Registration Dossier

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
21 October 2015 - 26 October 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Version / remarks:
Variation between the three identically treated negative control tissues exceeded the ≤ 18% standard deviation limit as given in the study protocol. The standard deviation for the negative control group in this study was 20%. However as there was a high confidence in the results, ie there were no borderline results in the negative control, positive control or test item treated groups a repeat of the study was considered unnecessary.
Deviations:
yes
Remarks:
This deviation was considered not to affect the purpose or integrity of the study.
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Version / remarks:
Variation between the three identically treated negative control tissues exceeded the ≤ 18% standard deviation limit as given in the study protocol. The standard deviation for the negative control group in this study was 20%. However as there was a high confidence in the results, ie there were no borderline results in the negative control, positive control or test item treated groups a repeat of the study was considered unnecessary.
Deviations:
yes
Remarks:
This deviation was considered not to affect the purpose or integrity of the study.
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Liquid
Details on test material:
Name: DAILUBE IS-30
Chemical Name: 1, 3-bis (2, 4, 4-trimethylpentan-2yl) trisulfane
Use: Lubricant Additive
Molecular Formula: CH3-C(CH3)2-CH2-C(CH3)2-SSS-C(CH3)2-CH2-C(CH3)2-CH3
Molecular Weight: 322 g/mol
Appearance Pale yellow
Physical State: Liquid
Batch Number: L002
Purity: 95.4% by weight
Storage: Room temperature, in the dark
Expiry Date: 2016/8/15
Specific details on test material used for the study:
Batch: L002
Purity: 95.4%
Physical state / Appearance: Pale yellow liquid
Expiry date: 15 August 2016
Storage Conditions: Room temperature in the dark

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: The main basal, supra basal, spinous and granular layers and a functional stratum corneum.
Source strain:
not specified
Justification for test system used:
Reconstructed human epidermis model showed evidence of being a reliable and relevant stand-alone test for predicting rabbit skin irritation when the endpoint is measured by MTT reduction and for being used as a replacement for the Draize Skin Irritation Test for the purpose of distinguishing between Irritating and Non-Irritating test items.
Vehicle:
unchanged (no vehicle)
Details on test system:
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 °C
- Temperature of post-treatment incubation (if applicable): 37 °C
DYE BINDING METHOD
- Dye used in the dye-binding assay:MTT
- Spectrophotometer: microplate reader
- Wavelength: 562 nm
- Filter:without
- Filter bandwidth: na
NUMBER OF INDEPENDENT TESTING RUNS / EXPERIMENTS TO DERIVE FINAL PREDICTION:
PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be irritant to skin if relative mean tissue viability is ≤50%
- The test substance is considered to be non-irritant to skin if relative mean tissue viability is >50%
Control samples:
yes, concurrent negative control
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
Amount/concentration applied:
10 µL (26.3 µL/cm2): the test item
10 µL of DPBS: negative control
10 µL of SDS 5% w/v: positive control
Duration of treatment / exposure:
15 minutes
Duration of post-treatment incubation (if applicable):
42 hours
Number of replicates:
Triplicate tissues

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Main
Value:
82.6
Vehicle controls validity:
not applicable
Negative controls validity:
not valid
Remarks:
There were no borderline results in the negative control, positive control or test item treated groups a repeat of the study was considered unnecessary.
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
Mean OD562 Values and Percentage Viabilities for the Negative Control Item, Positive Control Item and Test Item (Table 1).

Any other information on results incl. tables

Table 1: Mean OD562 Values and Percentage Viabilities for the Negative Control Item, Positive Control Item and Test Item

Item

OD562of tissues

Mean OD562of triplicate tissues

±SD of OD562

Relative individual tissue viability (%)

Relative mean viability (%)

± SD of Relative mean viability (%)

Negative Control Item

0.871

0.702

0.146

124.1

100*

20.8

0.619

88.2

0.617

87.9

Positive Control Item

0.042

0.084

0.041

6.0

12.0

5.9

0.124

17.7

0.085

12.1

Test Item

0.642

0.580

0.056

91.5

82.6

8.0

0.564

80.3

0.534

76.1


OD=Optical Density

SD=       Standard deviation

*=        The mean viability of the negative control tissues is set at 100%

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
UN GHS Not classified for Irritation (category 3 can not be determined).
Conclusions:
The test item was classified as non-irritant.
The following classification criteria apply:
EU DSD & CLP Not classified for Irritation.
UN GHS Not classified for Irritation (category 3 can not be determined).