Registration Dossier

Skin irritation (rat): not irritating [draft report, Kurth 1996]
Eye irritation (rabbit): severe irritant [draft report, Treher 1996]

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from January to February 1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

A combined study on acute dermal toxicity and on local tolerance was used to assess skin irritation/corrosion. The study design and reporting deviates from the current OECD TG 404, however, the study is considered sufficient for assessment of this endpoint.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

17 July 1992

Deviations:

yes

Remarks:

- exposure time 24 hrs instead of 4 hrs, 6 animals instead of 3, few details in test description and reporting

GLP compliance:

no

Species:

rat

Strain:

other: Han:WIST (SPF)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Schering AG, Germany
- Mean weight at study initiation: 104-106 g (males) or 98-105 g (females)
- Housing: 1 animal per cage
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 - 23
- rel. Humidity (%): 50 - 58
- Photoperiod (hrs dark / hrs light): 12 / 12

Type of coverage:

occlusive

Preparation of test site:

not specified

Vehicle:

other: 900 mg NaCl ad 100 mL bidist. water

Controls:

other: the untreated skin served as control

Amount / concentration applied:

208-212 mg/male animal or 196-210 mg/female animal

Duration of treatment / exposure:

24 hours

Observation period:

14 days

Number of animals:

3 males and 3 females

Details on study design:

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) On day 1 at application and after 24 h, 48 h and 72 h.

SCORING SYSTEM: According to Draize et al.

Irritation parameter:

erythema score

Basis:

animal: #1M; #3M, #4F, #5F, #6F

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

erythema score

Basis:

animal: #2M

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 11 days

Irritation parameter:

edema score

Basis:

animal: #1- #6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

2 out of 4 animals showed local irritations as folIows: animal no. 4F showed a very slight reddening (immediately after removal of the bandage and the substance), patchy on day 2, and animal no. 2M showed reddening, patehy (day 2 and 3), hardening of the skin, patehy (day 4) and skin neerosis (initial stage), spotty (day 5 to 10). All animals were without local findings trom day 11 onwards.

Interpretation of results:

GHS criteria not met

Conclusions:

A combined study on acute dermal toxicity and on local tolerance was conducted . The study design and reporting deviates from the current OECD TG 404, however, the study was considered sufficient for assessment of skin irritation/corrosion.A single dermal administration of the test substance for 24 hours to male and female rats at a dose of 2000 mg/kg was tolerated without any local irritations in 4/6 animals. Following grading at 24, 48 and 72 hours after patch removal 1/6 animals shows a mean value of 2 for erythema/eschar. All animals were without local findings trom day 11 onwards.

Executive summary:

In a primary dermal irritation study similar to OECD TG 404, young adult Wistar rats, female and male (3/sex) were dermally exposed to 208-212 mg/male animal or 196-210 mg/female animal of Triamine Dihydrochloride in physiological saline for 24 hours. Animals then were observed for 14 days. Irritation was scored by the method according to Draize et al..

Only in one male animal local irritations were observed at 24, 48, and 72 h. These irritations were reversible within 11 days. In this study, the test item is not a dermal irritant.

Endpoint conclusion:

no adverse effect observed (not irritating)

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from March to April 96

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

only one animal tested with severe irritating findings

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

24 February 1987

Deviations:

yes

Remarks:

despite of reversibility of effects no further animals tested to confirm irritation

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Schriever
- body weight: 2.9 kg
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 18 days
- Identification: tattoo numbers in the ear

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- rel. Humidity (%): 52 - 58
- Photoperiod (hrs dark / hrs light): 12 / 12

Vehicle:

unchanged (no vehicle)

Controls:

other: the left eye, which remained untreated, served as control.

Amount / concentration applied:

0.1 mL (corresponded to 51 mg substance)

Duration of treatment / exposure:

test substance remained in the eye (eye was not rinsed)

Observation period (in vivo):

13 days

Number of animals or in vitro replicates:

1 male- since severe intolerance reactions occurred in this animal, the other animals originally foreseen for this study were not exposed to the substance for humane reasons.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no washing

SCORING SYSTEM: According to Annex to EEC Directive 92/69/EWG, dated 31. Jul. 92, for the 17th adaptation of the EEC Directive 67/548/EWG, B. 5 Acute Toxicity (Eye irrtation) Off J EEC, L383, Luxembourg, 29 Dec. 92.

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Reversibility:

other: not applicable

Remarks on result:

not measured/tested

Remarks:

Chemosis refers to reddening of the eye lids and nictating membranes, only reddening of the conjunctivae was measured.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Remarks:

reddening

Basis:

animal #1

Time point:

24/48/72 h

Score:

3

Max. score:

3

Irritation parameter:

conjunctivae score

Remarks:

swelling

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Summary of local findings after single application of the test substance in the rabbit eye

Findings	Right eye 0.1 mL test item/eye; animal: 696M
Findings	A	X-Y
Outer eyelids
secretion with wetting of surrounding fur
+	1	1-1
++	2	1-2
+++	1	1-1
swelling of the edge of the eyelids (clue to severely edematous
(+)	1	7-7
swelling of conjunctiva palpebrae)
+	6	2-8
+++	3	1-1
reddening of the skin of the eyelids
(+)	1	7-7
+	7	1-9
++	3	1-3
incomplete eyelid closure
+	4	1-2
dacryorrhea, red-coloured	1	1-1
incrustation of secretoryproduct at the fur around the eye	5	2-6
red-coloured incrustation of secretory product at the fur around the eye	3	3-5
Conjunctiva palpebrae
reddening
(x)	2	11-12
+	2	9-10
++	5	4-8
+++	5	1-3
swelling, partly edematous
(+)	2	9-10
+	6	3-8
++	1	2-2
+++	3	1-1
injection of blood vessels
+	2	11-12
++	4	7-10
+++	8	1-6
hemorrhage
+	3	4-6
Membrana nicitans
reddening
+	2	7-8
++	8	1-6
swelling (partly edematous)
+	3	4-6
++	2	2-3
+++	3	1-1
injection of blood vessels
+	2	11-12
++	4	7-10
+++	7	1-6
hemorrhage
++	7	2-8
not visible due to bleeding of the mucosa	1	2-2
mucosa partly whitish	6	7-12
Conjunctiva sclerae
reddening
+	1	2-2
injection of blood vessels
+	4	3-8
++	1	2-2
could not be assessed due to the severe swelling of the conjunctiva palpebrae	3	1-1
injection of blood vessels
+	4	3-8
++	1	2-2
Cornea
opacity
+	3	1-1
bluntness
+	3	1-1
Iris
Iris only partly visible	3	1-1

A = summation of positive findings

x-y = first-last day of occurrence of a finding

(+) = very slight, + = slight, ++ = moderate, +++ = severe

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

In a local tolerance test on the rabbit eye according to OECD TG 405 in the initial test using one animal a single application of the test substance into the conjunctival sac provoked severe irritation which gradually subsided during the course of the 13 days observation period. The other animals originally foreseen for this study were not exposed to the substance for animal welfare reasons.
Despite of the reversibility of the effects the investigation of only one animal in this study is limitating the assessment. Since the structural analogue Z-Triamin-Dihydrochloride (CAS 221640-14-8) was found to be corrosive to the eye in a BCOP test (OECD TG 437) also for the Triamine-Dihydrochloride (CAS 221640-21-7) severe eye damage (corrosion) cannot be excluded.

Executive summary:

In a primary eye irritation study similar to OECD TG 405, 0.1 mL corresponding to 51 mg of Triamine Dihydrochloride was instilled into the conjunctival sac of the right eye of a young adult male New Zealand White rabbit and the animal was observed for 14 days. The eyes were not washed. Irritation was scored by the method described in the guideline.

A single application of 0.1 mL of the test substance into the conjunctival sac of the rabbit eye led to slight to severe secretion, reddening and swelling of the outer eyelids, moderate but predominantly severe reddening, swelling and vascular injection of the conjunctivae, incomplete eyelid closure accompanied by ectropium as well as slight opacity of the cornea on the application day. Moderate to severe reddening and vascular injection and slight to moderate swelling of the conjunctivae accompanied by hemorrhage into the mucous membrane of the conjunctivae were observed up to day 6. Afterwards the findings gradually subsided and had disappeared by day 13. With regard to the observed effects in this study, the test item is considered to be a severe eye irritant.

Endpoint conclusion:

adverse effect observed (irritating)

Endpoint conclusion:

no study available

Skin irritation:

A combined study on acute dermal toxicity and on local tolerance was conducted . The study design and reporting deviates from the current OECD TG 404, however, the study was considered sufficient for assessment of skin irritation/corrosion.A single dermal administration of the test substance for 24 hours to male and female rats at a dose of 2000 mg/kg was tolerated without any local irritations in 4/6 animals. Following grading at 24, 48 and 72 hours after patch removal 1/6 animals shows a mean value of 2 for erythema/eschar. All animals were without local findings trom day 11 onwards.

Eye irritation:

In a primary eye irritation study similar to OECD TG 405, 0.1 mL corresponding to 51 mg of Triamine Dihydrochloride was instilled into the conjunctival sac of the right eye of a young adult male New Zealand White rabbit and the animal was observed for 14 days. The eyes were not washed. Irritation was scored by the method described in the guideline.

A single application of 0.1 mL of the test substance into the conjunctival sac of the rabbit eye led to slight to severe secretion, reddening and swelling of the outer eyelids, moderate but predominantly severe reddening, swelling and vascular injection of the conjunctivae, incomplete eyelid closure accompanied by ectropium as well as slight opacity of the cornea on the application day. Moderate to severe reddening and vascular injection and slight to moderate swelling of the conjunctivae accompanied by hemorrhage into the mucous membrane of the conjunctivae were observed up to day 6. Afterwards the findings gradually subsided and had disappeared by day 13. With regard to the observed effects in this study, the test item is considered to be a severe eye irritant.

Despite of the reversibility of the effects the investigation of only one animal in this study is limitating the assessment. Since the structural analogue Z-Triamin-Dihydrochloride (CAS 221640-14-8) was found to be corrosive to the eye in a BCOP test (OECD TG 437) also for the Triamine-Dihydrochloride (CAS 221640-21-7) severe eye damage (corrosion) cannot be excluded.

Effects on eye irritation: corrosive

Skin irritation:

1/6 animals with a mean value of 2 for erythema/eschar (rat, male/female, 24 hours)

Based on the available study result (non-irritant) a classification according to regulation (EC) No 1272/2008, Annex I, is not warranted.

Eye irritation:

severe irritant response in the initial test using one animal and waiving of confirmatory test for reasons of animal welfare

Therefore, severe eye damage (corrosion) cannot be excluded for the test substance and according to regulation (EC) No 1272/2008, Annex I, the test substance shall be allocated to Category 1.