Diguanidinium carbonate

Endpoint summary

Description of key information

- Oral: LD50 rat:  1089 mg/kg (m) and 1045 mg/kg (f), generally compliant to OECD TG 401
- Dermal: LD50: > 2000 mg/kg in female rats, OECD TG 402

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: - meets generally accepted scientific principles - sufficient information for assessment reported

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

post exposure period only 7 d

GLP compliance:

no

Remarks:

study predates implementation of GLP

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: own breeding facility
- Age at study initiation: not reported
- Weight at study initiation: 150 - 200 g
- Fasting period before study: 2 - 3 hours prior to oral application until 2 - 3 hours after application, food was removed. Water was given immediately after application.
- Housing: 5 per cage, Makrolon R cage
- Diet (e.g. ad libitum): ALTROMIN R-R-pellets, ad libitum
- Water (e.g. ad libitum): not reported
- Acclimation period: not reported, but raised in own breeding facility

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 26 ± 1
- Humidity (%): 50 - 60
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): not reported

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

test substance applied as 13.3 % aqueous solution

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 13.3 %
- Amount of vehicle (if gavage): 4.45, 6.7, 10, 15 ml/kg bw
- Justification for choice of vehicle: The test item was known to be sufficiently water soluble

MAXIMUM DOSE VOLUME APPLIED: 15 mL/kg

Doses:

592, 889, 1333 and 2000 mg/kg bw,

No. of animals per sex per dose:

10

Control animals:

no

Details on study design:

- Duration of observation period following administration: 7 d
- Frequency of observations and weighing: not reported
- Necropsy of survivors performed: not reported
- Other examinations performed: clinical signs, body weight, organ weights, histopathology, other: not reported

Statistics:

Thompson-Weil (Thompson WRT and Weil CS, 1952, Biometrics 8:51-4; Weil CS, 1952, Biometrics 8:249-63).
Tables from this article were supplemented with r-values by the company's data processing centre.

Sex:

male

Dose descriptor:

LD50

Effect level:

1 089 mg/kg bw

Based on:

test mat.

95% CL:

>= 934 - <= 1 296

Sex:

female

Dose descriptor:

LD50

Effect level:

1 045 mg/kg bw

Based on:

test mat.

95% CL:

>= 914 - <= 1 197

Mortality:

see table 1 for details

Clinical signs:

other: not reported

Gross pathology:

not reported

- Table 1: Mortality in Sprague-Dawley rats

Dose in mg/kg bw	fatalities within 7 d post dosing
	females	males
592	0/10	0/10
889	2/10	1/10
1333	9/10	10/10
2000	10/10	10/10
LD50 in mg/kg bw	1045	1089
95 % confidence limits	(914 - 1197)	(934 - 1269)

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Guanidine carbonate was tested for its acute oral toxicity in Sprague-Dawley rats of both sexes after single oral treatment following a procedure generally equivalent to OECD TG 401. Doses used were 592, 889, 1333 and 2000 mg/kg bw. Post exposure observation period was 7 d.
The LD50 values for males and females are 1089 mg/kg bw (95 % CL: 934 – 1296) and 1045 mg/kg bw (95 % CL: 914 – 1197) respectively.

Executive summary:

Guanidine carbonate was tested for its acute oral toxicity in Sprague-Dawley rats of both sexes after single oral treatment following a procedure generally equivalent to OECD TG 401. Doses used were 592, 889, 1333 and 2000 mg/kg bw. Post exposure observation period was 7 d.

LD50 values were determined by the method of Thompson-Weil.

The LD 50 values for males and females are 1089 mg/kg bw (95 % CL: 934 – 1296 mg/kg bw) and 1045 mg/kg bw (95 % CL: 914 – 1197 mg/kg bw), respectively.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 045 mg/kg bw

Quality of whole database:

- meets generally accepted scientific principles
- sufficient information for assessment reported

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

exposure considerations

Justification for data waiving:

other:

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:

from 1998-06-24 to 1999-01-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline-conform study performed under GLP without deviations

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

from 1987

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

described in EC-Guideline 92/69

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: Sprague Dawley, Him:OFA, SPF.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Forschungsinstitut für Versuchstierzucht, A-2325 Himberg, Germany
- Age at study initiation: Approximately 8 weeks (males) and 12 weeks (females)
- Weight at study initiation: 252-282 g (male), 225 - 251 g (female)
- Housing: Single caging in Makrolon cages type III (39 cm x 23 cm x 15 cm). Wire mesh lids. Sanitization of cages once a week. Bedding material: Aspen wood chips, type "4 HV" (Finn Tapvei Oy, SF-73620 Kortteinen), autoclaved. Bedding material was changed weekly.
- Diet (e.g. ad libitum): Altromin 1314 forte, gamma irradiated with 25 kGy 60Co, ad libitum (Producer: Altromin GmbH, D-32791 Lage).
- Water (e.g. ad libitum): Tap water, offered in Makrolon bottles with stainless steel canules, ad libitum.
- Acclimation period: 5 respectively 6 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 (average)
- Humidity (%): 55 (average)
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Remarks:

The patch was soaked with deionised water to get optimal contact with the skin.

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsal thoracal region, 5 x 6 cm, labelled in a relaxed animal (corresponding to an area of about 50 cm2, when the skin is slightly stretched at administration)
- % coverage: about 10 % of the estimated body surface
- Type of wrap if used: dressing (Fixomull Stretch, Fa. Beiersdorf).

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Residual test substance was wiped off using wet cellulose tissue.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- For solids, paste formed: no

Duration of exposure:

24 hours

Doses:

Range-finding study: 400, 894 and 2000 mg/kg bw
Main test: 2000 mg/kg bw

No. of animals per sex per dose:

Range-finding study:
400 mg/kg bw: 1 male, 1 female
894 mg/kg bw: 1 male, 1 female
2000 mg/kg bw: 1 male 1 female

Main test:
2000 mg/kg bw: 4 males, 4 females
Both animals dosed with 2000 mg/kg in the preliminary test were included into the main study, leading to 5 males and 5 females treated with 2000 mg/kg in the main test.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Body weight: before administration, 7 and 14 days after administration
Clinical observations: at least once per day.
Necropsy: 14 days after administration
- Necropsy of survivors performed: yes
- Other examinations performed: mortality, clinical signs, body weight

Preliminary study:

In a range finding study three groups of one male and one female each were dosed with 400, 894 or 2000 mg per kg body weight. All animals survived for 14 days after administration. Therefore it was decided to perform a limit-test with one dose of 2000 mg per kg body weight. Both animals,
dosed with 2000 mg/kg in the preliminary test, were included into the main study.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

All animals survived until the scheduled termination of the study.

Clinical signs:

other: 3/5 males and 3/5 females were affected: The finding, observed earliest immediately after administration and latest 6 h after administration, was: Chromodacryorrhoea on the day of the test substance administration was attributed to the discomfort, caused

Gross pathology:

All animals in the main study were normal at terminal necropsy.

Other findings:

Sex differences: Effects on body weights were more pronounced in the females, who are considered therefore to be slightly more susceptible then the males.

Observations in life

A grade of severity was recorded where applicable (low - medium - high).

Findings	Dose (mg/kg), sex	No. of the affected animals	Observation time (after administration)   First / last	Maximum grade of severity
No findings at any time	2000, m	22, 25	- /-	-
	2000, f	27, 28	- /-	-
Chromodacryorrhoea	2000, m	21	0.5 h / 6 h	low
		23	1 h / 2 h	low
		24	1 h / 6 h	low
	2000, f	26	0.5 h / 2 h	low
		29	2 h / 2 h	low
		30	0.5 h / 1 h	low

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The LD50, dermal of Guanidine carbonate is higher than 2000 mg/kg body weight in rats.

Executive summary:

A valid GLP-study was performed to investigate acute toxic effects of the test substance after a single dermal administration to rats. Methods and investigations were performed in conformance with OECD TG 402, 1987.

Guanidine carbonate was administered once dermally under semiocclusive conditions on an area of approximately 5 x 6 cm on the dorsal thoracal region of 5 male and 5 female Him:OFA rats (Sprague Dawley). The duration of the exposure was 24 hours. The dose was 2000 mg per kg body weight.

Body weight was measured before administration, 7 and 14 days after administration. Clinical observations were made at least once per day. Necropsy was performed 14 days after administration.

All animals survived until the scheduled termination of the study. Body weight and body weight gain were inconspicuous during the whole study in all males. A slight body weight loss in the first week was noted in one female. Another female gained only 1 g body weight in the second week. 3/5 males and 3/5 females were affected by chromodacryorrhoea on the day of application, lasting until maximum 6 h after administration. This effect was attributed to the discomfort caused by the dressing. The same signs were observed in both sexes. All animals were normal at the post mortem examination. Effects on body weights were more pronounced in the females, who are considered therefore to be slightly more susceptible than the males.

Except that the body weight gain of the females was slightly reduced, no toxic effects of the test substance were noted in life at the dose of 2000 mg/kg body weight. No mortality occurred. The LD50 of Guanidine carbonate is higher than 2000 mg/kg body weight in rats of either sex.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Guideline-conform study performed under GLP without deviations

Additional information

Oral toxicity

A LD50 oral (gavage, Sprague-Dawley rats) of 1089 mg/kg bw (95 % CL 934 - 1269) in male and 1045 mg/kg bw (95 % CL 914 - 1197) in female rats is reported for Guanidine carbonate in the key study (Wendtlandt 1978). This result is supported by two supporting studies. Grambow et al. (2005) report a LD50 of 1020 mg/kg bw in the rat after single gavage application. Bergner et al. (1969) observed in a feeding study with Guanidine hydrogen carbonate a LD100 causing death in all rats 2 -3 days, which corresponds to ca. 2455 mg diguanidinium carbonate/kg bw (total dose).

Inhalation toxicity

No studies are available on the acute inhalation toxicity of Guanidine carbonate.

Dermal toxicity

A LD50 dermal (semiocclusive for 24 h, Sprague Dawley rats) of > 2000 mg/kg bw in male and female rats is reported for Guanidine carbonate in the key study (Ott 1999), with a reliability score of 1, conducted according to OECD TG 402. The test item was administered once dermally under semiocclusive conditions on an area of approximately 5 x 6 cm on the dorsal thoracal region of 5 male and 5 female Him:OFA rats (Sprague Dawley). The duration of the exposure was 24 hours. The dose was 2000 mg per kg body weight. No fatalities occurred, body weight development was normal in males while 2 females had intermittently reduced body weight increase. Chromodacryorrhoea on the day of application in 6 of 10 animals was attributed to the discomfort by the dressing. No further clinical signs were recorded throughout the complete observation period. All animals were normal at the post mortem examination. Accordingly the substance does not need to be classified for acute dermal toxicity.

Justification for selection of acute toxicity – oral endpoint
The study of Wendtlandt (1978) was performed similar to OECD TG 401. Sufficient details are given in the study report to allow an assessment of the study. Grambow et al. (2005) reports an similar acute oral LD50 value without further explanation. Bergner et al. (1969) tested only one concentration which does not allow for the determination of a LD50 value. The study of Wendtlandt was therefore selected as the key study. The other two studies are reported as supporting information.

Justification for selection of acute toxicity – dermal endpoint
The GLP-conform study of Ott (1999) was performed in accordance with OECD TG 402. It was therefore selected as the key study.

Justification for classification or non-classification

Oral toxicity

A LD50 rat of 1045 mg/kg bw was observed in a study performed similarly to OECD TG 401. The substance needs to be classified as acute oral Category 4 (Warning, H302: Harmful if swallowed) according to CLP (Regulation (EC) No 1272/2008).

Inhalation toxicity

No studies are available on the acute inhalation toxicity of Guanidine carbonate.

Dermal toxicity

Based on the above stated assessment of the acute dermal toxicity of Guanidine carbonate (absence of toxicity up to 2000 mg/kg bw) the substance does not need to be classified according to CLP (Regulation (EC) No 1272/2008).