Registration Dossier

Toxicological information

Skin irritation / corrosion

Currently viewing:

Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Study carried out between 09 February and 21 February 2018.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study performed according to OECD test guideline No. 431 and in compliance with GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Version / remarks:
29 July 2016
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Remarks:
French GLP Compliance Programme for chemical products (inspected on 30-31 January 2017 / signed on 27 April 2017)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Remarks:
Brown-yellow viscous liquid
Details on test material:
Test item: Fenugreek Absolute
Batch No.: 2709379
Identity: 100 % UVCB Substance
Appearance: Brown-yellow viscous liquid
Storage conditions: Ambient temperature (10 °C to 30 °C), in original aluminium bottle, dark
Expiry date: October 24, 2018
Specific details on test material used for the study:
Test item was identified under the following code number: PH-17/0759

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: Not precised
Source strain:
other: Not applicable
Details on animal used as source of test system:
Not applicable
Justification for test system used:
Following the REACH bottom-up strategy, as the result of the OECD test guideline No. 439 was positive, the epiCS® Reconstructed Human Epidermis Model method was used to assess skin corrosion as recommended in the OECD test guideline No. 431.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
The test item was applied, as supplied, at the dose of 50 µL to 2 living and 4 killed Human skin model surfaces during 3 minutes at room temperature and during 1 hour at 37°C ± 1°C, 5% ± 1% CO2.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
50 µL of test item
- Concentration (if solution):
Not applicable: undiluted

VEHICLE
not applicable

NEGATIVE CONTROL
- Amount(s) applied (volume or weight):
50 µL of distilled water
- Concentration (if solution):
Not applicable

POSITIVE CONTROL
- Amount(s) applied (volume or weight):
50 µL of potassium hydroxide 8N (8N KOH)
- Concentration (if solution): Not applicable
Duration of treatment / exposure:
3 min at room temperature or 1 hour at 37°C ± 1°C, 5% ± 1% CO2
Duration of post-treatment incubation (if applicable):
not applicable
Number of replicates:
2 tissues per condition
Triplicate measurement of OD570 for each tissue.

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3 min
Value:
73.28
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
The mean OD of negative control tissues after 3 minutes exposure was 0.691(between 0.233 and 0.933)
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1 hour
Value:
101.43
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
The mean OD of negative control tissues after 3 minutes exposure was 0.771 (between 0.233 and 0.933)
Positive controls validity:
valid
Remarks on result:
no indication of irritation

Any other information on results incl. tables

Table 7.3.1/1: Individual and average values after 3 minutes exposure

 

Skin

OD

Mean OD / disc (#)

Mean OD / product

Viability %

Mean viability %

SD

Negative control

1

0.669

0.688

0.701

0.686

0.691

99.35

100.0

1.3

2

0.715

0.652

0.717

0.695

100.65

Positive control

3

0.025

0.025

0.024

0.025

0.042

3.62

6.08

4.9

4

0.059

0.058

0.059

0.059

8.54

Test item

13

0.418

0.403

0.415

0.412

0.506

59.67

73.28

27.2

14

0.603

0.600

0.596

0.600

86.89

#: mean of 3 values

OD: optical density

Table 7.3.1/2: Individual and average values after 1 hour exposure

 

Skin

OD

Mean OD / disc (#)

Mean OD / product

Viability %

Mean viability %

SD

Negative control

15

0.856

0.834

0.835

0.892

0.771

109.28

100.0

18.6

16

0.710

0.676

0.711

0.699

90.72

Positive control

17

0.004

0.004

0.004

0.004

0.003

0.52

0.39

0.3

18

0.002

0.002

0.002

0.002

0.26

Test item

27

0.661

0.627

0.626

0.638

0.782

82.80

101.43

37.2

28

0.961

0.907

0.906

0.925

120.05

#: mean of 3 values

OD: optical density

Table 7.3.1/3: Conclusion

Mean viability (%)

3 min exposure

1 hour exposure

Conclusion

Positive control

6.08

0.39

Corrosive sub-category 1B/1C

Test item

73.28

101.43

Non Corrosive

Applicant's summary and conclusion

Interpretation of results:
other: non-corrosive
Conclusions:
In accordance with the Regulation (EC) No. 1272/2008, the results obtained under these experimental conditions enable to conclude that the test item FENUGREEK ABSOLUTE does not have to be
classified in Category 1 “Corrosive”. The hazard statement “H314: Causes severe skin burns and eye damage” with the signal word “Danger” are not required.
Executive summary:

The aim of the study was to evaluate the possible corrosive effects of the test item after topical administration on in vitro human reconstituted epidermis (epiCS®, CellSystems®).

The test item FENUGREEK ABSOLUTE was applied as supplied, at the dose of 50 μL, to 2 living Human skin model surfaces (epiCS®, CellSystems®) during 3 minutes and 1 hour. The application was

followed by a rinse with 20 mL of DPBS. Cell viability was then measured by enzymatic conversion of the vital dye MTT into a blue formazan salt that was quantitatively measured after extraction from

tissues. The experimental protocol was established in accordance with the O.E.C.D. Test Guideline No. 431 dated 28 July 2015 and the method B.40bis of the Council regulation No. 440/2008.

3 minutes and 1 hour after the test item application, the mean percent viability of the epidermis skins treated with the test item were 73.28% and 101.43% (considered as 100%) versus 6.03% and

0.39%, respectively; with the positive control item (potassium hydroxide 8N).

In accordance with the Regulation (EC) No. 1272/2008, the results obtained under these experimental conditions enable to conclude that the test item FENUGREEK ABSOLUTE does not have to be

classified in Category 1 “Corrosive”. The hazard statement “H314: Causes severe skin burns and eye damage” with the signal word “Danger” are not required.