N,N'-(ethoxymethylsilylene)bis[N-methylbenzamide]

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

November 15, 2016 - December 6, 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 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:

N,N'-(ethoxymethylsilylene)bis[N-methylbenzamide]

EC Number:

240-354-5

EC Name:

N,N'-(ethoxymethylsilylene)bis[N-methylbenzamide]

Cas Number:

16230-35-6

Molecular formula:

C19H24N2O3Si

IUPAC Name:

N,N'-[ethoxy(methyl)silanediyl]bis(N-methylbenzamide)

Test material form:

liquid

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage JANVIER LABS (F-53941 Le genest St. Isle)
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8-9 weeks old
- Weight at study initiation: Step 1: 203 ± 7.0; Step 2 and 3: 214.8 ± 6.3
- Fasting period before study: overnight
- Housing: Rats were housed by group of three in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid. Each cage contains sawdust bedding which was changed at least 2 times a week. Each cage was installed in conventional air conditioned animal husbandry.
- Diet: foodstuff (ENVIGO - 2016) ad libitum
- Water: tap-water from public distribution system ad libitum. Microbiological and chemical analyses of the water were carried out once every six months by Bureau Veritas-eurofins (France)
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19ºC - 25ºC
- Humidity (%): 30% - 70%
- Air changes (per hr): Ten changes per hour
- Photoperiod (hrs dark / hrs light): Twelve hours light (07.00 to 19.00) / twelve hours darkness.

IN-LIFE DATES: From: November 15, 2016 To: December 6, 2016

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: In first step, test item was administered corresponding to 2g/kg, according to the calculated density. In second and third step, 0.27 mL of the test item (corresponding to 300 mg) were added to 1.54 mL of olive oil.
- Amount of vehicle (if gavage): 1.81 mL/kg.
- Justification for choice of vehicle: it produced the most suitable formulation at the requested concentration.

MAXIMUM DOSE VOLUME APPLIED: 1.81 mL/kg.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Whitout preliminary information. The selected starting dose is 2000 mg/kg body weight.

Doses:

2000 mg/kg, 300 mg/kg

No. of animals per sex per dose:

3 (Step 1)
6 (Step 2)

Control animals:

yes

Remarks:

Study No. TAO423-2016-005 (see "Other information on results").

Details on study design:

- Duration of observation period following administration: 14 days .
- Frequency of observations and weighing: Systemic observations at 30 min, 1h, 3h, 4h, 24h, 48h after administration and daily during 14 days. Weighing: on Day 0 (just before administering the test item) then on Day 2, Day 7, and Day 14.
- Necropsy of survivors performed: yes.
At the end of the study, the animals were anaesthetised with sodium pentobarbital and administration continued to fatal levels. Macroscopic observations were entered on individual autopsy sheets. Only those organs likely to be modified in cases of acute toxicity were examined. No organ was removed and preserved in view to microscopic examinations.
- Other examinations performed:
Body weight and body weight gain: the animals were weighted on D0 (just before administering the test item) then on D2, D7, and D14.
Clinical signs: Spontaneous activity, Preyer's reflex (noise), Respiratory rate, Convulsions, Tremors, Body temperature, Muscle tone, Palpebral opening, Pupil appearance, Salivation, Lachrymation, Righting reflex, Back hair appearance, Mortality.
Gross pathology: On D14, the animals were anaesthetised with sodium pentobarbital and administration continued to fatal levels. Macroscopic observations were entered on individual autopsy sheets. Only those organs likely to be modified in cases of acute toxicity were examined. Parameters examined: Oesophagus, Stomach, Duodenum, Jejunum, Ileon, Caecum, Colon, Rectum, Spleen, Liver, Thymus, Trachea, Lungs, Heart, Kidneys, Urinary Bladder, Ovaries, Uterus, Treatment Area, Adrenals and Pancreas.

Results and discussion

Effect levels

Effect levels 1

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Effect levels 2

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

ca. 500 mg/kg bw

Based on:

test mat.

Mortality:

At the dose of 2000 mg/kg bw, all three animals died (3/3), two at 24 hours post dose and one other at 48 hours post dose.
No mortality occurred during the study at the dose of 300 mg/kg.

Clinical signs:

At 2000 mg/kg bw: the mortalities were preceded by an absence or decrease in spontaneous activity (3/3), muscles tones (3/3), righting reflex (3/3), Preyer's reflex (3/3), associated with bradypnea (3/3), dyspnea (1/3), hypothermia (3/3), eyes partly (2/3) or totally (2/3) closed, mydriasis (3/3), an increase of salivation (1/3) and piloerection (1/3). Rigor mortis were noted before the necropsy (3/3).
At 300 mg/kg bw: no clinical signs related to the administration of the test item were observed during the study.

Body weight:

At 300 mg/kg bw: the body weight evolution of the animals remained normal throughout the study.

Gross pathology:

At 2000 mg/kg bw: the macroscopic examination of the animals revealed a thinning of a forestomach and corpus (1/3), a swelling of the stomach (2/3), red coloration of a forestomach and corpus (2/3). Furthermore, cellular lysis of main organs in the abdomen was noted (2/3).
At 300 mg/kg bw: the macroscopic examination of the animals at the end of the study did not reveal treatment related changes.

Any other information on results incl. tables

Clinical observations:

Dose 2000 mg/kg bw: results in Tables 1 to 6.

Dose 300 mg/kg bw: all the observations were Normal during the first 4 hours after administration of the test item, and then 14 days after.

Body weight evolution:

Dose 2000 mg/kg bw: results in Table 7.

Dose 300 mg/kg bw: results in Table 8.

Macroscopical examinations:

Dose 2000 mg/kg bw: necropsy data sheets (Tables 9 and 10).

Dose 300 mg/kg bw: nothing to report.

Table 1. Test item at 2000 mg/kg bw. Observation data sheet.

OBSERVATIONS:	FEMALES
T0 + 30 minutes	Rf 0709	Rf 0710	Rf 0711
Spontaneous activity	D	0	D
Preyer’s réflex (noise)	D	0	D
Repiratory rate	N	N	N
convulsions	N	N	N
tremors	N	N	N
Body temperature	N	N	N
Muscle tone	N	0	D
Palpebral opening	N	N	N
Pupil appearance	N	Md	Md
Salivation	N	N	N
Lachrymation	N	N	N
Ringhting reflex	N	0	D
Back hair appearance	N	N	N
MORTALITY	0	0	0
Remarks	None

N: Normal / None (Convulsions, Tremors)

Md: Mydriasis

D: Decreased / limited (Righting reflex) / Hypothermia (Body temperature)

0: None

 

Table 2. Test item at 2000 mg/kg bw. Observation data sheet.

OBSERVATIONS:	FEMALES
T0 + 1 hour	Rf 0709	Rf 0710	Rf 0711
Spontaneous activity	0	0	0
Preyer’s réflex (noise)	0	0	0
Repiratory rate	B	B	B
convulsions	N	N	N
tremors	N	N	N
Body temperature	N	D	N
Muscle tone	D	0	D
Palpebral opening	N	Cc	Pc
Pupil appearance	Md	Md	Md
Salivation	N	N	N
Lachrymation	N	N	N
Ringhting reflex	D	0	D
Back hair appearance	N	N	N
MORTALITY	0	0	0
Remarks	None

N: Normal / None (Convulsions, Tremors)

B: Bradypnea

Cc: Eyes completely closed

Pc. Eyes partly closed

Md: Mydriasis

D: Decreased / limited (Righting reflex) / Hypothermia (Body temperature)

0: None

 

Table 3. Test item at 2000 mg/kg bw. Observation data sheet.

OBSERVATIONS:	FEMALES
T0 + 3 hours	Rf 0709	Rf 0710	Rf 0711
Spontaneous activity	0	0	0
Preyer’s réflex (noise)	0	0	0
Repiratory rate	B	B	B
convulsions	N	N	N
tremors	N	N	N
Body temperature	D	D	D
Muscle tone	0	0	0
Palpebral opening	Pc	Cc	Cc
Pupil appearance	Md	Md	Md
Salivation	N	N	N
Lachrymation	N	N	N
Ringhting reflex	0	0	0
Back hair appearance	N	N	N
MORTALITY	0	0	0
Remarks	None

N: Normal / None (Convulsions, Tremors)

B: Bradypnea

Cc: Eyes completely closed

Pc. Eyes partly closed

Md: Mydriasis

D: Decreased / limited (Righting reflex) / Hypothermia (Body temperature)

0: None

 

Table 4. Test item at 2000 mg/kg bw. Observation data sheet.

OBSERVATIONS:	FEMALES
T0 + 4 hours	Rf 0709	Rf 0710	Rf 0711
Spontaneous activity	0	0	0
Preyer’s réflex (noise)	0	0	0
Repiratory rate	B	B	B
convulsions	N	N	N
tremors	N	N	N
Body temperature	D	D	D
Muscle tone	0	0	0
Palpebral opening	Pc	Cc	Cc
Pupil appearance	Md	Md	Md
Salivation	N	N	N
Lachrymation	N	N	N
Ringhting reflex	0	0	0
Back hair appearance	N	N	N
MORTALITY	0	0	0
Remarks	None

N: Normal / None (Convulsions, Tremors)

B: Bradypnea

Cc: Eyes completely closed

Pc. Eyes partly closed

Md: Mydriasis

D: Decreased / limited (Righting reflex) / Hypothermia (Body temperature)

0: None

 

Table 5. Test item at 2000 mg/kg bw. Observation data sheet.

OBSERVATIONS:	FEMALES
D1	Rf 0709	Rf 0710	Rf 0711
Spontaneous activity	0
Preyer’s réflex (noise)	0
Repiratory rate	B
convulsions	N
tremors	N
Body temperature	D
Muscle tone	0
Palpebral opening	Pc
Pupil appearance	N
Salivation	A
Lachrymation	A
Ringhting reflex	0
Back hair appearance	Pi
MORTALITY	0	1	1
Remarks	Rf0710 & Rf0711: found dead at T0 + 22 hours and 39 minutes

N: Normal / None (Convulsions, Tremors)

A: Increased

B: Bradypnea

Cc: Eyes completely closed

Pc. Eyes partly closed

Pi: Piloerection

D: Decreased / limited (Righting reflex) / Hypothermia (Body temperature)

0: None

 

Table 6. Test item at 2000 mg/kg bw. Observation data sheet.

OBSERVATIONS:	FEMALES
D1	Rf 0709	Rf 0710	Rf 0711
Spontaneous activity
Preyer’s réflex (noise)
Repiratory rate
convulsions
tremors
Body temperature
Muscle tone
Palpebral opening
Pupil appearance
Salivation
Lachrymation
Ringhting reflex
Back hair appearance
MORTALITY	1
Remarks	Rf0709: found dead on day 2

 

Table 7. Test item at 2000 mg/kg bw. Body weight and weight gain in grams.

FEMALES	D0	D2			D2-D0	D7	D7-D0		D14	D14-D0
Rf 0709	208	†
Rf 0710	206	†
Rf 0711	195	†
MEAN	203.0
Standard deviation	7.0

†: animal found dead.

 

Table 8. Test item at 300 mg/kg bw. Body weight and weight gain in grams.

FEMALES	D0	D2			D2-D0	D7	D7-D0		D14	D14-D0
Rf 0721	214	224			10	266	52		282	68
Rf0722	213	220			7	250	37		278	65
Rf 0723	206	214			8	244	38		256	50
Rf 0738	220	235			15	251	31		275	55
Rf 0739	212	227			15	254	42		258	46
Rf 0740	224	242			18	269	45		279	55
MEAN	214.8	227.0			12.2	255.7	40.8		271.3	56.5
Standard deviation	6.3		10.2	4.4		9.8	7.3	11.3			8.5

 

Table 9. Necropsy data sheet of rats Rf0710 to Rf0711 (16 November 2016)

 

Found dead:   X	Euthanasia:	At term:
GENERAL APPEARANCE BEFORE AUTOPSY: Rigor mortis, found dead at T0 + 22 hours and 39 minutes
	Observed Organs	Observations
* OESOPHAGUS	X	N.t.R.
* STOMACH	X	Forestomach 6 corpus red colored, swollen stomach
* DUODENUM	-	Cell lysis
* JEJUNUM	-	Cell lysis
* ILEON	-	Cell lysis
* CAECUM	-	Cell lysis
* COLON	-	Cell lysis
* RECTUM	-	Cell lysis
* SPLEEN	-	Cell lysis
* LIVER	X	N.t.R.
* THYMUS	X	N.t.R.
* TRACHEA	X	N.t.R.
* LUNGS	X	N.t.R.
* HEART	X	N.t.R.
* KIDNEYS	X	N.t.R.
* URINARY BLADDER	X	N.t.R.
* OVARIES	X	N.t.R.
* UTERUS	X	N.t.R.
* TREATMENT AREA	-	-
* ADRENALS	X	N.t.R.
* PANCREAS	X	N.t.R.
PARTICULARS. None

N.t.R.: Nothing to report

 

Table 10. Necropsy data sheet of rat Rf0709 (17 November 2016)

 

Found dead:   X	Euthanasia:	At term:
GENERAL APPEARANCE BEFORE AUTOPSY: Rigor mortis, found dead on day 2
	Observed Organs	Observations
* OESOPHAGUS	X	N.t.R.
* STOMACH	X	Thickening of the forestomach and the corpus
* DUODENUM	X	N.t.R.
* JEJUNUM	X	N.t.R.
* ILEON	X	N.t.R.
* CAECUM	X	N.t.R.
* COLON	X	N.t.R.
* RECTUM	X	N.t.R.
* SPLEEN	X	N.t.R.
* LIVER	X	N.t.R.
* THYMUS	X	N.t.R.
* TRACHEA	X	N.t.R.
* LUNGS	X	N.t.R.
* HEART	X	N.t.R.
* KIDNEYS	X	N.t.R.
* URINARY BLADDER	X	N.t.R.
* OVARIES	X	N.t.R.
* UTERUS	X	N.t.R.
* TREATMENT AREA	-	-
* ADRENALS	X	N.t.R.
* PANCREAS	X	N.t.R.
PARTICULARS. Body weight: 194 g

N.t.R.: Nothing to report

 

Applicant's summary and conclusion

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The LD50 of the test item is higher than 300 mg/kg bw and lower than 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off of the test item may be considered as 500 mg/kg body weight by oral route in the rat.

Executive summary:

The acute oral toxicity of the test item has been tested in accordance with OECD 423 and EU method B.1 tris, following GLP. The test item was administered, as supplied, to a group of 3 female Sprague-Dawley rats at the dose of 2000 mg/kg and 6 female Sprague-dawley rats at the dose of 300 mg/kg body weitght by oral gavage. At the highest dose, all three animals died (3/3), two at 24 hours post dose and one other at 48 hours post dose. The mortalities were preceded by an absence or decrease in spontaneous activity (3/3), muscles tones (3/3), righting reflex (3/3), Preyer's reflex (3/3), associated with bradypnea (3/3), dyspnea (1/3), hypothermia (3/3), eyes partly (2/3) or totally (2/3) closed, mydriasis (3/3), an increase of salivation (1/3) and piloerection (1/3). Rigor mortis were noted before the necropsy (3/3). The macroscopic examination of the animals revealed a thinning of a forestomach and corpus (1/3), a swelling of the stomach (2/3), red coloration of a forestomach and corpus (2/3). Furthermore, cellular lysis of main organs in the abdomen was noted (2/3). At the dose of 300 mg/kg, no mortality occurred during the study and the body weight evolution of the animals remained normal during the study. Furthermore, no clinical signs related to the administration of the test item were observed during the study. The macroscopic examination of the animals at the end of the study did not reveal treatment related changes. The LD50 of the test item is higher than 300 mg/kg bw and lower than 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off of the test item may be considered as 500 mg/kg body weight by oral route in the rat. Based on the available information, the test item has to be classified in category 4 according to CLP Regulation (EC) no. 1272/2008.