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

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 September 2003 to 01 October 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:

2004

Report date:

2004

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: "Acute Dermal Toxicity" and JMAFF: Japanese Test Guidelines (2000).

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Details on test material:

Identification: HSY-2701
Description: Red powder
Batch: 1004-1
Purity: 99.8 %
Test substance storage: At room temperature in the dark
Stability under storage conditions: Stable
Expiry date: 12 February 2004

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, Sulzfield, Germany
- Age at study initiation: Approximately 12 weeks old.
- Weight at study initiation: Males 396 to 443 g and females 251 to 277 g (Body weight variation did not exceed ± 20 % of the sex mean).
- Fasting period before study: Not reported
- Housing: Individually housed in labelled Macrolon cages (type III, height 15 cm) with purified sawdust bedding material (SAWI, Jelu Werck, Rosenburg, Germany).
- Diet (e.g. ad libitum): Free access to standard pelleted laboratory animal diet (Altromin (code VRF 1), Lage, Germany).
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: At least five days prior to treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.0 ± 3.0 °C
- Humidity (%): 30-70% (34-73 % actual range).
- Air changes (per hr): 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 23 hours artificial fluorescent light and 12 hours darkness

Administration / exposure

Type of coverage:

occlusive

Vehicle:

propylene glycol

Details on dermal exposure:

TEST SITE
- Area of exposure: approximately 25 cm^2 for males, 18 cm^2 for females on the back of each animal.
- % coverage: 10 % of total body surface
- Type of wrap if used: The formulations were held in contact with the skin with a dressing consisting of a surgical gauze patch (Surgy 1D), successively covered with aluminium foil and Coban flexible bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The skin was cleaned of residual test substance using water.
- Time after start of exposure: 24 hours post application.

VEHICLE
- Amount(s) applied (volume or weight with unit): 10 mL/kg

Duration of exposure:

24 hours

Doses:

2000 mg/kg

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days (study terminated on day 15)
- Frequency of observations and weighing: Mortality/Viability observations were performed twice daily. Body weights were measured on days 1 (pre-administration), 8 and 15.
- Necropsy of survivors performed: yes

Statistics:

The results were evaluated according to the OECD Harmonised Integrated Hazard Classification System for Human Health and Environmental Effects of Chemical Substance (OECD 1998) and the EC criteria for classification and labelling of dangerous substances and preparations (Council Directive 67/548/EEC and all adaptations to technical progress and amendments of the Directive published in the Official Journal of the European Communities.

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 occurred during the course of the study.

Clinical signs:

other: Red or yellow staining, scales, scabs and/or maculate erythema were noted on the treated dorsal skin of all animals during the observation period. In addition, alopecia and or/scabs in the cheek region (three males; between days 1 and 13) or lethargy (one

Gross pathology:

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

Other findings:

No further observations were reported.

Any other information on results incl. tables

Individual results for clinical signs, body weight and mortality are tabulated in the attached document.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

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

Executive summary:

In a GLP compliant study the acute dermal toxicity of HSY-2701 was investigated in male and female Wistar rats. Animals were dosed with 2000 mg/kg bw in propylene glycol under an occlusive wrap. After 24 hours, residual test material was removed and the animals were observed for the following 14 days for signs of toxicity. The LD₅₀ of HSY-2701 was determined to be >2000 mg/kg bw.