Hydrocarbons, C14-C16, n-alkanes, isoalkanes,

Endpoint summary

Description of key information

An in vitro skin irritation study was conducted with Hydrocarbons, C14-C16, n-alkanes, isoalkanes, <2% aromatics according to OECD 439 and in compliance with GLP (WIL Research, 2014a). The relative mean tissue viability after 15 minutes exposure and 42 hours post incubation was >50% of the mean viability of the negative controls and the substance was therefore determined to be not irritating to skin.

 

An in vitro eye irritation study was conducted with Hydrocarbons, C14-C16, n-alkanes, isoalkanes, <2% aromatics according to OECD 437 and in compliance with GLP ( WIL Research, 2014b). The 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:

Time point: 15 minutes

Value:

85

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 skin irritation study was conducted with Hydrocarbons, C14-C16, n-alkanes, isoalkanes, <2% aromatics (GS250) 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, C14-C16, n-alkanes, isoalkanes, <2% aromatics compared to the negative control tissues was 85%. 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-2015

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): approx. 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:

15 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 minutes

Value:

0

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

No permeability measurement was performed with one of the three corneas treated with test substance. This cornea was excluded from the IVIS determination. Since the cornea shifted in the holder after the opacity measurement it was impossible to perform a reliable permeability measurement with Na-fluorescein. It was therefore not possible to calculate an IVIS value for this cornea. Since this cornea gave reliable results for opacity and the other two cornea treated with the test substance gave reliable results for both opacity and permeability, this deviation had no influence on the study results

Interpretation of results:

GHS criteria not met

Conclusions:

An in vitro eye irritation study was conducted with Hydrocarbons, C14-C16, n-alkanes, isoalkanes, <2% aromatics (GS250), 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 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, C14-C16, 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-SM^TM, for 15 minutes. The positive control had mean relative skin viability (the measure of irritancy) of 21%, and an optical density at 570 nm within the historical control data range. The mean relative skin viability of the test substance was 85%, so it was concluded that the test substance is non irritant under the conditions of this test.

An in vitro eye irritation study was conducted with Hydrocarbons, C14-C16, n-alkanes, isoalkanes, <2% aromatics according to OECD 437 and in compliance with GLP (WIL Research, 2014b). The substance did not indicate an irritancy potential based on either the opacity or permeability endpoints.

Justification for classification or non-classification

Based on in vitro data for the substance itself, Hydrocarbons, C14-C16, n-alkanes, isoalkanes, <2% aromatics does not need to be classified as irritant for skin and eye according to the criteria of Regulation (EC) No 1272/2008.