Amidinourea phosphate

Acute Toxicity: oral

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

Amidinourea phosphate

EC Number:

241-659-6

EC Name:

Amidinourea phosphate

Cas Number:

17675-60-4

Molecular formula:

C2H6N4O.xH3O4P

IUPAC Name:

diaminomethylideneurea;phosphoric acid

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

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

Test animals

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

Administration / exposure

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

Results and discussion

Preliminary study:

See any other information on results

Effect levels

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

Any other information on results incl. tables

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

Applicant's summary and conclusion

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.