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Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2007-11-27 to 2008-01-22
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
Principles of method if other than guideline:
not applicable
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: liquid/paste
Details on test material:
- Name of test material (as cited in study report): N-C12,14 alkyl-1,3-diaminopropane
- Physical state: liquid/ paste
- Purity test date: 2007-10-15
- Composition of test material, percentage of components:
N-Alkyl-1,3-diaminopropanes: 94%
N-Alkyl-amines: 6%

Total Primary amines: 5.7 %
Total Diamines: 94.3 %
Iodine number: 2 g I/100g (0-3)
Water: 0.42 % (0-0.5 %)

C12 N Primary amine: 4.3 %
C14 N Primary amine: 1.3 %
C16 N Primary amine: 0.1 %
C12 Diamine: 73.4 %
C14 Diamine: 20.2 %
C16 Diamine: 0.7 %

Acrylnitrile: <4 ppm
Fatty acids: <1 %
Fatty alkyl nitriles: <1 %

- Lot/batch No.: S000904
- Expiration date of the lot/batch: 2010-09-30
- Storage condition of test material: at RT
- pH: basic
- Colour: Max 3 Gardner
- Product name: Duomeen 1214D
- Melting Point: 24-34°C

Test animals

Species:
rat
Strain:
other: HsdRccHan: WIST (SPF)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Winkelmann GmbH, D-33178 Borchen
- Weight at study initiation:
Step 1: 168 g (1 animal)
Step 2: 189-200 g
Step 3: 133-173 g
Step 4: 180-214 g

- Fasting period before study: overnight and 3-4 hours after dosing
- Housing: macrolon cages on Altromin saw fibre bedding
- Diet: ad libitum (Altromin 1324 (TPF)
- Water: ad libitum (drinking water)
- Acclimation period: at least 5 days
- Other: marked individually by tail painting, SPF animals

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 55 +/- 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
cotton seed oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle:
Step 1: 1.0 g/ 5 mL
Step 2: 0.3 g/ 10 mL
Step 3: 0.05 g/ 10 mL
Step 4: 0.05 g/ 10 mL

- Amount of vehicle: 10 mL/ kg body weight
- Justification for choice of vehicle: non-toxic and lipophil
- Lot/batch no. : Lot 067K0116, Sigma Chemicals

MAXIMUM DOSE VOLUME APPLIED: 2.00 mL

DOSAGE PREPARATION : freshly mixed to homogeneity prior to administration

CLASS METHOD
- Rationale for the selection of the starting dose:
preliminary test with one animal performed (2000 mg/kg), due to immeadiate preacute lethality
the starting dose was choosen to be 300 mg/kg bw
Doses:
Step 1: 2000 mg/kg bw
Step 2: 300 mg/kg bw
Step 3: 50 mg/kg bw
Step 4: 50 mg/kg bw
No. of animals per sex per dose:
Step 1 (2000 mg/kg bw): 1 female rat
Step 2 (300 mg/kg bw): 3 female rats
Step 3 (50 mg/kg bw): 3 female rats
Step 4 (50 mg/kg bw): 3 female rats
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days

- Frequency of observations and weighing: A careful clinical examination was made several times on the day of dosing.
Parts of this were at least three observations within the first four hours post-dose.
Animals were observed once a day thereafter.
The animals were weighed prior to the administration and once a week thereafter.

- Necropsy of survivors performed: yes

- Other examinations performed: Cage side 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.
Statistics:
not applicable

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
approximate LD50
Effect level:
200 mg/kg bw
Remarks on result:
other: acute toxic class method; according to GHS (Globally Harmonized Classification System) the test item N-C12,14 alkyl-1,3-diaminopropane was classified into Category 3
Mortality:
Step 1 (2000 mg/kg bw, 1 female rat): within 1 h 30 min post-dose the animal was found dead
Step 2 (300 mg/kg bw): within 2 days post-dose all three animals were found dead
Step 3 (50 mg/kg bw): no deaths observed within the observation period
Step 4 (50 mg/kg bw): 1 animal was found dead 13 days post-dose
Clinical signs:
see remarks on results including tables and figures
Body weight:
Throughout the 14-day observation period weight loss was recorded for three animals. The weight gain of the other surviving animals was within
the expected range.
For further details see remarks on results including tables and figures.
Gross pathology:
Animal no. 1 of step 1 (2000 mg/ kg bw):
The stomach, small and large intestine were bloody.
Animal no.1 of step 2 (300 mg/ kg bw):
The stomach was bloody. The stomach and small intestines had a yellow content. The liver had a
dark colour.
Animal no.2 of step 2 (300 mg/ kg bw):
The animal was found in a face down position. The stomach mucosa was bloody and the stomach
content was yellow. The anus was smeared with faeces.
Animal no.3 of step 2 (300 mg/ kg bw):
The animal was found in a face down position. The stomach mucosa was
bloody and the stomach content was yellow.
Animal no. 2 of step 4 (50 mg/ kg bw):
Small and large intestine and the stomach were bloated. The spleen weighed 150 g.

Any other information on results incl. tables

Signs of toxicity related to dose level used, time of onset and duration:

Animal no. 1 of step 1(2000 mg/kg bw): 05 min post-dose: apathy, salivation. 20 min post-dose: moderately reduced spontaneous activity, salivation, diarrhoea, face down position. 1 h 30 min post-dose: this animal was found dead.

Animal no. 1 of step 2(300 mg/kg bw): 1 h 30 min post-dose: slightly reduced spontaneous activity, piloerection. 3 h 30 min post-dose: moderately reduced spontaneous activity, piloerection, diarrhoea. 22 h 30 min post-dose: moderately reduced spontaneous activity, piloerection, diarrhoea, sunken flank. 26 h 30 min: this animal was found dead.

Animal no. 2 of step 2 (300 mg/kg bw): 30 min as well as 3 h post-dose: slightly reduced spontaneous activity, apathy, piloerection. 24 h as well as 45 h post-dose: slightly reduced spontaneous activity, piloerection. 71 h as well as 94 h, 99 h post-dose: piloerection 117 h until the end of the observation period: no further symptoms were observed.

Animal No. 3 of step 2 (300 mg/kg bw): 45 min post-dose: slightly reduced spontaneous activity, piloerection, diarrhoea. 17 h 45 min post-dose: moderately reduced spontaneous activity, piloerection, diarrhoea. 41 h 15 min post-dose: this animal was found dead.

Animal no. 1 of step 3 (50 mg/kg bw): 45 min post-dose: slightly reduced spontaneous activity, piloerection. 17 h 45 min post-dose: moderately reduced spontaneous activity, piloerection, diarrhoea. 41 h 15 min post-dose: this animal was found dead.

Animal no. 2 of step 3 (50 mg/kg bw): 1 h, 20 h, 44 h post-dose: slightly reduced spontaneous activity, piloerection. 48 h post-dose: moderately reduced spontaneous activity, piloerection. 68 h, 92 h, 116 h post-dose: slightly reduced spontaneous activity, piloerection. 6 days post-dose until the end of the observation period: no further symptoms were observed.

Animal no.3 of step 3 (50 mg/kg bw): 2 h 30 min as well as 21 h 30 min, 46 h 30 min post-dose: piloerection. 4 h post-dose: slightly reduced spontaneous activity, piloerection. 3 days post-dose until the end of the observation period: no further symptoms were observed.

Animal no. 1 of step 4 (50 mg/kg bw): 3 h post-dose: slightly reduced spontaneous activity, piloerection. 21 h post-dose: slightly reduced spontaneous activity, piloerection, diarrhea. 45 h 30 min post-dose: slightly reduced spontaneous activity, piloerection. 69 h 30 min post-dose: piloerection. 97 h post-dose: slight piloerection, respiratory sounds. 119 h post-dose: piloerection. 6 days until the end of the observation period: no further symptoms were observed.

Animal no. 2 of step 4 (50 mg/kg bw): 3 h post-dose: slightly reduced spontaneous activity, piloerection. 21 h post-dose: slightly reduced spontaneous activity, piloerection, diarrhea. 45 h 30 min post-dose: slightly reduced spontaneous activity, piloerection. 69 h 30 min post-dose: piloerection. 93 h 30 min as well as 119 h post-dose: piloerection. 142 h 30 min post-dose: no symptoms observed. 11 days post-dose: slightly reduced spontaneous activity, piloerection. 12 days post-dose: slightly reduced spontaneous activity, piloerection, respiratory sounds. 13 days post-dose: this animal was found dead.

Animal no.3 of step 4 (50 mg/kg bw): 3 h as well as 69 h 30 min, 93 h 30 min, 119 h post-dose: piloerection. 3 h 30 min as well as 45 h 30 min: slightly reduced spontaneous activity, piloerection. 21 h post-dose: slightly reduced spontaneous activity, piloerection, diarrhea. 142 h 30 min post-dose until the end of the observation period: no further symptoms were observed.

Absolute body weights in g :

Animal No.

  Sex

Day 0

Day 7

Day 14

Step 1

(2000 mg/kg bw)

 

1

 female

168

This animal was found dead.

 

Step 2 (300 mg/kg bw)

1

 female

189

This animal was found dead.

2

 female

200

188- This animal was found dead.

.

3

 female

 199

192- This animal was found dead.

Step 3 (50 mg/kg bw)

 

 

 

 

1

 female

173

205

206

2

 female

133

162

177

3

 female

146

163

174

Step 4 (50 mg/kg bw)

 

 

 

 

1

 female

206

200

196

2

 female

180

182

This animal was found dead.

3

 female

217

252

255

Applicant's summary and conclusion

Interpretation of results:
Category 3 based on GHS criteria
Remarks:
Migrated information
Conclusions:
Following the reported data of this study, the substance needs to classified according to GHS criteria for acute oral toxicity into Category 3, with a LD50 cut-off of 200 mg/kg bw.
Executive summary:

The acute toxic class method for oral toxicity was performed on nulliparous female HsdRccHan:WIST rats, who received a single exposure of N-C12,14 alkyl-1,3-diaminopropane via oral gavage.

A careful clinical examination was made several times on the day of dosing. Parts of this were at least three observations within the first four hours post-dose. Animals were observed once a day thereafter for 14 days. Cage side 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 animals were sacrificed by an overdosage of pentobarbital. All animals were subjected to gross necropsy. All gross pathological changes were recorded.

In step 1 single female rat was dosed by oral gavage with 2000 mg N-C12,14 alkyl-1,3-diaminopropane/ kg body weight in cotton seed oil. Severe, immediate toxicity indicated by salivation, face down position, diarrhoea and moderately reduced spontaneous activity was observed in this animal, followed by death at 1 h and 30 min post-dose. In step 2 three female rats were dosed with 300 mg N-C12,14 alkyl-1,3-diaminopropane/ kg body weight. All three animals showed severe signs of toxicity indicated by piloerection, sunken flank, diarrhoea and reduced spontaneous activity and died within 2 days post-dose. In step 3 three female rats were dosed with 50 mg N-C12,14 alkyl-1,3-diaminopropane/ kg body weight. No mortality was observed in all three animals.

Observed signs of toxicity were reduced spontaneous activity, piloerection and diarrhoea. The animals had recovered within 3 (animal 2) to 6 days (animal 1 and 3) post-dose and showed no symptoms thereafter. In step 4 three additional female rats were dosed with 50 mg N-C12,14 alkyl-1,3-diaminopropane/ kg body weight. Mortality was observed in one animal. Observed signs of toxicity were reduced spontaneous activity, piloerection and diarrhoea. All three animals recovered within 6 days post-dose. Animal 1 and 3 of step 4 showed no further symptoms thereafter. Animal 2 of step 4 showed the same toxicity signs again from day 11 on and was found dead on day 13. According to the toxic class regime no further testing was required.

Following pathological changes were observed in the step 1 animal (2000 mg/kg bw): The stomach, small and large intestine were bloody. Animal 1 of step 2 (300 mg/kg bw) exhibited a bloody stomach. The stomach and small intestines had a yellow content. The liver had a dark colour. Animal 2 of step 2 (300 mg/kg bw) was found in a face down position and exhibited a bloody stomach mucosa and the stomach content was yellow. The anus was smeared with faeces. Animal 3 of step 2 (300 mg/kg bw) was found in a face down position and exhibited a bloody stomach mucosa and a yellow stomach content. In animal 2 of step 4 (50 mg/kg bw) the small and large intestine and the stomach were bloated. The spleen weight was markedly increased (150 g).

Considering the reported data of this toxicity test it can be stated that the test item N-C12,14 alkyl-1,3-diaminopropane showed acute oral toxic characteristics. On basis of the test results and in conformity with the criteria given in Annex VI of the Directive 2001/59/EC the substance should be classified as toxic. According to GHS (Globally Harmonized Classification System) the test item N-C12,14 alkyl-1,3-diaminopropane was classified into Category 3 (LD50 cut-off: 200 mg/kg bw).

The acute oral toxicity test was conducted under GLP according to the guidelines for the acute toxic class method OECD 423, EU method B1.tris and OPPTS 870.110 without deviation.