Ethyltriphenylphosphonium acetate

Endpoint summary

Description of key information

not irritating to skin: OECD TG 431: relative tissue viability 71% (3 min exposure), 21.9% (60 min exposure)

OECD TG 439: relative tiíssue viability 100.6%

irreversible effects to the eye: OECD TG 437, IVIS 79.1

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28 February 2018 - 09 March 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Version / remarks:

29 July 2016

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

The test item was ground to a fine powder before use.

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Justification for test system used:

recommended by guideline

Vehicle:

unchanged (no vehicle)

Remarks:

25 µL of sterile water was added for wetting of the test item to increase tissue surface contact.

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™ Reconstructed Human Epidermis Model Kit
- Tissue batch number(s): 25884
- Delivery date: 06 March 2018

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 °C
- Temperature of post-treatment incubation (if applicable): 37 °C, 5% CO2

REMOVAL OF TEST MATERIAL AND CONTROLS
- Rinsing was achieved by filling and emptying each tissue under a constant soft stream of DPBS to gently remove any residual test item. Excess DPBS was removed by blotting the bottom of the tissue insert with tissue paper and the tissue surface was swabbed. The rinsing procedure was then repeated once more to ensure the tissues were completely decontaminated.
- Observable damage in the tissue due to washing: no


MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg/mL
- Incubation time: 3 hours
- Spectrophotometer: Labtech LT-4500 microplate reader

NUMBER OF REPLICATE TISSUES: duplicates

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- killed tissues
- Procedure used to prepare the killed tissues (if applicable): freeze killed
- N. of replicates : duplicates
- Method of calculation used: True viability = mean OD tvt-(OD tkt-OD ukt)

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50%, or if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is less than 15%.
- The test substance is considered to be non-corrosive to skin if the viability after 3 minutes exposure is greater than or equal to 50% and the viability after 1 hour exposure is greater than or equal to 15%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 mg; 25 µL of sterile water was added for wetting of the test item to increase tissue surface contact.


NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL of sterile distilled water


POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL of 8.0 N Potassium Hydroxide

Duration of treatment / exposure:

3 min, 60 min

Duration of post-treatment incubation (if applicable):

3 h

Number of replicates:

2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 minute exposure

Value:

71

Negative controls validity:

valid

Positive controls validity:

valid

Remarks:

4.7% tissue viability

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 min exposure

Value:

21.9

Negative controls validity:

valid

Positive controls validity:

valid

Remarks:

4.1% tissue viability

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: no
- Direct-MTT reduction: The MTT solution containing the test item did not turn blue/purple. This was taken to indicate the test item did not reduce MTT.
- Colour interference with MTT: The solution containing the test item did not become colored. This was taken to indicate the test item did not have the potential to cause color interference.

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD570 for the negative control treated tissues was 1.750 for the 3 minute exposure period and 1.877 for the 60 minute exposure period. The negative control acceptance criteria were therefore satisfied.
- Acceptance criteria met for positive control: The relative mean tissue viability for the positive control treated tissues was 4.1% relative to the negative control following the 60 minute exposure period. The positive control acceptance criterion was therefore satisfied.
- Acceptance criteria met for variability between replicate measurements: In the range 20 to 100% viability the Coefficient of Variation between the two tissue replicates of each treatment group did not exceed 30%. The acceptance criterion was therefore satisfied

Tissue	Exposure Period	MeanOD570of individual tissues	Mean OD570of duplicate tissues	Standard Deviation	Coefficient of Variation (%)	Relative Mean Viability (%)
Negative Control	3 Minutes	1.672	1.750	0.110	6.3	100
		1.827
	60 Minutes	1.756	1.877	0.171	9.1
		1.998
Positive Control	3 Minutes	0.096	0.083	0.018	na	4.7
		0.070
	60 Minutes	0.066	0.078	0.016	na	4.1
		0.089
Test Item	3 Minutes	1.438	1.242	0.277	22.3	71.0
		1.046
	60 Minutes	0.375	0.412	0.052	12.7	21.9
		0.449

 

Interpretation of results:

other: not corrosive

Conclusions:

The test item was considered to be non-corrosive to the skin.

Executive summary:

The purpose of this test is to evaluate the corrosivity potential of ETPPAAc using the EpiDerm™ Human Skin Model after treatment periods of 3 and 60 minutes. 

Corrosion is directly related to cytotoxicity in the EpiDerm™ tissue.  Cytotoxicity is determined by the reduction of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to formazan by viable cells in the test item treated tissues relative to the corresponding negative control.  The results are used to make a prediction of the corrosivity potential of the test item. 

Duplicate tissues were treated with the test item for exposure periods of 3 and 60 minutes.  Negative and positive control groups were treated for each exposure period.    At the end of the exposure period the test item was rinsed from each tissue before each tissue was taken for MTTloading.  After MTT loading each tissue was placed in 2 mL Isopropanol for MTT extraction.  

At the end of the formazan extraction period each well was mixed thoroughly and triplicate 200 [Symbol]L samples were transferred to the appropriate wells of a pre-labeled 96well plate.  The optical density (OD) was measured at 570 nm (OD₅₇₀). 

Data are presented in the form of percentage viability (MTT reduction in the test item treated tissues relative to negative control tissues). 

 

The relative mean viabilities for each treatment group were as follows: 

Exposure Period	Percentage Viability
	Negative Control	Positive Control	Test Item
3 minute	100*	4.7	71.0
60 minute	100*	4.1	21.9

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

 

The quality criteria required for acceptance of results in the test were satisfied. 

The test item was considered to be non-corrosive to the skin. 

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 April 2018 - 16 April 2018

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)

Version / remarks:

updated 28 July 2015

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Justification for test system used:

as recommended in OECD guideline 439

Vehicle:

unchanged (no vehicle)

Remarks:

5 µL of sterile distilled water was topically applied to the epidermal surface in order to improve contact between the test item and the epidermis

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN
- Tissue batch number(s): 18-EKIN-015
- Delivery date: 10 April 2018

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature
- Temperature of post-treatment incubation (if applicable): 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
- Rinsing was achieved by filling and emptying each tissue insert for approximately 40 seconds using a constant soft stream of DPBS to gently remove any residual test item.
- Observable damage in the tissue due to washing: no

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3 mg/mL
- Incubation time: 3 h
- Spectrophotometer: Labtech LT 4500 microplate reader
- Wavelength: 570 nm (without a reference filter)

NUMBER OF REPLICATE TISSUES: 3

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

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; 5 µL of sterile distilled water was topically applied to the epidermal surface in order to improve contact between the test item and the epidermis

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

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 10 µL of SDS 5% w/v

Duration of treatment / exposure:

15 minutes

Duration of post-treatment incubation (if applicable):

42 h

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Value:

100.6

Negative controls validity:

valid

Positive controls validity:

valid

Remarks:

3.7% tissue viability

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: no
- Direct-MTT reduction: The MTT solution containing the test item did not turn blue or purple which indicated that the test item did not directly reduce MTT.
- Colour interference with MTT: The solution containing the test item was an off-white color. This color was attributed to the intrinsic color of the test item itself. It was therefore unnecessary to run color correction tissues. After rinsing, no residual test item or staining was remaining on the tissues confirming no color interference had occurred.

It was considered unnecessary to perform IL-1alpha analysis as the results of the MTT test were unequivocal.

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD570 for the negative control treated tissues was 0.989 and the standard deviation value of the viability was 8.3%. The negative control acceptance criteria were therefore satisfied.
- Acceptance criteria met for positive control: The relative mean tissue viability for the positive control treated tissues was 3.7% relative to the negative control treated tissues and the standard deviation value of the viability was 1.0%. The positive control acceptance criteria were therefore satisfied.
- Acceptance criteria met for variability between replicate measurements: The standard deviation calculated from individual tissue viabilities of the three identically test item treated tissues was 6.1%. The test item acceptance criterion was therefore satisfied.

Item	OD570of tissues	Mean OD570of triplicate tissues	± SD of OD570	Relative individual tissue viability (%)	Relative mean viability (%)	± SD of Relative mean viability (%)
Negative Control Item	0.932	0.989	0.082	94.2	100	8.3
	1.083			109.5
	0.951			96.2
Positive Control Item	0.043	0.037	0.010	4.3	3.7	1.0
	0.042			4.2
	0.025			2.5
Test Item	0.962	0.995	0.061	97.3	100.6	6.1
	1.065			107.7
	0.958			96.9

Interpretation of results:

GHS criteria not met

Conclusions:

The test item was classified as non-irritant. 

Executive summary:

The purpose of this test was to evaluate the skin irritation potential of ETPPAAc using the EPISKIN^TMreconstructed human epidermis model after a treatment period of 15 minutes followed by a post‑exposure incubation period of 42 hours. The principle of the assay was based on the measurement of cytotoxicity in reconstructed human epidermal cultures following topical exposure to the test item by means of the colorimetric MTT reduction assay. Cell viability is measured by enzymatic reduction of the yellow MTT tetrazolium salt (3‑[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to a blue/purple formazan salt (within the mitochondria of viable cells) in the test item treated tissues relative to the negative controls. 

Triplicate tissues were treated with the test item for an exposure period of 15 minutes. At the end of the exposure period each tissue was rinsed before incubating for 42 hours. At the end of the post‑exposure incubation period each tissue was taken for MTT-loading. The maintenance medium from beneath each tissue was transferred to pre‑labeled micro tubes and stored in a freezer for possible inflammatory mediator determination. After MTT-loading a total biopsy of each epidermis was made and placed into micro tubes containing acidified isopropanol for extraction of formazan crystals out of the MTT‑loaded tissues. 

At the end of the formazan extraction period each tube was mixed thoroughly and duplicate 200 µL samples were transferred to the appropriate wells of a pre‑labeled 96‑well plate. The optical density was measured at 570 nm.

Data are presented in the form of percentage viability (MTT reduction in the test item treated tissues relative to negative control tissues).

The relative mean viability of the test item treated tissues was100.6% after the 15‑Minute exposure period and 42‑Hours post‑exposure incubation period.

The quality criteria required for acceptance of results in the test were satisfied.

The test item was classified as non-irritant. 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15 March 2018 - 15 March 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

cattle

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
Eyes from adult cattle (typically 12 to 60 months old) were obtained from a local abattoir as a by-product from freshly slaughtered animals. The eyes were excised by an abattoir employee after slaughter, and were placed in Hanks’ Balanced Salt Solution (HBSS) supplemented with antibiotics (penicillin at 100 IU/mL and streptomycin at 100 µg/mL). They were transported to the test facility over ice packs on the same day of slaughter. The corneas were refrigerated on arrival and used within 24 hours of receipt.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL

Duration of treatment / exposure:

10 min

Duration of post- treatment incubation (in vitro):

240 min

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
All eyes were macroscopically examined before and after dissection. Only corneas free of damage were used.
The cornea from each selected eye was removed leaving a 2 to 3 mm rim of sclera to facilitate handling. The iris and lens were peeled away from the cornea. The isolated corneas were immersed in a dish containing HBSS until they were mounted in Bovine Corneal Opacity and Permeability (BCOP) holders.
The anterior and posterior chambers of each BCOP holder were filled with complete Eagle’s Minimum Essential Medium (EMEM) without phenol red and plugged. The holders were incubated at 32 ± 1 ºC for 60 minutes. At the end of the incubation period each cornea was examined for defects. Only corneas free of damage were used.
A pre treatment opacity reading was taken for each cornea using a calibrated opacitometer. The average opacity for all corneas was calculated.
Three corneas were randomly allocated to the negative control. Three corneas were also allocated to the test item and three corneas to the positive control item.

NUMBER OF REPLICATES
3

NEGATIVE CONTROL USED
Sodium chloride 0.9% w/v

POSITIVE CONTROL USED
Imidazole

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period:
rinsed 3 times with fresh complete EMEM containing phenol red before a final rinse with complete EMEM without phenol red

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: calibrated opacitometer
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of microtiter plate reader (OD492)
- Others (e.g, pertinent visual observations, histopathology): No histopathology was required since a definitive result was achieved.

SCORING SYSTEM: In Vitro Irritancy Score (IVIS):
In Vitro Irritancy Score = mean opacity value + (15 x mean permeability OD492 value)

DECISION CRITERIA:
IVIS UN GHS EU CLP
≤ 3 No Category Not classified for irritation
>3; ≤ 55 No prediction can be made No prediction can be made
> 55 Category 1 Category 1
H318: Causes serious eye damage

Irritation parameter:

in vitro irritation score

Value:

79.4

Negative controls validity:

valid

Remarks:

IVIS 1.2

Positive controls validity:

valid

Remarks:

IVIS 108.0

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: The corneas treated with the test item were cloudy post treatment. The corneas treated with the negative control item were clear post treatment. The corneas treated with the positive control item were cloudy post treatment.

DEMONSTRATION OF TECHNICAL PROFICIENCY:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The negative control gave opacity of ≤2.4 and permeability ≤0.072. The negative control acceptance criteria were therefore satisfied.
- Acceptance criteria met for positive control: The positive control In Vitro Irritancy Score was within the range of 65.1 to 123.3. The positive control acceptance criterion was therefore satisfied.

Individual and Mean Corneal Opacity and Permeability Measurements

Treatment	Cornea Number	Opacity				Permeability (OD492)		In Vitro Irritancy Score
		Pre-Treatment	Post-Treatment	Post-Treatment-Pre‑Treatment	Corrected Value		Corrected Value
Negative Control	2	4	4	0		0.017
	3	3	4	1		0.013
	6	3	5	2		0.005
				1.0		0.012		1.2
Positive Control	7	4	92	88	87.0	2.130	2.118
	10	4	78	74	73.0	2.260	2.248
	11	3	76	73	72.0	1.785	1.773
					77.3		2.047	108.0
Test Item	12	3	73	70	69.0	0.011	0.000
	13	3	90	87	86.0	0.006	0.000
	14	4	88	84	83.0	0.019	0.007
					79.3		0.002	79.4

 

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

in this study ETPPAAc caused irreversible effects to the eye.

Executive summary:

The purpose of this test was to identify test items that can induce serious eye damage and to identify test items not requiring classification for eye irritation or serious eye damage. The Bovine Corneal Opacity and Permeability (BCOP) test method is an organotypic model that provides short‑term maintenance of normal physiological and biochemical function of the bovine cornea in vitro. In this test method, damage by the test item ETPPAAc is assessed by quantitative measurements of changes in corneal opacity and permeability.

The test method can correctly identify test items (both chemicals and mixtures) inducing serious eye damage as well as those not requiring classification for eye irritation or serious eye damage, as defined by the United Nations (UN) Globally Harmonized System of Classification and Labelling of Items (GHS) and EU Classification, Labelling and Packaging (CLP) of chemicals (Regulation (EC) No 1272/2008), and it was therefore endorsed as scientifically valid for both purposes. Test items inducing serious eye damage are classified as UN GHS and EU CLP Category 1. Items not classified for eye irritation or serious eye damage are defined as those that do not meet the requirements for classification as UN GHS/EU CLP Category 1 or 2 (2A or 2B), i.e. they are referred to as UN GHS/EU CLP No Category.

The test item was applied at a concentration of 20% w/v in sodium chloride 0.9% w/v for 240 minutes. Negative and positive control items were tested concurrently. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate an In Vitro Irritancy Score (IVIS). 

 

The In Vitro irritancy scores are summarized as follows:

Treatment	In VitroIrritancy Score
Test Item	79.4
Negative Control	1.2
Positive Control	108.0

 

The test item is classified as Category 1. UN GHS or EU CLP Causes serious eye damage.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin corrosion

The purpose of this test is to evaluate the corrosivity potential of ETPPAAc using the EpiDerm™ Human Skin Model after treatment periods of 3 and 60 minutes. 

Corrosion is directly related to cytotoxicity in the EpiDerm™ tissue.  Cytotoxicity is determined by the reduction of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to formazan by viable cells in the test item treated tissues relative to the corresponding negative control.  The results are used to make a prediction of the corrosivity potential of the test item. 

Duplicate tissues were treated with the test item for exposure periods of 3 and 60 minutes.  Negative and positive control groups were treated for each exposure period.    At the end of the exposure period the test item was rinsed from each tissue before each tissue was taken for MTTloading.  After MTT loading each tissue was placed in 2 mL Isopropanol for MTT extraction.  

At the end of the formazan extraction period each well was mixed thoroughly and triplicate 200 [Symbol]L samples were transferred to the appropriate wells of a pre-labeled 96well plate.  The optical density (OD) was measured at 570 nm (OD570). 

Data are presented in the form of percentage viability (MTT reduction in the test item treated tissues relative to negative control tissues). 

The relative mean viabilities for each treatment group were 71.0% (test item) and 4.7% (positive control) for the 3 minute exposure and 21.9% (test item) and 4.1% (positive control) for the 60 minute exposure. The mean viability of the negative control tissues is set at 100% 

The quality criteria required for acceptance of results in the test were satisfied. 

The test item was considered to be non-corrosive to the skin. 

 

Skin irritation

The purpose of this test was to evaluate the skin irritation potential of ETPPAAc using the EPISKINTM reconstructed human epidermis model after a treatment period of 15 minutes followed by a post‑exposure incubation period of 42 hours. The principle of the assay was based on the measurement of cytotoxicity in reconstructed human epidermal cultures following topical exposure to the test item by means of the colorimetric MTT reduction assay. Cell viability is measured by enzymatic reduction of the yellow MTT tetrazolium salt (3‑[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to a blue/purple formazan salt (within the mitochondria of viable cells) in the test item treated tissues relative to the negative controls. 

Triplicate tissues were treated with the test item for an exposure period of 15 minutes. At the end of the exposure period each tissue was rinsed before incubating for 42 hours. At the end of the post‑exposure incubation period each tissue was taken for MTT-loading. The maintenance medium from beneath each tissue was transferred to pre‑labeled micro tubes and stored in a freezer for possible inflammatory mediator determination. After MTT-loading a total biopsy of each epidermis was made and placed into micro tubes containing acidified isopropanol for extraction of formazan crystals out of the MTT‑loaded tissues. 

At the end of the formazan extraction period each tube was mixed thoroughly and duplicate 200 µL samples were transferred to the appropriate wells of a pre‑labeled 96‑well plate. The optical density was measured at 570 nm.

Data are presented in the form of percentage viability (MTT reduction in the test item treated tissues relative to negative control tissues).

The relative mean viability of the test item treated tissues was100.6% after the 15‑Minute exposure period and 42‑Hours post‑exposure incubation period.

The quality criteria required for acceptance of results in the test were satisfied.

The test item was classified as non-irritant. 

 

Eye irritation

The purpose of this test was to identify test items that can induce serious eye damage and to identify test items not requiring classification for eye irritation or serious eye damage. The Bovine Corneal Opacity and Permeability (BCOP) test method is an organotypic model that provides short‑term maintenance of normal physiological and biochemical function of the bovine corneain vitro. In this test method, damage by the test item is assessed by quantitative measurements of changes in corneal opacity and permeability.

The test item ETPPAAc was applied at a concentration of 20% w/v in sodium chloride 0.9% w/v for 240 minutes. Negative and positive control items were tested concurrently. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate an In Vitro Irritancy Score (IVIS).

The In Vitro irritancy scores were 79.1 (test item), 1.2 (negative control) and 108.0 (positive control). The test item is classified as Category 1 (UN GHS or EU CLP Causes serious eye damage).

Justification for classification or non-classification

Skin irritation

Based on the available data, the test substance ethyltriphenyl phosphonium acetate does not need to be classified for skin irritation/corrosion according to regulation (EC) 1272/2008.

 

Eye irritation

Based on the available data, the test substance ethyltriphenyl phosphonium acetate has to be classified for eye irritation according to regulation (EC) 1272/2008 as Category 1 and should be labelled with H318 "Causes serious eye damage" .