3-methyl-1,1-diphenylurea

Endpoint summary

Description of key information

Acute Toxicity- Oral point

Only one study is available. GLP study. Klimish score 1.

The testing was performed according to the Method B.1 tris: Acute Oral Toxicity - Acute Toxic Class Method, Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008.

LD50=> 300 mg/kg to < 2000 mg/kg (for female)

Acute Toxicity - Dermal point

Only one study is avalable. GLP study. Klimish score 1.

Testing was performed according to Method B.3 - Acute toxicity(Dermal), Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008.

LD50>2000 mg/kg/bw (for male/female)

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:

03.02.-19.02.2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: breeding farm VELAZ s.r.o., Únětice, Czech Republic, RČH CZ 21760152- Age at study initiation: 6-7 weeks- Weight at study initiation: 112,76 g-136,62 g- Fasting period before study: 20 hours before study- Housing: animal room with monitoring conditions – 3 animals of one sex in one plastic breeding cage Velaz T4, sterilized shavings of soft wood- Diet: complete pelleted standard diet for experimental animals, ad libitum- Water: drinking water, ad libitum- Acclimation period: 6 daysENVIRONMENTAL CONDITIONS- Temperature (°C): 22 ± 3°C, permanently monitored- Humidity (%): 30 – 70 %, permanently monitored- Photoperiod (hrs dark / hrs light): light period 12-hour light/12 hour dark Body weight: before application, 8th day and before euthanasia of animalsMortality: dailyClinical examination: dailyPathological examination: 15th day

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

VEHICLE- Olive oil (pharmaceutical quality)- Batch No.: 5523502- Expiration: 09/2015- Manufacturer: Dr.Kulich Pharma s.r.o., Piletická 178/61, 500 03 Hradec KrálovéDOSAGE PREPARATION- test substance was weighted and mixed in vehicle (olive oil)- single volume of administered suspension was 1ml/100g of animal body weight.CLASS METHOD (if applicable)- Rationale for the selection of the starting dose: On the basis of information about none toxic effect the starting dose of 2000 mg/kg was selected.

Doses:

2000, 300 mg/kg

No. of animals per sex per dose:

3 animals per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days - Frequency of weighing: before application, at the 8th day of study and at the 15th day, before euthanasia of animals.- Necropsy of survivors performed: yes- Other examinations performed: clinical signs, body weight,Nutritious status, body surface, body foramina, thoracic, abdominal and cranial cavity

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

yesThe test substance administered at the dose of 2000 mg/kg caused death of two females.Remaining one female had to be humanely sacrificed for moribund status in the morning 2nd day of the study.See tables No.:3 and 4

Clinical signs:

At the dose 2000 mg/kg: all three femalespiloerection, gibbous posture, decreased reaction to stimuli, tachypnoea, the lateral posture, ataxia (pulling body and pelvic legs), red discharge fromnostrils and salivation, the cachexy, fur stained by urine, cyanosis of skin and mucous membrane, haemorrhagic discharge from nostrils and eye, the dilatation and flatulence of stomach.At dose 300 mg/kg: all six femalesno clinical signs of intoxicationSee tables No.:3 and 4

Body weight:

At dose 2000 mg/kg: all animals lost weigtat dose 300 mg/kg: all animals gained weightSee tables No.: 1 and 2

Gross pathology:

At dose 2000 mg/kg: all three femalesBody surface: cachexy, fur stained by urine and cyanosis of skin and mucous membrane, heamorrhagic discharge (nostrils, eye)Stomach - dilatation and flutulance. At dose 300 mg/kg: all six femalesno macroscopic changes

Table No. 1: Individual body weight (g) – 2000 mg/kg (Step No. 1)

Dose   (mg/kg)  (Step No.)	Animal No.	Body weight (g)			Body weight decrease (g)
		Before application	2 days p.a.	15 days p.a.	1-2 days p.a.	8-15 days p.a.
2000 (1)	1	129.78	113.44	-	16.34	-
	2	121.76	110.00	-	11.76	-
	3	123.99	117.33	-	6.66	-
	mean	125.18	113.59	-	11.59	-
	SD	4.14	2.43	-	4.84	-

Table No. 2: Individual body weight (g) – 300 mg/kg (Steps No. 2 and No. 3)

Dose   (mg/kg)  (Step No.)	Animal No.	Body weight (g)			Body weight gain (g)
		Before application	8 days p.a.	15 days p.a.	1-8 days p.a.	8-15 days p.a.
300 (2)	4	136.62	167.91	189.85	31.29	21.94
	5	125.68	151.94	166.07	26.26	14.13
	6	135.96	161.27	176.30	25.31	15.03
	mean	132.75	160.37	177.41	27.62	17.03
	SD	6.14	8.02	11.93	3.21	4.27
300 (3)	7	129.08	158.97	190.80	29.89	31.83
	8	117.64	153.47	173.94	35.83	20.47
	9	112.76	138.62	154.75	25.86	16.13
	mean	119.83	150.35	173.16	30.53	22.81
	SD	8.38	10.53	18.04	5.02	8.11

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The test substance toxicity was evaluated on the basis of mortality, clinical signs of intoxication, body weight decrease or increment during the observation period and necropsy findings at the end of study. The test substance administered at the dose 2000 mg/kg caused death of two females and moribund status of one female. The serious clinical signs of intoxication were detected during the study in all three animals administered at the dose 2000 mg/kg.The test substance administered at the lower dose of 300 mg/kg caused no death of females. No clinical signs of intoxication and no macroscopic changes in two administered groups of three females were recorded. According to the study results the value of LD50 of the test substance, Akardit, for female rats is in the range > 300 mg/kg to < 2000 mg/kg.

Executive summary:

The aim of the study was to investigate acute toxic effects of the test substance Akardit,after a single oral administration to Wistar rats.

 

The testing was performed according to the Method B.1 tris: Acute Oral Toxicity - Acute Toxic Class Method, Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008.

 

The test substance was administered in a single dose as solution in vehicle (olive oil), given orally via gavage to female Wistar rats.

 

The dosing was performed sequentially in three groups of three females: group No. 1 (first step) using the starting dose of 2000 mg/kg body weight, group No. 2 (second step) and group No. 3 (third step) using a dose of 300 mg/kg body weight. The volume of administered solution was 1 ml/100 g body weight of animals.

      

The test substance administered at the dose of 2000 mg/kg caused death of two females. Remaining one female had to be humanely sacrificed for moribund status in the morning 2^ndday of the study.

The clinical signs of intoxication such as piloerection, gibbous posture, decreased reaction to stimuli and tachypnoea were observed immediately 30 minutes after application in all three animals. The lateral posture, ataxia (pulling body and pelvic legs), red discharge from nostrils and salivation appeared 3hours after application in all three females. The piloerection, tachypnoe and decreased reaction to stimuli were still persist.

Two females was found out dead the 2^ndday morning after application. At this time the clinical examination was performed only in one moribund female. The clinical signs of intoxication such ascyanotic skin and mucous membrane, lacrimation and salivation, red discharge from nostrils,fur stained by urine was observed in one moribund female and the symptoms of approaching deathsuch as tremor, apathy, ataxia (pulling body and pelvic legs), no reaction to stimuli were observed.

 

During pathological examination the cachexy, fur stained by urine, cyanosis of skin and mucous membrane, haemorrhagic discharge from nostrils and eye, were found out in all three females. In abdominal cavity the dilatation and flatulence of stomach was recorded in all three females.

 

The test substance administered at the dose of 300 mg/kg caused no death and no clinical signs in two administered groups of females.

No macroscopic changes were diagnosed during pathological examination.

 

According to the study results the value of LD₅₀of the test substance, Akardit,for female rats is in the range > 300 mg/kg to < 2000 mg/kg.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03.02.-20.02.2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: breeding farm VELAZ s.r.o., Únětice, Czech Republic, RČH CZ 21760152- Fasting period before study: no- Housing:animal room with monitoring conditions – 1 animals of one sex in one plastic breeding cage- Diet: complete pelleted standard diet for experimental animals, ad libitum- Water: drinking water, ad libitum- Acclimation period: 7 days- Identification of animals: colour marks 1 - 5 on tail of animals, each cage was marked with the number of study, sex and dose of the test substance- Health condition: certificate of good health condition – from breeding farm; no signs of diseases were observed at clinical check-in, during the acclimatisation period and before the start of study.ENVIRONMENTAL CONDITIONS- Temperature (°C): 22 ± 3°C, permanently monitored- Humidity (%): 30 – 70 %, permanently monitored- Photoperiod (hrs dark / hrs light): light period 12-hour light/12 hour dark TIME SCHEDULE OF OBSERVATION Body weight:before application (0th), 8th and 15th day of study Mortality:dailyClinical signs:dailyPathological examination:15th day of study

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE- Area of exposure: 6x6 cm on the back of animals- % coverage: 10% of body surface- Type of wrap if used: mull and held by plaster REMOVAL OF TEST SUBSTANCE- Washing: wiped off with water- Time after start of exposure: after 24 hoursTEST MATERIAL- The test substance in delivered form was applied on the depilated area of skin.

Duration of exposure:

24h

Doses:

2000 mg/kg

No. of animals per sex per dose:

5 females/2000 mg/kg5 males/2000 mg/kg

Control animals:

no

Details on study design:

COURSE STUDYThe study was performed as limit test: two groups of animals – 5 males and 5 females at the dose of 2000 mg/kg. The pre-test was performed with 1 male and 1 female of those groups of the five animals. The pre-test was started one day before the start of limit testCLINICAL OBSERVATION-changes in skin and fur, eyes, visible mucous membranes, behaviour of animals, somatomotor activity, reactions to stimuli, and presence of lacrimation, salivation and discharge from nostrils, function of respiratory, digestive and urogenital system-the first day: twice (30 minutes and 3 hours after application)-the second day: twice (in the morning and in the afternoon) and daily thereafter for 14 days.PATHOLOGICAL EXAMINATION-Nutritional state, body surface, body foramina, thoracic, abdominal and cranial cavity-sacrificed on the 15th day

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

no death of animals

Clinical signs:

No clinical signs of intoxication

Body weight:

In males and females, the weight increments were adequate to species and age of animals in the experiment (see table No.1 and No.2).

Gross pathology:

No macroscopic changes

Table No. 1: Individual body weight of animals – 2000 mg/kg – males

Animal No.	Before application (0th day)	8th day	15th day	Body weight gain (g)
				day 0-8 p.a.	day 8-15 p.a.
1 (pre-test)	231.80	279.90	313.8	48.10	33.90
2	237.30	269.90	304.8	32.60	34.90
3	254.80	293.20	338.6	38.40	45.40
4	244.90	275.60	312.5	30.70	36.90
5	248.70	284.50	313.9	35.80	29.40
Average	243.5	280.62	316.72	37.12	36.1

Table No. 2: Individual body weight of animals – 2000 mg/kg – females

Animal No.	Before application (0th day)	8th day	15th day	Body weight gain (g)
				day 0-8 p.a.	day 8-15 p.a.
1 (pre-test)	211.50	225.50	254.2	14.00	28.70
2	207.20	219.70	228.8	12.5	9.10
3	207.80	210.60	228.2	2.80	17.60
4	216.90	222.10	238.6	5.20	16.50
5	208.60	214.60	240.0	6.00	25.40
Average	210.4	218.5	237.96	8.1	19.46

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance toxicity was evaluated on the basis of mortality, body weight changes, and clinical signs of toxicity during the observation period and necropsy findings at the end of study. The test substance applied on skin at a dose of 2000 mg/kg of animal weight did not cause death of animals. No clinical signs of toxicity were observed during the whole study.No macroscopic changes were diagnosed during pathological examination of all animals. According to the results of study, the value of LD50 (dermal) of the test substance, Akardit, for rats of both sexes is higher than 2000 mg/kg of body weight.

Executive summary:

The test substance,Akardit, was tested for acute dermal toxicity using Wistar rats.

Testing was performed according to Method B.3 - Acute toxicity(Dermal), Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008.

 

The study was performed as limit test: two groups of animals – 5 males and 5 females at the dose of 2000 mg/kg of body weight. The pre-test was performed with 1 male and 1 female from each group. After end exposure test substance of these pilot animals, the other animals of the group were dosed.

The test substance in delivered form was applied on the shaved skin of the test animals for 24 hours.

The test animals were observed 14 days after exposure to test substance, afterwards they were sacrificed, and the necropsy for macroscopic examination of the organs was performed.

    

The test substance applied at the dose of 2000 mg/kg of body weight did not cause death of animals. No clinical signs of toxicity were observed during the whole study. No macroscopic changes were diagnosed during pathological examination.   

 

According to the results of study, the value of LD₅₀(dermal) of the test substance,
Akardit, for rats of both sexes is higher than 2000 mg/kg of body weight. 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Justification for classification or non-classification

Acute toxicity by oral route: Based on the available data the substance akardit is classified as Acute Tox. 4 (H302).

Acute toxicity by dermal route. Based on available data the substance Akardit is not classified.