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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
October 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

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

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
Principles of method if other than guideline:
N/A
GLP compliance:
yes

Test material

Constituent 1
Reference substance name:
Apple, Malus sylvestris, ext.
EC Number:
286-475-7
EC Name:
Apple, Malus sylvestris, ext.
Cas Number:
85251-63-4
IUPAC Name:
Apple, Malus sylvestris, ext.
Test material form:
solid: particulate/powder

In vitro test system

Test system:
human skin model
Source species:
other: Human reconstructed epidermis SkinEthic
Cell type:
other: N/A
Cell source:
other: N/A
Source strain:
other: N/A
Details on animal used as source of test system:
N/A
Vehicle:
unchanged (no vehicle)
Details on test system:
Inserts (filter + epidermis) were gently detached from the agar and if necessary the bottom of the insert was wiped on ana absorbent paper in order to avoid leaving agar pieces onto the polycarbonate membrane.
Inserts were placed then into wells (6 wells culture plate) previously filled with 1 ml of growth medium at room temperature. Cultures were incubated overnight at 37°C 5% CO2.
Th epidermis was previously moistened with 10 µl deionised sterile wtare in order to unsure good contact with the test item and then 16 mg+/- 2 mg of test item were applied to the surface of the tissue.
16µL+/- 0.5 of the reference item were deposited with a positive displacement micropipette on the surface of the epidermises and a 7.5 mm diameter nylon mesh is gently applied on the surface of the epidermis using tweezers.
The epidermises were incubated in a 0.3 ml of maintenance medium (24 wells plate) at room temperature for 42 min +/-1 minute.
Nylon mesh were removed and the epidermises were rinsed with 25 ml of PBS by epidermis. the epidermises were incubated in a 2 mlof growth medium in 6 wells plate at 37°C, 5%CO2 for 42 hours +/- 1hour.
At the end of incubation period, the plates were stired approximatively 2 mins at 300 rotations/mins in order to homogenize the released mediators or enzymes in the culture medium.
all epidermises were incubated in a 0.3 ml of maintenance medium at 1 mg/ml MTT in 24 wells plate.
After 3 hours incubation at 37°C 5CO2, outside of inserts was rinsed with 2 ml of PBS.
Extraction was performed by placing the epidermises into wells filled with 0.8 ml of isopropanol then covered with 0.7 ml isopropanol for 2 hours +/- 5 mins under gentle agitation at room agitation at room temperature. The inserts were pierced and removed from wells then extraction solution was homogenized by pipeting. The absorbances were measured in triplicate on 200µl MTT extract in 96 wells plate. Absorbances are measured at 540 nm against isopropanol as blank.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
16 mg +/- 2 mg of the test item pur as well as the reference items were tested on three epidermises
Duration of treatment / exposure:
42 minutes
Duration of post-treatment incubation (if applicable):
42 hours
Number of replicates:
3

Test animals

Species:
other: N/A
Strain:
other: N/A

Test system

Type of coverage:
other: N/A
Preparation of test site:
other: N/A
Vehicle:
other: N/A

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1
Value:
ca. 99.1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
2
Value:
ca. 99.8
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3
Value:
ca. 96.2
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the retained experimental conditions and according to CLP regulation, the test item tested pure is classified non irritant.
Executive summary:

Under the retained experimental conditions and according to CLP regulation, the test item tested pure is classified non irritant.