Poly[oxy(methyl-1,2-ethanediyl)], .alpha.-[2-[bis(phosphonomethyl)amino]methylethyl]-.omega.-[2-[bis(phosphonomethyl)amino]methylethoxy]-, sodium salt (1:?)

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was performed between 23 June 2011 and 08 July 2011.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

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.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines for Toxicology Studies, 12 NohSan No. 8147, amended 10 December 2002

Deviations:

no

Test guideline 5

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Health and Welfare, 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Poly[oxy(methyl-1,2-ethanediyl)], alpha-[2-[bis(phosphonomethyl)amino]methylethyl]-omega-[2-[bis(phosphonomethyl)amino]methylethoxy]-sodium salt

EC Number:

937-739-1

Cas Number:

1128123-94-3

Molecular formula:

(C3H6O)n. C10H28N2O13P4. xNa,

IUPAC Name:

Poly[oxy(methyl-1,2-ethanediyl)], alpha-[2-[bis(phosphonomethyl)amino]methylethyl]-omega-[2-[bis(phosphonomethyl)amino]methylethoxy]-sodium salt

Constituent 2

Reference substance name:

Sodium chloride

EC Number:

231-598-3

EC Name:

Sodium chloride

Cas Number:

7647-14-5

Molecular formula:

ClNa

IUPAC Name:

sodium chloride

Constituent 3

Reference substance name:

Unknown

Molecular formula:

Unknown

IUPAC Name:

Unknown

Test material form:

solid

Details on test material:

Test Item Identification: Poly[oxy(methyl-1,2-ethanediyl)], alpha-[2-[bis(phosphonomethyl)amino]methylethyl]-omega-[2-[bis(phosphonomethyl)amino]methylethoxy]-sodium salt

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Three New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Harlan Laboratories UK Ltd., Leicestershire, UK.
- Age at study initiation: Twelve to twenty weeks old.
- Weight at study initiation: 2.40 to 2.69 kg
- Housing: The animals were individually housed in suspended cages.
- Diet (e.g. ad libitum): Free access to food (2930 Teklad Global diet) was allowed throughout the study.
- Water (e.g. ad libitum): Free access to mains drinking water was allowed throughout the study.
- Acclimation period: At least five days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): set to achieve limits of 17 to 23°C
- Humidity (%): set to achieve limits of 30 to 70%
- Air changes (per hr): At least fifteen changes per hour.
- Photoperiod (hrs dark / hrs light): lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

other: clipped free of fur

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
For the purpose of the study, the test item was ground to a powder before use.
The absorption of the test item was not determined.

At each test site a quantity of 0.5 g of the test item, moistened sufficiently with 0.5 ml of distilled water, was introduced.

Duration of treatment / exposure:

3 minutes (initial animal), 1 hour (initial animal) and 4 hours (all animals).

Observation period:

72 hours plus an additional observation was made at one treated skin site on Day 7.

Number of animals:

3

Details on study design:

MEASUREMENT OF PH:
The pH of the test item was determined prior to commencement of the study and found to be as follows:
10% w/w aqueous preparation of the test item: pH 4.5 (immediately and after 10 minutes).

PROCEDURE:
On the day before the test each rabbit was clipped free of fur from the dorsal/flank area using veterinary clippers. Only animals with a healthy intact epidermis by gross observation were selected for the study.

One rabbit was initially treated. Three suitable sites were selected on the back of the rabbit. At each test site a quantity of 0.5 g of the test item, moistened sufficiently with 0.5 ml of distilled water, was introduced sequentially under a 2.5 cm x 2.5 cm cotton gauze patch. Each patch was secured in position with a strip of surgical adhesive tape. To prevent the animal interfering with the patches, the trunk of the rabbit was wrapped in an elasticated corset for the duration of the exposure period.

One patch was removed at each of three time points: 3 minutes, 1 hour and 4 hours after application. Any residual test item was removed by gentle swabbing with cotton wool soaked in distilled water.

After consideration of the skin reactions produced in the first animal, an additional two animals were treated sequentially with 0.5 g of test item, moistened sufficiently with 0.5 ml of distilled water. One patch was applied to the back of each rabbit and was allowed to remain in contact with the skin for a period of four hours.

SCORING SYSTEM:
Immediately following removal of the patches and approximately 1, 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the Draize scale (see evaluation of skin reactions below).

Any other skin reactions and clinical signs of toxicity, if present, were also recorded.

An additional observation was made at one treated skin site on Day 7 to assess the reversibility of skin reactions.

Individual bodyweights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Remarks:

(Animal 1 - 70541)

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Result of 4 hour exposure

Results 2

Irritation parameter:

edema score

Basis:

mean

Remarks:

(Animal 1 - 70541)

Time point:

24/48/72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: Result of 4 hour exposure

Results 3

Irritation parameter:

erythema score

Basis:

mean

Remarks:

(Animal 2 - 70668)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Result of 4 hour exposure

Results 4

Irritation parameter:

edema score

Basis:

mean

Remarks:

(Animal 2 - 70668)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Result of 4 hour exposure

Results 5

Irritation parameter:

erythema score

Basis:

mean

Remarks:

(Animal 3 - 70669)

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Remarks on result:

other: Result of 4 hour exposure

Results 6

Irritation parameter:

edema score

Basis:

mean

Remarks:

(Animal 3 - 70669)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Remarks on result:

other: Result of 4 hour exposure

Irritant / corrosive response data:

Skin Reactions:

3-Minute Exposure Period:
The individual scores for erythema/eschar and oedema are given in Table 1.
No evidence of skin irritation was noted during the study.

1-Hour Exposure Period:
The individual scores for erythema/eschar and oedema are given in Table 1.
No evidence of skin irritation was noted during the study.

4-Hour Exposure Period:
The individual scores for erythema/eschar and oedema are given in Table 2.

Well-defined erythema and very slight oedema were noted at one treated skin site with very slight erythema and very slight oedema at one other treated skin site immediately and one hour after patch removal. Very slight erythema, with or without very slight oedema, was noted at these two treated skin sites at the 24-Hour observation. Very slight erythema and very slight oedema were noted at one of these treated skin sites at the 48-Hour observation with very slight erythema noted at the 72-Hour observation.

One treated skin site appeared normal at the 48-Hour observation and one other treated skin site appeared normal at the 7-Day observation.

No evidence of skin irritation was noted at one treated skin site during the study.

The individual mean scores for erythema and oedema required for classification according to the Globally Harmonised System of Classification and Labelling of Chemicals were as follows:
Animal 1 (70541):
Mean score for erythema/eschar formation: 1.0
Mean score for oedema formation: 0.7

Animal 2 (70668):
Mean score for erythema/eschar formation: 0.0
Mean score for oedema formation: 0.0

Animal 3 (70669):
Mean score for erythema/eschar formation: 0.3
Mean score for oedema formation: 0.0

Other effects:

Bodyweight:
Individual bodyweights and bodyweight changes are given in Table 3.
All animals showed expected gain in bodyweight during the study.

Any other information on results incl. tables

Table 1: Individual Skin Reactions Following 3 -Minute and 1 -Hour Exposures:

Skin Reaction	Observation Time (following patch removal)	Individual Scores – Rabbit Number and Sex
		70541 Male
		3-Minute Exposure	1-Hour Exposure
Erythema/Eschar Formation	Immediately	0	0
	1 Hour	0	0
	24 Hours	0	0
	48 Hours	0	0
	72 Hours	0	0
	7 Days	0	0
Oedema Formation	Immediately	0	0
	1 Hour	0	0
	24 Hours	0	0
	48 Hours	0	0
	72 Hours	0	0
	7 Days	0	0

Table 2: Individual Skin Reactions Following 4 -Hour Exposure

Skin Reaction	Observation Time (following patch removal)	Individual Scores – Rabbit Number and Sex			Total
		70541 Male	70668 Male	70669 Male
Erythema/Eschar Formation	Immediately	2	0	1	(3)
	1 Hour	2	0	1	(3)
	24 Hours	1	0	1	2
	48 Hours	1	0	0	(1)
	72 Hours	1	0	0	1
	7 Days	0	-	-	(0)
Oedema Formation	Immediately	1	0	1	(2)
	1 Hour	1	0	1	(2)
	24 Hours	1	0	0	1
	48 Hours	1	0	0	(1)
	72 Hours	0	0	0	0
	7 Days	0	-	-	(0)
Sum of 24 and 72-Hour Readings (S): 4
Primary Irritation Index (S/6): 4/6 = 0.7
Classification: mild irritant

( ) = Total values not used for calculation of primary irritation index.

- = Not applicable.

Table 3: Individual Bodyweights and Bodyweight Changes

Rabbit Number and Sex	Individual Bodyweight (kg)		Bodyweight Change (kg)
70541 Male	Day 0	Day 7	0.14
	2.54	2.68
70668 Male	Day 0	Day 3	0.08
	2.40	2.48
70669 Male	Day 0	Day 3	0.04
	2.69	2.73

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test item produced a maximum individual mean score of 1.0 and therefore did not meet the criteria for classification according to the Globally Harmonised System of Classification and Labelling of Chemicals.

Executive summary:

The study was performed to assess the irritancy potential of the test item to the skin of the New Zealand White rabbit. The method was designed to be compatible with the following:

- OECD Guidelines for the Testing of Chemicals No. 404 “Acute Dermal Irritation/Corrosion” (adopted 24 April 2002)

- Method B4 Acute Toxicity (Skin Irritation) of Commission Regulation (EC) No. 440/2008

- United States Environmental Protection Agency Health Effects Test Guidelines OPPTS 870.2500 Acute Dermal Irritation August 1998

- Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines for Toxicology Studies, 12 NohSan No. 8147, amended 10 December 2002

- Japanese Ministry of Health and Welfare, 1992

3-minute and 1-hour semi-occluded applications of the test item to the intact skin of one rabbit produced no evidence of skin irritation. A single 4-hour, semi-occluded application of the test item to the intact skin of three rabbits produced very slight to well-defined erythema and very slight oedema. No evidence of skin irritation was noted at one treated skin site.

The test item produced a maximum individual mean score of 1.0 and therefore did not meet the criteria for classification according to the Globally Harmonised System of Classification and Labelling of Chemicals.