Nonanoic acid, mixed diesters, with oxybis[propanol] and dodecanoic acid

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 10 November 2020 to 27 November 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP guideline study (OECD 420) without deviations

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Version / remarks:

Adopted 17th December 2001

Deviations:

no

Principles of method if other than guideline:

not applicable

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Nonanoic acid, mixed diesters, with oxybis[propanol] and dodecanoic acid

EC Number:

946-682-1

Cas Number:

2166089-27-4

Molecular formula:

Non applicable (UVCB)

IUPAC Name:

Nonanoic acid, mixed diesters, with oxybis[propanol] and dodecanoic acid

Test material form:

liquid

Details on test material:

- Appearance: Limpid liquid
- Storage condition of test material: Keep container tightly closed. Preferably store in the original packaging. Store at room temperature, protect from humidity.

Specific details on test material used for the study:

Storage conditions: at ambient temperature (15 to 25°C) in the dark

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Crl:CD (SD), SPF.

Sex:

female

Details on test animals or test system and environmental conditions:

Rationale for species: According to The Guidelines for Testing of Chemicals, the preferred rodent species was the rat, normally females are used. This is because literature surveys of conventional LD50 tests show that differences are observed, females are generally slightly more sensitive. Therefore, female SD rats were used in this study.

TEST ANIMALS
- Source: Beijing Vital River Laboratory Animal Technology Co., Ltd.
- Weight at study initiation: 201 g～211 g.
- Housing: Rats were reared in a plastic cage with a volume of 420 mm x 270 mm x 200 mm (length x width x height), and the cage was covered with sterilized shavings pad. Male and female animals were reared in different cages, and no more than 5 animals per cage.
- Diet: Breeding Rodent Diet., ad libitum
- Water: City drinking water, ad libitum
- Acclimation period: According to the health screening instructions provided by the veterinarian during the quarantine period, the experimental animals used in this test were in good health. During the adaptation period, there were no obvious abnormalities in the cage observation, body weight, food intake of the animals, indicating that the batch of animals can be used for experiments.

The animals care and use in this study were also in compliance with the Guide for the Care and Use of Laboratory Animals (2011) issued by The National Academies Press. The facility had passed the accreditation of Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC).
All procedures in this protocol were in compliance with the 3R principle (Reduction, Replacement and Refinement), the study did not unnecessarily duplicate any previous study. The animal use of this study had been reviewed and approved by Institutional Animal Care and Use Committee (IACUC) of the facility. The IACUC Number of this study was IACUC-20-205.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.7°C to 25.6°C
- Humidity (%): 40% to 64.8%.
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): A 12-hour light/12-hour dark cycle, 8:00 a.m. light and 8:00 p.m. dark.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

The test substance formulations were administered by gavage needle. All rats were fasted overnight prior to administration.

MAXIMUM DOSE VOLUME APPLIED: 5.45 mL/kg bw.

DOSAGE PREPARATION
Use as supplied. The amount of test substance were stored in EP tube before exposure, sealed with parafilm and stored at ambient temperature.

Doses:

Single dose of 5000 mg/kg bw. A period of 1 day was allowed as time interval.

No. of animals per sex per dose:

1 females in sighting study and another 4 females in main study.

Control animals:

no

Details on study design:

RATIONALE FOR DOSE SELECTION :
According to the Guidelines for Testing of Chemicals (No. 420. Acute Oral Toxicity-Fixed Dose Procedure), only when justified by specific regulatory needs, the use of an additional upper fixed dose level of 5000 mg/kg may be considered. Recognising the need to protect animal welfare, testing at 5000 mg/kg is discouraged and should only be considered when there is a strong likelihood that the results of such a test would have a direct relevance for protecting animal or human health. This test is technical supplementary information required by regulatory agencies. The results of the test substance: Acute Oral Toxicity Study in the Rat (Study Number: TS67MJ) which provided by Sponsor showed that the acute median lethal oral dose (LD50) to rats of test item was demonstrated to be greater than 2000 mg/kg body weight. The results of repeated dose 90-day oral toxicity study of the test substance in rats (Study Number: 18HXCD026Rat) which done by Center for Drug Safety Evaluation and Research of ZJU showed that SD rats were administered with the test substance (Lot No.:S18-0144) daily for 90 days at the dose level of 100, 300, 1000 mg/kg. No changes related to the exposure of the test substance were noted. The NOAEL was 1000 mg/kg.
For information on the acute toxicity of oral test LD50> 2000 mg/kg. This study selected 5000 mg/kg for starting dose, specific reference to the starting dose of 5000 mg/kg of test procedure, as shown in below, A sighting study starting dose of 5000 mg/kg is used to 1 female which out come no toxicity and the selection of 5000 mg/kg as the main study starting dose were allowed. A main study starting dose of 5000 mg/kg was used to 4 females for observing toxicity and death.

EXAMINATIONS:
The following parameters and end points were evaluated in this study: mortality and clinical signs, body weights and gross necropsy findings.

CLINICAL OBSERVATIONS
- Each Animal was observed individually after administration within 30 minutes, and at 1 h, 2 h, 3 h, 4 h, and continue to observe daily for 14 days.
- Observations should include changes in skin, fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous system, physical activity, and behavior pattern. Particular attention was directed to observations of tremors, convulsions, salivation, diarrhea, diarrhea, sleep and coma. Abnormal findings were recorded, including severity, occurrence, duration and reversibility.

MORTALITY
- During the study, all animals were observed once daily for mortality. The number of death animal and the time of death should be recorded. Animals which die during the study were necropsy duly to figure out the causes of death.

BODY WEIGHT
- Individual body weight of animals was weighted the day of exposure (D0) and on D1, D8, and D14.

GROSS NECROSPY
The survival animals were anesthetized by intraperitoneal injection of 3% pentobarbital sodium solution (45 mg/kg) on the day of planned dissection (D14). After anesthesia, the abdominal aorta was exudated and sacrificed and complete and systematic necropsy and gross pathological examinations were performed and recorded, including appearance, natural channel, chest cavity, abdominal cavity and its contents.

Any of tissues or organs had no obvious injury or lesions, therefore, the fixation of the tissues and organs was not performed. No tissue or organ was preserved, so histopathological examination was not performed.

Statistics:

Not applicable/relevant

Results and discussion

Preliminary study:

1 female with 5000 mg/kg bw: no clinical signs

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality and moribund Status were observed.

Clinical signs:

other: During the study, animals were administrated with the test substance at a dose of 5000 mg/kg, no toxic reation was noted on the day of exposure (D0) and the following observation period.

Gross pathology:

At the end of observation period, 5 rats at dose of 5000 mg/kg were necropsied, and no obvious gross pathological changes were found in the tissues and organs.

Other findings:

None

Any other information on results incl. tables

Table 1. Clinical observations

Dose (mg/kg)	Sex	Animal No.	Clinical observations
			The day of exposure (D0)	Symptom observation period (D1-D14)
5000	Female	1	No obvious abnormality	No obvious abnormality
		2	No obvious abnormality	No obvious abnormality
		3	No obvious abnormality	No obvious abnormality
		4	No obvious abnormality	No obvious abnormality
		5	No obvious abnormality	No obvious abnormality

 

Table 2. Individual Body Weight of Animals

Dose (mg/kg)	Sex	Animal No.	Weight (g)
			The day of exposure (D0)	D1	D8	D14
5000	Female	1	201	215	239	257
		2	202	215	238	253
		3	203	217	240	252
		4	206	220	250	258
		5	211	224	263	273

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the oral LD50 of the test substance is > 5000 mg/kg bw in rats. Therefore it is not classified according to the Regulation (EC) N° 1272-2008 and according to the GHS. No signal word or hazard statement is required.

Executive summary:

Introduction. The purpose of this study was to assess the toxic potential of the test item following a single oral dose in the rat using the Fixed Dose Method. The study complies with the requirements of OECD Guideline for the Testing of Chemicals No 420, adopted 17th December 2001.

The method provides information on the hazardous properties and allows the substance to be ranked and classified according to the Globally Harmonized System (GHS).

 

Method. 5 healthy female SD rats were employed in acute oral toxicity test, and acute toxic fixed dose method was adopted. For information on the acute toxicity of oral test LD50 > 2000 mg/kg.This study selected 5000 mg/kg for starting dose. A sighting study starting dose of 5000 mg/kg is used to 1 female which outcome no toxicity and the selection of 5000 mg/kg as the main study starting dose were allowed. A main study starting dose of 5000 mg/kg was used to 4 females for observing toxicity and death, no mortality was observed in main study, the animal experiment was completed. Each animal was observed individually within 30 minutes and at 1 h, 2 h, 3 h, 4 h, and continue to observe daily for 14 days.All surviving animals were weighted on the day of exposure (D0) and on D1, D8, and D14. At the end of the observation period, complete and systematic necropsy of all surviving animals was performed.

Results. 5 female rats were administrated with the test substance at a dose of 5000 mg/kg, no toxic reation was noted on the day of exposure (D0) and the following observation period. No abnormality in body weight was noted in the rest animals. At the end of the observation period, all rats were necropsied and had no obvious pathological changes in tissues and organs.

 

Conclusion. Under the test conditions, the oral LD50 of the test substance is > 5000mg/kg bw in rats. Therefore it is not classified according to the Regulation (EC) N° 1272-2008 and according to the GHS. No signal word or hazard statement is required.