C16-18-(even numbered)-alkyl fatty acid, compound with C16-18-(even numbered)-alkylamine

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 December 2012 - 21 February 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Compliant to GLP and testing guidelines; adequate consistence between data, comments and conclusions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

C16-18-(even numbered)-alkyl fatty acid, compound with C16-18-(even numbered)-alkylamine

EC Number:

800-984-9

Cas Number:

1428547-35-6

Molecular formula:

RNH3+, R’COO- where R = C16-18-alkyl and R' = C15-17-alkyl

IUPAC Name:

C16-18-(even numbered)-alkyl fatty acid, compound with C16-18-(even numbered)-alkylamine

Test material form:

other: solid

Details on test material:

- Other name: N-(hydrogenated tallow alkyl) amine, N-(hydrogenated tallow alkyl) fatty acid salt without free fatty acid
- Physical state: light yellow solid
- CAS number : 68551-29-1
- Batch number: EP 4756
- Analytical purity: 100%
- Expiry date: 10 January 2015
- Storage condition: at room temperature.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: breeder: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: approximately 8 weeks old on the day of treatment
- Mean body weight at study initiation: 200 g (range: 196 g to 206 g)
- Fasting period before study: yes, during the night before treatment
- Housing: the animals were housed by three from the same group in polycarbonate cages with stainless steel lids
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: at least 5 days before the beginning of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h.

IN-LIFE DATES: 29 January 2013 to 20 February 2013.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Maximum dose-volume applied: 10 mL/kg

DOSAGE PREPARATION:
The test item was administered as a suspension in the vehicle. The test item was ground to a fine powder, using a mortar and pestle, and then mixed progressively with the required quantity of vehicle. The dose formulations were then kept under magnetic stirring for at least 30 minutes, in a bath water at approximately 40°C.
Fresh dose formulations were prepared by the CiToxLAB France Pharmacy on the day of each administration in brown flasks prior to administration. Dose formulations were transferred to the dosing room in a tray containing hot water (starting at approximately 40°C).The temperature of the hot water was not continuously monitored.

CLASS METHOD:
- Rationale for the selection of the starting dose: the dose-levels were selected in agreement with the Sponsor following the results of a previous oral toxicity study in rats for 14 days where the NOAEL was established at 150 mg/kg/day. No death was observed in this study in rats treated with 50, 150 and 450 mg/kg/day. Clinical signs as piloerection and hunched posture were observed on the last day of treatment in 2/4 females given 450 mg/kg/day. Lower body weight, correlating with lower food consumption, was observed throughout the treatment period in both sexes given 450 mg/kg/day.
Therefore, 2000 mg/kg/day was chosen as the starting dose-level.

Doses:

2000 mg/kg.

No. of animals per sex per dose:

3 females per treatment step.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Clinical observations: frequently during the hours following treatment; then, at least once a day.
- Body weight: just before treatment on day 1; then on days 8 and 15.
- Necropsy of survivors performed: yes (macroscopic).

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No unscheduled deaths occurred during the study.

Clinical signs:

other: No clinical signs were observed in any animals.

Gross pathology:

There were no test item-related macroscopic findings.
The macroscopic observation correlated with common histological finding in control rats, and were thus was considered to be incidental.

Any other information on results incl. tables

 

Sex		Female
Group	CiToxLAB France Historical control data	1	2
Dose-level (mg/kg)	0	2000	2000
Body weight (mean (± SD))
. Day 1	208 (± 11.7)	198 (± 2.1)	202 (± 4.5)
. Day 8^a	246 (± 12.7)	231 (± 3.5)	239 (± 1.0)
. Day 15	266 (± 14.0)	244 (± 5.1)	259 (± 2.5)
Body weight change (mean (± SD))
. Days 1-8^b	+39 (± 5.1)	+33 (± 2.3)	+38 (± 3.5)
. Days 8-15^c	+20 (± 6.3)	+12 (± 4.0)	+20 (± 2.6)
. Days 1-15	+58 (± 5.8)	+45 (± 3.1)	+58 (± 3.8)

^a: day 9 for group 2,^b: days 1-9 for group 2,^c: days 9-15 for group 2.

SD: standard deviations.

Applicant's summary and conclusion

Interpretation of results:

other: not classified as toxic by oral route

Remarks:

Criteria used for interpretation of results: other: CLP Regulation

Conclusions:

The oral LD50 of the test item was higher than 2000 mg/kg in rats.
Therefore, the test item is not classified as toxic by oral route according to the criteria of CLP Regulation.

Executive summary:

The objective of this study was to evaluate the potential acute toxicity of the test item following a single oral administration (gavage) to rats.

 

Methods

The test item was administered once by oral route (gavage) to two groups of three fasted female Sprague-Dawley rats under a dosage-volume of 10 mL/kg. The test item was prepared in corn oil.

Based on available test item toxicity data, the starting dose-level of 2000 mg/kg was chosen. After the first assay, as no toxicity was observed, the results were confirmed in other females.

Each animal was observed at least once a day for mortality and clinical signs for 15 days. Body weight was recorded on day 1 and then on day 8 or 9 and on day 15.

On completion of the observation period, the animals were sacrificed and then submitted for a macroscopic post-mortem examination. Macroscopic lesions were preserved in buffered formalin then destroyed at the finalization of the study report as no microscopic examination was performed.

 

Results

No unscheduled deaths occurred during the study and no clinical signs were observed in any animals.

Body weight gain was unaffected by the test item treatment, when compared to CiTox LAB historical control data.

At necropsy, there were no macroscopic findings attributed to the test item administration.

 

Conclusion

The oral LD₅₀of the test item was higher than 2000 mg/kg in rats.

Therefore, the test item is not classified as toxic by oral route according to the criteria of CLP Regulation.