9-Octadecen-1-ol, (Z)-, phosphate

Endpoint summary

Description of key information

Skin irritation in vitro:
The results obtained from this in vitro skin irritation test using the EPISKIN model, indicated that the test item reveals no skin irritation potential under the utilised testing conditions. According to the current OECD Guideline No. 439, Reaction product of oleyl alcohol with polyphosphoric acid is considered as non-irritant to skin and is therefore not classified.
Eye irritation in vitro:
In this In vitro eye corrosives and severe irritants study, using the Isolated Chicken Eyes model with Reaction product of oleyl alcohol with poly-
phosphoric acid, according to the guideline OECD 438 ocular corrosion and severe irritation potential was observed. Thus, according to Regulation (EC) No 1272/2008 Reaction product of oleyl alcohol with polyphosphoric acid causes serious damage to the eye and has to be classified into 
Category 1.

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:

2013-02-06 to 2013-02-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: CLP and Guideline compliant study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

, 22 July 2010

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

, 06 July 2012

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: not applicable (reconstituted human epidermis)

Strain:

other: not applicable (reconstituted human epidermis)

Details on test animals or test system and environmental conditions:

EpiSkinTM Small Model (EpiSkinTMSM), EPISKIN SNC Lyon, France, is a three-dimensional human epidermis model. Adult human-derived epidermal keratinocytes are seeded on a dermal substitute consisting of a collagen type I matrix coated with type IV collagen. A highly differentiated and stratified epidermis model is obtained after 13-day culture period comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum. Its use for skin irritation testing involves topical application of test materials to the surface of the epidermis, and the subsequent assessment of their effects on cell viability.

Source: SKINETHIC Laboratories; 4, rue Alexandre Fleming, 69007 – LYON, France.
Batch No.: 13-EKIN-004
Expiry date: 11 February 2013

Type of coverage:

other: not applicable (reconstituted human epidermis)

Preparation of test site:

other: not applicable (reconstituted human epidermis)

Vehicle:

water

Controls:

not required

Amount / concentration applied:

The test item is a viscous liquid therefore exact weighing of treatment volume was not performed. A sufficient amount of the test item was used that evenly covered the whole epidermal surface. The test substance was gently spread onto the skin with a flat curved spatula.

Duration of treatment / exposure:

15 min

Observation period:

Not applicable (reconstituted human epidermis)

Number of animals:

Not applicable (reconstituted human epidermis). However, in this assay 3 replicates per test item and 3 replicates negative controls + 3 replicates positive controls were used. Furthermore 3 replicates of additional control for coloured test item (i.e. for non-specific OD evaluation) were used.

Details on study design:

Pre-incubation (day [-1]):
The “maintenance medium” was pre-warmed to 37 °C. The appropriate number of assay plate wells were filled with the pre-warmed medium (2 mL per well). The epidermis units were placed above the media in a separately prepared well. Contact of the epidermis units with the media was assured. The well was then incubated overnight at 37 °C in an incubator with 5 % CO2.

Application (day 0):
The test item is a viscous liquid therefore exact weighing of treatment volume was not performed. A sufficient amount of the test item was used that evenly covered the whole epidermal surface. The test substance was gently spread onto the skin with a flat curved spatula.

Positive and negative control
Test Item
A volume of 20 μL positive control (SDS 5 % aq.) or negative control (1 x PBS) was applied to the skin surface by using a suitable pipette. Chemicals were spread gently with the pipette tip in order to cover evenly all the epidermal surface if necessary.

Exposure (day 0):
The plates with the treated epidermis units were incubated for the exposure time of 15 minutes at room temperature.

Rinsing (day 0):
After the incubation time, the EPISKIN-SM units were removed and rinsed thoroughly with PBS 1 x solution to remove all of the test material from the epidermal surface. The rest of the PBS was removed from the epidermal surface with a suitable pipette tip linked to a vacuum source (care was taken to avoid damaging to the epidermis).

Post-incubation (day 0-2):
After rinsing the units were placed into the plate wells filled with fresh pre-warmed “Maintenance Medium” (2 mL/well) below them and then incubated for 42 hours at 37 °C in an incubator with 5 % CO2.

MTT test after 42 hours incubation (day 2):
After the 42 hours incubation the EPISKIN-SM units were transferred into the MTT solution filled wells (2 mL of 0.3 mg/mL MTT per well) and then incubated for 3 hours at 37 °C in an incubator with 5 % CO2 protected from light.

Formazan extraction (day 2):
At the end of incubation with MTT a formazan extraction step was undertaken:
A defined disk of epidermis from each replicate was cut from the unit (this involves the maximum area of the disk) using a biopsy punch (supplied as part of the kit). The epidermis was separated with the aid of forceps and both parts (epidermis and collagen matrix) were placed into a tube of 500 μL acidified isopropanol (one tube corresponding to one well of the tissue culture plate).

The capped tubes were thoroughly mixed by using a Vortex Mixer to achieve a good contact of all of the material to the acidified isopropanol. Then the mixture was incubated for about two hours at room temperature protected from light with gentle agitation (~150 rpm) for formazan extraction.

Cell viability measurements (day 2):
Following the formazan extraction, 2 × 200 μL samples from each tube were placed into the wells of a 96-well plate (labelled appropriately). The OD’s of each well were recorded using a 96-well plate spectrophotometer (Thermo Scientific; Multiscan FC) at a wavelength of 570 nm while using an acidified isopropanol solution blank (6 × 200 μL).

Irritation / corrosion parameter:

other: other: Optical density (OD) values

Value:

1.115

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 47 hours. (migrated information)

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The results obtained from this in vitro skin irritation test using the EPISKIN model, indicated that the test item reveals no skin irritation potential under the utilised testing conditions. According to the current OECD Guideline No. 439, Reaction product of oleyl alcohol with polyphosphoric acid is considered as non-irritant to skin and is therefore not classified.

Executive summary:

In this in vitro skin irritation test using the EPISKIN model, the test item Reaction product of oleyl alcohol with polyphosphoric acid did not show significantly reduced cell viability in comparison to the negative control. All obtained test item viability results were above 50 % when compared to the viability values obtained from the negative control. Therefore the test item was considered to be non-irritant to skin.

Positive and negative controls showed the expected cell viability values within acceptable limits. The experiment was considered to be valid.

The results obtained from this in vitro skin irritation test, using the EPISKIN model, indicated that the test item reveals no skin irritation potential under the utilised testing conditions. According to the current OECD Guideline No. 439, Reaction product of oleyl alcohol with polyphosphoric acid is considered as non-irritant to skin and is therefore not classified.

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:

2012-12-05 to 2012-12-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: according to GLP, OECD and EC guidelines

Qualifier:

according to guideline

Guideline:

OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants)

Version / remarks:

adopted 07 September 2009

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Commission Regulation (EU) No 1152/2010 of 08 December 2010 amending, Regulation (EC) No 440/2008 (REACH) B.48

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: not applicable: Isolated chicken eyes (ROSS 308)

Strain:

other: not applicable

Details on test animals or tissues and environmental conditions:

Chicken heads collection and transport
Species of chicken: ROSS 308
Source: TARAVIS KFT. 9600 Sárvár, Rábasömjéni út. 129. Hungary
The age and weight of the chickens used for this study were that of spring chickens traditionally processed by a poultry slaughterhouse
(i.e., approximately 7 weeks old, 1.5 – 2.5 kg). Head collection was performed by a slaughter house technician. The heads were transported to TOXI-COOP ZRT. at the earliest convenience for use approximately within 2 hours from collection. All eyes used in the assay were from the same groups of eyes collected on one specific day. After collection, the heads were inspected for appropriate quality and wrapped with paper moistened with saline, then placed in a plastic box that can be closed (4-5 heads/box).

Vehicle:

unchanged (no vehicle)

Controls:

other: not applicable; however, eyes for positive and negative control were used

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.2 g
Test item was spread evenly on a small piece (approximately 2 cm2) of flexible plastic film to be applied directly onto the cornea.

Duration of treatment / exposure:

After carefully covering the whole cornea with the test substance an exposure period of 10 seconds was applied.
Subsequently, the plastic film was removed from surface of the cornea carefully, taking care not to damage or touch the cornea with the application
equipment.

Observation period (in vivo):

The control and test eyes were evaluated pre-treatment and at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse.
Minor variations within ±5 minutes were considered acceptable.
The cornea thickness and cornea opacity were measured at all time points. Fluorescein retention was measured on two occasions, at base line (t=0) and 30 minutes after the posttreatment rinse.

Number of animals or in vitro replicates:

The treatment group and the concurrent positive control consisted of three eyes. The negative control group consisted of one eye.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: cornea surface was rinsed thoroughly with 20 mL saline solution at ambient temperature,
- Time after start of exposure: 10 seconds

SCORING SYSTEM:
Evaluation:
The endpoints evaluated were corneal opacity, swelling, fluorescein retention, and morphological effects (e.g., pitting or loosening of the epithelium).
Results from corneal opacity, swelling, and fluorescein retention were evaluated separately to generate an Isolated Chicken Eye (ICE) class for each
endpoint. The ICE classes for each endpoint were then combined to generate an Irritancy Classification for each test substance (regulatory EC/GHS
classification).
Severity of effects:
The effects were divided into four categories:
I = none
II = slight
III = moderate
IV = severe O

TOOL USED TO ASSESS SCORE: The cornea thickness and cornea opacity were measured at all time points. Fluorescein retention was measured on
two occasions, at base line (t=0) and 30 minutes after the post-treatment rinse.

Irritation parameter:

cornea opacity score

Value:

4

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Irritation parameter:

fluorescein retention score

Value:

3

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Irritation parameter:

percent corneal swelling

Remarks:

at up to 75 min

Value:

1

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Irritation parameter:

percent corneal swelling

Remarks:

at up to 240 min

Value:

2

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Irritation parameter:

other: corneal opacity score

Basis:

mean

Remarks:

ìsolated chicken eyes

Time point:

other: 30-75-120-180-240 min relative observation time

Score:

4

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

other: corneal thickness

Basis:

mean

Remarks:

isolated chicken eyes

Time point:

other: 30-75-120-180-240 min relative observation time

Score:

>= 0 - <= 2

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

other: The values are given in "% change of instrumental units in comparison to timepoint 0"

Irritation parameter:

other: fluorescein retention

Basis:

mean

Remarks:

isolated chicken eyes

Time point:

other: 30 min relative observation time

Score:

3

Max. score:

3

Reversibility:

other: not applicable

Irritant / corrosive response data:

In this in vitro eye irritation study using the isolated chicken eyes test with Reaction product of oleyl alcohol with polyphosphoric acid, ocular
corrosion and severe irritation potential was observed. The test item was classified as corrosive/severely irritating, CLP Classification: Category 1.

Other effects:

none

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

In this In vitro eye corrosives and severe irritants study, using the Isolated Chicken Eyes model with Reaction product of oleyl alcohol with poly-
phosphoric acid, according to the guideline OECD 438 ocular corrosion and severe irritation potential was observed. Thus, according to Regulation (EC) No 1272/2008 Reaction product of oleyl alcohol with polyphosphoric acid causes serious damage to the eye and has to be classified into
Category 1.

Executive summary:

In this In vitro eye corrosives and severe irritants study, using the Isolated Chicken Eyes model with Reaction product of oleyl alcohol with poly- phosphoric acid, according to the guideline OECD 438 ocular corrosion and severe irritation potential was observed. Thus, according to Regulation (EC) No 1272/2008 Reaction product of oleyl alcohol with polyphosphoric acid causes serious damage to the eye and has to be classified into Category 1.

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation/corrosion

Two in vitro skin irritation tests were performed in order to verify the non-classification status of the test substance Reaction product of oleyl alcohol with polyphosphoric acid.

1. Skin irritation (Episkin model)

In this in vitro skin irritation test using the EPISKIN model, the test item Reaction product of oleyl alcohol with polyphosphoric acid did not show significantly reduced cell viability in comparison to the negative control. All obtained test item viability results were above 50 % when compared to the viability values obtained from the negative control. Therefore the test item was considered to be non-irritant to skin.

Positive and negative controls showed the expected cell viability values within acceptable limits. The experiment was considered to be valid.

The results obtained from this in vitro skin irritation test, using the EPISKIN model, indicated that the test item reveals no skin irritation potential under the utilised testing conditions. According to the current OECD Guideline No. 439, Reaction product of oleyl alcohol with polyphosphoric acid is considered as non-irritant to skin and is therefore not classified.

2. Skin corrosion (Corrositex® model)

The results obtained from this in vitro membrane barrier test, using the Corrositex® model, indicated that the test item reveals no skin corrosion

potential under the utilised testing conditions. According to the current OECD Guideline No. 435, Reaction product of oleyl alcohol with

polyphosphoric acid is considered as non-corrosive to skin and therefore not classified.

Eye irritation in vitro

In this In vitro eye corrosives and severe irritants study, using the Isolated Chicken Eyes model with Reaction product of oleyl alcohol with poly- phosphoric acid, according to the guideline OECD 438 ocular corrosion and severe irritation potential was observed. Thus, according to Regulation (EC) No 1272/2008 Reaction product of oleyl alcohol with polyphosphoric acid causes serious damage to the eye and has to be classified into Category 1.

Justification for selection of skin irritation / corrosion endpoint:
Most reliable study

Justification for selection of eye irritation endpoint:
Most reliable study

Effects on respiratory irritation: highly irritating

Effect level: empty Endpoint conclusion: Adverse effect observed

Justification for classification or non-classification

Reaction product of oleyl alcohol with polyphosphoric acid causes serious damage to the eye and has to be classified into Category 1

according to Regulation (EC) No 1272/2008.

Based on skin irritation data available for reaction product of oleyl alcohol with polyphosphoric acid

no classification and labelling is required according to Regulation No (EC) 1272/2008 (CLP) or Directive 67/548/EEC (DSD) criteria.