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Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
March to April 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019
Report date:
2019

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
2,6-bis[[bis(2-hydroxyethyl)amino]methyl]-4-nonylphenol
EC Number:
243-500-6
EC Name:
2,6-bis[[bis(2-hydroxyethyl)amino]methyl]-4-nonylphenol
Cas Number:
20073-51-2
Molecular formula:
C25H46N2O5
IUPAC Name:
2,6-bis[[bis(2-hydroxyethyl)amino]methyl]-4-nonylphenol
Test material form:
liquid: viscous

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Justification for test system used:
This study addresses the human health endpoint skin irritation. It makes use of reconstructed
human epidermis (RhE) (obtained from human derived non-transformed epidermal keratinocytes)
which closely mimics the histological, morphological, biochemical and physiological properties
of the upper parts of the human skin, i.e., the epidermis. The use of reconstructed human
epidermis (RhE) is also recommended by the OECD and other regulatory authorities. SkinEthic
RHE model has been validated and is part of OECD validated reference methods (VRMs) and is
also a recommended model for conducting in vitro skin irritation studies. The results of the study
are believed to be of value in predicting the potential of inducing skin irritation by the test item in
humans.

Details on test system:
The test system used for the in vitro skin irritation test was reconstructed human epidermis RHE model
consists of normal human keratinocytes cultured for 17-days on an insert of 0.5 cm
polycarbonate filter at the air-liquid interface in a chemically defined growth medium. The cells
form a multilayered, highly differentiated and stratified epidermis model of the human epidermis
that consists of main basal, supra basal, spinous and granular layers and a functional stratum
corneum.
RHE model, were procured from SkinEthic Laboratories, Episkin 4, Rue Alexander Fleming, 69366 Lyon
Cedex 07, France; were used in the study (Lot N°19-RHE-027, APPENDIX 5).
Control samples:
yes, concurrent MTT non-specific colour control
Amount/concentration applied:
16 µL/0.5 cm2
Duration of treatment / exposure:
42 min.
Duration of post-treatment incubation (if applicable):
42 min.

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Value:
ca. 1.9
Negative controls validity:
valid
Positive controls validity:
valid

Applicant's summary and conclusion

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
Based on the results of this study, the classification for POLIOL MB 600 is as mentioned below:

Globally Harmonized System of Classification and Labelling of Chemicals: Category 2 (Skin Irritant)


Executive summary:

This study was performed to evaluate the non-irritant and irritant potential of

POLIOL MB 600 using reconstructed human epidermis (RHE) tissue in accordance with the United Nations 

Globally Harmonized System of Classification and Labelling of Chemicals (UN GHS).  


Tissues were exposed to the negative control (Dulbecco’s Phosphate Buffered Saline (DPBS), positive 

control (sodium dodecyl sulfate, 5% aqueous (SDS)) and POLIOL MB 600  in triplicate for 42 minutes at 

room temperature.  


The mean cell viability in tissues treated with the test item was 1.9% after 42 minutes exposure. A 

significant reduction in percent cell viability was observed in treated tissues when compared with that of the 

concurrent negative control.  

 

The Optical density (OD) values for the negative control replicates were between 1.633 and 1.853, against 

the guideline requirement of ≥ 0.8 and ≤ 3.0 (≥ 1.2 as per SkinEthic SOP). The OD of the blank was 

between 0.040 and 0.044 which met the guideline requirement of OD < 0.1. The positive control showed 

a 1% cell viability, against the acceptance criteria of <40% for the SkinEthic RHE model, when compared 

with that of the concurrent negative control. Variation between tissue replicates (i.e., CV% value) was 

17.00% for positive control and 6.46% for negative control against the guideline requirement of ≤ 18%, 

which demonstrate the efficiency of the test system, SkinEthic 

replicates (i.e., CV% value) was 20.21% for the test item group. All criteria for a valid study were met as 

describe in the study plan.  


Based on results of this study, the classification for POLIOL MB 600 is as follows:  

Globally Harmonized System of Classification and Labelling of Chemicals   :  


Category 2 (Skin Irritant)