Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1981
Report date:
1981

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Principles of method if other than guideline:
OECD guideline 402: at least 5 animals of the same sex per dose group. In this study 4 animals (2 males, 2 females) were used per dose group.
OECD guideline 402: care should be taken to avoid abrading the skin, which could alter its permeability. In this study the skin of the rabbits were abraded by making 4 epidermal incisions with a clean needle through the stratum corneum.
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Piperazine
EC Number:
203-808-3
EC Name:
Piperazine
Cas Number:
110-85-0
Molecular formula:
C4H10N2
IUPAC Name:
piperazine
Specific details on test material used for the study:
- Name of test material (as cited in study report): 4236-42-15
- Physical state: light yellow liquld
- Analytical purity: no data
- Lot/batch No.: J-82
- Stability under test conditions: no apparent change in the physical state of the test article during administration
- Storage condition of test material: no data

Test animals

Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Perfection Breeders, Inc., Douglassville, Pennsylvania
- Age at study initiation: adult
- Weight at study initiation: 2.010-2.250 kg
- Housing: individually in cages sized in accordance with the "Guide for the Care of Laboratory Animals" of the Institute of Laboratory Resources,
National Research Council.
- Diet (e.g. ad libitum): Wayne Rabbit Ration, ad libitum
- Water (e.g. ad libitum): fresh tap water, fit for human consumption, ad libitum,
- Acclimation period: 5d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3°C
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12h dark/12h light

Administration / exposure

Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: trunk (clipped free of fur)
- % coverage: min. 10%
- Type of wrap if used: rubber dam and an ace bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): washing was done
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 8 g/kg, 7, 9 and 10 g/kg
Duration of exposure:
24h
Doses:
Single dose: 8000 mg/kg;
LD50 doses: 7000, 9000 and 10000 mg/kg
No. of animals per sex per dose:
4 animals per dose (2 male, 2 female)
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 30 min, 2 and 4 hours after 24 h period of exposure and twice daily thereafter for 14 days.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, dermal signs
Statistics:
According to the method of Litchfield and Wilcoxon (1949) JPET 96: 99-114.

Results and discussion

Preliminary study:
In the preliminary study using 10 animals and dosing at 8000 mg/kg bw of the test material 3/10 animals died.
Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
8 300 mg/kg bw
95% CL:
7 300 - 9 400
Mortality:
7000 mg/kg : 1/4 animal died
9000 mg/kg : 2/4 animals died
(8000 mg/kg: 3/10 animals died)
10000 mg/kg : 4/4 animals died
Clinical signs:
-cyanosis, diarrhea, salivation, mild convulsions, bleeding from mouth and nose, ataxia, loss of righting, abnormal stance, catatonia, ptosis, and decreased activity
-necropsy of dying animals on study revealed black skin discoloration at application site, with hemorrhage below skin layers and skin necrosis
Body weight:
normal weight gain observed in all animals
Gross pathology:
Terminal necropsy revealed a discoloration of the tissue at application site, mottled kidneys and patchy lungs ( no visible lesions except signs of skin destruction and hemorrhage at application site).

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Based upon the observations made in the Acute Dermal Toxicity Test in rabbits, the LD 50 was 8300 mg/kg bw with 95% confidence limits of 7300 to 9400 mg/kg.
Executive summary:

In an acute dermal toxicity study performed according to OECD 402, but with deviations in number of animals used and abrasion of the skin, an LD50 value of 8300 mg/kg was established.