N-C16-18-alkyl-(evennumbered) C18 unsaturated) propane-1,3-diamine

Endpoint summary

Description of key information

Acute LD50 between 500 and 1000 mg/kgbw, based on read-across.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

2007-11-16 to 2008-01-03

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

Principles of method if other than guideline:

not applicable

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

other: HsdRccHan:WIST (Full-Barrier)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Winkelmann GmbH, D-33178 Borchen
- Weight at study initiation:
Step 1: 154-166 g
Step 2: 171-191 g
Step 3: 172-179 g

- Fasting period before study: overnight
- Housing: Macrolon cages on Altromin saw fiber bedding
- Diet: ad libitum, Altromin 1324
- 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 g/ 5 mL and 2 g/10 mL cotton seed oil
Step 2: 0.3 g/ 10 mL cotton seed oil
Step 3: 0.3 g/ 10 mL cotton seed oil
- Amount of vehicle: 10 mL/ kg body weight
- Justification for choice of vehicle: non-toxic characteristic
- Lot/batch no.: Lot 067K0116, Sigma Chemicals

MAXIMUM DOSE VOLUME APPLIED: 1.91 mL

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: 2000 mg/ kg was selected as the starting dose

Doses:

Step 1: 2000 mg/ kg
Step 2 and 3: 300 mg/ kg body weight

No. of animals per sex per dose:

female nulliparous rats
2000 mg/ kg dose (Step 1): 3 animals
300 mg/ kg dose (Step 2 and 3): 3 animals + 3 animals

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 postdose.
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. 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.

Statistics:

not applicable

Preliminary study:

no data

Sex:

female

Dose descriptor:

approximate LD50

Effect level:

500 mg/kg bw

Remarks on result:

other: acute toxic class method; according to GHS (Globally Harmonized Classification System) the test item N-Oleyl-1,3-diaminopropane was classified into Category 4

Mortality:

Step 1 (2000 mg/kg bw): all 3 animals died within 24 h (one was euthanised due to animal welfare reasons, two were found dead in their cages)
Step 2 (300 mg/kg bw): 1 animal was found dead 10 days after treatment
Step 3 (300 mg/kg bw): no mortality observed

Clinical signs:

other: see remarks on results including tables and figures

Gross pathology:

Effect on organs (related to dose level):

Animal no. 1 of step 1 (2000 mg/ kg bw):
The animal was found dead in a lateral position. Rests of the test item were
found in the stomach. The lungs were infiltrated with blood.

Animal no. 2 of step 1 (2000 mg/ kg bw):
Rests of the test item were found in the stomach. The lungs were infiltrated
with blood.

Animal no. 3 of step 1 (2000 mg/ kg bw):
Rests of the test item were found in the stomach, which was bloated. The
small intestine was empty and of a yellow colour. This animal was
euthanised due to animal welfare reasons.

Animal no. 2 of step 2 (300 mg/ kg bw):
Stomach, small and large intestine bloated and empty.

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

Animal no. 1 of step 1 (2000 mg/kg bw): 30 min post-dose: salivation. 2 h post-dose: slightly reduced spontaneous activity, apathy, piloerection, salivation, half eyelid-closure. 4 h post-dose: moderately reduced spontaneous activity, piloerection, salivation, full eyelid-closure. 5 h as well as 9 h post-dose: moderately reduced spontaneous activity, apathy, piloerection, salivation, half eyelid-closure. 24 h post-dose the animal was found dead.

Animal no. 2 of step 1 (2000 mg/kg bw): 30 min post-dose: salivation. 2 h post-dose: slightly reduced spontaneous activity, apathy, piloerection, salivation, half eyelid-closure. 4 h post-dose: moderately reduced spontaneous activity, piloerection, salivation, full eyelid-closure. 5 h post-dose: moderately reduced spontaneous activity, apathy, piloerection, salivation, half eyelid-closure. 9 h post-dose: moderately reduced spontaneous activity, apathy, piloerection, salivation, half eyelid-closure, slight articulation. 24 h post-dose the animal was found dead.

Animal No. 3 of step 1 (2000 mg/kg bw): 30 min post-dose: salivation. 2 h post-dose: slightly reduced spontaneous activity, apathy, piloerection, salivation. 4 h post-dose: moderately reduced spontaneous activity, piloerection, salivation, full eyelid-closure. 5 h post-dose: moderately reduced spontaneous activity, apathy, piloerection, salivation, half eyelid-closure. 9 h post-dose: moderately reduced spontaneous activity, apathy, piloerection, salivation, half eyelid-closure, slight articulation. 24 h post-dose: moderately reduced spontaneous activity, piloerection, salivation, half eyelid-closure, severe articulation. 25 h post-dose: for animal welfare reasons this animal was euthanised.

Animal no. 1 of step 2 (300 mg/kg bw): 3 h 25 min as well as 7 h, 24 h, 30 h post-dose: piloerection. 48 h post-dose until the end of the observation period: no symptoms were observed.

Animal no. 2 of step 2 (300 mg/kg bw): 45 min as well as 2 h 45 min post-dose: slightly reduced spontaneous activity, piloerection, half eyelid-closure. 4 h 45 min post-dose: slightly reduced spontaneous activity, piloerection. 23 h 45 min post-dose: weight loss, stertorousness. 47 h 45 min as well as 71 h 45 min, 95 h 45 min, 119 h 45 min, 143 h 45 min, 167 h 45 min, 191 h 45 min post-dose: piloerection, weight loss, stertorousness. 215 h 45 min. post-dose: moderately reduced spontaneous activity, piloerection, weight loss, stertorousness. 10 days (238 h 45 min) post-dose: the animal was found dead.

Animal no. 3 of step 2 (300 mg/kg bw): 1 h 15 min post-dose: piloerection. 2 h 45 min as well as 4 h 45 min post-dose: slightly reduced spontaneous activity, piloerection. 23 h 45 min post-dose until the end of the observation period: no symptoms were observed.

Animal no. 1, 2 and 3 of step 3 (300 mg/kg bw): 15 min as well as 1 h 45 min post-dose: slightly reduced spontaneous activity, piloerection. 3 h 45 min post-dose: moderately reduced spontaneous activity, piloerection. 21 h 45 min as well as 45 h 45 min post-dose: slightly reduced spontaneous activity, piloerection. 69 h 45 min post-dose: piloerection. 93 h 45 min post-dose until the end of the observation period: no 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	154	This animal was found dead.
2	female	157	This animal was found dead.
3	female	166	This animal was found dead
Step 2 (300 mg/kg bw)
1	female	171	189	212
2	female	191	137	This animal was found dead.
3	female	173	187	200
Step 3 (300 mg/kg bw)
1	female	172	180	196
2	female	179	192	206
3	female	173	165	182

Interpretation of results:

harmful

Remarks:

Migrated information criteria given in Annex VI of the Directive 2001/59/EC Criteria used for interpretation of results: EU

Conclusions:

Considering the reported data of this toxicity test it can be stated that the test item N-Oleyl-1,3-diaminopropane showed acute oral toxic characteristics. According to GHS (Globally Harmonized Classification System) the test item N-Oleyl-1,3-diaminopropane was classified into Category 4 (LD50
cut-off: 500 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-Oleyl-1,3-diaminopropane (liquid) 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 three female rats were dosed by oral gavage with 2000 mg N-Oleyl-1,3-diaminopropane/ kg body weight in cotton seed oil. Severe toxicity indicated by salivation, apathy, piloerection and half eyelid-closure were observed in this group after the application. Two animals died within 24 h and one was euthanized 25 h post-dose for animal welfare reasons. In step 2 three female rats were dosed with 300 mg N-Oleyl-1,3-diaminopropane/ kg body weight. Two animals showed slight signs of toxicity indicated by piloerection and reduced spontaneous activity until 48 h post-dose and had recovered completely thereafter. One animal showed severe signs of toxicity indicated by reduced spontaneous activity, half eyelid-closure, piloerection, weight loss and stertorousness and was found dead after 10 days. Therefore three additional female rats were dosed with 300 mg N-Oleyl-1,3 -diaminopropane/kg body weight. Observed signs of toxicity were slightly reduced spontaneous activity and piloerection until 4 days post-dose. The animals showed no symptoms thereafter. According to the toxic class regime no further testing was required.

Following pathological changes were observed in the step 1 animals (2000 mg/kg bw): rests of the item were found in the stomach of all three rats, the lungs were infiltrated with blood in all three rats, one animal had a bloated stomach and an empty small intestine of a yellow colour. The animal of step 2 (300 mg/kg bw) that was found dead after 10 days had a bloated and empty stomach and small and large intestine.

Considering the reported data of this toxicity test it can be stated that the test item N-Oleyl-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 harmful. According to GHS (Globally Harmonized Classification System) the test item N-Oleyl-1,3-diaminopropane was classified into Category 4 (LD₅₀ cut-off: 500 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

Value:

500 mg/kg bw

Quality of whole database:

High. Consistent and reliable data over category.

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

A total of 14 studies were available for evaluation of the acute oral toxicity of Diamine C16-18, C18-unsaturated. Of these, 9 studies evaluated the acute toxicity of Diamine C16-18, C18-unsaturated itself. One of these, on Duomeen T (Kynoch 1984)is considered to be supportive, and resulted to an LD50 in rats of 945 mg/kg bw.

The other studies generally showed much lower toxicity, but the test substance is not clearly described and could possibly be formulations, these studies are not further considered.

 

Data more relevant for the classification is derived from series of highly reliable GLP studies performed together over the category of alkyl-diamines. (Also see ‘Justification in support of cross-reading between Diamines’, document Support Cross-reading - Diamines Final 20131220.pdf attached to IUCLID chapter 13)

 

Alkyl chain	Study	Physical state	Oral LD50cut-off mg/kg bw
Oleyl	RA from 7173-62-8, Acute toxicity: oral_liquid, Haist 2008	liquid	500
Oleyl	RA from 7173-62-8 Acute toxicity: oral_paste, Haist 2008	liquid/ paste	1000
C12-14	RA from 90640-43-0, Acute toxicity: oral. Haist 2008	liquid/ paste	200
HT	RA from 68603-64-5, Acute toxicity: oral. Haist, 2008	solid	2500
Coco	RA from 61791-63-7, Acute toxicity: oral. Haist 2008	liquid	300
Tallow	read across to Oleyl	paste	500-1000

 

For acute oral toxicity, a relation can be observed with chain length and level of unsaturation: acute toxicity become less with increasing lengths of alkyl chains, and related with increasing levels of unsaturation. The diamines based on relative short chains up to C₁₄, Cat.3 classification follows. The diamines based on higher alkyl chain lengths are classified as Cat.4 when also a high level of unsaturation is present. In HT-diamine with fully saturated longer alkyl chains of C_16-18, has a LD50 cut-off 2500, and falls in GHS Cat.5, but no classification is needed for CLP.

 

Diamine C16-18, C18-unsaturated can in this case best be compared to Oleyl diamine. Although the chain-length distribution is the same as for tallow, the presence of some unsaturated C18, makes cross-reading to Oleyl more appropriate.

 

Inhalation:

There is no study on inhalation toxicity available for N-C16-18-alkyl-(evennumbered) C18 unsaturated) propane-1,3-diamine.

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.Diamine C16-18, C18-unsaturatedis a paste with a vapour pressure less than 0.0015 Pa at 20°C (value based on read-across from shorter chain C_12-14-diamine). 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.

 

There is no risk for aspiration as with a mp around 37 °C, Diamine C16-18, C18-unsaturatedis a paste upon ingestion.

 

Dermal:

There is no dermal LD 50 value for acute skin toxicity of N-C16-18-alkyl-(evennumbered) C18 unsaturated) propane-1,3-diamine, and due to the corrosive properties 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. Low systemic toxicity via dermal route can therefore also be expected forDiamine C16-18, C18-unsaturated.

Use and handling of the pure substance is only industrial and professional (formulation), and the classification as irritant of the substance requires risk management methods which eliminate the risk of acute systemic toxicity from potential for skin contact.

Justification for classification or non-classification

An LD50 value of 500-1000 mg/kg will lead to CLP Category 4 for acute oral toxicity.

Dermal systemic toxicity is expected to be low, while severe irritating properties will limit likelihood of exposures.

Related to low vp and no inhalable particles, there will be low likelihood of exposures 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.

Diamine C16-18, C18-unsaturatedis a paste with mp of 37°C and a vapour pressure less than 0.0015 Pa at 20°C (value based on read-across from shorter chain C_12-14-diamine). 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.