3,4-dichlorobenzonitrile

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1997-02-11 - 1997-02-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

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

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

other: Directive 96/54/EC, Part B.1 tris "Acute Toxicity-Oral, Acute Toxic Class Method"

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

3,4-dichlorobenzonitrile

EC Number:

229-494-8

EC Name:

3,4-dichlorobenzonitrile

Cas Number:

6574-99-8

Molecular formula:

C7H3Cl2N

IUPAC Name:

3,4-dichlorobenzonitrile

Test material form:

solid: bulk

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Sulzfeld, Germany.
- Age at study initiation: approx. 6 weeks
- Weight at study initiation: within +/- 20 % of the sex mean
- Fasting period before study: food was withheld overnight prior to dosing until approximately 3-4 hours after administration of the test substance.
- Housing: group housing of 3 animals per sex per cage in labelled polycarbonate cages containing purified sawdust as bedding material (Woody SPF, supplied by B.M.I, Helmond, The Netherlands)
- Diet (e.g. ad libitum): free access to standard pelleted laboratory animal diet (from Carfil Quality BVBA, Oud-Turnhout, Belgium)
- Water (e.g. ad libitum): free access to tap-water
- Acclimation period: at least 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 50
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): lighting was 12 hours artificial flourescent light and 12 hours dark per day

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

- Method: Oral gavage, using a stainless steel stomach tube.
- Frequency: Once, on day 1.
- Animals: 3 animals of the same sex per dose group.

Doses:

200 mg/kg (females), 200 mg/kg (males) and 2000 mg/kg (females)

No. of animals per sex per dose:

three

Control animals:

no

Details on study design:

OBSERVATIONS:
- Mortality/Viability: Twice daily. The time of death was recorded as precisely as possible
- Body weights: Days 1 (pre-administration), 8 and 15 and at death (if found death after day 1).
- Clinical signs: At periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15. The time of onset, degree and duration were recorded.

- Necropsy of survivors performed: yes

Statistics:

not performed

Results and discussion

Effect levels

Effect levels 1

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 200 - < 2 000 mg/kg bw

Based on:

test mat.

Effect levels 2

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

> 200 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

All femals dosed at 2000 mg/kg b.w. were found dead within 24 hours of dosing.

Clinical signs:

other: 200 mg/kg: Hunched posture, uncoordinated movements. 2000 mg/kg: Lethargy, hunched posture, uncoordinated movements, piloerection. The surviving animals had recovered from symptoms between days 2 and 3. The surviving animals had recovered from the sympto

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Any other information on results incl. tables

The test substance was ranked within LD50 values ranges of 0 - 25, 25 - 200 or 200 - 2000 mg/kg b.w. or as exceeding 2000 mg/kg b.w.

No statistical analysis was performed (The method used is not intended to allow the calculation of a percise LD50 value).

The results were evaluated according to the EEC criteria for classification and labelling requirements for dangerous substances and preparations (Guidelines in Commission Directive 93/21/EEC, 27th april 1993).

Table 1: Body weights of test groups.

Group / Sex	Animal	Day 1 [g]	Day 2 [g]	Day 8 [g]	Day 15 [g]
Group 1 / females	1	126		179	202
	2	131		171	195
	3	122		161	170
	Mean	126		170	189
	St. Dev.	5		9	17
	N	3		3	3
Group 2 / males	4	185		249	283
	5	194		277	309
	6	186		239	277
	Mean	188		255	290
	St. Dev.	5		20	17
	N	3		3	3
Group 3 / females	7	138	137	-	-
	8	148	147	-	-
	9	143	137	-	-
	Mean	143		-	-
	St. Dev.	5		-	-
	N	3		0	0

Table 2: Observed clinical signs of test group 1 (females, 200 mg/kg)

Test day			1	1	1	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15
Time after treatment. hours			0	2	4
Animal number	Signs	max. grade
1	gait/motility: uncoordinated movements	(3)	-	1	2	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
2	gait/motility: uncoordinated movements	(3)	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
3	Posture: Hunched posture   gait/motility: uncoordinated movements	(1)     (3)	-     -	-     2	-     2	1     1	-     -	-     -	-     -	-     -	-     -	-     -	-     -	-     -	-     -	-     -	-     -	-     -	-     -	-     -

Table 3: Observed clinical signs of test group 2 (males, 200 mg/kg)

Test day			1	1	1	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15
Time after treatment. hours			0	2	4
Animal number	Signs	max. grade
4	gait/motility: uncoordinated movements	(3)	-	1	2	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
5	gait/motility: uncoordinated movements	(3)	-	1	2	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
6	gait/motility: uncoordinated movements	(3)	-	1	2	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

Table 4: Observed clinical signs of test group 3 (females, 2000 mg/kg).

Test day			1	1	1	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15
Time after treatment. hours			0	2	4
Animal number	Signs	max. grade
7	Behavior: Lethargy   Posture: Hunched posture   gait/motility: uncoordinated movements	(3)     (1)     (3)	-     -     -	-     -     2	1     1     2	+     +     +
8	Posture: Hunched posture   gait/motility: uncoordinated movements	(1)     (3)	-     -	-     2	1     2	+     +
9	Posture: Hunched posture   gait/motility: uncoordinated movements   Skin /FUR / Plumage: Piloerection	(1)     (3)     (1)	-     -     -	-     2     1	1     2     1	+     +     +

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The oral LD50 value of 3,4-Dichlorobenzonitrile in Wistar rats was established to be within the range of 200-2000 mg/kg body weight.
According to the EEC criteria for classification and labelling, 3,4-Dichlorobenzonitrile must be classified as harmful if swallowed.
Based on the test result and observation, it can be expected that the classificatoin is also suitable for H302 where 300mg/kg is the threshold.

Executive summary:

The study was carried out based on the guidelines described by oral gavage to three femal Wister rats at 200 mg/kg body weight. In a stepwise procedure additional groups of animals were dosed at 200 (males) and 2000 (females) mg/kg body weight. All animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed on the day of death or after terminal sacrifice 8 (day 15).

All three animals dosed at 2000 mg/kg b.w. were found dead within 24 hours of dosing. The surviving animals recovered from all clinical signs observed between days 2 and 3. The body weight gain shown by the surviving animals over the study period was considered to be normal. No abnormalities were found at macroscopic post mortem examination of the animals.

The oral LD50 value of 3,4 -Dihchlorobenzonitrile in Wistar rats was established to be within the range of 200 -2000 mg/kg body weight.