Methyl 4-(acetylamino)-5-bromo-o-anisate

Endpoint summary

Description of key information

Acute oral toxicity:

Key study: OECD 423 and EU method B.1 tris. Klimish score = 1. GLP study. The LD50 of the test item is higher than 2000 mg/kg body weight by oral route in the rat.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March 14, 2017 - March 29, 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

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

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Stability under test conditions: yes

Species:

rat

Strain:

Sprague-Dawley

Remarks:

SPF Caw

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: JANVIER LABS (53940 Le Genest St Isle - France)
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8 weeks old
- Weight at study initiation: 205.8 ± 10.1
- Fasting period before study: overnight
- Housing: 3 females 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 (e.g. ad libitum): Foodstuff (ENVIGO -2016) ad libitum
- Water (e.g. ad libitum): 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: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19ºC - 25ºC
- Humidity (%): 30% - 70%. A relative humidity lower than 30% was registered from 14 to 19 March 2017 and from 22 to 29 March 2017. The minimum value measured was 19%. The deviation is considered as without impact on the conclusion of the study.
- Air changes (per hr): At least 10 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours contimuous light (07.00 to 19.00) and twelve hours darkness.

IN-LIFE DATES: From: To: 14 March 2017 - 29 March 2017

Route of administration:

oral: gavage

Vehicle:

DMSO

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 2 g/ 10 mL
- Amount of vehicle (if gavage): 10 mL/Kg
- Justification for choice of vehicle: DMSO produced the most suitable formulation at the requested concentration.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/Kg

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Without preliminary information. The selected starting dose is 2000 mg/kg body weight.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

6

Control animals:

yes

Remarks:

Control group refer to a control study performed from 28 February 2017 to 14 March 2017

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 adminitrationand daily during 14 days. Weighing= D0 (just before administering the test item) then on D2, D7, and D14.
- Necropsy of survivors performed: yes. At termination, macroscopic observation was observed. 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.
- Other examinations performed:
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.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

act. ingr.

Mortality:

No mortality occurred during the study.

Clinical signs:

Myosis was noted at 30 minutes post dose in three treated animals (3/6). The animals recovered a normal activity at 1 hour post dose. No others signs of systemic toxicity were noted.

Body weight:

The body weight evolution of the animals remained normal during the study.

Gross pathology:

The macroscopic examination of the animals at the end of the study did not reveal treatment related changes.

Clinical observations: observation data sheets (Tables 1 to 4).

Body weight evolution: Table 5.

Macroscopical examinations: necropsy data sheets (Tables 5 and 6).

 

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

OBSERVATIONS:	FEMALES			FEMALES
T0 + 30 minutes	Rf 1251	Rf 1252	Rf 1253	Rf 1270	Rf 1271	Rf 1272
Spontaneous activity	N	N	N	N	N	N
Preyer’s réflex (noise)	N	N	N	N	N	N
Repiratory rate	N	N	N	N	N	N
convulsions	N	N	N	N	N	N
tremors	N	N	N	N	N	N
Body temperature	N	N	N	N	N	N
Muscle tone	N	N	N	N	N	N
Palpebral opening	N	N	N	N	N	N
Pupil appearance	Ms	Ms	Ms	N	N	N
Salivation	N	N	N	N	N	N
Lachrymation	N	N	N	N	N	N
Ringhting reflex	N	N	N	N	N	N
Back hair appearance	N	N	N	N	N	N
MORTALITY	0	0	0	0	0	0
Remarks	None			None

N: Normal / None (Convulsions, Tremors)

Md: Ms (Myosis)

 

 

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

OBSERVATIONS:	FEMALES			FEMALES
T0 + 1 hour T0 + 3 hours T0 + 4 hours	Rf 1251	Rf 1252	Rf 1253	Rf 1270	Rf 1271	Rf 1272
Spontaneous activity	N	N	N	N	N	N
Preyer’s réflex (noise)	N	N	N	N	N	N
Repiratory rate	N	N	N	N	N	N
convulsions	N	N	N	N	N	N
tremors	N	N	N	N	N	N
Body temperature	N	N	N	N	N	N
Muscle tone	N	N	N	N	N	N
Palpebral opening	N	N	N	N	N	N
Pupil appearance	Ms	Ms	Ms	N	N	N
Salivation	N	N	N	N	N	N
Lachrymation	N	N	N	N	N	N
Ringhting reflex	N	N	N	N	N	N
Back hair appearance	N	N	N	N	N	N
MORTALITY	0	0	0	0	0	0
Remarks	None			None

N: Normal / None (Convulsions, Tremors)

Md: Ms (Myosis)

 

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

OBSERVATIONS:	FEMALES			FEMALES
D1 to D14	Rf 1251	Rf 1252	Rf 1253	Rf 1270	Rf 1271	Rf 1272
Spontaneous activity	N	N	N	N	N	N
Preyer’s réflex (noise)	N	N	N	N	N	N
Repiratory rate	N	N	N	N	N	N
convulsions	N	N	N	N	N	N
tremors	N	N	N	N	N	N
Body temperature	N	N	N	N	N	N
Muscle tone	N	N	N	N	N	N
Palpebral opening	N	N	N	N	N	N
Pupil appearance	N	N	N	N	N	N
Salivation	N	N	N	N	N	N
Lachrymation	N	N	N	N	N	N
Ringhting reflex	N	N	N	N	N	N
Back hair appearance	N	N	N	N	N	N
MORTALITY	0	0	0	0	0	0
Remarks	None			None

N: Normal / None (Convulsions, Tremors)

 

Table 4. 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 1251	214	226			12	254	40		281	67
Rf1252	195	208			13	232	37		280	85
Rf 1253	204	212			8	238	34		274	70
Rf 1270	220	230			10	256	36		278	58
Rf 1271	207	227			20	250	43		275	68
Rf 1272	195	206			11	234	39		258	63
MEAN	205.8	218.2			12.3	244.0	38.2		274.3	68.5
Standard deviation	10.1		10.7	4.1		10.6	3.2	8.5			9.1

 

Table 5. Necropsy data sheet of rats Rf1251 to Rf1253 (28 March 2017)

 

Found dead:	Euthanasia:    X	At term:
GENERAL APPEARANCE BEFORE AUTOPSY: Normal
	Observed Organs	Observations
* OESOPHAGUS	X	N.t.R.
* STOMACH	X	N.t.R.
* 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. None

N.t.R.: Nothing to report

 

Table 6. Necropsy data sheet of rats Rf1270 to Rf1272 (29 March 2017)

 

Found dead:	Euthanasia:  X	At term:
GENERAL APPEARANCE BEFORE AUTOPSY: Normal
	Observed Organs	Observations
* OESOPHAGUS	X	N.t.R.
* STOMACH	X	N.t.R.
* 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: None

N.t.R.: Nothing to report

 

 

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 of the test item is higher than 2000 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 to a group of 6 female Sprague-Dawley rats at the dose of 2000 mg/kg body weight by oral gavage. No mortality occurred during the study and the body weight evolution of the animals remained normal during the study. Myosis was noted at 30 minutes post dose in three treated animals (3/6). The animals recovered a normal activity at 1 hour post dose. No others signs of systemic toxicity were noted. 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 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off of the test item may be considered to be higher than 5000 mg/kg body weight by oral route in the rat.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

5 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute oral toxicity:

Key study: 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 to a group of 6 female Sprague-Dawley rats at the dose of 2000 mg/kg body weight by oral gavage. No mortality occurred during the study and the body weight evolution of the animals remained normal during the study.Myosis was noted at 30 minutes post dose in three treated animals (3/6). The animals recovered a normal activity at 1 hour post dose. No others signs of systemic toxicity were noted.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 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off of the test item may be considered to be higher than 5000 mg/kg body weight by oral route in the rat.

Justification for classification or non-classification

Based on the available information (LD50 > 2000 mg/kg bw), the substance is not classified for acute toxicity according to CLP Regulation (EC) no. 1272/2008