Diphosphoric acid, compd. with piperazine (1:1)

Endpoint summary

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:

3 March to 6 March 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Species: SPF bred New Zealand White albino rabbits
Supplier: Centre Lagro, France
Sex and age: males, young adult, ca 9 weeks old
Date of arrival: 26 February 2003
Quarantine/acclimatization: 5 days
Body weights at start of study: 1716, 1778 and 1895 g
Body weights at termination: 1775, 1850 and 1937 g
Caging: individually in stainless steel cages with perforated floor
Lighting: 12 hours light/12 hours dark cycle
Temperature during testing: 20± 3 °C.
Relative humidity during testing: 30-70%. Upper limit incidentally up to 99.9%, because of meteorological circumstances and/or wet cleaning of the animal room.
Ventilation: ca 10 air changes/hour
Diet/water: standard laboratory diet ad libitum. Each batch of this diet is analysed by the supplier (SDS Special Diets services, Whitham, England) for the nutrients and contaminants and the results are kept available in the archives. Tap water (N.V. Hydron MiddenNederland) ad libitum. Results of routine physical, chemical and microbiological examination of drinking water as conducted by the supplier are kept available in the archives. The results of diet and water analyses were considered acceptable for this study.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

no

Amount / concentration applied:

0.5 g of the test substance and moistened with 0.5 ml of water.

Duration of treatment / exposure:

4 h

Observation period:

72 h

Number of animals:

3

Details on study design:

Upon arrival, the rabbits were checked for overt signs of ill health and anomalies. Three days prior to the start of the study, the hair was removed from the back and flanks of the animals using electric clippers in a way as to avoid abrasions. The clipping was repeated if considered necessary. The study was carried out with three rabbits. The rabbits were treated as follows: just prior to application, a skin site suitable for application was selected on the clipped flank of the animal. A plastic cup with a diameter of circa 2.5 cm was loaded with an amount of 0.5 g of the test substance and moistened with 0.5 ml of water. The cup loaded with the test mixture was fixed to the selected application site by means of semi-occlusive, adhesive tape (Leukopor, BDF, Germany). Subsequently, the entire trunk of the rabbit was wrapped with a self-adhesive-gauze (Fixomull, BDF, Germany) to maintain the cup in position and to retard evaporation of volatile substances. The gauze was additionally fastened with two strips of tape (Leukoflex, BDF, Germany). After a 4-hour exposure period, the test substance and cup were removed and the test site was cleaned with a tissue moistened with water. One hour after treatment, the resulting skin reactions were evaluated by the method of Draize et al. (J. Pharmacol. Exp. Ther. 82 (1944) 377-390). Further skin readings were made at approximately 24, 48, and 72 h after treatment.

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 1, 24, 48 & 72h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 1, 24, 48 & 72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

At 1, 24, 48 and 72 h after removal, no signs of skin irritation (erythema, edema) were observed in any of the three rabbits.

Table 1: Skin irritation scores of T-1063FM, after a single 4-hour dermal exposure

Animal no.	Hours after removal of the test material:
	1h	24h	48h	72h
	A-B	A-B	A-B	A-B
2	0-0	0-0	0-0	0-0
4	0-0	0-0	0-0	0-0
6	0-0	0-0	0-0	0-0

A=erythema (including ischemia, haemorrhages and incrustation); B = oedema       

 

Table 2: Mean values for erythema an oedema of each rabbit scored at 24, 28 and 72h after treatment with T-1063FM

Animal no.	Erythema	Oedema
2	0.0	0.0
4	0.0	0.0
6	0.0	0.0

 

Interpretation of results:

not classified

Remarks:

Criteria used for interpretation of results: EU

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:

3 March to 17 March 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Species: SPF bred New Zealand White albino rabbits
Supplier: Centre Lagro, France
Sex and age: 3 males, young adult, ca 9 weeks old
Date of arrival: 26 February 2003
Quarantine/acclimatization: 5 days
Body weights at start of study: 1880, 1613 and 1993 g
Body weights at termination: 2100, 1804 and 2250 g
Caging: individually in stainless steel cages with perforated floor
Lighting: 12 hours light/12 hours dark cycle
Temperature during testing: 20 ± 3 °C.
Relative humidity during testing: 30-70%. Upper limit incidentally up to 99.9%, because of meteorological circumstances and/or wet cleaning of the animal room.
Ventilation: ca 10 air changes/hour
Diet/water: standard laboratory diet ad libitum. Each batch of this diet is analysed by the supplier (SDS Special Diets services, Whitham, England) for the nutrients and contaminants and the results are kept available in the archives. Tap water (N.V. Hydron MiddenNederland) ad libitum. Results of routine physical, chemical and microbiological examination of drinking water as conducted by the supplier are kept available in the archives. The results of diet and water analyses were considered acceptable for this study.

Vehicle:

not specified

Controls:

yes, concurrent no treatment

Amount / concentration applied:

ca 0.1 ml of the test substance (0.035 g)

Duration of treatment / exposure:

Single treatment

Observation period (in vivo):

14 days

Number of animals or in vitro replicates:

3

Details on study design:

The in vitro/ex vivo Non-GLP Chicken Enucleated Eye Test did not show severe signs of irritation. Therefore, it was decided to proceed with the in vivo test. Both eyes of the rabbits selected were examined just before testing. Only rabbits without observable eye defects were used. The study was carried out with three rabbits and each rabbit was treated as follows: An amount of ca 0.1 ml of the test substance (0.035 g) was instilled in the conjunctival cul-de-sac of one of the eyes. After administration, the upper and lower eye lid were carefully closed and subsequently held together for at least one second before releasing, to prevent loss of material. The other eye remaining untreated, served as a control. The reactions of the test eyes were judged at circa one, 24, 48, 72 hand at 7 and 14 days after treatment using the scoring scale.

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.7

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2.7

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

fully reversible within: 14 days

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24, 48, 72h

Score:

0 - 1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24, 48, 72h

Score:

0 - 1

Max. score:

2

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

mean

Time point:

other: 24, 48, 72h

Score:

2.7 - 3

Max. score:

3

Reversibility:

fully reversible within: 14 days

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24, 48, 72h

Score:

1 - 2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritant / corrosive response data:

At 1 h after treatment, moderate redness and moderate swelling of the conjunctivae and moderate ocular discharge were observed in the three rabbits.
At 24, 48 and 72 h after treatment, slight opacity and slight iritis were observed in one rabbit, while moderate or severe redness and slight or moderate swelling of the conjunctivae and slight, moderate or severe ocular discharge were observed in the three rabbits.
At 7 days after treatment slight redness and moderate swelling of the conjunctivae were still observed in the third rabbit (no. 36).

Other effects:

At one hour after treatment, amounts of the test substance were still present in the conjunctival cul-de-sac of the rabbits. These remnants were removed by rinsing of the eye with physiological saline.
In addition, a haemorrhage of the conjunctivae and ischemic necrosis (only animal no. 36) were observed in the rabbits at 48 and 72 h after treatment.
At 7 days after treatment, eye effects had cleared in two rabbits.
At 14 days after treatment, eye effects had cleared in the third rabbit as well.

Table 1: Individual scores awarded to the ocular lesions elicited by compound T-1063FM

Animal no.	Corneal effects		Iris	Conjunctivae		Discharge
	Score	Area		Redness	Swelling
1 HOUR
32	0	0	0	2	2	2
34	0	0	0	2	2	2
36	0	0	0	2	2	2
24 HOURS
32	0	0	0	2	1	1
34	0	0	0	3	2	1
36	1	4	1	3	2	2
48 HOURS
32	0	0	0	3*	1	1
34	0	0	0	3*	1	1
36	1	4	1	3*	2	3
72 HOURS
32	0	0	0	3*	1	0
34	0	0	0	3*	1	0
36	1	4	1	3*	2	3
7 Days
32	0	0	0	0	0	0
34	0	0	0	0	0	0
36**	0	0	0	1	2	0

*=haemorrhage of the lower conjunctivae

**= effects had cleared completely at 14 days after treatment

 

Table 2: Mean values for corneal opacity, iritis, redness and swelling of the conjunctivae of each rabbit scored at 24, 48 and 72h after treatment with T-1063FM

Animal no.	Opacity	Iritis	Redness	Swelling
32	0.0	0.0	2.7	1.0
34	0.0	0.0	3.0	1.3
36	1.0	1.0	3.0	2.0

 

Interpretation of results:

irritating

Remarks:

Criteria used for interpretation of results: EU

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Effects on eye irritation: irritating

Justification for classification or non-classification

In view of the absence of any effects during the skin irritation study the attained results do not necessitate any classification and labelling regarding skin irritation according to EU classification rules [REGULATION (EC) 1272/2008].

In view of the reversible effects observed in the eye irritation study the substance has to be classified and labelled as irritating to eyes category 2 according to REGULATION (EC) 1272/2008.