p-chlorophenylhydrazine hydrochloride

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from June 19 to July 23, 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

p-chlorophenylhydrazine hydrochloride

EC Number:

214-030-9

EC Name:

p-chlorophenylhydrazine hydrochloride

Cas Number:

1073-70-7

Molecular formula:

C6H7ClN2.ClH

IUPAC Name:

(4-chlorophenyl)hydrazine hydrochloride

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- batch No.of test material: 6108120253
- Expiration date of the batch: March 2013

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, 97633 Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8 - 10 weeks old
- Weight on the day of administration:
Step 1 / animals no. 1 - 3: 169 – 173 g; Step 2 / animals no. 4 - 6: 149 – 166 g; Step 3 / animals no. 7 - 9: 147 – 160 g
Step 4 / animals no. 10 - 12: 152 – 160 g.

Housing and Feeding Conditions
- Full barrier in an air-conditioned room
- Temperature: 22 3 °C
- Relative humidity: 55 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: 10 x / hour
- Free access to Altromin 1324 maintenance diet for rats and mice (lot no. 0715)
- Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- The animals were kept in groups in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. 261111)
- Certificates of food, water and bedding are filed at BSL BIOSERVICE
- Adequate acclimatisation period (at least five days) under laboratory conditions

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

cotton seed oil

Details on oral exposure:

At dosage of 2000 mg/kg the test item was suspended in the vehicle cottonseed oil at a concentration of 0.2 g/mL and administered at a dose volume of 10 mL/kg.
At dosage of 300 mg/kg the test item was suspended in the vehicle cottonseed oil at a concentration of 0.03 g/mL and administered at a dose volume of 10 mL/kg.
At dosage of 50 mg/kg the test item was suspended in the vehicle cottonseed oil at a concentration of 0.005 g/mL and administered at a dose volume of 10 mL/kg.

Doses:

2000, 300 and 50 mg/kg
The test item was administered at a single dose by gavage using a feeding tube.
The test item was administered at a dose volume of 10 mL/kg body weight.

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

The surviving animals were observed for 14 days after dosing for general clinical signs, morbidity and mortality.
The animals were weighed on day 1 (prior to the administration) and on days 8 and 15.
A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded. Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. All abnormalities were recorded.
Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern were examined. Particular attention was directed to observations of tremor, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Pathology
The animals which died spontaneously were necropsied as soon as they were found dead. The animal which had to be sacrificed for ethical reasons during the observation period was necropsied as soon as it was killed.
At the end of the observation period the surviving animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally at a dosage of approximately 8 mL/kg bw.
All animals were subjected to gross necropsy. All gross pathological changes were recorded and in case of findings the tissues were preserved for a possible histopathological evaluation. The preserved tissues of which no histopathological evaluation was made will be discarded 3 months after the release of the final report unless otherwise agreed upon with the sponsor.

Results and discussion

Effect levels

Effect levels 1

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

200 mg/kg bw

Based on:

test mat.

Effect levels 2

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 50 - < 300 mg/kg bw

Based on:

test mat.

Mortality:

All animals treated with the test item at a dose of 2000 mg/kg died spontaneously on the day of treatment. Two animals treated with the test item at a dose of 300 mg/kg died spontaneously on test day 2. One animal of this group had to be sacrificed for ethical reasons on test day 3. All remaining animals survived until the end of the study.

Clinical signs:

The most relevant clinical findings in the animals treated with the test item at a dose of 2000 mg/kg bw were recumbency, apathy, wasp waist, bradykinesia, cyanosis, and half eyelid-closure.
The most relevant clinical findings in the animals treated with the test item at a dose of 300 mg/kg bw were reduced spontaneous activity, apathy, recumbency, bradykinesia, cyanosis, piloerection, half eyelid-closure, kyphosis, haematuria, and hypothermia.
The most relevant clinical findings in the animals treated with the test item at a dose of 50 mg/kg bw were reduced spontaneous activity, cyanosis, and bradykinesia.

Body weight:

Throughout the 14-day observation period, the body weight gain of the surviving test animals was within the normal range of variation for this strain.

Gross pathology:

Macroscopic findings of animals died spontaneously or sacrificed for ethical reasons:
At necropsy, a dark liver and gall bladder and a white/fair lung were observed in all animals of steps 1 and 2.
Macroscopic findings of surviving animals:
At necropsy, no macroscopic findings were observed in any of the surviving animals.

Applicant's summary and conclusion

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

Under the conditions of the present study, a single oral application of the test item 4-CHLOROPHENYLHYDRAZINE HYDROCHLORIDE to rats at a doses of 2000 mg/kg body weight and 300 mg/kg body weight was associated with mortality. A single oral application of the test item 4-CHLOROPHENYLHYDRAZINE HYDROCHLORIDE to rats at a dose of 50 mg/kg body weight was associated with signs of toxicity, but no mortality.
The median lethal dose of 4-CHLOROPHENYLHYDRAZINE HYDROCHLORIDE after a single oral administration to female rats, observed over a period of 14 days is:
LD50 cut-off (rat): 200 mg/ kg bw

On the basis of the test results and in conformity with the criteria given in Annex I of Regulation (EC) 1272/2008 [8], the substance should be classified as toxic into category 3.
On the basis of the test results and in conformity with the criteria given in GHS (Globally Harmonized System of Classification and Labelling of Chemicals) [9], the substance should be classified into category 3.