Registration Dossier

Toxicological information

Eye irritation

Currently viewing:

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reason / purpose for cross-reference:
read-across: supporting information

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report date:
2017

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
October 2015

Test material

Constituent 1
Chemical structure
Reference substance name:
(+)-pin-2(3)-ene
EC Number:
232-087-8
EC Name:
(+)-pin-2(3)-ene
Cas Number:
7785-70-8
Molecular formula:
C10H16
IUPAC Name:
(1R,5R)-2,6,6-trimethylbicyclo[3.1.1]hept-2-ene
Constituent 2
Chemical structure
Reference substance name:
(-)-pin-2(3)-ene
EC Number:
232-077-3
EC Name:
(-)-pin-2(3)-ene
Cas Number:
7785-26-4
Molecular formula:
C10H16
IUPAC Name:
(1S,5S)-2,6,6-trimethylbicyclo[3.1.1]hept-2-ene
impurity 1
Chemical structure
Reference substance name:
(-)-pin-2(10)-ene
EC Number:
242-060-2
EC Name:
(-)-pin-2(10)-ene
Cas Number:
18172-67-3
Molecular formula:
C10H16
IUPAC Name:
(1S,5S)-6,6-dimethyl-2-methylenebicyclo[3.1.1]heptane
impurity 2
Chemical structure
Reference substance name:
(1S)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane
EC Number:
227-337-8
EC Name:
(1S)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane
Cas Number:
5794-04-7
Molecular formula:
C10H16
IUPAC Name:
(1S,4R)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane
impurity 3
Chemical structure
Reference substance name:
(1R)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane
EC Number:
227-336-2
EC Name:
(1R)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane
Cas Number:
5794-03-6
Molecular formula:
C10H16
IUPAC Name:
(1R,4S)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane
impurity 4
Chemical structure
Reference substance name:
1,7,7-trimethyltricyclo[2.2.1.02,6]heptane
EC Number:
208-083-7
EC Name:
1,7,7-trimethyltricyclo[2.2.1.02,6]heptane
Cas Number:
508-32-7
Molecular formula:
C10H16
IUPAC Name:
1,7,7-trimethyltricyclo[2.2.1.0~2,6~]heptane
impurity 5
Chemical structure
Reference substance name:
(1R,5R)-6,6-dimethyl-2-methylenebicyclo[3.1.1]heptane
Cas Number:
19902-08-0
Molecular formula:
C10H16
IUPAC Name:
(1R,5R)-6,6-dimethyl-2-methylenebicyclo[3.1.1]heptane
impurity 6
Chemical structure
Reference substance name:
7,7-dimethyl-2-methylene-, (1S,4R)-bicyclo[2.2.1]heptane
Cas Number:
7378-37-2
Molecular formula:
C10H16
IUPAC Name:
7,7-dimethyl-2-methylene-, (1S,4R)-bicyclo[2.2.1]heptane
impurity 7
Chemical structure
Reference substance name:
7,7-dimethyl-2-methylene-(1R,4S)-bicyclo[2.2.1]heptane
Cas Number:
116724-26-6
Molecular formula:
C10H16
IUPAC Name:
7,7-dimethyl-2-methylene-(1R,4S)-bicyclo[2.2.1]heptane
Test material form:
liquid
Details on test material:
Batch No. : FAB-01072015
Purity : 96.1% (sum of the two main constituents)
Name of test material (as cited in study report): alpha-pinene multiconstituent
Physical state: colourless liquid
Storage Conditions: +2°C to +8°C, under nitrogen and protected from light
Expiry Date: 30 June 2016

Test animals / tissue source

Species:
human
Strain:
other: reconstructed human Cornea-like Epithelium
Details on test animals or tissues and environmental conditions:
EpiOcular TM tissue Model OCL-212-ver2.0 supplied by MatTek

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
Test item alpha-pinene multiconstituent was applied as supplied at the dose of 50 µL.
Duration of treatment / exposure:
30 minutes
Duration of post- treatment incubation (in vitro):
A 12-minute post exposure immersion period at room temperature and a 1-hour and 58-minute post exposure incubation at standard culture conditions.
Number of animals or in vitro replicates:
2 replicates

Results and discussion

In vitro

Results
Irritation parameter:
other: percent tissue viability
Value:
80.39
Negative controls validity:
valid
Positive controls validity:
valid
Remarks:
percent tissue viability = 31.99%

Any other information on results incl. tables

Table 1:

INDIVIDUAL AND AVERAGE VALUES OF OD AFTER 30 MINUTES EXPOSURE

 

Tissue

OD

Mean OD/disc

Mean OD/ product

Viability %

Mean viability %

Difference of viability %

Negative control

1

1.035

1.023

0.961

1.006

1.079

93.28

100.00

13.44

2

1.158

1.128

1.167

1.151

106.72

Positive control

3

0.367

0.351

0.357

0.358

0.345

31.19

31.99

2.41

4

0.335

0.333

0.328

0.332

30.78

Test item

7

0.896

0.879

0.862

0.879

0.867

81.50

80.39

2.33

8

0.873

0.853

0.841

0.855

79.28

OD: optical density

 

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
alpha-Pinene multiconstituent does not need classification as eye irritant or serious eye damage according to Regulation (EC) n° 1272/2008 and UN GHS.
Executive summary:

In a GLP study conducted according to the OECD 492 guideline the irritant potential of alpha-pinene multiconstituent was evaluated. The method is based on viability of Reconstructed Human Cornea-like Epithelium (RhCE) when exposed to the test substance. The mean percent tissue viability of the RhCE replicates treated with test item alpha-pinene multiconstituent was 80.39% versus 31.99% in the positive control (Methyl acetate). In conclusion, in accordance with Regulation (EC) n° 1272/2008 and UN GHS, the test item does not require classification as eye irritant or serious eye damage. No hazard statement or signal word is required.