Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24 November 2016 - 28 December 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
May 2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
no
Qualifier:
according to
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
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
impurity
Type:
impurity
Test material form:
solid
Details on test material:
Identification: Nitrate amine salt of N-[3-dimethylaminopropyl]- C14-C20 amides, saturated, reaction products with ethylene oxide
Appearance: Light yellow lumps
Test item storage: At room temperature; Store in closed container

Test animals

Species:
rat
Strain:
other: Crl:WI (Han)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Young adult animals (approx. 8-9 weeks old)
- Weight at study initiation: 145 - 193 g
- Fasting period before study: 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.
- 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 to 24
- Humidity (%): 40 to 70
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Remarks:
Elix, Millipore S.A.S., Molsheim, France
Details on oral exposure:
GAVAGE METHOD: plastic feeding tubes.

Frequency: single dosage, on Day 1.

VEHICLE
- Justification for choice of vehicle: The vehicle was selected based on trial preparations performed at Charles River Den Bosch and on test item data supplied by the Sponsor. There was no information available regarding the stability in vehicle.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg body weight.

DOSAGE PREPARATION: The 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.
No correction was made for purity of the test item.
The concentration of the test item in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight.
In order to obtain homogeneity, the test item preparations were stirred for 30 minutes during preparation.

CLASS METHOD
- Rationale for the selection of the starting dose: 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 2000 mg/kg. 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.
Doses:
2000 and 300 mg/kg body weight
No. of animals per sex per dose:
2000 mg/kg: 3
300 mg/kg: 6 (2 groups of three females in a stepwise manner)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily. The time of death was recorded as precisely as possible.
Body weights: Days 1 (pre-administration), 8 and 15 and at death (if found dead after Day 1).
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: The animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities recorded.
- 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:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
At 2000 mg/kg bw, two animals were found dead at Day 2 and one animal was found dead at Day 3.
At 300 mg/kg bw, no mortality occurred.
Clinical signs:
At 2000 mg/kg, hunched posture, uncoordinated movements, piloerection, rales, hypersensitivity to touch, ptosis, yellow discoloration of the faeces, faeces containing mucus and/or diarrhoea were noted for all animals on Days 1 and/or 2.
At 300 mg/kg, hunched posture, uncoordinated movements, piloerection, salivation and/or quick breathing were noted for all animals on Days 1 and/or 2.
Body weight:
The mean 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.
Gross pathology:
At 2000 mg/kg, pale discoloration of the glandular mucosa of the stomach was noted for all animals and reddish discoloration of the thymus was noted for two animals.
At 300 mg/kg, no abnormalities were found at macroscopic post mortem examination of the animals.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
In an acute oral toxicity study with rats, performed according to OECD 423 guideline and GLP principles, an LD50 >300 but <2000 mg/kg bw was determined.
Executive summary:

The substance Nitrate amine salt of N-[3-dimethylaminopropyl]- C14-C20 amides, saturated, reaction products with ethylene oxide was tested in an acute oral toxicity study in rats, performed according to OECD 423 guideline and GLP principles.

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 2000 mg/kg bw. Based on the observed mortality a second group of 6 (2x3) females was treated at a dose level of 300 mg/kg bw. At 2000 mg/kg bw, two animals were found dead at Day 2 and one animal was found dead at Day 3. At 300 mg/kg bw, no mortality occurred.

At 2000 mg/kg, hunched posture, uncoordinated movements, piloerection, rales, hypersensitivity to touch, ptosis, yellow discoloration of the faeces, faeces containing mucus and/or diarrhoea were noted for all animals on Days 1 and/or 2. At 300 mg/kg, hunched posture, uncoordinated movements, piloerection, salivation and/or quick breathing were noted for all animals on Days 1 and/or 2.

At 2000 mg/kg, pale discoloration of the glandular mucosa of the stomach was noted for all animals and reddish discoloration of the thymus was noted for two animals. At 300 mg/kg, no abnormalities were found at macroscopic post mortem examination of the animals.

Based on the results of the study, the LD50 was established to be within the range of 300 - 2000 mg/kg body weight. The substance shall be classified as acute oral category 4 and shall be labeled as H302: Harmful if swallowed according to Regulation (EC) No 1272/2008.