2-({2-[4-(dimethylamino)benzoyloxy]ethyl}(methyl)amino)ethyl 4-(dimethylamino)benzoate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 April 2006 to10 May 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

December 17, 2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

466-080-8

EC Name:

-

Molecular formula:

C23H31N3O4

IUPAC Name:

466-080-8

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Name: LFC 2098
Product/common name: LFC 2098
Purpose: Industrial chemical
Colour: white
Physical state: solid, powder
Purity: 99.69%
Storage: at room temperature, protected from light
Molecular formula: C23H3104N3
Molecular weight: 413
Safety precautions: Routine hygienic procedures were sufficient to assure personnel health and safety.

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
HsdRccHan : WIST rats (Full-Barrier), Sex: females, non-pregnant, nulli-parous.
Step 1: Body weight at the commencement of the study: 166 - 211 g;
Step 2: Body weight at the commencement of the study: 190 - 244 g;
Three female animals were used for each step.
The animals were derived from a controlled full barrier maintained breeding
system (SPF).
Source: Harlan Winkelmann GmbH, D-33178 Borchen.
According to Art. 9.2, No.7 of the German Act on Animal Welfare the animals were bred for experimental purposes.

ENVIRONMENTAL CONDITIONS
The animals were barrier maintained (semi-barrier) in an air-conditioned room
Temperature: 22 ± 3 °C
Rel. humidity: 55 ± 10%
Artificial light, sequence being 12 hours light, 12 hours dark
Air change: 10 x / hour
Feeding ad libitum, ssniff R/M-H, 10 mm V1534-000 complete diet for rats/mice - maintenance, totally-pathogen-free (TPF)
Free access to tap water (drinking water, municipal residue control, microbiol. controlled periodically)
The animals were kept in Macrolon cages on Lignocel bedding
Certificates of food, water and bedding are filed at BSL Bioservice
Adequate acclimatization period

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

cotton seed oil

Details on oral exposure:

In the first step 1 g of the test item was dissolved with the vehicle ad 5 mL to gain a concentration of 2000 mg/kg body weight.
In the second step 2 g of the test item were dissolved with the vehicle ad 10 mL to gain a concentration of 2000 mg/kg body weight.
The test substance was freshly mixed prior to administration and stirred throughout dose administration to guarantee stability and homogeneity. The vehicle was chosen due to its non—toxic characteristics.

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

Group 1: 3 female rats
Group 2: 3 female rats

Control animals:

no

Details on study design:

In the first step 1 g of the test item was dissolved with the vehicle ad 5 mL to gain a concentration of 2000 mg/kg body weight.
In the second step 2 g of the test item were dissolved with the vehicle ad 10 mL to gain a concentration of 2000 mg/kg body weight.

Prior to the administration a detailed clinical observation was made of all animals.
Prior to administration of the test item animals were fasted by withholding food overnight. Following the period of fasting the animals were weighed and the test item was administered. Then the food was withheld for a further 3-4 hours.
The test item was administered in a single dose by gavage using an intubation cannula.
The test item was administered at a volume of 10 mL/kg body weight.
The starting dose (step 1) was selected to be 2000 mg/kg body weight. As two of three animals survived, the second step was performed with the same dose. According to the acute toxic class method regime no further testing was required since no compound-related mortality was found in any animals of step 2.
The surviving animals were observed for 14 days after dosing.
The animals were weighed prior to the administration and once a week thereafter.
A careful clinical examination was made several times on the day of dosing.
Part of this were at least three observations within the first four hours post-dose. Animals were observed once a day thereafter.
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.
At the end of the observation period the surviving animals were sacrificed by an overdosage of pentobarbital. All animals were subjected to gross necropsy. All gross pathological changes were recorded.
Individual reactions of each animal were recorded at each observation time.
Toxic response data were recorded by sex and dose level.
Nature, severity and duration of clinical observations were described,
Body weight changes were summarized in tabular form.
Necropsy findings were described.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

2 500 mg/kg bw

Based on:

test mat.

Mortality:

Animal No. l of step 1 showed slightly reduced spontaneous activity, apathy and piloerection 120, 180, 240, 300 as well as 330 min post-dose. 1 day post-dose this animal was found dead.

Clinical signs:

Animal No. l of step 1 showed slightly reduced spontaneous activity, apathy and piloerection 120, 180, 240, 300 as well as 330 min post-dose. 1 day post-dose this animal was found dead.
Animal No, 2 of step 1 showed slightly reduced spontaneous activity, apathy as well as piloerection 60 min. post-dose. 70, 90 as well as 150 min. post-dose slightly reduced spontaneous activity, apathy, piloerection and half eyelid-closure were recorded. 180 as well as 200 min. post-dose slightly reduced spontaneous activity, apathy as well as piloerection were observed 220 as well as 240 min. post-dose slightly reduced spontaneous activity as well as piloerection were recorded. 24 hours post-dose no symptoms were observed.
Animal No. 3 of step 1 showed slightly reduced spontaneous activity 240 as well as 300 min. post-dose. 400 min. post—dose no symptoms were recorded.
No other treatment related effects Were observed in any animals of step 1.

The second step was performed with the same dosage in the same manner to a further group of 3 female rats (HsdRccHan : WIST).
No treatment related effects were observed in any animals of step 2.

Therefore, according to OECD Guideline 423, a sufficient estimation of the acute oral toxicity of the test item is provided.

Body weight:

Throughout the 14-days observation period no weight loss was recorded in any surviving animals. The weight gain was within the expected range.

Gross pathology:

At the end of the observation period the surviving animals were sacrificed.
Necropsy of all animals was carried out to record gross pathological changes.
Animal No. 1 of step 1 was found in abdominal position. It showed foamy discharge on the nose, which was already in crystalline appearance. Liquid stomach contents were recorded.
No other treatment related effects were observed in any animals of steps 1 and 2.

The euthanized animals showed acute injection of blood vessels in the abdominal region, which is due to the euthanasia injection.

Any other information on results incl. tables

Step	sex	dose (mg/kg)	number of animals	number of deaths
1	F	2000	3	1
2	F	2000	3	0

Step 1 (2000 mg/kg bw)
Animal no.	Weight (g)
	Day 0	Day 7	Day 14
1	174	Found dead 1 day post dose
2	166	183	185
3	211	216	225
Step 2 (2000 mg/kg bw)
Animal no.	Weight (g)
	Day 0	Day 7	Day 14
1	206	234	238
2	244	265	273
3	190	202	218

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

LD50 cut-off: 2500 mg/kg body weight according to the conditions of the test

Executive summary:

In this guideline (OECD 423) study, conducted with GLP certification, the test material (EC 466-080-8) was shown to have an LD50 >2500 mg/kg bodyweight. In conformity with the criteria given in Annex VI of the Directive, the substance should be: not classified