Phosphinate1030

Endpoint summary

Description of key information

The substance was tested for its skin irritant properties using the three-dimensional human skin model Episkin-SM. The study was performed according to OECD Guideline 439. 10 mg of the test item and 10 µL distilled water 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).

The substance was tested for its eye irritant properties in 3 New Zealand White rabbits.The study was performed according to OECD Guideline 405. Effects on conjunctivae (redness and swelling) were observed in all animals one hour after application. These signs decreased significantly within 72 hours and were fully reversible within 14 days. According to the findings in this study the substance does not meet the criteria for classification laid down in the EU Classification Labelling and Packaging Regulation (1272/2008/EC).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

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

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 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

Species:

other: EpiSkin reconstructed human epidermis model (SkinEthic Laboratories)

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

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).

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".

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Apr - May 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: charles River Deutschland GmbH, Kißlegg, Germany
- Weight at study initiation: 2.3 - 3.7 kg
- Housing: in separate cages
- Diet (e.g. ad libitum): ssniff K-H (V2333), ad libitum and hay (approx. 15 g daily)
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: 1 week under study conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 +/- 3 °C
- Humidity (%): 50 +/- 20 %
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES:
From: 27.04. To: 18.05.2004

Vehicle:

unchanged (no vehicle)

Controls:

other: The eye remained untreated and was used for control purposes.

Amount / concentration applied:

0.1 g

Duration of treatment / exposure:

24 h after administration the treated eyes were washed out with isotonic saline at approx. 37°C

Observation period (in vivo):

1 hour and 24, 48 and 72 hours as well as 7 and 14 days following treatment
At 24 and 72 hours and after 7 and 14 days, the eyes were further examined for corneal lesions under UV light after instillation of one drop of 0.01 % Fluorescein-sodium

Number of animals or in vitro replicates:

3

Details on study design:

Test Item Administration:
The test item was applied as a weight of 0.1 g/animal, the dose specified in the test guidelines for solid test items.

An initial test was performed using one animall.

As neither serious eye damage nor severe effect were observed in the initial test a confirmatory test with two further animals was performed.

About 24 h before the stert of the study both eyes of all animals were examined under UV light for corneal lesions after instillation of one drop of a 0.01% fluorescein-sodium solution. Only animals without ocular abnormalies were used for the study. 0.1 g of the test item was administered once to the conjuntival sac of the left eye of three rabbits. In each case the untreated eye served as control.

24 h after administration the treated eyes were washed out thoroughly with isotonic saline at approx. 37°C. The eyes were also washed out at designated examination times at which discharge was observed or a corneal examination with fluorescein was performed.

Rationale: No study on eye irritation was available. The test item is not classified as corrosive or irritating to skin.

Observations

Viability / Mortality: Daily
Clinical Signs: Daily
Eye Reactions: See below
Body Weights: At start of acclimatization, on test day 1 and at termination of observation.

Assessment of Eye Reactions:

The eye reactions were assessed according to the numerical scoring system listed in the Commission Regulation (EC) No 440/2008, B.5, at approximately 1, 24, 48 and 72 hours, as well as 7 days after administration. Scleral reddening and ocular discharge were also assessed. Eye examinations were made with a Varta Cliptrix diagnostic-lamp (Roth AG, 4153 Reinach / Switzerland)

Data was summarized in tabular form, showing for each animal the irritation scores for the designated observation time, a description of the degree and nature of irritation, the presence of serious lesions and non-ocular effects. The scores of each animal at the following reading times (24, 48, 72 hours) were used in calculating the respective mean values (with the exception of the sclerae) for each type of lesion.

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

1.44

Max. score:

3

Reversibility:

fully reversible within: 14 days

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.22

Max. score:

4

Reversibility:

not fully reversible within: 48 h

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritant / corrosive response data:

From one hour up to 7 days after administration two animals showed some definitely injected up to crimson colored blood vessels. Slight swelling with partial up to half closed eyelids was observed till 24 hour after application. The third animal showed only one hour up to 24 hours after treatment some hyperaemic up to crimson colored vessels in the conjunctiva as well as obvious swelling with partial eversion of lids one hour after application. Additionally, serous colorless eye discharge was noted one hour after administration in all animals.
14 days after administration all signs of irritation had disappeared.

Other effects:

No clinical signs of systemic toxicity were observed.

Interpretation of results:

other: not irritating

Conclusions:

According to the findings in this study the substance does not meet the criteria for classification laid down in the EU Classification Labelling and Packaging Regulation (1272/2008/EC).

Executive summary:

The substance was tested for its eye irritant properties in 3 New Zealand White rabbits. The study was performed according to OECD Guideline 405. Effects on conjunctivae (redness and swelling) were observed in all animals one hour after application. These signs decreased significantly within 72 hours and were fully reversible within 14 days.

According to the findings in this study the substance does not meet the criteria for classification laid down in the EU Classification Labelling and Packaging Regulation (1272/2008/EC).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation: 1 key study available: not irritating

Eye irritation: 1 key study available: not irritating

 

There is one fully reliable study available on the skin irritancy potential. The substance was tested for its skin irritant properties using the three-dimensional human skin model Episkin-SM. The study was performed according to OECD Guideline 439. 10 mg of the test item and 10 µL distilled water 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). In an additionally performed skin corrosivity assay using the Epiderm model (according to OECD Guideline 431) the substance proved to be non corrosive.

There is one fully reliable study available on the eye irritancy potential. The substance was tested for its eye irritant properties in 3 New Zealand White rabbits. The study was performed according to OECD Guideline 405. Effects on conjunctivae (redness and swelling) were observed in all animals one hour after application. These signs decreased significantly within 72 hours and were fully reversible within 14 days.

Justification for classification or non-classification

With reference to the reported results of an in vitro skin irritation study the substance has not to be classified as irritant to the skin according to the criteria laid down in the EU Classification Labelling and Packaging Regulation (1272/2008/EC).

With reference the reported scores and the reversibility of the observed effects the substance does not have to be classified as irritant to the eyes according to the criteria laid down in the EU Classification Labelling and Packaging Regulation (1272/2008/EC).