Phosphoric acid, mono- and di-C6-10-alkyl esters

Endpoint summary

Description of key information

Acute dermal irritation: The test article was assessed for irritant or corrosive effects according to OECD Guideline 404. The test article was determined to be a dermal irritant under the conditions of this study.

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:

Experimental start date: 18 Jul 2011, Experimental termination date: 08 Aug 11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP and in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not effect the quality of the relevant results.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Covance Research Products Inc., Denver, PA on 01 Jun 2011, 08 Jun 2011 and 16 Jun 2011
- Age at study initiation: the animals were born on 08 Jan 2011, 22 Jan 2011 and 19 Feb 2011
- Weight at study initiation: 3.0- 3.3 kg
- Housing: individually housed in suspended cages. Adsorbent paper bedding was placed beneath the cages and changed at least 3 times/week.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): temperature controlled
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): the test article was dosed by volume, 0.5 mL/site

Duration of treatment / exposure:

Exposure periods of 3 minutes (for initial animal), 1 hour (for initial animal) and 4 hours (for all animals).

Observation period:

Up to 14 days

Number of animals:

3 in total.

Details on study design:

TEST SITE
- Area of exposure: Each dose site was approximately 6 cm2.

DOSING
Initially, one rabbit was dosed sequentially on sites #1, 2 and 3. The test article was dosed by volume, 0.5 ml/site. A 2 x 3 cm gauze patch was placed on the skin and the test article was applied to the gauze. Gentle pressure was applied to aid in the distribution of the test article over the prepared site. The rabbit was gently held in place and a piece of porous dressing was secured with non-irritating tape over dose site #1 (semi-occlusive) for the 3 minute exposure period. The torso was covered with a piece of porous dressing large enough to cover dose sites 2 &3 with at least 5 cm square to spare on all sides of the gauze patch. Porous, non-irritating tape was used to encircle the trunk of the animal.

The dressing and test article patch covering site #1 were removed at 3 minutes postdose, over site #2 at 1 hour postdose and the torso wrappings and patch covering site #3 at 4 hours postdose. All sites were gently washed with distilled water to remove residual test material.

Since no evidence of a corrosive or severely irritating effect was observed in the initial animal, two additional animals were added to the study. The animals were dosed at site #3 with 0.5 ml of the test article. After an exposure period of 4 hours, the wrappings and patches were removed and the sites gently washed with distilled water.

TYPE AND FREQUENCY OF OBSERVATIONS:
Initial animal: Site #1 was scored for dermal irritation immediately and at 60 minutes following patches removal for the 3 minute exposure. Site #2 was scored at 60 minutes following the 1 hour exposure.

All animals: Site #3 was scored at 60 minutes and at at 24,48 and 72 hours and on Days 7 and 14 following patch removal following the 4 hour exposure.

Erythema and edema were scored according to the numerical Draize technique. The skin was also evaluated for ulceration and necrosis or any evidence of tissue destruction. Any additional signs were described.

Draize scoring scheme:

Erythema & Eschar
No erythema: 0
Very slight erythema (barely perceptible): 1
Well defined erythema: 2
Moderate to severe erythema: 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth): 4

Edema
No edema: 0
Very slight edema (barely perceptible): 1
Slight edema (edges of area well-defined by definite raising): 2
Moderate edema (raised approximately 1.0 mm): 3
Severe edema (raised more than 1.0 mm, extending beyond the area of exposure): 4

Body weights of all animals were recorded pre-test, 72 hours and at termination.

Animals were observed for toxicological and pharmacological effects at each dermal observation period and observed for mortality daily. All animals were sacrificed using CO2 following study termination.

Irritation parameter:

erythema score

Basis:

mean

Remarks:

(Animal H4552)

Time point:

other: Mean of scores at 24, 48 and 72 h

Score:

3.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Reults from 4 hour exposure

Irritation parameter:

edema score

Basis:

mean

Remarks:

(Animal H4552)

Time point:

other: Mean of scores at 24, 48 and 72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Results from 4 hour exposure

Irritation parameter:

erythema score

Basis:

mean

Remarks:

(Animal H4525)

Time point:

other: Mean of scores at 24, 48 and 72 h

Score:

2.67

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: Results from 4 hour exposure

Irritation parameter:

edema score

Basis:

mean

Remarks:

(Animal H4525)

Time point:

other: Mean of scores at 24, 48 and 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Results from 4 hour exposure

Irritation parameter:

erythema score

Basis:

mean

Remarks:

(Animal H4520)

Time point:

other: Mean of scores at 24, 48 and 72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: Results from 4 hour exposure

Irritation parameter:

edema score

Basis:

mean

Remarks:

(Animal H4520)

Time point:

other: Mean of scores at 24, 48 and 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Results of 4 hour exposure

Dermal Observations

Initial animal:

3 minute exposure: No erythema and no edema was observed immediately following the 3 min exposure. At 60 min after patch removal following the 3 min exposure, very slight erythema and no edema was observed.

1 hour exposure: Well defined erythema and very slight edema was observed at 60 min after patch removal following the 1 hour exposure.

4 hour exposure: Moderate erythema and slight edema was observed at 1 hour. Severe erythema and slight edema was observed at 24 and 48 h. At 72 hours, erythema was moderate and the skin around the dose site felt thickened. Edema was slight. At 7 and 14 days following the 4 hours exposure, erythema and edema were absent with flaking skin noted on Day 7.

2 additional animals:

4 hour exposure: Erythema was very slight 1 hour after patch removal and edema was very slight to slight. At 24 hour, erythema was very slight to well defined and edema was very slight. At 48 and 72 hour, erythema was well defined to moderate and edema was very slight. On Day 7, erythema was severe. On 1 animal, the skin felt as though it had separated from the underlying tissue. The other animal had a shiny area and flaking skin. By Day 14, no erythema or edema was observed. Flaking skin was noted on both animals and 1 animal had a shiny area. Hair regrowth was noted on both animals.

Systemic observations and body weights:

There were no abnormal physical signs noted during the observation period.

Body weights stayed the same in 2 out of the 3 animals. The remaining animal gained weight. 

Dermal observations, body weights and systemic observations

Exposure time	Time after patch removal	Rabbit Eartag	H4552	H4525	H4520
		Sex	F	F	F
		Pretest body weight- kg	3.0	3.3	3.0
		72 h body weight- kg	3.0	3.3	3.0
		Terminal body weight- kg	3.2	3.3	3.0
Site #1- 3 min	Immediately following patch removal	Erythema	0	N/A	N/A
		Edema	0	N/A	N/A
		Systemic observation	A	N/A	N/A
	60 min	Erythema	1	N/A	N/A
		Edema	0	N/A	N/A
		Systemic observation	A	N/A	N/A
Site #2- 1 hr	60 min	Erythema	2	N/A	N/A
		Edema	1	N/A	N/A
		Systemic observation	A	N/A	N/A
Site #3- 4 hr	60 min	Erythema	3	1	1
		Edema	2	2	1
		Systemic observation	A	A	A
	24 h	Erythema	4	2	1
		Edema	2	1	1
		Systemic observation	A	A	A
	48 h	Erythema	4	3	2
		Edema	2	1	1
		Systemic observation	A	A	A
	72 h	Erythema	3a	3	2
		Edema	2	1	1
		Systemic observation	A	A	A
	7 d	Erythema	0f	4b	4s, f
		Edema	0	0	0
		Systemic observation	A	A	A
	14 d	Erythema	0	0s, f, g	0f, g
		Edema	0	0	0
		Systemic observation	A	A	A

A= normal, N/A= not applicable, a= skin on and around dose site felt thick, b= skin felt separated from underlying tissue, s= shiny areas, f= flaking skin, g= hair regrowth

Interpretation of results:

Category 2 (irritant)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test article was assessed for irritant or corrosive effects according to OECD Guideline 404. The test article was determined to be a dermal irritant under the conditions of this study.

Executive summary:

Objective: To determine the irritant or corrosive effects, if any, of a test article when applied to the skin of a rabbit. This study was designed to comply with the standards set forth in OECD Guideline for Testing of Chemicals, Number 404 Acute Dermal Irritation/ Corrosion, adopted April 24, 2002.

Method synopsis: Since the test article was suspected to be a dermal irritant/ corrosive substance, 1 healthy female New Zealand White rabbit was dosed dermally with the test article. The test article was applied dermally to 3 intact sites for an exposure period of 3 minutes on site #1, 1 hour on site #2 and 4 hours on site #3. Erythema and edema was scored immediately after patch removal following the 3 minutes exposure and at 60 minutes after each patch removal for 1 and 4 hour exposure. Site #3 was scored again at 24, 48 and 72 hours. Reactions were scored again on Days 7 and 14 following patch removal. The skin was also evaluated for ulceration and necrosis or any evidence of tissue destruction at these time periods.

Since the reaction of the initial animal did not produce a corrosive or severely irritating effect, 2 additional female rabbits were added to the study. The 2 animals were dosed at site #3 for a 4 hour exposure. Erythema and edema were scored at 60 minutes, and at 24, 48 and 72 hours and on Days 7 and 14 following patch removal. Animals were observed for toxicological and pharmacological effects at each dermal observation period and observed for mortality daily. Body weights of all animals were recorded pretest, 72 hours and at termination.

Summary of Results

Initial animal:

3 minute exposure: No erythema and no edema was observed immediately following the 3 min exposure. At 60 min after patch removal following the 3 min exposure, very slight erythema and no edema was observed.

1 hour exposure: Well defined erythema and very slight edema was observed at 60 min after patch removal following the 1 hour exposure.

4 hour exposure: Moderate erythema and slight edema was observed at 1 hour. Severe erythema and slight edema was observed at 24 and 48 h. At 72 hours, erythema was moderate and the skin around the dose site felt thickened. Edema was slight. At 7 and 14 days following the 4 hours exposure, erythema and edema were absent with flaking skin noted on Day 7.

2 additional animals:

4 hour exposure: Erythema was very slight 1 hour after patch removal and edema was very slight to slight. At 24 hour, erythema was very slight to well defined and edema was very slight. At 48 and 72 hour, erythema was well defined to moderate and edema was very slight. On Day 7, erythema was severe. On 1 animal, the skin felt as though it had separated from the underlying tissue. The other animal had a shiny area and flaking skin. By Day 14, no erythema or edema was observed. Flaking skin was noted on both animals and 1 animal had a shiny area. Hair regrowth was noted on both animals.

There were no abnormal physical signs noted during the observation period.

Body weights stayed the same in 2 out of the 3 animals. The remaining animal gained weight. 

Conclusion

The test article is a dermal irritant.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation/ corrosion: OECD 404 Acute Dermal Irritation/ Corrosion

Objective: To determine the irritant or corrosive effects, if any, of a test article when applied to the skin of a rabbit. This study was designed to comply with the standards set forth in OECD Guideline for Testing of Chemicals, Number 404 Acute Dermal Irritation/ Corrosion, adopted April 24, 2002.

Method synopsis: Since the test article was suspected to be a dermal irritant/ corrosive substance, 1 healthy female New Zealand White rabbit was dosed dermally with the test article. The test article was applied dermally to 3 intact sites for an exposure period of 3 minutes on site #1, 1 hour on site #2 and 4 hours on site #3. Erythema and edema was scored immediately after patch removal following the 3 minutes exposure and at 60 minutes after each patch removal for 1 and 4 hour exposure. Site #3 was scored again at 24, 48 and 72 hours. Reactions were scored again on Days 7 and 14 following patch removal. The skin was also evaluated for ulceration and necrosis or any evidence of tissue destruction at these time periods.

Since the reaction of the initial animal did not produce a corrosive or severely irritating effect, 2 additional female rabbits were added to the study. The 2 animals were dosed at site #3 for a 4 hour exposure. Erythema and edema were scored at 60 minutes, and at 24, 48 and 72 hours and on Days 7 and 14 following patch removal. Animals were observed for toxicological and pharmacological effects at each dermal observation period and observed for mortality daily. Body weights of all animals were recorded pretest, 72 hours and at termination.

Summary of Results

Initial animal:

3 minute exposure: No erythema and no edema was observed immediately following the 3 min exposure. At 60 min after patch removal following the 3 min exposure, very slight erythema and no edema was observed.

1 hour exposure: Well defined erythema and very slight edema was observed at 60 min after patch removal following the 1 hour exposure.

4 hour exposure: Moderate erythema and slight edema was observed at 1 hour. Severe erythema and slight edema was observed at 24 and 48 h. At 72 hours, erythema was moderate and the skin around the dose site felt thickened. Edema was slight. At 7 and 14 days following the 4 hours exposure, erythema and edema were absent with flaking skin noted on Day 7.

2 additional animals:

4 hour exposure: Erythema was very slight 1 hour after patch removal and edema was very slight to slight. At 24 hour, erythema was very slight to well defined and edema was very slight. At 48 and 72 hour, erythema was well defined to moderate and edema was very slight. On Day 7, erythema was severe. On 1 animal, the skin felt as though it had separated from the underlying tissue. The other animal had a shiny area and flaking skin. By Day 14, no erythema or edema was observed. Flaking skin was noted on both animals and 1 animal had a shiny area. Hair regrowth was noted on both animals.

There were no abnormal physical signs noted during the observation period.

Body weights stayed the same in 2 out of the 3 animals. The remaining animal gained weight. 

Conclusion

The test article is a dermal irritant.

Eye irritation:

Based on the results of the an in-vivo skin irritation study it was considered appropriate not to conduct an eye irritation study. Based on the severe irritation observed in the skin irritation study, it is assumed that the substance will also cause serious eye irritation.

Respiratory irritation:

Based on the irritant properties of the substance, it can be anticipated that respiratory irritation may occur if significant inhalation of the substance occurred, however this is not anticipated.

Justification for selection of skin irritation / corrosion endpoint:
The study has been conducted according to OECD Guideline 404 and GLP and is adequately reported. The study has been assigned a reliability 1.

Effects on skin irritation/corrosion: irritating

Justification for classification or non-classification

Skin irritation/ corrosion: OECD 404 Acute Dermal Irritation/ Corrosion

The results from an acute dermal irritation study were assessed against the CLP criteria for skin corrosion/irritation.

The study determined that the test item produced a mean value of ≥2.3 -≤4 for erythema in 2 of the 3 animals tested from gradings at 24, 48 and 72 h after patch removal (one animal produced a mean score of 3.67 for erythema and one animal produced a mean score of 2.67 for erythema).

In accordance with the CLP regulations, the substance is therefore classified as a Cateogory 2 irritant, and the hazard statement H315; Causes skin irritation, shall apply.

Eye Irritation

Based on the results of the skin irritation study, it is also assumed that the substance will cause serious eye irritation. Therefore, the substance will be classificed as a Category 2 eye irriant, and the hazard statement H319: Causes serious eye irritation will apply.

Categories Display