Reaction product with n-butanol of high-boiling stream resulting from the hydroxylation, oxidative cleavage and hydrolysis of vegetable oil saturated and unsaturated C16-C22 triglycerides.

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Recent GLP compliant study, according to international guidelines with detailed test report.

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 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

FAV ES (FAV BUTILATO)

IUPAC Name:

FAV ES (FAV BUTILATO)

Test material form:

liquid: viscous

Details on test material:

Name:
FAV ES (FAV BUTILATO)

Batch no.:
P23_11_12 + P25_07_13

Appearance:
dark brown opaque viscous oil

Composition:
Reaction product of high-boiling residues resulting from the hydroxylation, oxidative cleavage and hydrolysis of vegetable oil saturated and unsaturated C16-C22 triglycerides with n-butanol

Molecular formula:
CxHyOz (aliphatic low molecular weight polyester family)

Purity:
> 99%

Production date:
23. Nov. 2012

Expiry date:
May 2014

Storage:
protected from light and stored in a dry place

Test animals / tissue source

Species:

other: in vitro

Strain:

other: in vitro

Details on test animals or tissues and environmental conditions:

not applicalbe: in vitro test

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

750 µL of liquid test substance was tested undiluted.

Duration of treatment / exposure:

Exposition time on the corneas was 10 min at 32 ± 1 °C. After thorough rinsing with cMEM with phenol red and final rinsing with cMEM without phenol red, the anterior chamber was filled with cMEM without phenol red, and the corneas were stored for an additional 2 hours at 32 ± 1°C (post-incubation).

Details on study design:

Test vessels:
All vessels used are made of glass or sterilizable plastic. They were sterilised before use by heating to 180 °C (two hours) or autoclavation.
The following vessels were used: Schott-bottles, glass vials, and culture flasks for solutions and media

Negative Control
Sodium chloride solution: 0.9% NaCl (CAS-No. 7647-14-5), dissolved in deionised water.

Positive Control
Dimethyl formamide, DMF, CAS-No. 68-12-2, undiluted

Test System:
Bos primigenius Taurus (Fresh bovine corneas). Fresh bovine eyes were obtained from the slaughterhouse Müller Fleisch GmbH, Enzstr. 2.4, 75217 Birkenfeld, Germany, on the day of the test. The cattle were between 12 and 60 months old. The eyes were transported to the test facility in Hank’s balanced salt solution (supplemented with 0.01% streptomycin and 0.01% penicillin). Then the corneas were dissected and incubated in medium at 32 ± 1 °C in an incubation chamber for 1 hour.

After having carefully cleaned and sterilised the cornea holders, they were kept in the incubation chamber at 32 °C.
On the day of the assay, the MEM without phenol red was supplemented with sodium bicarbonate, L-glutamine and 1% fetal calf serum (= complete MEM) and stored in a water bath at 32 °C ± 1 °C. The same was performed with the MEM with phenol red. After the arrival of the corneas they were examined and only corneas which were free from defects were used. The corneas were excised with a scalpel and cut from the globe with a 2-3 mm ring of sclera around the outside. Each cornea was transferred to a cornea holder in which pre-warmed cMEM without phenol red was filled. The holders were then incubated for one hour in the incubation chamber at 32 °C.

After the initial incubation, the medium was changed and the baseline opacity for each cornea was recorded. None of the corneas showed tissue damage; therefore, all corneas were used. The baseline opacity was measured by placing the holder with the cornea in a spectral photometer and recording the absorption at 570 nm.
For each treatment group (negative control solution, test item and positive control), three replicates were used. After removal of the pre-incubation medium, 750 µl negative control solution resp. test item resp. positive control were applied to each replicate.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

>= 0.623

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

In vivo

Irritant / corrosive response data:

Not irritating

Other effects:

The test item FAV ES (FAV BUTILATO) showed no effects on the cornea of the bovine eye.

Any other information on results incl. tables

Table 1: Absorption and Opacity Values Negative Control

Parameter	Negative Control
Absorption before exposition	0.1642	0.1553	0.1469
Absorption after exposition	0.2101	0.1917	0.1901
Opacity before exposition	1.4595	1.4299	1.4025
Opacity after exposition	1.6222	1.5549	1.5492
Opacity Difference	0.1627	0.1250	0.1467

Mean opacity difference of the negative control is 0.1448.

 

Table 2 Absorption and Opacity Values Test Item and Positive Control

Parameter	Test ItemFAV ES (FAV BUTILATO)			Positive Control
Absorption before exposition	0.1455	0.1680	0.1284	0.1276	0.1759	0.1385
Absorption after exposition	0.2263	0.3010	0.3215	1.8965	1.9475	1.7109
Opacity before exposition	1.3980	1.4723	1.3440	1.3415	1.4993	1.3756
Opacity after exposition	1.6838	1.9999	2.0965	78.7952	88.6135	51.3925
Opacity Difference	0.2859	0.5275	0.7525	77.4537	87.1142	50.0169

Table 3 IVIS

Test Group	IVIS	Mean IVIS	Relative Standard Deviation IVIS
Negative Control 0.9% NaCl	0.253	0.187	32.5
	0.133
	0.177
Test Item	0.639	0.623	10.5%
	0.551
	0.678
Positive Control DMF undiluted	92.102	86.251	22.3%
	101.837
	64.815

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The substance resulted not irritating

Executive summary:

The eye damaging potential of the substance was assessed via an in vitro test according to OECD and GLP guidelines, using the BCOP test.

Bovine corneas were used. They were collected from slaughtered cattle which were between 12 and 60 months old.

The test item was brought onto the cornea of a bovine eye which had been incubated with cMEM without phenol red at 32 ± 1 °C for one hour and whose opacity had been measured. The test item was incubated on the cornea for 10 minutes at 32 ± 1 °C. After removal of the test item and two hours post-incubation, opacity and permeability values were measured. Physiological sodium chloride solution was used as negative control. The negative control showed no irritating effect on the cornea. DMF undiluted was used as positive control. The positive control induced a severe irritation on the cornea. The test item showed no effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) is 0.623.

 

According to OECD Guideline no. 437 (Jul. 2013), a substance with an IVIS ≤ 3 requires no classification for eye irritation or serious eye damage.