Amines, N-(C16-18 and C18-unsatd. alkyl)-N,N',N'-trimethyltrimethylenedi-

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March 2016 - April 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

2002

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000, including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Date certificate 3 November 2015

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

695-101-5

Cas Number:

1275611-65-8

Molecular formula:

Not applicable

IUPAC Name:

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

Test material form:

liquid

Details on test material:

- Name of test material (as cited in study report): Diamine methylated
- Substance type:Amber liquid
- Physical state:Liquid
- Storage conditions of test material:At room temperature
- Content (certified): 100% (UVCB–Substance)
- Batch/Lot number:211401A Morris
- Expiration date of the lot/batch:24 March 2018

Test animals

Species:

rat

Strain:

other: Crl:WI (Han)

Sex:

female

Details on test animals or test system and environmental conditions:

- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Young adult animals (approx. 8-9 weeks old)
- Weight at study initiation: Body weight variation was within +/- 20% of the sex mean (141 - 190 grams).
- Fasting period before study: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test item.
- Housing: Group housing of 3 animals per cage in labeled Macrolon cages containing sterilized sawdust as bedding material and paper as cage-enrichment.
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
set to maintain:
- Temperature (°C): 18 – 24
- Humidity (%): 40 - 70
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 22 March 2016 to 21 April 2016

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: no vehicle (undiluted) at 300 mg/kg bw, corn oil (50 mg/kg bw)

Details on oral exposure:

GAVAGE METHOD: plastic feeding tubes.

Frequency: single dosage, on Day 1.

VEHICLE
The vehicle was selected based on trial preparations performed at WIL Research Europe and on test item data supplied by the Sponsor. There was no information available regarding the solubility or stability in vehicle.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION:
The test item (preparations) (w/w) were kept at room temperature and were dosed within 4 hours after adding the vehicle to the test item. Homogeneity was assessed by visual inspection of the solutions and the formulations were stirred during dosing, which ensures homogeneity sufficient for these kinds of studies. Adjustment was made for specific gravity of the vehicle and test item. No correction was made for purity of the test item.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Based on anticipated mortality at the next higher dose 300 mg/kg bw was selected as the starting dose.

Doses:

300 mg/kg bw
50 mg/kg bw

No. of animals per sex per dose:

300 mg/kg bw: 3
50 mg/kg bw: 6 (2 groups of three females in a stepwise manner)

Control animals:

no

Details on study design:

The toxicity of the test item was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 300 mg/kg bw. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.

Duration of observation period following administration: 14 days
Frequency of observations and weighing:
- Mortality/Viability: Twice daily. Animals showing pain, distress or discomfort, which was considered not transient in nature or was likely to become
more severe, were sacrificed for humane reasons based on OECD guidance document on humane endpoints (ENV/JM/MONO/2000/7). The time of death was recorded as precisely as possible.
- Body weights: Days 1 (pre-administration), 8 and 15 and at death.
- Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy performed: yes, all animals
- Other examinations performed: none.

Statistics:

No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 50 - < 300 mg/kg bw

Based on:

test mat.

Mortality:

At 300 mg/kg bw, on Day 3, one animal was found dead and two animals were sacrificed for humane reasons.
At 50 mg/kg bw, no mortality occurred.

Clinical signs:

other: At 300 mg/kg bw lethargy, hunched posture, slow and shallow breathing, piloerection, diarrhoea, chromodacryorrhoea in both eyes, lean appearance and/or ptosis were noted for the animals between Days 1 and 3. At 50 mg/kg bw, hunched posture was noted for a

Gross pathology:

At 300 mg/kg bw, abnormalities of the spleen (reduced in size), thymus (reduced in size) and/or GI-tractus (contents: gelatinous) were noted for the animals, at macroscopic post mortem examination.
At 50 mg/kg bw, macroscopic examination did not reveal any abnormalities.

Other findings:

Autolysis was noted for the animal found dead. This was considered not toxicologically relevant.

Applicant's summary and conclusion

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

In an acute oral toxicity study with Diamine methylated in rats, performed according to OECD/EC test guidelines, an LD50 within the range of 50-300 mg/kg bw was determined, with a LD50 cut-off value of 200 mg/kg bw.

Executive summary:

Assessment of acute oral toxicity with Diamine methylated in the rat (Acute Toxic Class Method) was performed according to OECD/EC guidelines and in accordance with GLP principles. Initially, Diamine methylated was administered by oral gavage to three female Wistar rats at 300 mg/kg bw. In a stepwise procedure two additional groups of three females were dosed at 50 mg/kg bw.

At 300 mg/kg bw, on Day 3, one animal was found dead and two animals were sacrificed for humane reasons. At 50 mg/kg bw, no mortality occurred. At 300 mg/kg lethargy, hunched posture, slow and shallow breathing, piloerection, diarrhoea, chromodacryorrhoea in both eyes, lean appearance and/or ptosis were noted for the animals between Days 1 and 3. At 50 mg/kg bw, hunched posture was noted for all animals on Day 1. The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain. At 300 mg/kg bw, abnormalities of the spleen (reduced in size), thymus (reduced in size) and/or GI-tractus (contents: gelatinous) were noted for the animals, at macroscopic post mortem examination. At 50 mg/kg bw, macroscopic examination did not reveal any abnormalities.

The oral LD50 value of Diamine methylated in Wistar rats was established to be within the range of 50-300 mg/kg bw. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 200 mg/kg bw.

Based on these results:

- according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments), Diamine methylated should be classified as: Toxic if swallowed (Category 3) for acute toxicity by the oral route;

- according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments), Diamine methylated should be classified as Category 3 and should be labeled as H301: Toxic if swallowed.