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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-05-03 to 2017-06-16
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2019

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Principles of method if other than guideline:
- Principle of test:
The study was designed to determine, in a preliminary phase, the maximum tolerated dose (MTD) and, successively, the toxicity of the test item over a period of 14 consecutive days of treatment (main phase).

- Short description of test conditions:
In the preliminary phase (Phase 1), 2 male and 2 female rats/groupwere initially administered at a dose level of 300mg/kg/day (Group 1) and then at 2000mg/kg/day (Group 2), each followed by a 7-day observation period.
In the main phase (Phase 2), 4 animals/sex/dose were administered once daily at 100, 500 and 1000mg/kg/day for at least 14 consecutive days.

- Parameters analysed / observed:
– preliminary phase: clinical signs, body weight, gross macroscopic observations;
– main phase: clinical signs, body weight, food consumption, clinical pathology investigations (including thyroid hormones determination), terminal body weight, organ weights and macroscopic observations.
GLP compliance:
no
Remarks:
However, study was carried out in a GLP compliant facility (DRF for OECD 422 study)
Test type:
other: dose - range finder
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Octadecanoic acid, reaction products with triethylenetetramine
EC Number:
270-171-6
EC Name:
Octadecanoic acid, reaction products with triethylenetetramine
Cas Number:
68412-15-7
Molecular formula:
C24H54N4O2
IUPAC Name:
N'-[2-(2-aminoethylamino)ethyl]ethane-1,2-diamine;octadecanoic acid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS srl, San Pietro al Natisone (UD), Italy.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 6-7 weeks
- Weight at study initiation: males: 150 to 174 g
- Fasting period before study: Phase 1: overnight fast prior to each dosing day; Phase 2: the diet was offered ad libitum throughout the study
- Housing: up to 5 of one sex to a cage, in clear polysulfone solid bottomed cages
- Diet: commercially available laboratory rodent diet (4 RF 21, Mucedola S.r.l.) ad libitum
- Water: drinking water via water bottles ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
22 +/- 2 °C
- Humidity (%): 55 +/- 15 %
- Air changes (per hr): 15 to 20 air changes per hour
- Photoperiod (hrs dark / hrs light):
artificaial light for 12 hours each

IN-LIFE DATES: From: 2017-05-03 to 2017-06-16

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Remarks:
0.5 % aqueous solution
Details on oral exposure:
VEHICLE
- Concentration in vehicle: Phase 1: 30 and 200 mg/L
Phase 2: 10, 50 and 100 mg/mL
- Amount of vehicle (if gavage): 0.5 % aqueous solution


MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg


Doses:
Phase 1: 300 and 2000 mg/kg single oral dose
Phase 2: 0, 100, 500 and 1000 mg/kg treatment for 14 consecutive days
No. of animals per sex per dose:
Phase 1: 2 animals/sex/group
Phase 2: 4 animals/sex/group
Control animals:
no
Details on study design:
The following investigations were performed:
– preliminary phase: clinical signs, body weight, gross macroscopic observations;

– main phase: clinical signs, body weight, food consumption, clinical pathology investigations
(including thyroid hormones determination), terminal body weight, organ weights and macroscopic observations.

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred during any phases of the study.
Clinical signs:
other: No clinical signs were observed during the preliminary phase (Phase 1) and main phase (Phase 2) of the study.
Gross pathology:
Animals of both sexes treated at 300 and 2000 mg/kg/day killed at termination did not show any macroscopic changes.

Any other information on results incl. tables

Haematology

Phase 2

Some statistically significant differences between control and treated animals were observed, such as: reduction of mean corpuscular volume in animals of both sexes dosed

at 500 mg/kg/day and in males receiving 1000 mg/kg/day (4% to 7% below controls), reduction of mean corpuscular haemoglobin in males treated at 500 mg/kg/day (4%) and increase

of mean corpuscular haemoglobin concentration in females from the same treated group (5%). Due to the minimal severity and/or the absence of dose-relation, these findings were

considered of no toxicological relevance.

Clinical chemistry

Phase 2

Glucose was increased in males dosed at 500mg/kg/day and in animals of both sexes receiving 1000mg/kg/day. Compared with controls, changes were 50% to 64%, with no doserelation.

In addition, transaminases enzymes were increased in a number of animals, including controls, therefore this finding was not attributed to treatment. The statistically

significant decrease of alanine aminotransferase recorded in males dosed at 500mg/kg/day (40% below controls) was not dose-related, therefore it was considered incidental.

Thyroid hormone determination

Phase 2

No relevant changeswere recorded. Thyroid stimulatinghormonewas decreased infemales underlined dosed at 1000mg/kg/day (43% belowcontrols).

This condition (lowserumthyroid stimulating hormone levels, normal T4 and T3 levels) was subclinical and asymptomatic, therefore it was considered to be not adverse.

Terminal body weight and organ weights

Phase 2

Terminal body weight did not show any differences among treated groups and controls.

No changes of toxicological significance were observed in the absolute and relative weight of organs.

Macroscopic observations

Phase 2

No treatment-related changes were noted in treated animals of both sexes sacrificed at termination, when compared to controls. Sporadic changes as adhesion of thymus to lungs and heart in one female treated at 100 mg/kg/day (no. E0149021 - low dose) were not considered attributable to treatment.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
For acute oral toxicity of Octadecanoic acid, reaction products with triethylenetetramine a LD50 value of > 2000 mg/kg can be established.
Executive summary:

The toxicity of Octdecanoic acid, reaction products with triethylenetetramine after oral administration was investigated in a study similar to OECD guideline 423 by gavage to Sprague Dawley rats.

The study was designed to determine, in a preliminary phase, the maximum tolerated dose (MTD) and, successively, the toxicity of the test item over a period of 14 consecutive days of treatment (main phase) in order to select dose levels for the subsequent OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test) in rats.

 

In the preliminary phase (Phase 1), 2 male and 2 female rats/group were initially administered at a dose level of 300 mg/kg/day (Group 1) and 2000 mg/kg/day (Group 2), each followed by a 7-day observation period.

No toxicity was observed at each dose level investigated.

 

On the basis of results obtained in Phase 1, in the main phase (Phase 2), 4 animals/sex/dose were administered once daily at 100, 500 and 1000mg/kg/day for at least 14 consecutive days.

 

No mortality and no clinical signs were recorded in main phase animals; body weights as well as terminal body weights were not affected by treatment.

Minimal alterations were observed in some male and/or female animals at the haematological and clinical chemistry evaluation.

However, due to the minimal severity and/or absence of dose-relation, these findings were considered of no toxicological relevance.

No treatment-related findings were reported at post mortem macroscopic observations of all treated animals.

On the basis of the results obtained in this preliminary study, it can be concluded that the test item Octadecanoic acid, reaction products with triethylenetetramine, showed no toxic effects when administered daily at dose levels of 100, 500 and 1000 mg/kg/days for at least 14 consecutive days.

For acute oral toxicity of Octadecanoic acid, reaction products with triethylenetetramine a LD50value of > 2000 mg/kg can be established.