Registration Dossier

Administrative data

Description of key information

In an acute oral toxicity study with rats, performed according to OECD/EC test guidelines and GLP principles, an LD50 between 300 and 2000 mg/kg bw  for Guanidinium phosphate (1:1)was determined.
In an acute dermal toxicity study with rats, performed according to OECD/EC test guidelines, an LD50 >2000 mg/kg bw for guanidinium phosphate (1:1) was determined.

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:
03JAN2013 to 24JAN2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 2011, including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
other: Crl:WI (Han)
Sex:
female
Details on test animals and environmental conditions:
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Young adult animals (approx. 8-12 weeks old)
- Weight at study initiation: Body weight variation was within +/- 20% of the sex mean (150-203g)
- Fasting period before study: yes, overnight
- Housing: Group housing of 3 animals per cage in labeled Macrolon cages
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany)
- Water: Free access to tap water
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 – 24
- Humidity (%): 40 - 70
- Air changes (per hr): approx 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 03JAN2013 to 24JAN2013
Route of administration:
oral: gavage
Details on oral exposure:
GAVAGE METHOD: plastic feeding tubes.

Frequency: single dosage, on Day 1.

VEHICLE: water
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor.

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg (10 mL/kg) body weight.

DOSAGE PREPARATION: The formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. No correction was made for purity of the test substance. The concentration of the test substance in vehicle was varied to allow constant dosage volume in terms of mL/kg body weight.
Doses:
2000 or 300 mg/kg body weight


No. of animals per sex per dose:
2000 mg/kg bw: 3
300 mg/kg bw: 6 (2 groups of three females in a stepwise manner)
Control animals:
no
Details on study design:
The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 2000 mg/kg. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily
Body weights: Days 1 (pre-administration), 8 and 15
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: The animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide procedure. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities recorded
- Other examinations performed: none
Statistics:
No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Three female were found dead on Day 1. No further mortality occurred.
Clinical signs:
At a dose level of 2000 mg/kg bw the following clinical signs were noted: Lethargy, tremor, abnormal posture, flat and/or hunched posture, abnormal gait, piloerection, restless and fearful behavior and/or chromodacryorrhoea were noted on Day one (day of death).
At a dose level of 300 mg/kg bw, three animals showed hunched posture and piloerection on Day 1.
Body weight:
The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.
Gross pathology:
Macroscopic post mortem examination of the animals at 2000 mg/kg (that were all found dead on Day 1) revealed in two animals: several reddish foci in the glandular mucosa of the stomach and/or gastro-intestinal tract distended with gas.
No abnormalities were found at macroscopic post mortem examination of the animals at 300 mg/kg.
Interpretation of results:
harmful
Remarks:
Migrated information cat. 4 Criteria used for interpretation of results: EU
Conclusions:
In an acute oral toxicity study with rats, performed according to OECD/EC test guidelines and GLP principles, an LD50 between 300 and 2000 mg/kg bw for Guanidinium phosphate (1:1) was determined.
Executive summary:

An acute oral toxicity study was performed with rats according to OECD/EC test guidelines and GLP principles following the acute toxic class method. Three rats were exposed to 2000 mg/kg bw guanidinium phosphate (1:1), all three died on day 1. Lethargy, tremor, abnormal posture, flat and/or hunched posture, abnormal gait, piloerection, restless and fearful behavior and/or chromodacryorrhoea were noted on the day of death. Macroscopic post mortem examination of these animals revealed in two animals several reddish foci in the glandular mucosa of the stomach and/or gastro-intestinal tract distended with gas.

Two groups of three rats were exposed to 300 mg/kg bw, no further mortality occurred. Three animals showed hunched posture and piloerection on Day 1, no abnormalities were found at necropsy. The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

Based on these data, the LD50 for guanidinium phosphate (1:1) was determined to be between 300 and 2000 mg/kg bw, the substance is classified for oral toxicity in category 4 according to Regulation (EC) 1272/2008.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
300 mg/kg bw
Quality of whole database:
The study has been performed according to OECD and/or EC guidelines and according to GLP principles (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:
10JAN2013 to 24JAN2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, April 2011; including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: Wistar strain, Crl:WI (Han)
Sex:
male/female
Details on test animals and environmental conditions:
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Young adult animals (approx. 10 weeks old)
- Weight at study initiation: Body weight variation was within +/- 20% of the sex mean: males 261-286g, females 185-215g
- Housing: Individually housed in labeled Macrolon cages
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany)
- Water: Free access to tap water
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 – 24
- Humidity (%): 40 - 70
- Air changes (per hr): approx 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 10JAN2013 to 24JAN2013
Type of coverage:
occlusive
Vehicle:
water
Details on dermal exposure:
One day before exposure (Day -1) an area of approximately 5x7 cm on the back of the animal was clipped.

The test substance formulation was applied in an area of approx. 10% of the total body surface, i.e. approx. 25 cm² for males and 18 cm² for females. The test substance formulation was held in contact with the skin with a dressing, consisting of a surgical gauze patch, successively covered with aluminum foil and elastic bandage. A piece of tape was additionally used for fixation of the bandages in females only.

Frequency: Single dosage, on Day 1.

Washing: Following application, dressings were removed and the skin cleaned of residual test substance using tap water.
Duration of exposure:
24 hours.
Doses:
2000 mg/kg


No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
VEHICLE water
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at WIL Research and on test substance data supplied by the sponsor

Dose volume: 2000 mg/kg (10 mL/kg) body weight

DOSAGE PREPARATION: \\ tekst onder Preparation toevoegen

Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily
Body weights: Days 1 (pre-administration), 8 and 15
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy.
- Other examinations performed: none.
Statistics:
None.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Piloerection was noted in five males and three females on Day 1. In addition, one male and one female showed chromodacryorrhoea on that day.
Body weight:
The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Other findings:
Scales and/or macular erythema were seen in the treated skin-area of all five females during the observation period.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an acute dermal toxicity study with rats, performed according to OECD/EC test guidelines, an LD50 >2000 mg/kg bw for guanidinium phosphate (1:1) was determined.
Executive summary:

An acute dermal toxicity study was performed with rats, according to OECD/EC test guidelines. Five male and five female rats were dermally exposed to guanidinium phosphate at 2000 mg/kg bw. No mortality occurred, and minor clinical symptoms were noted ( piloerection in five males and three females; one male and one female with chromodacryorrhoea, all on day 1 only). Scales and/or macular erythema were seen in the treated skin-area of all five females during the observation period. No unexpected changes in body weight gain occurred and no abnormalities were found at necropsy.

Based on these data, an LD50 >2000 mg/kg bw for guanidinium phosphate (1:1) was determined. Therefore, the substance is not classified for dermal toxicity according to Regulation (EC) No 1272/2008.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The study has been performed according to OECD and/or EC guidelines and according to GLP principles (reliability 1).

Additional information

An acute oral toxicity study was performed with rats according to OECD/EC test guidelines and GLP principles following the acute toxic class method. Three rats were exposed to 2000 mg/kg bw guanidinium phosphate (1:1), all three died on day 1. Lethargy, tremor, abnormal posture, flat and/or hunched posture, abnormal gait, piloerection, restless and fearful behavior and/or chromodacryorrhoea were noted on the day of death. Macroscopic post mortem examination of these animals revealed in two animals several reddish foci in the glandular mucosa of the stomach and/or gastro-intestinal tract distended with gas.

Two groups of three rats were exposed to 300 mg/kg bw, no further mortality occurred. Three animals showed hunched posture and piloerection on Day 1, no abnormalities were found at necropsy. The body weight gain shown by the surviving animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

An acute dermal toxicity study was performed with rats, according to OECD/EC test guidelines. Five male and five female rats were dermally exposed to guanidinium phosphate at 2000 mg/kg bw. No mortality occurred, and minor clinical symptoms were noted ( piloerection in five males and three females; one male and one female with chromodacryorrhoea, all on day 1 only). Scales and/or macular erythema were seen in the treated skin-area of all five females during the observation period. No unexpected changes in body weight gain occurred and no abnormalities were found at necropsy.


Justification for selection of acute toxicity – oral endpoint
Only one study available.The study has been performed according to OECD and/or EC guidelines and according to GLP principles (reliability 1).

Justification for selection of acute toxicity – dermal endpoint
Only one study available.The study has been performed according to OECD and/or EC guidelines and according to GLP principles (reliability 1).

Justification for classification or non-classification

Based on the available data, guanidinium phosphate (1:1) is classified cat.4 for oral toxicity, and is not classified for dermal toxicity or toxicity via inhalation according to Regulation (EC) No 1272/2008.