Benzoic acid, 2-[[(2'-cyano[1,1'-biphenyl]-4-yl)methyl]amino]-3-nitro-, methyl ester

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Date of start of the experimental period: 27 May 2004. Date of end of the experimental period: 10 Jun. 2004.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

methyl 2-[[4-(2-cyanophenyl)phenyl]methylamino]-3-nitrobenzoate

EC Number:

604-135-1

Cas Number:

139481-28-0

Molecular formula:

C22H17N3O4

IUPAC Name:

methyl 2-[[4-(2-cyanophenyl)phenyl]methylamino]-3-nitrobenzoate

Test material form:

solid: particulate/powder

Details on test material:

batch No. February 2004

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

CHS (Controlled Health Status) Sprague-Dawley Ico: OFA-Sd (IOPS Caw)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratoires France, Domaine des Oncins, 69592 L'Arbresle Cedex, France.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8-12 weeks at the time of administration.
- Housing: Daily observations were performed at the time of delivery of the animals and during the period of acclimatization. Animals were housed in cages of standard dimensions with sawdust bedding (or equivalent). Cages were cleaned at least once per week.
The animals were placed in an air-conditioned (19-23° C) animal house kept at relative humidity between 45% and 65% in which non-recycled filtered air was changed approximately 10 times per hour. The artificial day/night cycle involved 12 hours light and 12 hours darkness with light on at 7.30 a.m.

- Diet (e.g. ad libitum): RM1 (E)-SQC SDS/DIETEX (quality controlled/radiation sterilised) was available ad libitum. The criteria for acceptable levels of contaminants in the feed supply were within the limits of the analytical specifications established by the diet manufacturer.

- Water (e.g. ad libitum): Drinking water was available ad libitum in polycarbonate feeder bottles with a stainless steel nipple. A specimen of water is obtained every 6 months and sent to the Laboratoire Départemental d'Analyse du Cher- 216, Rue Louis Mallet- 18014 Bourges Cedex, France, for analysis. The criteria for acceptable levels of contaminants in the water supply were within the limits of the analytical specifications.

- Acclimation period: Minimum of five days before the treatment in the laboratory animal house where the experiment took place.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23 °C
- Humidity (%): between 45% and 65%
- Air changes (per hr): 10 times per hour.
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Type of coverage:

other: CAN 3 was applied to each animal on a piece of absorbant gauze measuring 30 cm2 (6 cm x 5 cm).

Vehicle:

other: CAN 3 was moistened with sterile water in order to ensure good contact with the skin.

Details on dermal exposure:

Timing, frequency and duration of administration
CAN 3 was applied to the back of the animal on a previously shaved area. The absorbent gauze was held in place for 24 hours using an elastic band. At the end of the exposure period, any remaining CAN 3 was removed by washing with sterile water (without success) following by ethanol 70°.

Application of the test substance
CAN 3 was applied to the skin in the dorsal region of each animal without prior fasting. For this purpose, on the day prior to application, the dorsal region of each rat was carefully clipped on an area of approximately 50 cm2 using an electric clipper, avoiding damage to the skin.
Any animals found to have damaged skin at the time of application was removed from the study. On the day of study, test substance was applied to the prepared area to expose at least 10% of the total body surface. CAN 3 was applied to each animal on a piece of absorbant gauze measuring 30 cm2 (6 cm x 5 cm).
Remark: the application area was the same for all the animals. The total body area has been calculated according to the following formula:
A= 0.1 PVexp0.685
A= total body area in m2 - PV: Body weight of the group in kg.

Duration of exposure:

24 hours.

Doses:

2000 mg/kg body weight.

No. of animals per sex per dose:

A single group of ten animals (5 males and 5 females) was dosed at the dose of 2000 mg/kg body weight.

Control animals:

not required

Details on study design:

- Duration of observation period following administration: Animals were examined clinically twice on the day of treatment (between 30 and 90 minutes after
administration and then again between 3 and 4 hours post-dose). Thereafter they were examined clinically at least once daily for 14 days.
- Frequency of observations and weighing: On D1 between 30 and 90 minutes after administration and on D7 the animals were submitted to a full clinical examination outside the housing cage, including functional and neurobehavioural tests.
- Necropsy of survivors performed: yes
- Other examinations performed: mortality, clinical and skin observations, body weight, macroscopic findings during necropsy.

Results and discussion

Preliminary study:

A dose of 2000 mg/kg caused no mortality and no dermal reactions during a 14-day period, in the male and female Sprague-Dawley rats.

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study.

Clinical signs:

other: No clinical signs were observed during the course of the study.

Gross pathology:

No organ or tissue gross findings were seen at necropsy.

Other findings:

No dermal reactions were observed during the course of the study.

Any other information on results incl. tables

Body weight and clinical signs are reported in the illustration attached below.

Applicant's summary and conclusion

Interpretation of results:

other: The substance is not classified for Acute dermal toxicity according to CLP Regulation criteria.

Conclusions:

Under the experimental conditions adopted, dermal application of the test substance CAN 3 (batch No. February 2004) at a dose of 2000 mg/kg caused no mortality and no dermal reactions during a 14-day period, in the male and female Sprague-Dawley rats.
Under the experimental conditions adopted, the LD50 of the test substance CAN 3 (batch No. February 2004) administered by the dermal route was higher than 2000 mg/kg body weight in the male and female Sprague-Dawley rats.

Executive summary:

Under the experimental conditions adopted, dermal application of the test substance CAN 3 (batch No. Febrnary 2004) at a dose of 2000 mg/kg caused no mortality and no dermal reactions during a 14 -day period, in the male and female Sprague-Dawley rats.

Under the experimental conditions adopted, the LD50 of the test substance CAN 3 (batch No. February 2004) administered by the dermal route was higher than 2000 mg/kg body weight in the male and female Sprague-Dawley rats.