N,N-bis(2-ethylhexyl)formamide

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 Mar - 01 Apr 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted in 2001

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

N,N-bis(2-ethylhexyl)formamide

EC Number:

860-695-9

Cas Number:

88247-41-0

Molecular formula:

C17H35NO

IUPAC Name:

N,N-bis(2-ethylhexyl)formamide

Test material form:

liquid

Test animals

Species:

rat

Strain:

other: Crl:CD(SD)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Hino Breeding Center, Charles River Laboratories, Japan
- Age at study initiation: 7 weeks
- Weight at study initiation: 191.3 - 216.7 g
- Fasting period before study: From 18 h before administration to 3 h after administration.
- Housing: In groups of 3/cage during the quarantine period and acclimatisation and individually thereafter. The animals were kept in hanging stainless steel cages with mesh-floor (260W x 380D x 180H mm) before the group allocation and in hanging stainless steel cages with mesh-floor (165W x 300Dx 150H mm) after the group allocation.
- Historical data: Historical control data comprising clinical signs, body weight and necropsy data were available for animals administered the vehicle (olive oil).
- Diet: pelleted diet MF (Lots 201008 and 201214, Oriental Yeast), ad libitum, autoclaved before use
- Water: chlorinated water (sodium hypochlorite was added at 3 - 5 ppm to maintain the chloric level), ad libitum
- Acclimation period: 6 days
- Microbiological status when known : SPF


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 25
- Humidity (%): 40 - 70
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 02 Mar 2021 To: 01 Apr 2021

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 3 and 20% (w/v)
- Amount of vehicle: 10 mL/kg bw
- Justification for choice of vehicle: Although the test substance did not dissolve and was not suspended to purified water at a concentration of 20.0 w/v%, the test substance was dissolved to olive oil at a concentration of 20.0 w/v%. The condition of the formulation such as color did not change at room temperature four hours after the preparation. Therefore, olive oil was selected as a vehicle.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION:
Dosing formulation was prepared on each administration day. The test substance was weighed and mixed with olive oil to be dissolved. The solution was filled up to the prescribed volume with olive oil to prepare the dosing formulation. The dosing formulation was carried to the animal room.

CLASS METHOD:
- Rationale for the selection of the starting dose: Since no toxic information of the test substance was available, the dose level of the 1st step was set at 300 mg/kg bw. Subsequent doses were set according to the test method.

Doses:

300 and 2000 mg/kg bw

No. of animals per sex per dose:

3 females per step per dose group

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: In each step, the animals were observed continuously for 10 min after the administration, and observed 30 min and 3 h after the administration on the administration day, and animals were observed once in the morning from 1 - 14 days after the administration.
- Frequency of weighing: Body weights were recorded prior to administration (Day 0) and on Days 1, 7 and 14 after administration.
- Necropsy of survivors performed: The animals were subjected to a gross necropsy 14 days after the administration. The animals were euthanized by bleeding from the abdominal aorta under isoflurane anesthesia. External surface of the body, all orifices, subcutis, cranial, thoracic, abdominal and pelvic cavities with their contents were observed.

Statistics:

Statistical analysis was not performed.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study period.

Clinical signs:

other: 300 mg/kg bw (1st and 2nd step): 2/6 animals showed mucous stool 3 h after administration. The animals fully recovered within 24 h post administration. There were no abnormalities observed thereafter. 2000 mg/kg bw (3rd and 4th step): No clinical signs of

Gross pathology:

Necropsy and pathological examination revealed no substance-related findings.

Any other information on results incl. tables

Table 1: Body weight development

Step	Dose (mg/kg)	Animal No.	Body weights (g)
			Day after administration
			Initial	1	7	14
1st	300	1	197.2	218.8	240.1	252.1
			ΔBW	21.6	21.3	12.0
		2	191.3	204.8	234.7	254.6
			ΔBW	13.5	29.9	19.9
		3	197.6	217.8	245.5	264.9
			ΔBW	20.2	27.7	19.4
2nd		4	199.7	222.4	239.3	251.5
			ΔBW	22.7	16.9	12.2
		5	201.2	223.8	253.5	260.7
			ΔBW	22.6	29.7	7.2
		6	203.3	222.1	253.7	280.7
			ΔBW	18.8	31.6	27.0
3rd	2000	7	216.7	215.9	246.6	266.6
			ΔBW	-0.8	30.7	20.0
		8	211.3	223.5	252.0	271.6
			ΔBW	12.2	28.5	19.6
		9	212.4	218.1	264.2	307.8
			ΔBW	5.7	46.1	43.6
4th		10	213.0	217.0	248.5	263.5
			ΔBW	4.0	31.5	15.0
		11	209.9	220.1	248.1	249.2
			ΔBW	10.2	28.0	1.1
		12	209.7	216.9	237.0	249.2
			ΔBW	7.2	20.1	12.2
Historical control data (n = 30)			mean ± SD	16.3 ± 4.2
			Range	7.4 - 27.9
ΔBW: Difference from previous body weight

Applicant's summary and conclusion

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008

Conclusions:

The substance was tested for acute oral toxicity in female rats according to OECD guideline 423, at dose levels of 300 and 2000 mg/kg bw. No mortality occured and the resulting LD50 was > 2000 mg/kg bw.