Phosphonic acid, P-2-propen-1-yl-, diethyl ester

Endpoint summary

Description of key information

The test substance is non-corrosive in the EPIDERM Human Skin Model.
The test substance is non-irritant in the EPISKIN reconstructed human epidermal model.
The outcome of the BCOP test with the substance shows that no prediction on the eye irritation of the substance can be made.

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:

13 Jan 2015 to 19 Jan 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: study performed according to the guideline and under GLP

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Principles of method if other than guideline:

exposure period 15 minutes with post-exposure incubation period of 42 hours

GLP compliance:

yes (incl. QA statement)

Details on study design:

The EpiSKIN assay uses reconstructed human epidermis to assess skin irritation. For the viability test enzymatic conversion of MTT to formazan is used.

In a pre-test direct interaction of the test substance with the detection chemical MTT is assessed. No interference was determined.

In the main study triplicate tissues were treated with 10 uL of the test substance during 15 minutes. In addition triplicate tissues were treated with a negative control (DPBS) and a positive control (5% SDS). After treatment tissues were rinsed, incubated in maintenance fluid for 42 hours at 37 °C (5% CO2) and treated with MTT during 3 hours (at 37 °C/5% CO2). After MTT loading a total biopsy of each epidermis (epidermis and collagen) was made, that was suspended in acidified isopropanol for 3 days and kept in the refrigerator at 1-10°C. Thereafter the optical density of the extracted MTT solutions was measured at 562 nm. Viability was expressed as percentage MTT conversion versus negative control
Tissue viability > 50% means non-irritant
Tissue viability ≤ 50% means irritant

Other effects:

The test substance did not interfere with the reduction of MTT to blue formazan salt

	Mean OD562± SD	Relative mean viability ± SD
Negative control:	0.827 ± 0.02	100 ± 2.9%
Test substance:	0.822 ± 0.05	99.4 ± 6.3%
SDS:	0.243 ± 0.06	29.4 ± 7.4%

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: other: tissue viability > 50%: non-irritant

Conclusions:

The test substance is non-irritant in the EPISKIN reconstructed human epidermal model.

Executive summary:

In this assay reconstructed human skin tissues were treated with 10 uL of the test substance in triplicate during 15 minutes. After treatment tissues were rinsed, kept for 42 hours and treated with MTT. Optical density of extracted MTT solutions was measured at 562 nm. The viability was 99.4%, which is indicative for non-irritant substances.

The test substance is non-irritant in the EPISKIN reconstructed human epidermal model.

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:

28 Jan 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: study according to the guideline under GLP. Study does not allow definitive conclusion

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

GLP compliance:

yes (incl. QA statement)

Vehicle:

other: 0.9% w/v sodium chloride solution

Amount / concentration applied:

Test item: as such
Positive control: ethanol > 98.8%
Negative control:0.9% w/v sodium chloride.

Duration of treatment / exposure:

10 min

Details on study design:

Bovine corneas were obtained from a local abatoir, preserved in HBBS and prepared on arrival in the laboratory. Corneas free of damage were mounted in a closed BCOP holder. Both chambers were filled with MEM and incubated for 60 minutes at 32 ± 1 ºC.
In the test undamaged corneas (3/treatment) were checked for opacity (zero measurement) and exposed to 0.75 mL the test substance, negative or positive control (anterior chamber) at 32 ± 1 ºC for 10 minutes. After exposure the chambers and corneas were rinsed and chambers were filled with fresh MEM. Corneas were checked visually and opacity was assessed after incubation for 120 min at 32 ± 1 ºC. In additon the corneas were exposed to sodium fluorin (4 mg/L) (anterior chamber) for 90 minutes. Optical density at 492 nm of the medium in the posterior chamber was measured to assess permeability.
Measurements for opacity were corrected for the initial opacity and the opacity measured in the vehicle controls.
Measurements for permeability were corrected for vehicle control.
All results were averaged.

Irritation parameter:

other: IVIS

Score:

25.4

Remarks on result:

other: mean opacity value + (15 x mean OD492 value)

	Opacity Post incubation – pre-treatment	Opacity corrected	Permeability	Permeability corrected	IVIS
Neg control	1		0.072
	2		0.043
	1		0.058
	Mean 1.3		Mean 0.058		2.2
Pos control	30	28.7	1.495	1.437
	27	25.7	1.490	1.432
	24	22.7	1.298	1.240
		Mean 25.7		Mean 1.370	46.2
Test substance	21	19.7	0.398	0.340
	15	13.7	0.385	0.327
	25	23.7	0.671	0.613
		Mean 19.0		Mean 0.427	25.4

 

Interpretation of results:

other: no prediction can be made

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

The outcome of the BCOP test with the substance shows that no prediction on the eye irritation of the substance can be made.

Executive summary:

The substance is tested in the in vitro BCOP assay. The calculated IVIS based on assessments of opacity and permeability for the test substance is 6.1. This does not allow a definitive conclusion on the irritant properties of the test substance, but indicates that the substance is not causing serious eye damage.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Justification for selection of skin irritation / corrosion endpoint:
guideline study under GLP

Justification for classification or non-classification

The available data on skin irritation potential of the test substance do not meet the criteria for classification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.

The available data on eye irritation potential of the test substance are insufficient to determine whetherclassification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC is necessary. One impurity that is included between 1.0 and 3.0 % is classified as Serious Eye Damage/Eye Irritation Category 1, and therefore the substance is classified as Serious Eye Irritant/Eye Irritation Category 2.