Amidinourea phosphate

Endpoint summary

Description of key information

Acute Oral Toxicity

A single oral dose of the test item to female rats at a dose of 2000 mg/kg bw associated with signs of toxicity and mortality. The signs of toxicity recovered within up to 1-day post-dose. After this time point no signs of toxicity were visible. Rats dosed at 300 mg/kg bw showed signs of toxicity but no mortality. The signs of toxicity recovered within up to 240 minutes post-dose.The median lethal dose observed over a period of 14 days is: LD50 cut-off (rat): 2000 mg/ kg bw According to Annex I of Regulation (EC) 1272/2008 guanylurea phosphate is proposed to be classified into Category 4.

Acute Dermal Toxicity

Under the conditions of the present study, a single dermal application of the test item to rats at a dose of 2000 mg/kg bw was associated with no mortality and no signs of toxicity or of irritation were evident. The dermal LD50 was determined to be > 2000 mg / kg bw.

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:

21-03-2017 to 24-08-2017

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)

Version / remarks:

OECD Guidelines for Testing of Chemicals, Section 4, No. 423, “Acute Oral Toxicity – Acute Toxic
Class Method” adopted 17 December 2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Specific details on test material used for the study:

Batch No.: 16VL8189
Expiry Date: 03 August 2017
Storage Conditions: room temperature, protected from light

Species:

rat

Strain:

Wistar

Remarks:

WISTAR rats Crl: WI(Han)

Sex:

female

Details on test animals or test system and environmental conditions:

Prior to the administration a detailed clinical observation was made of all animals. Only healthy animals were used, and food was withheld from the test animals for 16 to 19 hours (access to water was permitted). Following the period of fasting the animals were weighed and the test item was administered. Food was provided again approximately 4 hours post dosing.

Housing and Feeding Conditions:

Full barrier in an air-conditioned room
Temperature: 22 3 °C
Relative humidity: 55 10%
Artificial light, sequence being 12 hours light, 12 hours dark
Air change: 10 x / hour
Free access to Altromin 1324 maintenance diet for rats and mice
Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals).

The animals were kept in groups in IVC cages, type III H, polysulphone cages on Altromin saw
fibre bedding

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

Single oral dose at a dose volume of 10 mL/kg body weight.

Doses:

2000 mg/kg body weight and 300 mg/kg body weight

No. of animals per sex per dose:

3 per step, see results

Control animals:

no

Details on study design:

The starting dose was selected to be 2000 mg/kg body weight. Compound-related mortality was recorded for one animal of step 1. Based on these results and according to the acute toxic class method regime, a second step was performed at a dose of 2000 mg/kg body weight. Compoundrelated mortality was recorded for any animal of step 2. Based on these results and according to the acute toxic class method regime, a third step was performed at a dose of 300 mg/kg body weight. No compound-related mortality was recorded for any animal of step 3. Based on these results and according to the acute toxic class method regime, a fourth step was performed at a dose of 300 mg/kg body weight. No compound-related mortality was recorded for any animal of step 4. Based on these results and according to the acute toxic class method regime, no further testing was required.

The surviving animals were observed for 14 days after dosing for general clinical signs, morbidity and mortality.

The animals were weighed on day 1 (prior to the administration) and on days 8 and 15.

A careful clinical examination was made several times on the day of dosing (at least once during the first 30 minutes and with special attention given during the first 4 hours post-dose). As soon as symptoms were noticed they were recorded. Thereafter, the animals were observed for clinical signs once daily until the end of the observation period. All abnormalities were recorded. Cageside observations included changes in the skin and fur, eyes and mucous membranes. Also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern were examined. Particular attention was directed to observations of tremor, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

The animals which died spontaneously during the observation period were necropsied directly after onset of death. At the end of the observation period the surviving animals were sacrificed with an overdosage of pentobarbital injected intraperitoneally at a dosage of 250-400 mg/kg bw. All animals were subjected to gross necropsy. All gross pathological changes were recorded but not preserved for possible histopathological evaluation.

Statistics:

Results were interpreted according to OECD Guideline 423, Annex 2

Preliminary study:

See any other information on results

Key result

Sex:

female

Dose descriptor:

LD50

Remarks:

LD50 cut-off (rat): 2000 mg/ kg bw

Effect level:

>= 2 000 mg/kg bw

Based on:

test mat.

Mortality:

The test item showed mortality and other acute oral toxicity characteristics after a single dose administration of 2000 mg/kg body weight. The test item showed no mortality but other acute oral
toxicity characteristics after a single dose administration of 300 mg/kg body weight.

Clinical signs:

other: See any other information on results

Gross pathology:

See any other information on results

Clinical Signs - Individual Data
Step	Female No.	Dose (mg/kg bw)	Timepoint	Observations
1	1	2000	0-60 min	nsf
			60 - 240 min	Moderately reduced spontaneous activity, hunched posture, moderate piloerection, half eyelid closure, slight salivation
			240 -15 d	nsf
	2	2000	0 -120 min	nsf
			120 - 240 min	Moderately reduced spontaneous activity, hunched posture, moderate piloerection, half eyelid closure, slight salivation
			240 - 300 min	Died spontaneously
	3	2000	0 -120 min	nsf
			120 - 240 min	Moderately reduced spontaneous activity, hunched posture, moderate piloerection, half eyelid closure, slight salivation
			240 min -15 d	nsf
2	4	2000	0-30 min	Moving the bedding, moderate salivation
			30 - 60 min	Moderately reduced spontaneous activity, moving the bedding, hunched posture, slow movements, slight piloerection, half eyelid closure, slight salivation
			60 -120 min	Moderately increased spontaneous activity, hunched posture, slight piloerection, half eyelid closure, slow movements
			120 - 240 min	Moderately reduced spontaneous activity, hunched posture, moderate piloerection, half eyelid closure
			240 min	Moderately reduced spontaneous activity, hunched posture, moderate piloerection, eyes closed, slow movements.  Died spontaneously
	5	2000	0-30 min	Moving the bedding, moderate salivation
			30 - 60 min	Moving the bedding, moderately reduced spontaneous activity, hunched posture, slow movements, slight piloerection, half eyelid closure, slight salivation
			60 -120 min	Moderately reduced spontaneous activity, hunched posture, slow movements, slight piloerection, half eyelid closure
			120 -180 min	Moderately reduced spontaneous activity, hunched posture, moderate piloerection, half eyelid closure
			180 min	Died spontaneously
	6	2000	0-30 min	Moving the bedding, moderate salivation
			30 - 60 min	Moving the bedding, moderately reduced spontaneous activity, hunched posture, slow movements, slight piloerection, half eyelid closure, slight salivation
			60 -120 min	Moderately reduced spontaneous activity, hunched posture, slow movements, slight piloerection, half eyelid closure
			120 - 240 min	Moderately reduced spontaneous activity, hunched posture, moderate piloerection, half eyelid closure
			240 min	Moderately reduced spontaneous activity, hunched posture, slow movements, moderate piloerection, eyes closed. Died spontaneously
3	7	300	0-30 min	nsf
			30 - 60 min	Moderately reduced spontaneous activity, prone position, half eyelid closure
			60 -120 min	Slightly reduced spontaneous activity, half eyelid closure, slight salivation
			120 -180 min	Slightly reduced spontaneous activity, half eyelid closure, moderate piloerection
			180 - 240 min	Slightly reduced spontaneous activity,
			240 min - 15d	nsf
	8	300	0-30 min	nsf
			30 - 60 min	Moderately reduced spontaneous activity, prone position, half eyelid closure
			60 -120 min	Slightly reduced spontaneous activity, half eyelid closure
			120 -180 min	Slightly reduced spontaneous activity, half eyelid closure, moderate piloerection
			180 - 240 min	Slightly reduced spontaneous activity
			240 min- 15d	nsf
	9	300	0-30 min	nsf
			30 - 60 min	Moderately reduced spontaneous activity, prone position, half eyelid closure
			60 -120 min	Slightly reduced spontaneous activity, half eyelid closure
			120 -180 min	Slightly reduced spontaneous activity, half eyelid closure, moderate piloerection
			180 - 240 min	Slightly reduced spontaneous activity
			240 min-15 d	nsf
4	10	300	0 min - 15 d	nsf
	11	300	0 min - 15 d	nsf
	12	300	0 min - 15 d	nsf
d = day; min = minute(s); nsf = no specific findings

Body Weights (g) & Body Weight Gain %
Step	Female No.	Dose (mg/kg bw)	BW (g)			Body weight in comparison to day 1 (%)
			Day 1	Day 8	Day 15	15
1	1	2000	146	166	175	20
	2	2000	146	Died spontaneously
	3	2000	150	165	174	16
2	4	2000	159	Died spontaneously
	5	2000	162
	6	2000	158
3	7	300	140	172	184	31
	8	300	140	170	199	42
	9	300	130	162	182	40
4	10	300	142	173	191	35
	11	300	140	170	183	31
	12	300	149	176	174	17
Day 1 = day of administration; bw = body weight

Findings at Necropsy - Individual Data
Step	Female No.	Starting Dose (mg/kg bw)	Organ	Macroscopic Findings
1	1	2000	-	nsf
	2		stomach	Test item residues
	3		-	nsf
2	4		stomach	Test item residues
	5
	6
3	7	300	-	nsf
	8		-	nsf
	9		-	nsf
4	10		-	nsf
	11		-	nsf
	12		-	nsf
bw = body weight; nsf = no specific findings

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

LD50 cut-off (rat): 2000 mg/ kg bw

Executive summary:

A single oral application of the test item guanylurea phosphate to female rats at a dose of 2000 mg/kg body weight was associated with signs of toxicity and mortality. The signs of toxicity recovered within up to 1-day post-dose. After this time point no signs of toxicity were visible. A single oral application of the test item guanylurea phosphate to female rats at a dose of 300 mg/kg body weight was associated with signs of toxicity but no mortality. The signs of toxicity recovered within up to 240 minutes post-dose. After this time point no signs of toxicity were visible.

The median lethal dose observed over a period of 14 days is: LD50 cut-off (rat): 2000 mg/ kg bw. According to Annex I of Regulation (EC) 1272/2008 guanylurea phosphate is proposed to be classified into Category 4.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Reliability 1

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

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:

20-03-2017 to 29-06-2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

OECD Guidelines for Testing of Chemicals, Section 4, No. 402, “Acute Dermal Toxicity” adopted 24 Feb, 1987

Deviations:

yes

Remarks:

For technical reasons all raw data were recorded on paper.

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Specific details on test material used for the study:

Batch No.: 16VL8189
Expiry Date: 03 August 2017
Storage Conditions: room temperature, protected from light

Species:

rat

Strain:

Wistar

Remarks:

WISTAR Crl: WI(Han) rats

Sex:

male/female

Details on test animals or test system and environmental conditions:

Age at the beginning of the study: males: 9-10 weeks old; females: 9-10 weeks old
Body weight on the day of administration: males: 265 – 289 g females: 200 – 215 g

Housing and Feeding Conditions:

Full barrier in an air-conditioned room
Temperature: 22 3 °C
Relative humidity: 55 10%
Artificial light, sequence being 12 hours light, 12 hours dark
Air change: 10 x / hour
Free access to Altromin 1324 maintenance diet for rats and mice
Free access to tap water, sulphur acidified to a pH value of approximately 2.8 (drinking water,
municipal residue control, microbiological controls at regular intervals)
The animals were kept individually in IVC cages, type III H, polysulphone cages on Altromin saw
fibre bedding
Adequate acclimatisation period (at least five days) under laboratory conditions

Preparation of the Animals:

The animals were marked for individual identification by tail painting.
Approximately 25 (for males) and 24 (for females) hours before the test, the fur was removed from
the dorsal area of the trunk using an electric clipper. Care was taken to avoid abrading the skin, and
only animals with healthy intact skin were used.
No less than 10% of the body surface was cleared for the application.
Prior to the application a detailed clinical observation was made of all animals. Only healthy animals
were used.

Type of coverage:

occlusive

Vehicle:

water

Details on dermal exposure:

Approximately 25 (for males) and 24 (for females) hours before the test, the fur was removed from the dorsal area of the trunk using an electric clipper. Care was taken to avoid abrading the skin, and only animals with healthy intact skin were used.

No less than 10% of the body surface was cleared for the application.

Prior to the application a detailed clinical observation was made of all animals. Only healthy animals were used.

Duration of exposure:

24 hours

Doses:

Single dose of 2000 mg/kg body weight

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

OECD 402: All animals were observed for 14 days after dosing.

Statistics:

Signs of erythema and oedema were assessed using the scoring system (Table 2) laid down in OECD Guideline 404

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No test substance related deaths

Clinical signs:

other: No signs of toxicity for both sexes. No skin irritation for both sexes.

Gross pathology:

No significant findings.

Absolute Body Weights in g and Body Weight Change in %

Dose: 2000 mg/kg body weight
Animal No. / Sex	^g Day 1	g Day 8	^g Day 15	% Day 1-15
21 / male	265	281	304	15
22 / male	273	291	317	16
23 / male	273	299	326	19
24 / male	289	307	350	21
25 / male	271	289	322	19
26 / female	206	211	217	5
27 / female	201	205	210	4
28 / female	205	203	224	9
29 / female	215	219	233	8
30 / female	200	210	214	7

Conclusions:

The dermal LD50was determined to be > 2000 mg GUANYLUREA PHOSPHATE / kg body weight.

Executive summary:

Under the conditions of the present study, single dermal application of the test item GUANYLUREA PHOSPHATE to rats at a dose of 2000 mg/kg body weight was associated with no mortality and neither signs of toxicity nor signs of irritation. The dermal LD50 was determined to be > 2000 mg GUANYLUREA PHOSPHATE / kg body weight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Reliability 1

Justification for classification or non-classification

Acute Oral Toxicity

According to Annex I of Regulation (EC) 1272/2008 guanylurea phosphate is proposed to be classified into Category 4.

Acute Dermal Toxicity

According to Annex I of Regulation (EC) 1272/2008 guanylurea phosphate no classification is proposed.