Registration Dossier

Diss Factsheets

Administrative data

Description of key information

- skin irritation: not irritating, 2 studies available, OECD TG 404 
- eye irritation: Category 1 (irreversible effects on the eye), 2 studies available, OECD TG 405
- respiratory irritation: no studies available

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 1998-07-21 to 1999-01-27
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline-conform study performed under GLP
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: J. Schitkovits, A-2620 Natschbach.
- Age at study initiation: not reported
- Weight at study initiation: 2.5 - 2.9 kg
- Housing: Individual caging in metal wire cages, Ehret GmbH, D-79312 Emmendingen, type KK 016R, 79 cm x 59 cm x 35 cm.
- Diet (e.g. ad libitum): Altromin 2023 diet for rabbits, ad libitum. Random samples of the feed are analysed for contaminants by Altromin, D-32791 Lage.
- Water (e.g. ad libitum): Tap water ad libitum
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 (average)
- Humidity (%): 60 (average)
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Preparation of test site:
other: The hair was clipped on the dorsal area of the trunk one day before the application.
Vehicle:
water
Remarks:
Samples of the test item were moistened with distilled water.
Controls:
other: Surrounding untreated skin was taken as a control area.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g

VEHICLE
- Amount(s) applied (volume or weight with unit): Samples of the test item were moistened with distilled water.
Duration of treatment / exposure:
4 hours
Observation period:
3 days. Observations were made 1, 24, 48 and 72 hours after patch removal.
Number of animals:
3 females
Details on study design:
TEST SITE
- Area of exposure: dorsal area of the trunk
- Type of wrap if used: The test item samples were placed on cellulose patches (pure Zellin swap, Hartmann, Wiener Neudorf) and fixed marginally (Blenderm R surgical tape, hypoallergenic 3M). The application sites were covered semi-occlusive by dressing (self adhesive non woven fabric, hypoallergenic, Beiserdorf)

REMOVAL OF TEST SUBSTANCE
- Washing: Residual test item was removed with wet Kleenex tissues, if necessary
- Time after start of exposure: After 4h exposure (Thus the duration of the exposure was 4 hours.)

SCORING SYSTEM:
Erythema / Eschar formation
0: No erythema.
1: Very slight erythema (barely perceptible).
2: Well-defined erythema.
3: Moderate to severe erythema.
4: Severe erythema (beet redness) or eschar formation (injuries in depth).
Oedema formation
0: No oedema.
1: Very slight oedema (barely perceptible).
2: Slight oedema (edges of area well defined by definite raising).
3: Moderate oedema (raised approximately 1 mm).
4: Severe oedema (raised more than 1 mm and extending beyond area of exposure).
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: mean of scores at 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
edema score
Basis:
mean
Time point:
other: mean of scores at 24, 48 and 72 hours
Score:
0
Max. score:
4

No general toxic effects were noted and all areas treated with the test substance and all control areas were normal before the

application and at each observation time.

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test substance Guanidine carbonate did not cause any adverse skin reactions in this study.
Executive summary:

The aim of this study was to investigate possible irritation or corrosion by the test substance following a single application to the intact skin of New Zealand White rabbits. Methods and investigations were performed in conformance with the OECD TG 404.

Approximately 0.5 g Guanidine carbonate, moistened with distilled water, was applied via a patch to an area of about 2.5 cm x 2.5 cm of the intact skin of each of 3 rabbits and covered by semi-occlusive dressing. The duration of the exposure was 4 hours.

The body weight was measured at the start and at the termination of the test. The animals were observed for general signs of toxicity once daily and the skin was examined 1, 24,48 and 72 h after patch removal.

No general toxic effects were observed. All exposed skin sites were normal at each examination term. In all animals mean scores for erythema as well as for oedema formation, calculated from the examinations performed 24,48 and 72 hours after patch removal, were "0", i.e. normal. The test substance did not cause any adverse skin reactions in this study.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 1998-07-21 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 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: J. Schitkovits, A-2620 Natschbach
- Age at study initiation: not reported
- Weight at study initiation: 2.9 - 3.2 kg
- Housing: Individual caging in metal wire cages, Ehret GmbH, D-79312 Emmendingen, type KK 016R, 79 cm x 59 cm x 35 cm.
- Diet (e.g. ad libitum): Altromin 2023 diet for rabbits, ad libitum.
- Water (e.g. ad libitum): Tap water from an automatic watering system, ad libitum.
- Acclimation period: 4 days (animal No. 51), 11 days (animals No. 52 and No. 53).

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 (average)
- Humidity (%): 60 (average)
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
other: The left eyes remained untreated and served as negative controls.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): The equivalent of 0.1 mL of the test substance was administered. The weights of the administered doses were: 85 mg, 98 mg and 95 mg.
Duration of treatment / exposure:
single dose (no washing)
Observation period (in vivo):
Both eyes of the animals were examined within 24 hours before instillation and approximately 1, 24,48 and 72 hours p.a.
Additional examinations were performed 6, 8, 10, 13, 15, 17 and 21 days after the instillation, ending individually for each animal.
Number of animals or in vitro replicates:
3 females
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): not done

SCORING SYSTEM:
CORNEA
Opacity : degree of density (area most dense area is taken for reading).
0 No ulceration or opacity.
1 Scattered or diffuse areas of opacity (except for slight dulling of normal lustre), details of iris clearly visible.
2 Easily discernible translucent area, details of iris slightly obscured.
3 Nacreous area, no details of iris visible, size of pupil barely discernible.
4 Opaque cornea, iris not discernible through the opacity.

IRIS
0 Normal.
1 Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or injection; any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive).
2 No reaction to light, haemorrhage, gross destruction (any of all these or all together).

CONJUNCTIVAE
Redness: (refers to the most severe effect of palpebral and bulbar conjunctivae in comparison to the control eye).
0 Blood vessels normal.
1 Some blood vessels definitely hyperaemic (injected).
2 Diffuse crimson colour, individual vessels not easily discernible.
3 Diffuse beefy red.

Chemosis: lids and/or nictating membranes.
0 No swelling.
1 Any swelling above normal (includes nictating membranes).
2 Obvious swelling with partial eversion of lids.
3 Swelling with lids about half closed.
4 Swelling with lids more than half closed.


TOOL USED TO ASSESS SCORE: otoscope lamp
Irritation parameter:
cornea opacity score
Basis:
animal #1
Remarks:
(animal No. 51)
Time point:
other: mean of scores at 24, 48 and 72 h
Score:
1.7
Max. score:
4
Reversibility:
not fully reversible within: 15 d
Remarks on result:
other: cornea was not examinable on day 17 and 21 due to severe chemosis
Irritation parameter:
cornea opacity score
Basis:
animal #2
Remarks:
(animal No. 52)
Time point:
other: mean of scores at 24, 48 and 72 h
Score:
2.7
Max. score:
4
Reversibility:
not reversible
Remarks:
until death on day 14
Remarks on result:
other: euthanized on day 14 for humane reasons
Irritation parameter:
cornea opacity score
Basis:
animal #3
Remarks:
(animal No. 53)
Time point:
other: mean of scores at 24, 48 and 72 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Remarks:
until death on day 14
Remarks on result:
other: found dead on day 14
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
(animal No. 51)
Time point:
other: mean of scores at 24, 48 and 72 hours
Score:
0.3
Max. score:
2
Reversibility:
not fully reversible within: 72 h
Remarks on result:
other: iris was not discernible from day 6 on due to the opacity of the cornea.
Irritation parameter:
iris score
Basis:
animal #2
Remarks:
(animal No. 52)
Time point:
other: mean of 24 and 48 h
Score:
1
Max. score:
2
Reversibility:
not fully reversible within: 48 h
Remarks on result:
other: iris was not discernible from 72 h on due to the opacity of the cornea.
Irritation parameter:
iris score
Basis:
animal #3
Remarks:
(animal No. 53)
Time point:
other: mean of scores at 24, 48 and 72 h
Score:
0
Max. score:
2
Reversibility:
fully reversible within: 10 d
Remarks on result:
other: iris was not discernible on day 13 due to the opacity of the cornea.
Irritation parameter:
conjunctivae score
Basis:
animal #1
Remarks:
(animal No. 51)
Time point:
other: mean scores from 24, 48 and 72 h
Score:
3
Max. score:
3
Reversibility:
not reversible
Remarks:
until end of test (day 21)
Remarks on result:
other: mucopurulent ocular discharge from 24 h p.a. until terminal examination.
Irritation parameter:
conjunctivae score
Basis:
animal #2
Remarks:
(animal No. 52)
Time point:
other: mean scores from 24, 48 and 72 h
Score:
3
Max. score:
3
Reversibility:
not reversible
Remarks:
until death on day 14
Remarks on result:
other: mucopurulent ocular discharge from 24 h p.a. until terminal examination and swelling dorsal of the instilled eye from 48 h until 14 d p.a.
Irritation parameter:
conjunctivae score
Basis:
animal #3
Remarks:
(animal No. 53)
Time point:
other: mean scores of 24, 48 and 72 hours
Score:
2
Max. score:
2
Reversibility:
not reversible
Remarks:
until death on day 14
Remarks on result:
other: mucopurulent ocular discharge from 24 h p.a. until terminal examination.
Irritation parameter:
chemosis score
Basis:
animal #1
Remarks:
(animal No. 51)
Time point:
other: mean of scores at 24, 48 and 72 h
Score:
4
Max. score:
4
Reversibility:
not reversible
Remarks:
until end of test (day 21)
Irritation parameter:
chemosis score
Basis:
animal #2
Remarks:
(animal No. 52)
Time point:
other: mean of scores at 24, 48 and 72 h
Score:
4
Max. score:
4
Reversibility:
not reversible
Remarks:
until death on day 14
Irritation parameter:
chemosis score
Basis:
animal #3
Remarks:
(animal No. 53)
Time point:
other: mean of scores at 24, 48 and 72 h
Score:
3
Max. score:
4
Reversibility:
not reversible
Remarks:
until death on day 14
Irritant / corrosive response data:
Cornea: In all animals corneal abnormalities were noted. Diffuse areas of opacity (score " 1 ") raised to easily discernible translucent area, details of iris slightly obscured (score "2") and to nacreous area (score "3") and opaque cornea (score "4") were the maximum grade of severity reached. The whole corneae were affected most of the time.

Iris: In animal No. 51 score "1" was noted 24 h. p.a. followed by 2 normal findings. Animal No. 52 reached score "2" 48 h p.a. and animal No. 53 score "1" was seen from 6 to 10 d p.a. Later on, no examinations were possible due to the opaque cornea in all animals.

Conjunctivae, redness: In animals No. 51 and No. 52 a diffuse beefy red (score "3") was noted from 1 h p.a. onwards. In animal No. 53 a diffuse crimson colour (score "2") from 1 h to 10 d raised also to a diffuse beefy red 13 d p.a.

Conjunctivae, chemosis: In animals No. 51 and No. 52 swelling with lids more than half closed (score "4") was present at each reading time. In animal No. 53 changes from an obvious swelling to closed eyes (scores "2", "3" and "4") were noted from 1 h onwards. A mucopurulent discharge from the instilled eye of all animals was noted from 24 p.a. until the end of the examinations.
Other effects:
Deaths: Animal No. 53 was found dead 14 days p.a. A moderate fibinous to fibrous pericarditis and foci of enteritis in the small intestine were noted post mortem. Severe ocular lesions may have worsened a pre-existing poor health condition and therefore have contributed to the death. No direct influence of the test substance is considered to have been the cause of death.
In animal No. 52 a swelling dorsal of the instilled eye was noted from 48 h until 13 d p.a. The nature of the swelling was not identified. In the same animal a total destruction of the cornea was noted 14 d p.a. after rinsing of the eye. This animal was subsequently euthanized for humane reasons.

Scores of the test eyes. Examination of cornea and iris.

Time after

instillation

(p.a.)

Cornea

Iris

Animal No.

Animal No.

51

52

53

51

52

53

l h

0

1a

0

0

0

0

24 h

1a

1a

1a

1

0

0

48 h

2a

3a

1b

0

2

0

72 h

2a

4a

1b

0

c

0

Mean (24 – 72 h)

1.7

2.7

1.0

0.3

c

0.0

6 d

4b

4a

2a

c

c

1

8 d

4b

4a

3a

c

c

1

10 d

4b

4a

3a

c

c

1

13 d

4b

4a

4b

c

c

c

14 d

-

4a

-

-

c

-

15 d

4b

-

-

c

-

-

17 d

h

-

-

c

-

-

21 d

h

-

-

c

-

-

a) 1/2 of the cornea affected.

b) whole cornea affected.

c) iris was not discernible due to the opacity of the cornea.

h) cornea was not examinable due to severe chemosis

-) not examined.

Scores of the test eyes. Examination of conjunctivae.

Time after

instillation

(p.a.)

Conjunctivae (Redness)

Conjunctivae (Chemosis)

Animal No.

Animal No.

51

52

53

51

52

53

l h

3

3de

2

4

4

2

24 h

3e

3ef

2e

4

4

3

48 h

3e

3ef

2e

4

4

3

72 h

3e

3ef

2e

4

4

3

Mean(24 – 72 h)

3.0

3.0

2.0

4.0

4.0

3.0

6 d

3e

3ef

2e

4

4

3

8 d

3e

3ef

2e

4

4

4

10 d

3e

3ef

2e

4

4

4

13 d

3e

3ef

3e

4

4

4

14 d

-

3ef

-

-

4

-

15 d

3e

-

-

4

-

-

17 d

3e

-

-

4

-

-

21 d

3e

-

-

4

-

-

-) not examined.

d) turbid ocular discharge 24 h p.a.

e) mucopurulent ocular discharge from 24 h p.a. until terminal examination. swelling dorsal of the instilled eye from 48 h until 14 d p.a.

f) swelling dorsal of the instilled eye from 48 h until 14 d p.a.

Interpretation of results:
Category 1 (irreversible effects on the eye)
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test substance Guanidine carbonate is proposed to be classified with R41 Risk of serious damage to eyes) according to Council Directive 2001/59/EC (28th ATP of Directive 67/548/EEC) and Category 1 (Danger; H318: Causes serious eye damage) according to CLP (Regulation (EC) No 1272/2008).
Executive summary:

The aim of this study was to investigate possible irritating or corrosive effects of the undiluted test substance following a single administration into a conjunctival sac of the eye of New Zealand White rabbits. Methods and investigations were performed in conformance with the OECD TG 405.

The approximate equivalent of 0.1 mL Guanidine carbonate was instilled into the conjunctival sac of one eye of each of 3 rabbits. The body weight was measured at the start and at the termination of the test. The animals were examined for general signs of toxicity once daily. Eye examinations were performed 1, 24, 48,7 2 h and 6, 8, 10, 13, 15, 17, 21 d after the administration (p.a.) of the test substance (ending individually).

One animal died 14 days p.a. A moderate fibinous to fibrous pericarditis and foci of enteritis in the small intestine were noted post mortem. A direct influence of the test substance on the death is not likely. In 2/3 animals no general toxic effects were observed.

In all animals irreversible corneal alterations were observed with the maximum score of 4 with the whole cornea affected. The irises were affected in varying severities at the beginning. Later on, an examination was made impossible by opacity of the cornea in all animals. Pronounced redness was observed in all animals with the maximum score of 3. An obvious to a severe swelling was seen in all animals, reaching the maximum score of 4. A mucopurulent ocular discharge was present in all animals from 24 h p.a. until the end of the examinations. In 1/3 animals additionally a swelling dorsal of the test eye was noted from 48 h until 13 d p.a. In the same animal a total destruction of the cornea was noted 14 d p.a. after rinsing of the eye. This animal was subsequently euthanized for humane reasons.

The following mean scores were calculated from the individual examinations performed 24 h, 48 h and 72 h p.a.:

 

Animal No.

 

51

52

53

Cornea:

1.7

2.7

1.0

Iris:

0.3

*

0.0

Conjunctivae, redness:

3.0

3.0

2.2

Conjunctivae, chemosis:

4.0

4.0

3.3

*Iris was not discernible at all examination times

Ocular lesions were not reversible in all animals.

The classification of R41 (Risk of serious damage to eyes) according to Council Directive 2001/59/EC (28th ATP of Directive 67/548/EEC) and Category 1 (Danger; H318: Causes serious eye damage) according to CLP (Regulation (EC) No 1272/2008) is proposed.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation/corrosion

One fully reliable in vivo study is available (Ott 1999) conducted according to GLP and OECD TG 404. In addition one study that predates the implementation of OECD guidelines and GLP is available (Wendtlandt 1978).

The experiment reported by Ott (1999) was conducted using semiocclusive exposure of 0.5 g of water-moistened Guanidine carbonate to 6.25 cm² of shaved skin of New Zealand White rabbits for 4 h. No irritating effects or signs of general toxicity were seen at any observation time point.

Therefore, the test item considered not to be irritating or corrosive.

The study of Wendtlandt (1978) was conducted by applying 0.5 mL of a 25 % (w/w) solution of Guanidine carbonate in water to the skin of three young adult New Zealand White rabbits. The test site was not covered and the test substance was not washed off. Results are not delivered in numerical form but only as qualitative verbal description of effects. The reported effects suggest that the substance is not irritating to the skin.

It can be summarized that both in vivo tests agree in that Guanidine carbonate is not irritating to the skin.

Eye irritation/corrosion

One fully reliable in vivo study is available (Ott 1999) conducted according to OECD TG 405 and GLP. In addition, one study that predates the implementation of OECD guidelines and GLP is available (Wendtlandt 1978).

Te study of Ott (1999) was conducted by applying 0.1 mg of the test item to the conjunctival sac of three New Zealand White rabbits. One animal died 14 days p.a. A moderate fibinous to fibrous pericarditis and foci of enteritis in the small intestine were noted post mortem. A direct influence of the test substance on the death is not likely. In 2/3 animals no general toxic effects were observed. Drastic effects on the eye were seen resulting in irreversible corneal alterations (score of 4), effects on the iris (scoring impeded by opacity of the cornea), pronounced redness (score of 3), obvious to severe swelling (score of 4) and mucopurulent ocular discharge in all animals. In 1/3 animals additionally a swelling dorsal of the test eye was noted from 48 h until 13 d post administration. In the same animal a total destruction of the cornea was noted 14 d p.a. after rinsing of the eye. This animal was subsequently euthanized for ethical reasons.

The test of Wendtlandt (1978) was conducted by applying 0.1 mL of a 25 % (w/w) solution of Guanidine carbonate in water to the conjunctival sac of three young adult New Zealand White rabbits. Results are not delivered in numerical form but only as qualitative verbal description of effects. After 24 post dosing an intensive reddening of the conjunctivae was found. At 72 h post dosing, additionally chemosis in all animals and discharge in 4 of 6 animals was reported. All effects were reversed within 1 week in all animals except 1 which still showed slight conjunctivae reddening 1 week post dosing. As this test was conducted only with a 25 % (w/w) solution of the test item in water it is not relevant for the classification.

It can be summarized based on the only fully reliable study (Ott 1999) that Guanidine carbonate has irreversible effects when applied to rabbit eyes.

Respiratory irritation

No data on respiratory irritation is available.

 


Justification for selection of skin irritation / corrosion endpoint:
The study of Ott (1999) was performed under GLP in accordance with OECD TG 404. The study of Wendtlandt (1978) differs in some aspects from the guideline. Ott (1999) was therefore selected as the key study and Wendtlandt (1978) is reported as supporting information.

Justification for selection of eye irritation endpoint:
The study of Ott (1999) was performed under GLP in accordance with OECD TG 404. The study of Wendtlandt (1978) differs in some aspects from the guideline. Ott (1999) was therefore selected as the key study and Wendtlandt (1978) is reported as supporting information.

Effects on eye irritation: corrosive

Justification for classification or non-classification

Skin irritation/corrosion

Guanidine carbonate was found to be not irritating to the skin in a valid GLP study performed according to OECD TG 404. Therefore, the substance does not need to be classified according to CLP (Regulation (EC) No 1272/2008).

 

Eye irritation/corrosion

Irreversible effects to the rabbit eye were observed in a valid GLP study performed according to OECD TG 405. Guanidine carbonate has therefore to be classified as Category 1 (Danger; H318: Causes serious eye damage) according to CLP (Regulation (EC) No 1272/2008).

 

Respiratory irritation

As no data on respiratory irritation is available for Guanidine carbonate a classification is not possible according to CLP (Regulation (EC) No 1272/2008).

Categories Display