C16-18-(even numbered, C18-unsaturated)-alkylamines acetates

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study according to GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

C16-18-(even numbered, C18-unsaturated)-alkylamines acetates

EC Number:

817-198-7

Cas Number:

1273322-47-6

Molecular formula:

R-NH2xHOOCCH3 R = alkyl, mainly C16-18-(even numbered) and C18-unsaturated

IUPAC Name:

C16-18-(even numbered, C18-unsaturated)-alkylamines acetates

Test material form:

other: solid

Details on test material:

Physical State at RT:solid
Colour:white-yellow
Storage:at room temperature

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Species/strain: healthy rats, WISTAR rats Crl: WI(Han) (full barrier)
Source: Charles River, 97633 Sulzfeld, Germany
Sex: female, non-pregnant, nulliparous
Number of animals: 3 per step
Age at the beginning of the study: 8 - 11 weeks old
Body weight
on the day of administration: Step 1 / animals no. 1 - 3: 165 – 192 g;
Step 2 / animals no. 4 - 6: 150 – 163 g;
Step 3 / animals no. 7 - 9: 153 – 181 g
Step 4 / animals no. 10 - 12: 153 – 172 g.
The animals were derived from a controlled full barrier maitained breeding system(SPF). According to Art. 9.2, No. 7 of the German act on animal welfare the animals were bred for experimental purposes.

Environmental Conditions:
Full barrier in an air-conditioned room
Temperature: 22 +/-3 °C
Relative humidity: 55 +/-10%
Artificial light, sequence being 12 hours light, 12 hours dark
Air change: 10 x / hour
-Free access to Altromin 1324 maintenance diet for rats and mice (lot no. 1455)
-Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
-The animals were kept in groups in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding (lot no. 060411)
-Certificates of food, water and bedding are filed at BSL BIOSERVICE
-Adequate acclimatisation period (at least five days) under laboratory conditions

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: Sesame Oil

Details on oral exposure:

The test item was administered at a single dose by gavage using a feeding tube
The test item was administered at a dose volume of 10 mL/kg body weight
Preparation of Dose Formulation:
The test item was weighed out into a tared plastic vial on a precision balance. Finally it was homogenised for 1 to 2 minutes using a homogeniser.

Doses:

Step Sex/No. Dose (mg/kg)
1 female/1-3 2000
2 female/4-6 2000
3 female/7-9 300
4 female/10-12 300
The starting dose was selected to be 2000 mg/kg body weight. Compound-related mortality was recorded for 1 animal of step 1. Based on these results and according to the acute toxic class method regime, a second step was performed at the same dose. Compound-related mortality was recorded for 2 animals of step 2. Based on these results and according to the acute toxic class method regime, a third step was performed at a dose of 300 mg/kg body weight. No compound-related mortality was recorded for any animal of step 3. Based on these results and according to the acute toxic class method regime, a fourth step was performed at the same dose. No compound-related mortality was recorded for any animal of step 4. Based on these results and according to the acute toxic class method regime no further testing was required

No. of animals per sex per dose:

3 Animals per step (4 steps were used)

Control animals:

no

Details on study design:

Preparation of Animals:
The animals were marked for individual identification by tail painting.
Prior to the administration a detailed clinical observation was made of all animals.
Prior to the administration food was withheld from the test animals for 16 to 19 hours (access to water was permitted). Following the period of fasting the animals were weighed and the test item was administered. Food was provided again approximately 4 hours post dosing.
Opservation Period:
All animals were observed for 14 days after dosing for general clinical signs, morbidity and mortality. The animals were weighed on day 1 (prior to the administration) and on days 8 and 15.

Clinical Examination: A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded. Thereafter, the animals were observed for clinical signs once daily until the end of the observation period.

Pathology: At the end of the observation period the animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally (Narcoren®, Merial; lot no.: 212041; expiry date: 04/2014) at a dosage of approximately 8 mL/kg bw. All animals were subjected to gross necropsy. All gross pathological changes were recorded.

Evaluation of Results:
Results were interpreted according to OECD Guideline 423, Annex 2 (see also flow charts in the appendix of the study plan).
Individual reactions of each animal were recorded at each time of observation.
Toxic response data were recorded by dose level.
Nature, severity and duration of clinical observations were described.
Body weight changes were summarised in tabular form.
Necropsy findings were described.
On the basis of the test results, the test substance may be classified in any of the following classes in conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC :
• Very toxic
Substances and preparations shall be classified as very toxic, and assigned the symbol “T+” and indication of danger “very toxic” in accordance with the criteria specified below:
R28 Very toxic if swallowed
- LD50 oral, rat  25 mg/kg
- less than 100% survival at 5 mg/kg oral, rat by the fixed dose procedure, or
- high mortality at doses  25 mg/kg oral, by the acute toxic class method.
• Toxic
Substances and preparations shall be classified as toxic, and assigned the symbol “T” and indication of danger “toxic” in accordance with the criteria specified below. Risk phrases shall be assigned in accordance with the following criteria:
R25 Toxic if swallowed
- LD50 oral, rat 25 < LD50  200 mg/kg
- discriminating dose, oral rat 5 mg/kg: 100% survival but evident toxicity, or
- high mortality in the dose range > 25 to  200 mg/kg oral, rat, by the acute toxic class method.
• Harmful
Substances and preparations shall be classified as harmful, and assigned the symbol “Xn” and indication of danger “harmful” in accordance with the criteria specified below. Risk phrases shall be assigned in accordance with the following criteria:
R22 Harmful if swallowed
- LD50 per oral, rat 200 < LD50  2000 mg/kg
- discriminating dose, oral rat, 50 mg/kg: 100% survival but evident toxicity,
- less than 100% survival at 500 mg/kg, rat oral by the fixed dose procedure, or
- high mortality in the dose range > 200 to  2000 mg/kg oral, rat, by the acute toxic class method.
On the basis of the test results, the following risk phrases may be assigned in conformity with the criteria given in Annex I of Regulation (EC) 1272/2008 :
Category 1: LD50  5 mg/kg. DANGER. Skull and crossbones in diamond. Fatal if swallowed.
Category 2: LD50 > 5 mg/kg  50 mg/kg. DANGER. Skull and crossbones in diamond. Fatal if swallowed.
Category 3: LD50 > 50 mg/kg  300 mg/kg. DANGER. Skull and crossbones in diamond. Toxic if swallowed.
Category 4: LD50 > 300 mg/kg  2000 mg/kg. WARNING. Exclamation point in diamond. Harmful if swallowed.
On the basis of the test results, the following risk phrases may be assigned in conformity with the criteria given in GHS - Globally Harmonized System of Classification and Labelling of Chemicals, third revised edition, July 2009 :
Category 1: LD50  5 mg/kg
DANGER. Skull and crossbones in diamond. Fatal if swallowed.
Category 2: LD50 > 5 mg/kg  50 mg/kg
DANGER. Skull and crossbones in diamond. Fatal if swallowed.
Category 3: LD50 > 50 mg/kg  300 mg/kg.
DANGER. Skull and crossbones in diamond. Toxic if swallowed.
Category 4: LD50 > 300 mg/kg  2000 mg/kg.
WARNING. Exclamation point in diamond. Harmful if swallowed.
Category 5: LD50 > 2000 mg/kg  5000 mg/kg.
WARNING. No symbol. May be harmful if swallowed.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

other: LD50 cut-off

Effect level:

2 000 mg/kg bw

Based on:

test mat.

Mortality:

One Aniaml from step 1 and 2 animals from step 2 at 2000mg/kg were found dead.
No Mortalitiy was observed in animals of step 3 and 4 at 300mg/kg dose.

Clinical signs:

other: The most relevant clinical findings in the animals treated with the test item at a dose of 2000 mg/kg bw were reduced spontaneous activity, piloerection, nasal discharge and chromodacryorrhea, diarrhoea, body weight loss, half and complete eyelid-closure,

Gross pathology:

Macroscopic findings of surviving animals:
Macroscopic findings of animals not having survived until the end of the observation period:
Necropsy revealed autolysis on all organs, and a discoloured, bloated caecum.
With the exception of acute injection of blood vessels in the abdominal region, which is due to the euthanasia injection, no specific gross pathological changes were recorded for any other animal.

Any other information on results incl. tables

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Applicant's summary and conclusion

Interpretation of results:

harmful

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the present study, a single oral application of the test item C16-18-(even numbered, C18-unsaturated)-alkylamines acetates to rats at a dose of 2000 mg/kg body weight was associated with signs of toxicity and mortality.
Under the conditions of the present study, a single oral application of the test item C16-18-(even numbered, C18-unsaturated)-alkylamines acetates to rats at a dose of 300 mg/kg body weight was associated with signs of toxicity but no mortality.
The median lethal dose of C16-18-(even numbered, C18-unsaturated)-alkylamines acetates after a single oral administration to female rats, observed over a period of 14 days is:
LD50 cut-off (rat): 2000 mg/ kg bw
In conformity with the criteria given in Annex VI to Commission Directive 2001/59/EC , the test item C16-18-(even numbered, C18-unsaturated)-alkylamines acetates has obligatory labelling requirement for toxicity and is classified as harmful.
According to Annex I of Regulation (EC) 1272/2008 , the test item C16-18-(even numbered, C18-unsaturated)-alkylamines acetates has obligatory labelling requirement for toxicity and is classified into Category 4.
According to GHS (Globally Harmonized Classification System) , the test item C16-18-(even numbered, C18-unsaturated)-alkylamines acetates has obligatory labelling requirement for toxicity and is classified into Category 4.
For details of the classification criteria see Evaluation of Results.

Executive summary:

Two groups, each of three female WISTAR Crl: WI(Han) rats, were treated with the test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The test item was suspended in the vehiclesesame oilat a concentration of 0.2 g/mL and administered at a dose volume of 10 mL/kg.

Two further groups, each of three female WISTAR Crl: WI(Han) rats, were treated with the test item by oral gavage administration at a dosage of 300 mg/kg body weight. The test item was suspended in the vehicle sesame oil at a concentration of 0.03 g/mL and administered at a dose volume of 10 mL/kg.

The animals were observed on delivery, on inclusion in the study and before administration for mortality/morbidity and other clinical signs. All animals were examined for clinical signs several times on the day of dosing and once daily until the end of the observation period. Their body weights were recorded on day 1 (prior to the administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

Three animals treated with the test item at a dose of 2000 mg/kg were found dead 7 or 8 days after dosing (one animal after step 1, two animals after step 2). All remaining animals treated with the test item at a dose of 2000 mg/kg survived until the end of the study.

All animals treated with the test item at a dose of 300 mg/kg survived until the end of the study.

The most relevant clinical findings in the animals treated with the test item at a dose of 2000 mg/kg bw were reduced spontaneous activity, piloerection, nasal discharge and chromodacryorrhea, diarrhoea, body weight loss, half and complete eyelid-closure, moving and eating the bedding. The clinical signs persisted for up to 13 days after dosing in the surviving animals.

The most relevant clinical findings in the animals treated with the test item at a dose of 300 mg/kg bw were reduced spontaneous activity, piloerection and prone position on the day of dose administration.

Body weight loss was recorded during the first week of the observation period for all animals treated with the test item at a dose of 2000 mg/kg bw. During the second week the body weight gain of the surviving animals was within the normal range of variation for this strain.

For all animals treated with the test item at a dose of 300 mg/kg bw, the body weight gain of the all animals was within the normal range of variation for this strain throughout the 14-day observation period. At necropsy, no macroscopic findings were observed in any animal surviving until the end of the observation period of any step. Macroscopy of animals not having survived until the end of the observation period revealed autolysis on all organs, and a discoloured, bloated caecum.

Under the conditions of the present study, a single oral application of the test item to rats at a dose of 2000 mg/kg body weight was associated with reversible signs of toxicity, but no mortality. The median lethal dose (LD50 cut-off) of the registration substance observed over a period of 14 days was 2000 mg/kg body weight.