2,6,10-trimethylundec-9-enal

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 June 2014 to 15 December 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

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)

Deviations:

no

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)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2,6,10-trimethylundec-9-enal

EC Number:

205-460-8

EC Name:

2,6,10-trimethylundec-9-enal

Cas Number:

141-13-9

Molecular formula:

C14H26O

IUPAC Name:

2,6,10-trimethylundec-9-enal

Test material form:

liquid

Specific details on test material used for the study:

Batch SC00010875
Expiration date 09 April 2015

Test animals / tissue source

Species:

other: Bovine eyes

Strain:

other: Corneas

Details on test animals or tissues and environmental conditions:

The eyes were removed after slaughter, completely immersed in Hanks’ Balanced Salt Solution (containing penicillin at 100 IU/mL and streptomycin at 100 μg/mL) in a suitably sized container and transported on the same day to the testing facility.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 750 µL
- Concentration: Undiluted

Duration of treatment / exposure:

10 minute incubation at 32°C ± 1°C.

Duration of post- treatment incubation (in vitro):

After the 1st incubation (10 minute incubation at 32°C ± 1°C) with exposure (750 µL) , each cornea was washed with media containing phenol red followed by media without phenol red and the corneas assessed for opacity. The corneas were then incubated (horizontally) for 2 hours ± 10 minutes after which, corneal opacity was measured.

Number of animals or in vitro replicates:

Three corneas

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

1.03

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

Corneal Opacity :
The mean corrected opacity reading for the test article was 1.0.
The mean corrected opacity reading for the negative control was 0.0.
The mean corrected opacity reading for the positive control was 35.3.

Corneal Permeability:
The mean group corrected optical density for the test article was 0.002.
The mean group corrected optical density for the negative control was 0.000.
The mean group corrected optical density for the positive control was 0.183

The assay was considered valid as the assay acceptance criteria were met.

Any other information on results incl. tables

Table 1: Corneal Opacity

Test substance	Cornea No.	Initial Opacity	Post Incubation Opacity	Change in Opacity	Mean Change in Opacity	Corrected Opacity	Mean Corrected Opacity
Test material	2	3	4	1	N/A	-0.3	1.0
	5	3	7	4		2.7
	15	3	5	2		0.7
NaCl (negative control)	6	2	3	1	1.33	-0.3	0.0
	17	2	4	2		0.7
	21	2	3	1		-0.3
DMF (positive control)	16	3	52	49	N/A	47.7	35.3
	18	3	22	19		17.7
	24	3	45	42		40.7

 

Table 2: Corneal Permeability

Test substance	Cornea No.	Mean Blank OD490	OD490	Corrected OD490	Mean Corrected OD490	Final Corrected OD490
Test material	2		0.022	0.022	N/A	0.003	0.002
	5		0.021	0.021		0.002
	15		0.019	0.019		0.000
NaCl (negative control)	6	0.000	0.021	0.021	0.019	0.002	0.000
	17		0.016	0.016		-0.003
	21		0.019	0.019		0.000
DMF (positive control)	16		0.163	0.163	N/A	0.144	0.183
	18		0.168	0.168		0.149
	24		0.273	0.273		0.254

 

Table 3: IVIS scores

Test substance	IVIS
Test material	1.03
NaCl (negative control)	0.00
DMF (positive control)	38.07

Applicant's summary and conclusion

Interpretation of results:

not classified

Conclusions:

The test article, Adoxal, produced an In Vitro Irritancy Score (IVIS) of 1.03 and does not require classification for eye irritation.

Executive summary:

In the in vitro study (Dreher, 2014), the potential of the test material to cause serious eye irritation was investigated in vitro in a study conducted in accordance with the standardised guidelines OECD 437 and EU Method B.47 under GLP conditions using bovine corneas.

750 µL of undiluted test material was applied to each of three corneas and incubated for 10 minutes at 32 °C ± 1 °C. After incubation, each cornea was washed with media containing phenol red followed by media without phenol red and incubated horizontally for 2 hours ± 10 minutes and then measured for opacity.

Permeability of the corneas was measured by replacing the media in the anterior chamber with 1 mL of 4 mg/mL sodium fluorescein. The posterior chamber was filled with fresh media. The corneas were incubated vertically for 95 minutes at 32 °C ± 1 °C. The posterior chambers were the removed and three 350 µL aliquots of the media were taken for analysis of optical density at 490 nm.

The mean corrected opacity reading for the test material was determined to be 1.0. The mean corrected opacity reading for negative and positive controls was 0.0 and 35.3, respectively. The corneas treated with the positive control were cloudy following treatment.

The mean corrected optical density for the test material was determined to be 0.002. The mean corrected optical densities for negative and positive controls were 0.000 and 0.183, respectively.

Under the conditions of the test, the test material was determined to not require classification for eye irritation. The test material produced an in vitro irritancy score (IVIS) of 1.03 in bovine corneas.