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Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Remarks:
The Reconstructed human Epidermis (RhE) Test Method
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Experimental Phase scheduled: 01. Apr. 2019 - 17. May. 2019 Final Report: Jun. 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
OECD 423 / EU B.1 trans

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)
Version / remarks:
OECD (2015)
Deviations:
not applicable
Remarks:
Interruption of the power supply. Temperature dropped to 32°C and CO2 conc. dropped to 3%. This is uncritical: values outside the range were short term - no abnormalities. The deviation was assessed and signed by the study director on 10. Apr. 2019.
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
Sodium phenylphosphinate
EC Number:
224-305-5
EC Name:
Sodium phenylphosphinate
Cas Number:
4297-95-4
Molecular formula:
C6H7O2P.Na
IUPAC Name:
sodium phenylphosphinate
Test material form:
solid: particulate/powder
Specific details on test material used for the study:
Test Item
Designation in Test Facility: 19022101G
Date of Receipt: 21. Feb. 2019
Specification
The following information concerning identity and composition of the test item is provided by the sponsor.
Name Sodium phenylphosphinate
Batch no. 20190104
Appearance white powder
Composition approx. 98 % Sodium phenylphosphinate, approx. 1.5 % phenylphosphonic acid, approx. 0.5 % others
Purity 98%
Homogeneity homogeneous
Expiry date 15. Jan. 2020
Storage Room Temperature (20 ± 5°C)

The following additional information, also provided by the sponsor, is relevant to the con-duct of the study, according to OECD 439:
CAS No. 4297-95-4
EC-No. 224-305-5
Condition at Receipt Room temperature, in proper conditions
Stability in solvents H2O: not stated; EtOH: not stated; acetone: not stated; CH3CN: not stated; DMSO: not stated
Solubility H2O: not stated; EtOH: not stated; acetone: not stated; CH3CN: not stated; DMSO: not stated

Structural Formula/
SMILES Code not stated
Storage
The test item will be stored in the test facility in a closed vessel at room temperature (20±5°C).
Negative Control
“Dulbecco’s Phosphate Buffered Saline” (DPBS buffer without CaCl2 and without MgCl2).
Procured from MatTek In Vitro Life Science Laboratories and/or prepared by LAUS GmbH.
Composition:
KCl 0.2 g
KH2PO4 0.2 g
NaCl 8.0 g
Na2HPO4 * 7H2O 2.16 g
H2O ad 1 L
Note: different salts (crystal water) may be used; molar composition will be the same.
Positive Control
Sodium dodecyl sulphate, (SDS), CAS-No. 151-21-3, solution in H2O demin., concentra-tion 50 g/L.

In vitro test system

Test system:
human skin model
Remarks:
EpiDermTM procured by MatTek
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: MatTek In Vitro Life Science Laboratories, Mlynské Nivy 73, Bratislava - Slovakia
Source strain:
not specified
Vehicle:
unchanged (no vehicle)
Details on test system:
EpiDermTM tissues were procured from MatTek In Vitro Life Science Laboratories, Brati-slava.
Designation of the kit: EPI-200-SIT
Day of delivery: 02. Apr. 2019
The test system is a commercially available EpiDermTM-Kit, procured by MatTek.
The EpiDermTM tissue consists of human-derived epidermal keratinocytes which have been cultured to form a multi-layered, highly differentiated model of the human epidermis. It con-sists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers representing main lipid classes analogous to those found in vivo. The EpiDermTM tissues are cultured on spe-cially prepared cell culture inserts
Batch no.: 28690
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 μL
- Concentration (if solution):


NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 μL
- Concentration (if solution):

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 μL
- Concentration (if solution):5%
Duration of treatment / exposure:
35 minutes
Duration of post-treatment incubation (if applicable):
42 hours and 20 minutes
Number of replicates:
3

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Test consists of a topical exposure of the neat test item to RhE model followed by cell viability test. Cell viability is measured by dehydrogenase conversion of MTT into a blue formazan salt that is quantitatively measured after extraction from tissues.
Value:
87.2
Vehicle controls validity:
not specified
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
All validity criteria were met.
Values for negative control and for positive control were within the range of historical data of the test facility.
Therefore, the experiment is considered valid.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Therefore, the test item Sodium phenylphosphinate is considered
non-irritant to skin in the Reconstructed human Epidermis (RhE) Test Method.
Executive summary:

Three tissues of the human skin model EpiDermTM were treated with the test item for 60 minutes.

The test item was applied directly to each tissue and spread to match the tissue size (0.63 cm2; as indicated by the supplier).

DPBS-buffer was used as negative control and 5% SDS solution was used as positive con-trol.

After treatment with the negative control, the mean absorbance value was within the re-quired acceptability criterion of 0.8 ≤ mean OD ≤ 2.8, OD was 1.9.

The positive control showed clear irritating effects. The mean value of relative tissue viability was reduced to 2.6% (required: 20%).

The variation within the tissue replicates of negative control, positive control and test item was acceptable (required: ≤ 18%).

The mean value of relative tissue viability of the test item was reduced to 87.2% after the treatment. This value is above the threshold for skin irritation (50%). Therefore, the test item is considered as non-irritant to skin.

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