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Description of key information

Available studies result to GHS classification Category 3 for acute oral toxicity (LD50 between 50 and 300 mg/kg bw) with a LD50 cut-off of 200 mg/kg bw. No data available on acute toxicity via inhalation or dermal route.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
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
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
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

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
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.

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

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.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
200 mg/kg bw
Quality of whole database:
The study is GLP compliant and has Klimisch score 1

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Oral

There are two studies available for evaluation.

 

In the first, the toxic effects following an acute oral dose of N-C12,14 alkyl-1,3-diaminopropane (CAS 90640-43-0, also referred to as C12-14-diamine) was determined according to the guidelines for the acute toxic class method (OECD 423) performed under GLP.

In step 1 a single female rats were dosed by oral gavage with 2000 mg N-(hydrogenated tallow alkyl) trimethylenediamine/kg bw in cotton seed oil. Within 1 h 30 min post-dose the animal was found dead. In step 2 three female rats were dosed with 300 mg/kg bw. 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/kg bw. No mortality occurred. Observed signs of toxicity were reduced spontaneous activity, piloerection and diarrhoea. The animals had recovered within 3 to 6 days. In step 4 three additional female rats were dosed with 50 mg/kg bw. 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. According to GHS classification, the substance need to classified for acute oral toxicity into Category 3, with a LD50 cut-off of 200 mg/kg bw.

 

The second, also GLP compliant, study evaluated the acute oral toxicity of N-Dodecyl-1,3-diaminepropane (CAS 5538-95-4, also referred as C12-Diamine) according to OECD 401.

Test substance was dissolved in vegetable oil administered to groups of 5 male and 5 female rats at levels of 2000, 1000, and 315 mg/kg bw, and additionally in groups of 5 females only at 200 and 100 mg/kg bw.

At 2000 and 1000 mg/kg bw all animals died. At 315 mg/kg bw all 5 female animals died and 3/5 male animals. At 200 and 100 mg/kgbw none of the animals died. Consequently, LD50 was determined to be between 200 mg/kg bw (0% mortality) and 315 mg/kg (80% mortality). Based on these results classification into GHS Category 3 for acute oral toxicity seems appropriate.

 

Inhalation:

There is no study on inhalation toxicity available for N-C12,14 alkyl-1,3-diaminopropane.

REACH stipulates that testing by the inhalation route is appropriate if exposure of humans via inhalation is likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size. REACH guidance R.7.a, chapter. 7.4 Acute toxicity, indicates that in principle no inhalation studies are needed when vp < 0.1 Pa at 20°C or particle size > 100 µm. C12-14-diamine is a liquid/paste with no inhalable particles and a vapour pressure less than 0.0015 Pa at 20°C. Also the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route will be unlikely to occur, and no acute inhalation test was performed.

 

N-C12,14 alkyl-1,3-diaminopropane is a liquid with pasty elements, and a melting point of 27°C and thus is not a fluid at ambient temperatures around 20°C and lower. No information on viscosity is available at 40°C, and hence the hazard for aspiration cannot be adequately assessed.

 

Dermal:

There is no dermal LD 50 value for acute skin toxicity of N-C12,14 alkyl-1,3-diaminopropane, and due to the corrosive nature of the substance it is not ethical to carry out this animal study.

A study evaluating the acute dermal toxicity N-Coco-1,3-diaminopropane according has shown mortality in 1 out of 10 rabbits at a limit dose of 2000 mg/kg bw, indicating low systemic toxicity via dermal route.

Justification for classification or non-classification

Available studies result to GHS classification Category 3 for acute oral toxicity (LD50 between 50 and 300 mg/kg bw) based on LD50 cut-off of 200 mg/kg bw. Testing for dermal toxicity is not required in view of its corrosive properties, but information from acute dermal toxicity study performed on the comparable N-Coco-1,3-diaminopropane indicates a low systemic toxicity following dermal application, for which no classification would be required.

There is no data on toxicity following exposures via inhalation. Related to low vp and no inhalable particles, no inhalation studies are required in view of limited exposure via inhalation.

No classification STOT-SE Cat.3 needed:

The active substance is not structurally related to any known class of neurotoxic chemicals. In addition, repeated dose studies did not show indications of specific neurotoxicity, in specific neurotoxicity measures as sensory activity, grip strength, and motor activity assessment.