis were 1/4, 0/2, 1/3 and 0.78/4 respectively.

The fluorescein tests performed on each day of observation revealed corneal staining (grade 1) in all animals up to 3/4 of the surface.

All findings observed had disappeared in animal nos. 1 and 3 until 5 days, in animals no. 2 until 8 days after instillation.

The irises were not affected by instillation of the test item.

There were no systemic intolerance reactions concerning behaviour, body weight and food consumption.

According to the EC-Commission directive 67/548/EEC and its subsequent amendments on the approximation of the laws, regulations and administrative provision relating to the classification, packaging and labelling of dangerous substances and the results obtained under the present test conditions, Fatty acids, C16-18 and C18-unsatd., compds. with triethanolamine is non- irritating to eyes, hence, no labelling is required.

According to the EC Regulation 1272/2008 and subsequent regulations, the test item required labelling with 'Warning' and 'H319: Causes serious eye irritation'. According to the classification of the Globally Harmonized Classification System (GHS) the test item should be assigned to the hazard category 2A and labelled with an exclamation mark, 'Warning' and 'Causes serious eye irritation'.