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Diss Factsheets

Administrative data

Endpoint:
eye irritation
Remarks:
other: SkinEthic Reconstructed Human Corneal Epithelial Model
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study was conducted by a GLP accredited laboratory using the SkinEthic Reconstructed Human Corneal Epithelial Model, as described in the scientific article "Prevalidation of a new in vitro reconstituted human cornea model to assess the eye irritating potential of chemicals" by Van Goethem & al., Toxicology in Vitro 20 (2006) 1–17. The study was conducted on Gas oil (polymer-derived), thermal cracked, full range, from which the registered substance is derived via steam stripping, and which is compositionally similar to the registered substance.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: SkinEthic Reconstructed Human Corneal Epithelial Model
Deviations:
yes
Remarks:
Freeze killed tissues not used for quantitative correction of results, and dosing occurred on day 8 instead of day 7. Deviations were not considered to have affected the study.
Principles of method if other than guideline:
The experimental design of the study consists of a test for direct reduction of MTT (3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) by the test item followed by the main test.

For the main test, triplicate SkinEthic tissues were treated with 30 µL of the test item for 10 minutes. Triplicate tissues treated with 30 µL of Solution A (supplied by SkinEthic) served as the negative control and triplicate tissues treated with 30 µL of 2% w/v Sodium Dodecyl Sulphate (SDS) served as the positive control.

At the end of the exposure period each SkinEthic tissue was rinsed. The rinsed tissues (two per group) were taken for MTT loading. The remaining tissues were retained for possible histopathology. Following MTT loading the reduced MTT was extracted from the tissues.

After extraction the absorbency of triplicate aliquots of the extracted MTT solution for each SkinEthic tissue was measured. The optical density was measured at 540 nm (OD540). Data are presented in the form of percentage viability (MTT conversion relative to negative controls).

The test item was classified according to the following criteria:
I) If the percentage relative mean tissue viability was ≥60% the test item was considered to be non irritant (NI).
II) If the percentage relative mean tissue viability was <60% the test item was considered to be irritant (I).
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
Gas oil (polymer derived) thermal-cracked, full range
IUPAC Name:
Gas oil (polymer derived) thermal-cracked, full range
Constituent 2
Reference substance name:
700-882-3
EC Number:
700-882-3
IUPAC Name:
700-882-3
Constituent 3
Reference substance name:
Not assigned
IUPAC Name:
Not assigned
Test material form:
other: Liquid
Details on test material:
Name of test material: Gas oil (polymer derived) thermal-cracked, full range (Sample Ref TSM13/02)
Substance type: UVCB
Physical state: Liquid
Storage condition of test material: Room temperature in the dark

Test animals / tissue source

Species:
other: SkinEthic Reconstructed Human Corneal Epithelial Model
Strain:
other: SkinEthic Reconstructed Human Corneal Epithelial Model
Details on test animals or tissues and environmental conditions:
SkinEthic Reconstructed Human Corneal Epithelial Model (HCE, SkinEthic Laboratories, Lyon, France), received 22 May 2013

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: Triplicate tissues with 30 µL of Solution A served as the negative control and triplicate tissues treated with 30 µL of 2% w/v Sodium Dodecyl Sulphate (SDS) served as the positive control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 µL
Duration of treatment / exposure:
10 minutes
Number of animals or in vitro replicates:
N/A
Details on study design:
REMOVAL OF TEST SUBSTANCE
Washing (if done): Dulbecco's Phosphate Buffered Saline
Time after start of exposure: 10 minutes

SCORING SYSTEM: Optical density at 540 nm

TOOL USED TO ASSESS SCORE: filter photometer (Anthos 2001 microplate reader)

Results and discussion

In vivo

Results
Irritation parameter:
other: Relative Mean Viability (%)
Basis:
mean
Time point:
other: 10 minutes
Score:
108.8
Reversibility:
not specified
Irritant / corrosive response data:
The relative mean viability of the test item treated tissues after a 10 minute exposure period was 108.8%.
Other effects:
It was considered unnecessary to proceed with tissue histopathology

Any other information on results incl. tables

Item  OD(540) of Individual Tissue Mean OD(540) Relative Mean Viability (%)
Negative Control 0.902 0.927 100
0.952
Positive Control 0.073 0.065 7
0.057
Test Item 1.039 1.009 108.8
0.978

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: other: According to study plan
Conclusions:
The relative mean viability of the test item treated tissues after a 10 minute exposure period was 108.8%. According to the study plan followed the test item was considered to be non-irritant.
Executive summary:

The in vitro eye irritation of the test substance was determined in accordance with the SkinEthic Reconstructed Human Corneal Epithelial Model, as described in the scientific article "Prevalidation of a new in vitro reconstituted human cornea model to assess the eye irritating potential of chemicals" by Van Goethem & al., Toxicology in Vitro 20 (2006) 1–17. The experimental design of the study consists of a test for direct reduction of MTT (3 [4,5 dimethylthiazol-2 -yl]-2,5 -diphenyl-tetrazolium bromide) by the test item followed by the main test. The relative mean viability of the test item treated tissues after a 10 minute exposure period was 108.8%. According to the study plan followed the test item was considered to be non-irritant.

Gas oil (polymer-derived), thermal-cracked, full-range and Thermal cracking oil from blends of rubber, fuel oils and paraffin waxes, steam-stripped are two very closely related substances. They consist of hydrocarbons having carbon numbers predominantly in the same range (C7 to C32) and boil over a very similar temperature range. The oils contain a relatively large proportion of substituted aromatic hydrocarbons (between 62 and 70%), particularly 1-ring aromatic hydrocarbons. Detailed analytical characterisation shows that the oils have a significant number of individual components in common. Both oils are produced by thermal cracking of the same range of hydrocarbon feedstocks at a temperature of 450-500°C. Thermal cracking oil from blends of rubber, fuel oils and paraffin waxes, steam-stripped differs from Gas oil (polymer-derived), thermal-cracked, full-range in that it has been through a steam-stripping process that removes a proportion of the lower boiling components. In addition to their composition and similar manufacturing process, testing has revealed that the two oils have very similar physico-chemical parameters, especially their solubility in water which is in the region of 3 mg/l suggesting that the two substances might be expected to behave similarly in biological and environmental aqueous environments.

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