3,3'-(1,1,3,3-tetramethyldisiloxane-1,3-diyl)bispropylamine

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 November 2015 to 14 December 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Version / remarks:

2013

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

3,3'-(1,1,3,3-tetramethyldisiloxane-1,3-diyl)bispropylamine

EC Number:

219-588-7

EC Name:

3,3'-(1,1,3,3-tetramethyldisiloxane-1,3-diyl)bispropylamine

Cas Number:

2469-55-8

Molecular formula:

C10H28N2OSi2

IUPAC Name:

3,3'-(1,1,3,3-tetramethyldisiloxane-1,3-diyl)bispropylamine

Constituent 2

Reference substance name:

3,3’-(1,1,3,3-tetramethyldisiloxane-1,3-diyl)bispropylamine

IUPAC Name:

3,3’-(1,1,3,3-tetramethyldisiloxane-1,3-diyl)bispropylamine

Test material form:

other: liquid

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature
- Stability under test conditions: Unstable after repeated contact to air.
- Solubility and stability of the test substance in the solvent/vehicle: The test item was applied undiluted.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: not applicable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item was applied undiluted. No treatment of test material prior to testing was performed.
- Preliminary purification step (if any): None
- Final dilution of a dissolved solid, stock liquid or gel: not applicable
- Final preparation of a solid: not applicable

FORM AS APPLIED IN THE TEST (if different from that of starting material): not applicable

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: not specified

Source strain:

other: not applicable

Details on animal used as source of test system:

Not applicable

Justification for test system used:

EpiDermTM reconstructed human epidermis model consists of human keratynocytes and therefore represents in vitro the target organ of the species of interest and closely mimics the biochemical and physiological properties of the upper parts of the human, i.e. the epidermis.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDermTM Reconstructed Human Epidermis.
- Tissue batch number(s): 23306
- Production date: not specified
- Shipping date: not specified
- Delivery date: not specified
- Date of initiation of testing: 23 November 2015

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: After dosing, all tissues and plates were tranferred to the incubator for 35 min at 37°C. After 35 min, all plates were removed from the incubator and placed under the sterile flow until the 60 +/- 1 min incubation time of the first dosed tissue were over.
- Temperature of post-treatment incubation (if applicable): at 37°C for 24 +/- 2 hours.

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: After the 60 minutes incubation the tissues were washed intensively with DPBS.
- Observable damage in the tissue due to washing: None reported
- Modifications to validated SOP: not applicable

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 300 µL
- Incubation time: 3 h at 37°C
- Spectrophotometer: plate specrophotometer
- Wavelength: OD measured at 570 nm
- Filter: not specified
- Filter bandwidth: not specified
- Linear OD range of spectrophotometer: without reference wavelength

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: 1.179+/-0.156, acceptance criteria OD (540-570 nm) [1.0 - 3.0]
- Barrier function: 6.26, acceptance criteria ET-50 [4.77-8.72 hrs]
- Morphology: not specified
- Contamination: No contamination detected
- Reproducibility: not specified

NUMBER OF REPLICATE TISSUES: Triplicate

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Fresh tissues / killed tissues: Since the test item showed clear irritant potential, no correction was performed.
- Procedure used to prepare the killed tissues (if applicable): not applicable
- N. of replicates : not applicable
- Method of calculation used: not applicable

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: One

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be irritant to skin if the viability after 60 minutes exposure is less than 50%
- The test substance is considered to be non-irritant to skin if the viability after 60 minutes exposure is greater than or equal to 50%
- Justification for the selection of the cut-off point(s) if different than recommended in TG 431 and 439: not applicable

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Concentration (if solution): 30 µL

NEGATIVE CONTROL
- Concentration (if solution): 30 µL DPBS

POSITIVE CONTROL
- Concentration (if solution): 30 µL 5% SDS solution

Duration of treatment / exposure:

60 min

Duration of post-treatment incubation (if applicable):

42 hours

Number of replicates:

Triplicate

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 min treatment and 42 hours post-incubation

Value:

< 50

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: None damage reported
- Direct-MTT reduction: The mixture of 30 µL test item per 1 mL MTT medium showed reduction of MTT compared to the solvent. The mixture turned blue/purple.
- Colour interference with MTT: The mixture of 30µL of the test item per 300 µL aqua dest. showed no colouring detectable by unaided eye assessment.

DEMONSTRATION OF TECHNICAL PROFICIENCY:
- Mean absolute OD 570nm NC: Value 1.931, cut off 0.8 < NC < 2.8
- Relative Viability [%] PC: value 4.7, cut off =< 20%
- SD Viability [%]: value 0.6-14.8, cut off < 18%

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes
- Range of historical values if different from the ones specified in the test guideline: not applicable

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Conclusions:

In the in vitro skin irritation study for 3,3'-(1,1,3,3-tetramethyldisiloxane-1,3-diyl)bispropylamine using EpiDermTM, reported mean relative tissue viability (% negative control) of =< 50 % (5.3%) after 60 min treatment and 42 hours post incubation.