4-[4-(diethylamino)benzylidene]-5-ethoxy-2-phenyl-2,4-dihydro-3H-pyrazol-3-one

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14 January 2003 to 30 January 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

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:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Details on test material:

Identification: HSY-2701
Description: Red powder
Batch: 02J-20
Purity: 99.99 wt%
Test substance storage: At room temperature protected
Stability in vehicle: Not indicated
Expiry date: 29 June 2003

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Approximately 9 weeks old.
- Weight at study initiation: 182 to 209 g for females and 255 to 274 g for males. (Body weight variation did not exceed ± 20 % of the sex mean)
- Fasting period before study: Food was withheld overnight (for a maximum of 20 hours) prior to dosing until 3-4 hours after administration of the test substance.
- Housing: Group housing of 3 animals per sex cage in labelled Macrolon cages (type IV; height 18 cm) containing purified sawdust as bedding material (SAWI, Jelu Werk, Rosenberg, Germany).
- Diet (e.g. ad libitum): Free access to standard pelleted laboratory animal diet (from Altomin (code VRF 1), Lage, Germany)
- 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 ± 3 °C
- Humidity (%): 30-70 % RH
- Air changes (per hr): 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light and 12 hours dark per day.

All females were nulliparous and non-pregnant

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed by the testing laboratory

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3 animals per sex per dose (6 in total, 3 males and 3 females)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality/viability observations were performed twice daily. Bodyweights were recorded on day 1 (pre-administration), 8 and 15. Clinical signs were recorded at periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15. The symptoms were graded according to fixed scales and the time of onset, degree and duration were recorded:
Maximum grade 4 - grading slight (1) to very severe (4)
Maximum grade 3 - grading slight (1) to severe (3)
Maximum grade 1: presence is scored (1)
- Necropsy of survivors performed: yes, macroscopic examinations were recorded

Statistics:

The test substance substance was ranked within LD50 value ranges of 0-25, 25-200, 200-2000 or exceeding 2000 mg/kg body weight. No statistical analysis was performed (The method used was not intended to allow the calculation of a precise LD50 value).
The results were evaluated according to the EC criteria for classification and labelling of dangerous substances and preparations (Council Directive 67/548/EEC and all adaptations to technical progress and ammendments of the Directive published in the Official Journal of the European Communities).

Results and discussion

Preliminary study:

not applicable to test methodology.

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: Red discolouration of the faeces was noted.

Gross pathology:

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

Other findings:

No further observations were included.

Any other information on results incl. tables

Table 1: Clinical Signs

Test Day		1	1	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	Max grade
Hours after Treatment		0	2	4
Females 2000 mg/kg
Animal 1
Skin/Fur
Red Discolouration (Faeces)		-	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	(1)
Animal 2
Posture
Hunched Posture		-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	-	-	(1)
Skin/Fur
Red Discolouration (Faeces)		-	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	(1)
Animal 3
Skin/Fur
Red Discolouration (Faeces)		-	-	-	-	1	-	-	-	-	-	-	-	-	-	-	-	-	(1)
Males 2000 mg/kg
Animal 4
Skin/Fur/Plumage
Red Discolouration (Faeces)		-	-	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	(1)
Animal 5
Skin/Fur
Red Discolouration (Faeces)		-	-	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	(1)
Animal 6
Skin/Fur
Red Discolouration (Faeces)		-	-	-	1	1	-	-	-	-	-	-	-	-	-	-	-	-	(1)

Table 2: Bodyweights (Gram)

SEX/DOSE LEVEL	ANIMAL	DAY 1	DAY 8	DAY 15
Females 2000 mg/kg
	1	182	219	227
	2	208	242	266
	3	209	255	265
	Mean	200	239	253
	Std. Dev.	15	18	22
	N	3	3	3
Males 2000 mg/kg
	1	256	312	334
	2	274	328	357
	3	255	316	347
	Mean	262	319	346
	Std. Dev.	11	8	12
	N	3	3	3

 

Table 3: Macroscopic findings

Animal	Finding	Day of Death
Females 2000 mg/kg
1	No findings noted	Scheduled necropsy Day 15 after treatment
2	No findings noted	Scheduled necropsy Day 15 after treatment
3	No findings noted	Scheduled necropsy Day 15 after treatment
Males 2000 mg/kg
1	No findings noted	Scheduled necropsy Day 15 after treatment
2	No findings noted	Scheduled necropsy Day 15 after treatment
3	No findings noted	Scheduled necropsy Day 15 after treatment

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LD50 of HSY-2701 in Wistar rats was established to exceed 2000 mg/kg body weight.

Executive summary:

In a GLP compliant study, the acute toxicity of HSY-2701 was determined in accordance with OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method). 6 Wistar rats (3 male and 3 females) were dosed with 2000 mg/kg bw in corn oil. The LD₅₀ of HSY-2701 was determined to be >2000 mg/kg bw.