N1,N3-diallylpropane-1,3-diamine dihydrochloride

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2020-04-16 to 2020-08-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity: Fixed Dose Procedure)

Version / remarks:

adopted 09th October 2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

(prop-2-en-1-yl)({3-[(prop-2-en-1-yl)amino]propyl})amine dihydrochloride

EC Number:

812-745-6

Cas Number:

205041-15-2

Molecular formula:

C9 H20 Cl2 N2

IUPAC Name:

(prop-2-en-1-yl)({3-[(prop-2-en-1-yl)amino]propyl})amine dihydrochloride

Test material form:

solid

Details on test material:

- CAS: 205041-15-2
- Batch: LWDDP00072
- Colour: white
- Purity (GC): 100%
- Expiry date: 2020-08-11
- Storage conditions: room temperature
- Stability: stable under recommended storage conditions

Specific details on test material used for the study:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item was used as delivered by the sponsor. In order to ensure good skin contact, it was moistened with 1 mL of aqua ad injectionem (Deltamedica, lot 910137, expiry date: September 2022). This vehicle was chosen due to its non-irritating characteristics.

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, 97633 Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 9-12 weeks
- Weight at study initiation: 217-232 g
- Housing: The animals were kept in groups except during exposure and unless there were reasons to house individually (e.g. if there is concern that contact with other animals could increase stress due to the nature and severity of the signs of toxicity, or could result in exacerbation of local skin effects) in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding
- Diet: Altromin 1324 maintenance diet for rats and mice, ad libitum
- Water: Tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals), ad libitum
- Acclimation period: Adequate acclimatisation period (at least five days) under laboratory conditions
- Microbiological status when known : The animals were derived from a controlled full-barrier maintained breeding system (SPF).

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

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsal area of the trunk
- % coverage: The test item was applied at a single dose, uniformly over an area which was approximately 10% of the total body surface.
- Type of wrap if used: The test item was held in contact with the skin by a dressing throughout a 24-hour period. This consisted of a semi-occlusive dressing made of a porous gauze and non-irritating tape and was fixed with an additional dressing in a suitable manner.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period the residual test item was removed using aqua ad injectionem (sterile water).
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied: 2000 mg/kg bw
- For solids: The test item was used as delivered by the sponsor. In order to ensure good skin contact, it was moistened with 1 mL aqua ad injectionem.

Duration of exposure:

24 hours

Doses:

- 2000 mg/kg bw

No. of animals per sex per dose:

3 females

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: Signs of erythema and oedema were assessed using the scoring system laid down in OECD Guideline 404 (for details see Table 1 in box "Any other information on materials and methods incl. tables") at 24, 48 and 72 hours after patch removal.
- Frequency of weighing: The animals were weighed on day 1 (prior to the application), on days 8 and 15.
- Necropsy of survivors performed: At the end of the observation period the animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally at a dosage of 250–400 mg/kg bw. All animals were subjected to gross necropsy and examined macroscopically for gross pathological changes. In absence of gross pathological changes no tissues were preserved for a possible histopathological evaluation
- Clinical signs: 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 6 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. All abnormalities were recorded. 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. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

Statistics:

n.a.

Results and discussion

Preliminary study:

One animal was administered a dose of 2000 mg/kg bw. As the animal survived without showing any signs of systemic toxicity, this dose was used as a starting dose for the main study.

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed after treatment with the test material.

Clinical signs:

other: No clinical signs were observed after treatment with the test material during the whole observation period.

Gross pathology:

No specific gross pathological changes were recorded for any animal.

Other findings:

No signs of acute dermal toxicity were observed after treatment with the test material during the whole observation period, but mild signs of irritation were observed in the main experiment which were fully reversible after Day 3 or Day 4 after patch removal (for details see Table 2 in section "Any other information on results incl. tables").

Any other information on results incl. tables

Table 2: Skin Irritation at Application Site - Individual Data – DRF and Main Study

Time after Patch Removal	Dose Range Finding Study		Main Study
	Animal No. 1*		Animal No. 2**		Animal No. 3**
	E/O	C	E/O	C	E/O	C
Day 2 (0±2 h)	0/0	nsf	1°/0	nsf	1°/0	nsf
Day 3 (24±2 h)	0/0	nsf	1°/0	nsf	1°/0	nsf
Day 4 (48±2 h)	0/0	nsf	0/0	nsf	1°/0	nsf
Day 5 (72±2 h)	0/0	nsf	0/0	nsf	0/0	nsf
Day 6 - 15	0/0	nsf	0/0	nsf	0/0	nsf

Table 2: Absolute Body Weights [g] and Body Weight Change [%] - DRF and Main Study

Animal No.	Absolute Body Weights [g]			Body Weight Change [%]
	Day 1	Day 8	Day 15	Day 1-15
1*	217	226	236	9
2**	223	225	231	4
3**	232	224	240	3

* = dose range finding study animal; ** = main study animal

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In an acute dermal toxicity study conducted according to OECD 402, N1,N3-diallylpropane-1,3-diamine dihydrochloride was dermally administered to rats for 24 hours. The application was associated with no mortality and neither signs of toxicity, except for mild signs of irritation which were reversible within 5 days. Based on the results, the dermal LD50 can be considered to be greater than 2000 mg/kg bw.

Executive summary:

In an acute dermal toxicity study conducted according to OECD 402, three female Wistar rats were dermally exposed to N1,N3-diallylpropane-1,3-diamine dihydrochloride (100% purity) under semi-occlusive conditions for 24 hours to approximately 10% of the total body surface at a dose of 2000 mg/kg bw. Animals were then observed for 14 days. There were no treatment related mortalities, no changes in body weight, no clinical signs or other signs of acute dermal toxicity, except mild signs of dermal irritation which were fully reversible within five days. Based on the results, the dermal LD50 can be considered to be greater than 2000 mg/kg bw and no classification according to CLP Regulation 1272/2008 is warranted.