Phosphinate1030

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Nov 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

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

Test material information

Constituent 1

Test material form:

other: solid

In vitro test system

Test system:

human skin model

Source species:

human

Details on animal used as source of test system:

n.a.

Justification for test system used:

This test uses the EPISKIN-SM™ reconstructed human epidermis model (SkinEthic) which consists of normal human epidermal keratinocytes (NHEK) 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 skin, i.e. the epidermis.

Vehicle:

unchanged (no vehicle)

Details on test system:

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 +/- 1°C
- Temperature of post-treatment incubation (if applicable): 37 +/- 1°C
REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps: 1
DYE BINDING METHOD
- Dye used in the dye-binding assay: MTT
- Wavelength: 550 nm
NUMBER OF INDEPENDENT TESTING RUNS / EXPERIMENTS TO DERIVE FINAL PREDICTION: 3
-The test item is considered to be irritant to skin in accordance with regulation EC 1272/2008 and UN GHS “Category 2” [16], if the tissue viability after 15 min of exposure and 42 h of post-incubation is less or equal to 50%. The test substance may be considered as non-irritant to skin in accordance with UN GHS “No Category” if the tissue viability after exposure and post-treatment incubation is higher than 50%.
Negative control: PBS
Positive control: 5% SDS
:

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 mg

VEHICLE
- Amount(s) applied (volume or weight with unit): moistened with 10 µL distilled water

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 10 µL PBS

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 10 µL SDS solution
- Concentration (if solution): 5%

Duration of treatment / exposure:

15 min

Duration of post-treatment incubation (if applicable):

42 +/- 1 h

Number of replicates:

3

Test animals

Species:

other: EpiSkin reconstructed human epidermis model (SkinEthic Laboratories)

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

mean of three replicates

Value:

102.3

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: none
- Direct-MTT reduction: none
- Colour interference with MTT: none

DEMONSTRATION OF TECHNICAL PROFICIENCY:

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

In vivo

Irritant / corrosive response data:

The test item showed no irritant effects. The mean relative tissue viability (% negative control) was > 50% (102.3%) after 15 min. treatment and 42 h post incubation.
The controls confirmed the validity of the study. The mean OD550 of the six blank values was < 0.1. The mean absolute OD550 of the three negative control tissues was ≥ 0.6 and ≤ 1.5. The mean relative tissue viability (% negative control) of the positive control was ≤ 40% (6.1%). The maximum standard deviation of viability of replicate tissues of all dose groups was < 18% (1.7% - 6.6%).



If mean tissue viability is > 50% relative to the mean negative control, the test item is classified as non-irritant (EU CLP and UN GHS: No Category).
If mean tissue viability is <= 50% relative to the mean negative control, the test item is classified as irritant (EU CLP and UN GHS: Category 2).
In the in vitro skin irritation test using the EpiSkin human epidermis model 10 mg test item + 5 µL A. dest. were applied topically for 15 minutes. After 42 h post-incubation cytotoxic effects were determined via MTT reduction assay.
The mean relative tissue viability (% negative control) was > 50%.
The test item is therefore classified as "non-irritant" (EU CLP and UN GHS: No Category).

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: OECD 439

Conclusions:

No irritant effects of the substance were observed in this in vitro-study, therefore the test item is classified as "non-irritant" in accordance with UN GHS and EU CLP "No Category".

Executive summary:

The skin irritation potential of the substance was assessed in an in vitro test using the EPISKIN-Standard Model. Under the given test conditions, the substance showed no irritant effects. The relative mean tissue viability after 15 min. of exposure and 42 h post incubation was >= 50 %. The substance is therefore classified as "non-irritant" in accordance with UN GHS and EU-CLP "No Category".