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

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 March to 25 March 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

457-330-7

EC Name:

-

Cas Number:

66034-17-1

Molecular formula:

Hill formula: C4H10N2:H4O7P2 CAS formula: C4H10N2:H4O7P2

IUPAC Name:

(phosphonooxy)phosphonic acid; piperazine

Test material form:

solid: particulate/powder

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Species: Wistar outbred rat; Crl:(WI) WU BR
Supplier: Charles River, Germany
Sex and age: males and females, ca 7-8 weeks old upon arrival
Date of arrival: 26 February 2003
Quarantine/acclimatization: 13 days
Caging: a maximum of five animals per macrolon cage; individual housing during dermal exposure
Lighting: 12 hours light/12 hours dark cycle
Temperature during testing: 22 ± 3°C.
Relative humidity during testing: 30-70%. Upper limit incidentally up to 100%, 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.

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

At the end of the acclimatization period, the hair was removed from the back and flanks of the animals using electric clippers in a way as to avoid abrasions. The next day, the test material was applied to the clipped area of 5 male and 5 female rats by weight in one dose level of 2000 mg of the test substance per kg body weight. For this purpose, the animals were dosed with a 4 ml/kg body weight of a 500 mg/ml aqueous suspension of the test substance in order to obtain the 2000 mg/kg dose level. The weight variation of the animals did not exceed plus or minus 20% of the mean body weight at the start of the study. The amount of the test material was calculated for each animal individually and distributed in a thin layer over an area of intact skin, measuring at least 4 x 5 cm. For each animal, the exposure area was at least 20 cm2, i.e. at least 10% of the total body surface of the animals. The exposed area was covered by a gauze, which was fixed to the application site by means of adhesive tape and the entire trunk of the rat was wrapped with a tape to maintain the test patch in position and to retard evaporation of volatile substances. During the exposure period the animals were housed individually. After an exposure period of circa 24 hours the materials and residues of the material applied were removed with water. The animals were observed for mortality up to 14 days after treatment.

Duration of exposure:

24 h

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 males and 5 females

Control animals:

no

Details on study design:

Observation and measurements
Clinical signs: All visible reactions to treatment were recorded, including type, severity, onset and duration. Observations were made within 1 hour and within 4 hours after dosing, and subsequently in surviving animals at least once daily throughout an observation period of 14 days.
Body weights: The body weight of each animal was recorded immediately before dosing on day 0, and of the surviving animals on days 3, 7 and 14 of the study.
Dermal reactions: The skin reactions were evaluated by the method of Draize et al. (J. Pharmacol. Exp. Ther. 82 (1944) 377-390). Skin readings were made on day 1 after substance removal, and in surviving animals on days 3, 7, and 14 of the study.
Termination of the study: At the end of the observation period, on day 14 of the study, all surviving animals were killed with carbon dioxide and examined for external changes. Next, the abdomen and the thorax of each animal was opened and examined for gross pathological changes.

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed during the 14-day study period.

Clinical signs:

other: No clinical signs were observed during the 14-day study period.

Gross pathology:

Examination at necropsy of the animals did not reveal distinct treatment-related gross alterations.

Other findings:

No signs of skin irritation were observed during the 14-day observation period.

Any other information on results incl. tables

Table 1

Acute dermal toxicity of T-1063FM in rats; individual and mean bodyweights, dose amounts applied, and mortality

Animal no.	Dose (ml) applied^1	Bodyweights (g) recorded on day:				Motrality^2
		0	3	7	14
2000 mg/kg Males
2	1.1	267	268	285	317	-
4	1.1	274	274	287	319	-
6	1.0	261	263	276	298	-
8	1.1	265	259	273	300	-
10	1.1	272	275	286	318	-
Mean bodyweight		268	268	281	310	0/5
Females
1	0.80	201	202	210	216	-
3	0.74	185	185	192	212	-
5	0.74	185	185	192	205	-
7	0.77	192	187	195	227	-
9	0.78	194	192	197	213	-
Mean bodyweight		191	190	197	215	0/5

¹2000 mg/kg; dose of 4 ml/kg bodyweight of a 500 mg/ml aqueous suspension of the test suspension

²- = animal survived; + = animal died

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

Since all animals survived the 2000 mg/kg dose level, the dermal LD50 of T-1063FM is considered to be higher than 2000 mg/kg body weight.