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Description of key information

An in vitro skin irritation was conducted on Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics according to OECD TG 439 and in compliance with GLP (WIL Research, 2014a). The mean relative skin viability of the test substance was 91%, so it was concluded that the test substance is not irritant under the conditions of this test.

A bovine corneal opacity in vitro assay (BCOP test) was conducted on Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics according to OECD TG 437 and in compliance with GLP (WIL Research, 2014b). The endpoints assessed in this assay are opacity and permeability; negative and positive controls gave expected results. The test substance did not indicate an irritancy potential based on either the opacity or permeability endpoints.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
21-Jul-2014 to 28-Jul-2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 µl

NEGATIVE CONTOL:
- Amount(s) applied (volume or weight with unit): 10 µl Phosphate buffered saline

POSITIVE CONTROL
- Amount(s) applied (volume or weight with unit): 10 µl
- Concentration (if solution): 5% (aq) Sodium dodecyl sulphate
Duration of treatment / exposure:
Exposure:15 minutes
Post incubation period: 42 hours
Details on study design:
TEST SITE
- Area of exposure: human epidermis model
- % coverage: 0.38 cm2

REMOVAL OF TEST SUBSTANCE
- Washing (if done): phosphate buffered saline
- Time after start of exposure: 15 minutes

POST INCUBATION PERIOD
- 42 hours

SCORING SYSTEM:
- After a 42 hour incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment. Cell viability was calculated for each tissue as a percentage of the mean of the negative control tissues.
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
15-minute exposure
Value:
94
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
The absolute mean of the negative control was slightly below the historical control data range. It was concluded that the deviation had no effect on the study results.
Positive controls validity:
valid
Remarks:
A clear toxic effect was observed with the positive control. The low absolute mean OD570 value of the positive control, which was in the lower range of the historical control data range, resulted in a good measurement window.
Remarks on result:
no indication of irritation
Interpretation of results:
GHS criteria not met
Conclusions:
An in vitro skin irritation study was also conducted with Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics (GS270) in accordance with OECD Test Guideline 439 and in compliance with GLP. The in vitro assay did not indicate any skin irritation potential.
Executive summary:

Skin irritation is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after 15 minutes treatment with Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics compared to the negative control tissues was 94%. Since the mean relative tissue viability for the test substance was above 50% after 15 minutes treatment the test substance is considered to be non-irritant.

 

The positive control had a mean cell viability of 21% after 15 minutes exposure. The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was slightly below laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 7%, indicating that the test system functioned properly.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15-Jul-2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
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 (incl. QA statement)
Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µl per cornea

NEGATIVE CONTROL
- Amount(s) applied (volume or weight with unit): 750 µl of physiological saline per cornea

POSITIVE CONTROL
Amount(s) applied (volume or weight with unit): 750 µl per cornea
Concentration (if solution): 10% (w/v) Benzalkonium Chloride


Duration of treatment / exposure:
10 minutes
Details on study design:
TEST SITE
- Isolated bovine cornea

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 10 minutes

SCORING SYSTEM:
- After exposure the cornea was thoroughly rinsed to remove the test substance and incubated for 2 hours with fresh medium followed by opacity measurement and the permeability of the corneas was determined after a 90 minutes incubation period with sodium fluorescein.

- The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:

In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).


TOOL USED TO ASSESS SCORE:
- opacitymeter and microplate reader

DATA EVALUATION:
In vitro score range UN GHS
≤ 3 No Category
> 3; ≤ 55 No prediction can be made
>55 Category 1

Irritation parameter:
in vitro irritation score
Run / experiment:
10-minute exposure
Value:
0.1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Interpretation of results:
GHS criteria not met
Conclusions:
An in vitro eye irritation study was also conducted with Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics (GS270), in accordance with OECD test guideline 437 and in compliance with GLP. The BCOP test did not indicate an irritancy potential based on either the opacity or permeability endpoints.
Executive summary:

The negative control responses of the opacity and permeability values were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas.The mean in vitro irritancy score of the positive control (10% (w/v) Benzalkonium Chloride) was 139 and within the historical positive control data range. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

The test substance did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of 0.1 after 10 minutes of treatment.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics has been tested in an in vitro assay conducted according to OECD TG 439 and in compliance with GLP (WIL Research, 2014a). The test substance was applied to a human three-dimensional epidermal model, EPISKIN-SMTM, for 15 minutes. The positive control had mean relative skin viability (the measure of irritancy) of 21%, and an optical density at 570 nm very slightly below the historical control data range. The mean relative skin viability of the test substance was 91%, so it was concluded that the test substance is not irritant under the conditions of this test.

Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics has been tested in a bovine corneal opacity in vitro assay (BCOP test) conducted according to OECD TG 437 and in compliance with GLP (WIL Research, 2014b). The endpoints assessed in this assay are opacity and permeability; negative and positive controls gave expected results. The test substance did not induce ocular irritation as assessed by both endpoints.

Justification for classification or non-classification

As all reliable studies show no irritation effect or irritation effects slight enough not to classify the test materials, Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics does not need to be classified as irritant for skin and eye according to the criteria of Annex VI of Regulation (EC) No 1272/2008.