2-Propenoic acid, 2-methyl-, 2-hydroxyethyl ester, reaction products with phosphorus oxide

Endpoint summary

Description of key information

The test substance was not irritating to the skin, but caused corrosive effects on the eyes.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From March 16, 2004 to March 26, 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000, including the most recent partial revisions

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

- Name of the test substance as cited in the report: PARAD Substance 139.
- Batch: 31104251.
- Molecular formula: C6H10O3.xH3O4P.
- Molecular weight: ~250.
- Description: Slightly yellowish viscous liquid.
- Purity: Not available (mixture).
- Test substance storage: At room temperature in the dark.
- Stability under storage conditions: Stable.
- Expiry date: December 31, 2004.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

- Source: Charles River Deutschland, Kisslegg, Germany
- Age and body weight: Animals used within the study were at least 6 weeks old and body weights were at least 1 kg.
- Identification: Earmark
- Conditions: Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per h, a temperature of 21.0±3.0°C (actual range: 17.0 – 24.1°C), a relative humidity of 30-70% (actual range: 33 - 67%) and 12 h artificial fluorescent light and 12 h darkness per day.
- Accommodation: Individually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 56x44x37.5 cm).
- Diet: Standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Lage, Germany) approximately 100 g per d. In addition, hay (BMI, Helmond, the Netherlands) was provided at least three times a week.
- Water: ad libitum
- Acclimatisation period: At least 5 d before start of treatment under laboratory conditions.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

other: Adjacent areas of the untreated skin of each animal served as controls

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.5 mL
- Concentration (if solution): Undiluted

Duration of treatment / exposure:

4 h

Observation period:

72 h

Number of animals:

3 males

Details on study design:

A single rabbit was tested at the initial step. Two other rabbits were treated in a similar manner one week later, after considering the degree of skin irritation observed in the first rabbit.

Approximately 24 h before treatment, the dorsal fur was clipped with electric clippers, exposing an area of approximately 150 square centimeters (10x15 cm²). Whenever considered necessary the treated skin areas were re-clipped at least 3 h before the observations, to facilitate scoring.

A health inspection was performed prior to the commencement of treatment, to ensure that the animals were in a good state of health. Special attention was paid to the skin to be treated, which was intact and free from abnormalities. Each animal was treated by dermal application of 0.5 mL of the test substance. The test substance was applied to the skin of one flank, using a metalline patch of 2x3 cm. The patch was mounted on Micropore tape, which was wrapped around the abdomen and secured with Coban elastic bandage. Four hours after the application, the dressing was removed and the skin cleaned of residual test substance using water.

Observations:
Mortality/Viability: Twice daily.
Toxicity: At least once daily.
Body Weight: Day of treatment (prior to application) and at termination.
Irritation: The skin reactions were assessed at approximately 1, 24, 48 and 72 h and / or 7 d after the removal of the dressings and test substance. The irritation scores and a description of all other (local) effects were recorded. Adjacent areas of the untreated skin of each animal served as controls.

Numerical scoring system:
Erythema and eschar formation:

No erythema ................................................................................................... 0
Very slight erythema (barely perceptible) ...................................................1
Well-defined erythema ....................................................................................2
Moderate to severe erythema ........................................................................3
Severe erythema (beet redness) *.................................................................. 4
*. Where signs of necrosis or corrosion (injuries in depth) prevent erythema scoring, the maximum grade for erythema (= 4) is given.

Oedema formation:
No oedema ........................................................................................................ 0
Very slight oedema (barely perceptible) ....................................................... 1
Slight oedema (edges of area well-defined by definite raising) ................ 2
Moderate oedema (raised approximately 1 mm) ......................................... 3
Severe oedema (raised more than 1 mm and extending beyond the area of exposure) 4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.3

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

other: The skin irritation had resolved within 7 d after exposure.

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

other: The skin irritation had resolved within 72hrs after exposure.

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.7

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

other: The skin irritation had resolved within 72hrs after exposure.

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible

Remarks:

The skin irritation had resolved within 48hrs after exposure.

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible

Remarks:

The skin irritation had resolved within 72hrs after exposure.

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible

Remarks:

The skin irritation had resolved within 48hrs after exposure.

Irritant / corrosive response data:

Irritation:
Four hours exposure to 0.5 mL of test substance resulted in well-defined erythema and in slight or moderate oedema in the treated skin-areas of the three rabbits. The skin irritation had resolved within 72 h after exposure in two animals and within 7 d after exposure the remaining animal. Scaliness was noted in one animal at 72 h after exposure.

Corrosion:
There was no evidence of a corrosive effect on the skin.

Colouration / remnants:
No staining of the treated skin by the test substance was observed and no test substance remnants were seen.

Other effects:

Toxicity / mortality:
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Table 1: Mean value irritation scores (24, 48 and 72 h)

 

Animal number	Mean 24 - 72 h
	Erythema	Oedema
1	1.3	0.3
2	1.0	1.0
3	0.7	0.3
Primary irritation index: 1.6

 

Table 2: Mean value irritation scores (24 and 72 h)

 

Animal number	Mean 24 - 72 h
	Erythema	Oedema
1	1.5	0.5
2	1.0	1.0
3	0.5	0.5
Primary irritation index: 1.7

 

Interpretation of results:

other: CLP criteria not met

Remarks:

does not have to be classified

Conclusions:

Under the study conditions, the test substance was considered to be not irritating to the skin.

Executive summary:

A study was conducted to assess the skin irritation potential of the test substance in rabbits according to OECD Guideline 404, EU Method B.4, US EPA 870.2500 and JMAFF Japanese test Guidelines, 2000, in compliance with GLP. A total of 0.5 mL of the test substance was applied onto the clipped skin of three animals for 4 h using a semi-occlusive dressing. The test substance was applied to the skin of one flank, using a metalline patch of 2x3 cm. After exposure for 4 h, the dressing was removed and residues of substance were carefully rinsed off with distilled water. Observations were made 1, 24, 48 and 72 h and / or 7 d after exposure. Exposure to test substance resulted in well-defined erythema and in slight or moderate oedema in the treated skin areas of the three rabbits. The skin irritation had resolved within 72 h after exposure in two animals and within 7 d after exposure in the remaining animal. Scaliness was noted in one animal at 72 h after exposure only. Based on the 24 and 72 h readings, the primary irritation index was calculated to be 1.7. Under the study conditions, the substance was considered to be not irritating to the skin (van Huygevoort AHBM, 2004).

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:

From March 22, 2004 to March 22, 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

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000, including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

- Name of the test substance (as cited in the report): PARAD Substance 139.
- Batch: 31104251.
- Molecular formula: C6H10O3.xH3O4P.
- Molecular weight: ~250.
- CAS number: 52628-03-2.
- Description: Slightly yellowish viscous liquid.
- Purity: Not available (mixture).
- Storage: At room temperature in the dark.
- Stability under storage conditions: Stable
- Expiry date: December 31, 2004.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

- Age and body weight: The animal used within the study was at least 6 weeks old and its body weight was between 1.0 and 3.5 kg.
- Identification: Earmark.
- Conditions: Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per h, a temperature of 21.0±3.0°C (actual range: 16.6 - 21.6°C), a relative humidity of 30 - 70% (actual range: 33 - 62%) and 12 h artificial fluorescent light and 12 h darkness per day.
- Accommodation: Individually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 56x44x37.5 cm). Acclimatisation period was at least 5 d before start of treatment under laboratory conditions.
- Diet: Standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Lage, Germany) approximately 100 g per d. In addition, hay (BMI, Helmond, the Netherlands) was provided at least three times a week.
- Water: ad libitum.

Vehicle:

unchanged (no vehicle)

Controls:

other: The other eye remained untreated and served as the reference control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.1 mL

Duration of treatment / exposure:

1 h

Observation period (in vivo):

1 h

Number of animals or in vitro replicates:

1

Details on study design:

- A health inspection was performed prior to commencement of treatment, to ensure that the animal was in a good state of health. Special attention was paid to the eyes, which were free from any abnormality.

- On test Day 1, 0.1 mL of the test substance as the maximum required amount, was instilled in the conjunctival sac of an eye of one animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control. Based on the severity of the ocular lesions observed during the study, the animal was sacrificed for ethical reasons immediately after the 1 h observation and the two further rabbits assigned to the study were not treated.

- Observations:
Mortality/Viability: Twice daily.
Toxicity: At least once daily.
Body Weight: Day of treatment (prior to instillation).
Irritation: The eyes of the animal were examined approximately 1 h after instillation of the test substance. The irritation scores and a description of all other (local) effects were recorded.

- The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

CORNEAL IRRITATION:
Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity (may include slight dulling of normal lustre) ............................................ 0
Scattered or diffuse areas of opacity, details of iris clearly visible ................................................. 1
Easily discernible translucent area, details of iris slightly obscured .............................................. 2
Nacreous area, no details of iris visible, size of pupil barely discernible ....................................... 3
Opaque cornea, iris not discernible through the opacity .............................................................. 4

Area of cornea involved:
No ulceration or opacity ............................................................................................................... 0
One quarter or less but not zero .................................................................................................. 1
Greater than one quarter, but less than half ................................................................................. 2
Greater than half, but less than three quarters ............................................................................. 3
Greater than three quarters, up to whole area .............................................................................. 4

IRIS:
Normal ........................................................................................................................................ 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia, or injection, any of these or combination thereof, iris still reacting to light (sluggish reaction is positive) ....................................................................................................... 1
No reaction to light, hemorrhage, gross destruction (any or all of these) ...................................... 2

CONJUNCTIVAL IRRITATION:
Redness (refers to palpebrae and sclera, excluding cornea and iris):
Blood vessels normal .................................................................................................................. 0
Some blood vessels definitely hyperaemic (injected) ................................................................... 1
Diffuse, crimson color, individual vessels not easily discernible .................................................... 2
Diffuse beefy red .......................................................................................................................... 3

Chemosis (refers to lids and/or nictitating membranes):
No swelling ………………………………………… ........................................................................ 0
Any swelling above normal (includes nictitating membranes) ...................................................... 1
Obvious swelling with partial eversion of lids ................................................................................ 2
Swelling with lids about half closed ............................................................................................. 3
Swelling with lids more than half closed ....................................................................................... 4

Discharge:
No discharge (may include small amounts observed in inner canthus of normal animals) ........... 0
Any amount different from normal and/or lacrimation ................................................................... 1
Discharge with moistening of the lids and hairs just adjacent to lids ............................................. 2
Discharge with moistening of the lids and hairs (considerable area around the eye) ................... 3
Where standard lighting was considered inadequate for observing minor effects, eye examinations were performed using an ophthalmic examination lamp.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: 1 hr

Score:

ca. 3

Max. score:

3

Reversibility:

not reversible

Remarks on result:

other: Based on the degree of the ocular injury, it was concluded that ocular corrosion had occurred into the treated eye of the animal. Based on animal welfare reasons, no other animals were tested.

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: 1 hr

Score:

ca. 1

Max. score:

1

Reversibility:

not reversible

Remarks on result:

other: Based on the degree of the ocular injury, it was concluded that ocular corrosion had occurred into the treated eye of the animal. Based on animal welfare reasons, no other animals were tested.

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: 1 hr

Score:

ca. 3

Max. score:

3

Reversibility:

not reversible

Remarks on result:

other: Based on the degree of the ocular injury, it was concluded that ocular corrosion had occurred into the treated eye of the animal. Based on animal welfare reasons, no other animals were tested.

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 1 hr

Score:

ca. 2

Max. score:

2

Reversibility:

not reversible

Remarks on result:

other: Based on the degree of the ocular injury, it was concluded that ocular corrosion had occurred into the treated eye of the animal. Based on animal welfare reasons, no other animals were tested.

Irritant / corrosive response data:

Irritation and corrosion:
Instillation of 0.1 mL of test substance into an eye of one rabbit resulted in effects on the cornea, iris and conjunctivae. The corneal injury consisted of opacity (maximum grade 3). Iridial irritation grade 1 was observed. The irritation of the conjunctivae consisted of redness, chemosis and discharge. Gray/white discoloration (signs of necrosis) was noted on the eyelids and nictitating membrane, 1 h after instillation of the test substance. Based on the severity of the ocular lesions observed during the study, the animal was sacrificed for ethical reasons immediately after the 1 h observation.

Colouration / remnants:
No staining of (peri) ocular tissues by the test substance was observed and no test substance remnants were seen.

Other effects:

Toxicity / mortality:
No symptoms of systemic toxicity were observed in the animal during the test period and no mortality occurred.

Table 1: Individual eye irritation scores

 

Time after dosing	Cornea			Iris	Conjunctivae
	Opacity	Area	Fluor area (%)		Redness	Chemosis	Discharge	Comments
1 h	3	2	-	1	3	2	2	a, b

 

Fluor area (%): green staining (percentage of total corneal area) after fluorescein treatment.

Comments:

(a) Signs of necrosis were noted on the eyelids and nictitating membrane.

(b) Based on the severity of the ocular lesions observed during the study, the animal was sacrificed for ethical reasons immediately after the 1 h observation.

Interpretation of results:

other: Category 1

Remarks:

as per CLP criteria

Conclusions:

Under the study conditions, the test substance was found to cause corrosive effects on the eyes.

Executive summary:

A study was conducted to assess the eye irritancy potential of the test substance in rabbit according to OECD Guideline 405, EU Method B.5, US EPA OPPTS 870.2400 and JMAFF Guideline, 2000, in compliance with GLP. A single dose of 0.1 mL test substance was instilled into the sac of one eye. The lids were then gently held together for approximately one second to prevent loss of the test substance. Gray/white discoloration (signs of necrosis) was noted on the eyelids and nictitating membrane, 1 h after instillation of the test substance. Based on the severity of the ocular lesions observed during the study, the animal was sacrificed for ethical reasons immediately after the 1 h observation. The remaining two rabbits assigned to the study were not treated. Based on the degree of the corneal injury, it was concluded that ocular corrosion had occurred by instillation of test substance into the rabbit eye in the animal. Hence, the substance was found to cause corrosive effects on the eyes (van Huygevoort AHBM, 2004).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation:

A study was conducted to assess the skin irritation potential of the test substance in rabbits according to OECD Guideline 404, EU Method B.4, US EPA 870.2500 and JMAFF Japanese test Guidelines, 2000, in compliance with GLP. A total of0.5 mL of the test substance was applied onto the clipped skin of three animals for 4 h using a semi-occlusive dressing. The test substance was applied to the skin of one flank, using a metalline patch of 2x3 cm. After exposure for 4 h, the dressing was removed and residues of substance were carefully rinsed off with distilled water. Observations were made 1, 24, 48 and 72 h and / or 7 d after exposure. Exposure to test substance resulted in well-defined erythema and in slight or moderate oedema in the treated skin areas of the three rabbits. The skin irritation had resolved within 72 h after exposure in two animals and within 7 d after exposure in the remaining animal. Scaliness was noted in one animal at 72 h after exposure only. Based on the 24 and 72 h readings, the primary irritation index was calculated to be 1.7. Under the study conditions, the substance was considered to be not irritating to the skin (van Huygevoort AHBM, 2004).

Eye irritation:

A study was conducted to assess the eye irritancy potential of the test substance in rabbit according to OECD Guideline 405, EU Method B.5, US EPA OPPTS 870.2400 and JMAFF Guideline, 2000, in compliance with GLP.A single dose of 0.1 mL test substance was instilled into the sac of one eye. The lids were then gently held together for approximately one second to prevent loss of the test substance. Gray/white discoloration (signs of necrosis) was noted on the eyelids and nictitating membrane, 1 h after instillation of the test substance. Based on the severity of the ocular lesions observed during the study, the animal was sacrificed for ethical reasons immediately after the 1 h observation. The remaining two rabbits assigned to the study were not treated. Based on the degree of the corneal injury, it was concluded that ocular corrosion had occurred by instillation of test substance into the rabbit eye in the animal. Hence, the substance was found to cause corrosive effects on the eyes(van Huygevoort AHBM, 2004).

Justification for classification or non-classification

Skin irritation:

Based on the results of an in vivo skin irritation study, the test substance does not need to be classified according to CLP criteria (EC 1272/2008).

Eye irritation:

Based on the results of an in vivo eye irritation study, the test substance is considered to be corrosive with a classification of Eye Damage 1 - H318 - causes serious eye damage according to CLP criteria (EC 1272/2008).