C10-13-alkyl(branched, unsaturated)-succinic acid-4-(3-methyl-butyl)-ester

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-02-08 to 2016-02-29

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline and GLP conform well documented scientific study report.

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)

Version / remarks:

2012

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Version / remarks:

1998

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Tetrapropylene succinic acid monoisobutylester

IUPAC Name:

Tetrapropylene succinic acid monoisobutylester

Constituent 2

Reference substance name:

921-522-3

EC Number:

921-522-3

IUPAC Name:

921-522-3

Test material form:

other: Light brown, viscous liquid

Details on test material:

Identification: Tetrapropylene succinic acid monoisobutylester
Appearance: Light brown, viscous liquid
Batch: ESD0018639
Purity/Composition: 93.0% (w/w)
Test item storage: At room temperature
expiry date: Stable under storage conditions until 30 November 2016

Specific details on test material used for the study:

Identification Tetrapropylene succinic acid monoisobutylester
Appearance Light brown, viscous liquid
Batch ESD0018639
Purity/Composition 93.0% (w/w)
Test item storage At room temperature
Stable under storage conditions until 30 November 2016 (expiry date)

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Species: Albino rabbit, New Zealand White, (SPF-Quality). Recognized by international guidelines as the recommended test system (e.g. EC, OECD)
Source: Charles River France, L’Arbresle Cedex, France
Number of animals: 1 Male
Age and body weight: At start of dosing, the animal was 18 weeks old and the body weight was at least 1.5 kg.
Identification: Earmark
Health inspection: At least prior to dosing. It was ensured that the animal was healthy and that eyes were free from any abnormality.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

instillation of 0.1 mL of the undiluted test item

Duration of treatment / exposure:

The animal was treated by instillation of 0.1 mL of the test item, in the conjunctival sac of one of the
eyes after gently pulling the lower lid away from the eyeball. The lids were then gently held together for
about one second to prevent loss of the test item. The other eye remained untreated and served as
the reference control.

Observation period (in vivo):

1, 24, 48, 72 hours after treatment and 7, 14 and 21 days

Number of animals or in vitro replicates:

The study was performed in a stepwise manner and was started by treatment of a single rabbit
(sentinel). Based on the duration of the ocular lesions of the first animal, the two further rabbits
assigned to the study were not treated.

Details on study design:

The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel).

Preemptive Pain management:
One hour prior to instillation of the test item, buprenorphine (Buprenodale®, Dechra Ltd., Stoke-on-Trent, United Kingdom) 0.01 mg/kg was administered by subcutaneous injection in order to provide a therapeutic level of systemic analgesia.

Five minutes prior to instillation of the test item, two drops of the topical anesthetic alcaine 0.5% (SA Alcon-Couvreur NV, Puurs, Belgium) were applied to both eyes.

Treatment
The animal was treated by instillation of 0.1 mL of the test item, in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test item. The other eye remained untreated and served as the reference control.

Immediately after the 24-hour observation, a solution of 2% fluorescein (Merck, Darmstadt, Germany) in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. This procedure was repeated to assess recovery. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.

Immediately after fluorescein examination on Day 2, in order to provide a continued level of systemic analgesia, buprenorphine 0.01 mg/kg and meloxicam (Metacam®, Boehringer Vetmed GmbH, Ingelheim/Rhein, Germany) 0.5 mg/kg were administered by subcutaneous injection.

Additional injections were supplied during the observation period to reduce pain and distress

After the final observation, the animal was sacrificed by intra-venous injection of Euthasol® 20% (AST Farma BV, Oudewater, The Netherlands).

Based on the duration of the ocular lesions of the first animal, the two further rabbits assigned to the study were not treated.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: 24, 48, 72 hours after application

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 14 days

Results 2

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: 24, 48, 72 hours after application

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 14 days

Results 3

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

other: 24,48, 72 hours after application

Score:

3

Max. score:

3

Reversibility:

not fully reversible within: 21 days after application

Results 4

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 24, 48, 72 hours after application

Score:

1.7

Max. score:

2

Reversibility:

fully reversible within: 14 days after application

Any other information on results incl. tables

The study was performed in a stepwise manner and was started by treatment of a single rabbit (sentinel). Based on the duration of the ocular lesions of the first animal, the two further rabbits assigned to the study were not treated.

Table1: Individual eye irritation scores

							Cornea							Iris				Conjunctivae									Comments
animal		Time after dosing					Opacity (0-4)		Area (0-4)		Fluor area (%)^1			(0-2)				Redness (0-3)			Chemosis (0-4)		Discharge (0-3)
994		1 hour					1		1					1				2			2		3				-
		24 hours					1		2		50			1				3			2		2				-
		48 hours					1		1					1				3			2		2				f
		72 hours					1		2		50			1				3			1		1				f
		7 days					1		1		25			1				2			1		1				f, p
		14 days					0		0		0			0				1			0		1				f
		21 days					0		0					0				1			0		0				-

 ¹  Green staining after fluorescein treatment (percentage of total corneal area).

 

Comments:

p         Pannus, neovascularization of the cornea.

f          Reduced elasticity of the eyelids.

Table 2:       Mean value eye irritation scores

Animal		Mean 24, 48 and 72 hours
		Corneal		Iris		Conjunctivae
		opacity				Redness		Chemosis
994		1.0		1.0		3.0		1.7

 

 

Table 3:       Animal specifications

Animal		Sex		Age at start		Body weights (grams)
				(weeks)		prior to application	after the final observation
994		♂		18		3201	3512

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on the persistence of the ocular lesions until 21 days after instillation the test substance is considered to be irritant to the eye.

Executive summary:

Instillation of 0.1 mL of Tetrapropylene succinic acid monoisobutylester (unchanged) into an eye of one rabbit resulted in severe effects on the cornea, iris and conjunctivae. The corneal injury consisted of opacity and epithelial damage. As a result of the corneal injury, pannus (neovascularization of the cornea) was apparent 8 days after instillation. The corneal injury resolved within 15 days. Iridial irritation was observed and also resolved within 15 days. The irritation of the conjunctivae consisted of redness, chemosis and discharge. Chemosis and discharge had completely resolved within 15 days and 22 days, respectively. Redness was still present at 21 days after instillation.

There was no evidence of ocular corrosion. No staining of (peri) ocular tissues by the test item was observed and no test item remnants were seen. No signs of systemic toxicity were observed in the animal during the test period and no mortality occurred. Based on the persistence of the ocular lesions until 21 days after instillation the test substance is considered to be irritant to the eye.