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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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:
27 November 2019 - 20 January 2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2020

Materials and methods

Test guideline
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)

Test material

Constituent 1
Reference substance name:
Extract from the seeds of Trigonella foenum-graecum (Fabaceae) obtained by extraction with polar solvents
EC Number:
950-727-0
Molecular formula:
Fenugreek extract
IUPAC Name:
Extract from the seeds of Trigonella foenum-graecum (Fabaceae) obtained by extraction with polar solvents
Specific details on test material used for the study:
Identification/Synonym FENUGREEK EXTRACT
Substance type UVCB substance
EC No. 950-727-0
Storage conditions room temperature, protected from light

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
foreskin from multiple donors
Source strain:
not specified
Details on animal used as source of test system:
Multi-layered, highly differentiated epidermis consisting of organized basal, spinous and granular layers, and a multilayered stratum corneum
Vehicle:
unchanged (no vehicle)
Details on test system:
SKIN DISC PREPARATION
EPISKIN™ - 0.38 cm2
- Quality control for skin discs: Histology scoring, magnitude of viability and barrier function (IC50 determination).

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37°C, 5 %CO2
- Temperature of post-treatment incubation: 37°C, 5 %CO2

REMOVAL OF TEST MATERIAL AND CONTROLS
- Washing: 25mL of sterile D-PBS

DYE BINDING METHOD
- Dye used in the dye-binding assay: MTT
- Wavelength: 595 nm

Study Acceptability Criteria
The assay was considered valid if the following criteria were met:
– Blank controls: mean OD value < 0.1.
– Negative controls: mean OD value ≥ 0.6 and ≤ 1.5, SD of % viability ≤ 18.
– Positive controls: mean viability expressed as percentage of the negative control
≤ 40% and SD of % viability ≤ 18.
– Test item: SD of % viability ≤ 18

Criteria Classification
Mean relative viability ≤ 50% UN GHS Category 2 or 1
Mean relative viability > 50% UN GHS No Category (for member states that do not adopt optional Category 3)
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied: 20 ± 2mg

VEHICLE
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:

NEGATIVE CONTROL
- Amount(s) applied: 20 µL
- Concentration: 5% D-PBS

POSITIVE CONTROL
- Amount(s) applied: 20 µL
- Concentration: 5% (w/v) SDS
Duration of treatment / exposure:
15 ± 0.5 minutes
Duration of post-treatment incubation (if applicable):
Recovery 42 ± 1 hour
MTT staining 3 hours
Number of replicates:
Live tissue 3 per conditions
Killed tissue at least 2 per conditions

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Live Tissue
Value:
94
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:
Killed Tissue
Value:
1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
not applicable
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
Study Acceptability Criteria
The assay was considered valid if the following criteria were met:
– Blank controls: mean OD value < 0.1.
– Negative controls: mean OD value ≥ 0.6 and ≤ 1.5, SD of % viability ≤ 18.
– Positive controls: mean viability expressed as percentage of the negative control
≤ 40% and SD of % viability ≤ 18.
– Test item: SD of % viability ≤ 18

Criteria Classification
Mean relative viability ≤ 50% UN GHS Category 2 or 1
Mean relative viability > 50% UN GHS No Category (for member states that do not adopt optional Category 3)

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The mean cell viability of the test item treated tissues, after the blank subtraction, was 94%. Based on the results obtained, the test item is classified as non-irritant to the skin (UN GHS No Category).