Registration Dossier

Oral, Braun (1978):

Under the conditions of the study the acute oral LD50 of the test material was determined to be greater than 20 g/kg, the highest dose tested.

Dermal, Calandra (1976): Under the conditions of the study the acute dermal LD50 was greater than 2000 mg/kg.

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Not specified

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

no

Remarks:

Study pre-dates GLP

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 200 to 295 g
- Fasting period before study: nineteen hours prior to test material administration
- Housing: collectively housed
- Diet: ad libitum
- Water: ad libitum

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 % (w/v) solution in tap water

Doses:

10.0 and 20.0 g/kg

No. of animals per sex per dose:

5 animals per sex per dose

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Observations for mortality and overt signs of effect were made at 0-2 hours, and at 4-6 hours following dosing and daily thereafter for fourteen days.
- Body weights were recorded initially and terminally.
- A gross necropsy was performed on spontaneous deaths.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 20 000 mg/kg bw

Based on:

test mat.

Mortality:

One animal at 20.0 g/kg died during this study.

Clinical signs:

other: Common in-life signs of effect noted during the fourteen day study included: ataxia, coarse and fine tremors, red nasal and red oral discharges, urinary staining of the abdomen, piloerection, motor activity decrease, animal thinness, abdominal griping, ir

Gross pathology:

Necropsy observations for the animal that died during the study were: clear oral discharge; red nasal discharge; urinary staining of the abdomen; stomach and intestines: distended with gas; liver: mottled; thoracic cavity contains red congealed substance.

Table 1: Summary of In-life Observations

Dose level (g/kg)	Observation	Incidence (number observed)
		Hours		Days
		0-2	4-6	1	2	3	4	7	14
10	Surviving animals	10	10	10	10	10	10	10	10
	Red nasal discharge	0	2	0	0	0	0	0	0
	Chromodacryorrhea	0	0	0	0	0	0	0	1
	Piloerection	6	3	4	2	0	0	2	0
	Motor activity decrease	9	8	8	9	0	0	1	0
	Animal thinning	0	0	3	3	0	0	0	0
	NOA	0	2	1	1	10	10	7	9
20	Surviving animals	10	10	10	10	10	10	9	9
	Ataxia	0	0	1	0	0	1	0	0
	Fine tremors	0	2	0	0	0	0	0	0
	Coarse tremors	0	0	3	0	0	1	0	0
	Red nasal discharge	0	4	1	1	2	0	1	0
	Clear nasal discharge	0	0	0	0	0	1	0	0
	Laboured breathing	0	1	0	0	0	0	0	0
	Respiratory rate decrease	0	0	1	0	0	0	0	0
	Red oral discharge	0	1	0	1	2	0	0	0
	Urinary staining	0	3	0	0	0	1	0	0
	Faecal staining	0	2	2	0	0	0	0	0
	Red staining on abdomen	0	0	1	0	0	0	0	0
	Soft stool	1	1	0	0	0	0	0	0
	White stool	0	0	6	1	0	0	0	0
	Pale stool	0	0	0	0	2	1	0	0
	Piloerection	3	5	5	0	2	3	1	1
	Motor activity decrease	10	10	8	3	3	2	3	2
	Motor activity increase	0	0	0	0	0	0	1	1
	Animal thinning	0	0	0	0	0	0	1	0
	Irritability	0	0	1	0	2	0	1	0
	Abdominal griping	0	0	0	0	2	2	0	0
	Ears reddened	0	0	0	0	0	1	0	0
	NOA	0	0	1	7	7	7	4	5

Interpretation of results:

other: Not classified in accordance with EU Criteria

Conclusions:

Under the conditions of this study the acute oral LD50 of the test material was determined to be greater than 20 g/kg, the highest dose tested.

Executive summary:

The acute oral toxicity of the test material was investigated in a study similar to OECD 401.

During the study, ten animals (5/sex) were dosed at 10.0 and 20.0 g/kg. A rangefinding study was performed to arrive at the appropriate dose levels. The test material was administered by oral intubation as a 30 % w/v solution in tap water. Observations for mortality and overt signs of effect were made at 0-2 hours, and at 4-6 hours following dosing and daily thereafter for fourteen days. Body weights were recorded initially and terminally and a gross necropsy was performed on spontaneous deaths.

Common in-life signs of effect noted during the fourteen day study included: ataxia, coarse and fine tremors, red nasal and red oral discharges, urinary staining of the abdomen, piloerection, motor activity decrease, animal thinness, abdominal griping, irritability, and white stool. Signs of effect persisted throughout the study. A loss of weight or a failure to exhibit normal weight gain was noted in 4 of 10 surviving animals at 10.0 g/kg and 1 of 9 surviving animals at 20.0 g/kg.

One animal at 20.0 g/kg died during this study. Necropsy observations for this animal were: clear oral discharge; red nasal discharge; urinary staining of the abdomen; stomach and intestines: distended with gas; liver: mottled; thoracic cavity contains red congealed substance.

Under the conditions of this study the acute oral LD50 of the test material was determined to be greater than 20 g/kg, the highest dose tested.

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

20 000 mg/kg bw

Reference

Endpoint:: acute toxicity: inhalation
Data waiving:: study scientifically not necessary / other information available
Justification for data waiving:: the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size
Justification for type of information:: JUSTIFICATION FOR DATA WAIVING
In accordance with REACH, Annex VIII, column 2 of information requirement 8.5, in addition to the oral route, for substances other than gases, the information required under 8.5.2 to 8.5.3 shall be provided for at least one other route. The choice for the second route will depend on the nature of the substance and the likely route of human exposure.

Information is provided for the dermal route which is deemed to be most appropriate as inhalation of the substance is unlikely. Testing by the inhalation route is therefore omitted.

Endpoint conclusion:

no study available

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Not specified

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

Young adult albino rabbits of the New Zealand strain were used as test animals. Twenty-four hours prior to the dermal applications, the backs of the rabbits were shaved free of hair with electric clippers. A range of several dose levels was studied using 1 rabbit per level. The test site of each animal was covered by wrapping the trunk with impervious plastic sheeting which was securely taped in place. The test material remained in contact with the skin for 24 hours. At the end of this period, the plastic sheeting and all residual test material were removed. The test sites were examined for local skin reactions and the animals were returned to their cages. Observations for mortality, local skin reactions and behavioural abnormalities were continued for a total of 14 days following the skin applications. Initial, 7- and 14-day body weights were recorded. A necropsy examination was conducted on all animals.

GLP compliance:

no

Remarks:

study pre-dates GLP

Test type:

standard acute method

Limit test:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: young
- Weight at study initiation: 2.86 - 2.98 g
- Housing: The rabbits were housed individually in suspended, wire-bottomed cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 7 days

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Remarks:

the test material was applied as an aqueous slurry

Details on dermal exposure:

TEST SITE
- Area of exposure: Back
- % coverage: 30%
- Twenty-four hours prior to the dermal applications, the backs of the rabbits were shaved free of hair with electric clippers. The shaved area on each animal constituted about 30 percent of the total body surface area. The animals were then returned to their cages to await testing on the following day. The 24-hour waiting period allowed recovery of the stratum corneum from the disturbance which accompanied the close-clipping procedure and permitted healing of any microscopic abrasions possibly produced during the process.
- Type of wrap if used: The test site of each animal was covered by wrapping the trunk with impervious plastic sheeting which was securely taped in place. This plastic wrap insured close contact of the epidermis and test material. To prevent oral ingestion of the test material, each animal was fitted with a lightweight, flexible plastic collar which was worn throughout the observation period.

REMOVAL OF TEST SUBSTANCE
- The test material remained in contact with the skin for 24 hours. At the end of this period, the plastic sheeting and all residual test material were removed.

TEST MATERIAL
- Applied as an aqueous slurry to abraded skin sites

Duration of exposure:

24 hours

Doses:

200, 500 2000 mg/kg

No. of animals per sex per dose:

1 animal per dose

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Observations for mortality, local skin reactions and behavioural abnormalities were continued for a total of 14 days following the skin applications.
- Initial, 7- and 14-day body weights were recorded.
- A necropsy examination was conducted on all animals.

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the observation period.

Clinical signs:

other: No pharmacotoxic symptoms were observed in the rabbits following dermal exposure to the test material. The test material was non-irritating to the skin of the albino rabbit.

Gross pathology:

Necropsy examination revealed a cyst on the liver and a haemorrhage in the lung in rabbit 2-M. These lesions appeared to be associated with naturally occurring disease. No other gross pathologic alterations were found.

Interpretation of results:

other: Not classified in accordance with EU Criteria.

Conclusions:

Under the conditions of this study the acute dermal LD50 was greater than 2000 mg/kg.

Executive summary:

The acute dermal toxicity of the test material was investigated using young adult male New Zealand White rabbits.

Twenty-four hours prior to the dermal applications, the backs of the rabbits were shaved free of hair with electric clippers. A range of several dose levels was studied using 1 rabbit per level. The test site of each animal was covered by wrapping the trunk with impervious plastic sheeting which was securely taped in place. The test material remained in contact with the skin for 24 hours. At the end of this period, the plastic sheeting and all residual test material were removed. The test sites were examined for local skin reactions and the animals were returned to their cages. Observations for mortality, local skin reactions and behavioural abnormalities were continued for a total of 14 days following the skin applications. Initial, 7- and 14-day body weights were recorded. A necropsy examination was conducted on all animals.

No mortality occurred during the observation period. No pharmacotoxic symptoms were observed in the rabbits following dermal exposure to the test material. The test material was non-irritating to the skin of the albino rabbit.

Necropsy examination revealed a cyst on the liver and a haemorrhage in the lung in rabbit 2-M. These lesions appeared to be associated with naturally occurring disease. No other gross pathologic alterations were found.

Under the conditions of this study the acute dermal LD50 was greater than 2000 mg/kg.

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute Oral

Braun (1978)

The acute oral toxicity of the test material was investigated in a study similar to OECD 401. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).

During the study, ten animals (5/sex) were dosed at 10.0 and 20.0 g/kg. A rangefinding study was performed to arrive at the appropriate dose levels. The test material was administered by oral intubation as a 30 % w/v solution in tap water. Observations for mortality and overt signs of effect were made at 0-2 hours, and at 4-6 hours following dosing and daily thereafter for fourteen days. Body weights were recorded initially and terminally and a gross necropsy was performed on spontaneous deaths.

Common in-life signs of effect noted during the fourteen day study included: ataxia, coarse and fine tremors, red nasal and red oral discharges, urinary staining of the abdomen, piloerection, motor activity decrease, animal thinness, abdominal griping, irritability, and white stool. Signs of effect persisted throughout the study. A loss of weight or a failure to exhibit normal weight gain was noted in 4 of 10 surviving animals at 10.0 g/kg and 1 of 9 surviving animals at 20.0 g/kg.

One animal at 20.0 g/kg died during this study. Necropsy observations for this animal were: clear oral discharge; red nasal discharge; urinary staining of the abdomen; stomach and intestines: distended with gas; liver: mottled; thoracic cavity contains red congealed substance.

Under the conditions of this study the acute oral LD50 of the test material was determined to be greater than 20 g/kg, the highest dose tested.

Calandra (1976)

The acute oral toxicity of the test material was investigated in a study which followed closely to the methods of the standardied guideline OECD 401. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).

During the study, Sprague-Dawley derived rats received a single oral dose of test material, by gavage. Several doses were tested with one animal per level at 30, 100, 300, 1000, 3000 and 1000 mg/kg. After oral administration, the animals were observed for 14 days. Inital and final bodyweights were recorded and a necropsy examination was performed on all animals.

Hypoactivity was noted within 1 minute post-oral administration of the test material in animal 6. This symptom had subsided by the following day. No pharmaco-toxic symptoms were observed in any of the other animals following dosing. All animals gained weight during the study.

No mortality was observed during the observation period and necropsy examination of the surviving animals did not reveal any gross pathologic alterations.

Under the conditions of this study the acute oral LD50 of the test material was greater than 10000 mg/kg.

Pelikan & Cerny (1970)

The acute oral toxicity of the test material was investigated by administering test material to mice, by gavage, at a single dose of 4000 mg/kg bw. The animals were sacrificed after 24 hours and any clinical signs, or findings at necropsy, were noted.

A preliminary study preceded the main study. In the prelmiinary study, mice received test material, as a single oral dose, by gavage, at test concentrations of 200; 400; 800; 1200; 1600; 2400; 3200; 4000; 6000 mg/kg bw. The mice in the two control groups received by the same method and on the same day a single dose of 0.2 mL of Oleum helianthi or of pure water.

In the preliminary study, the 48-hour LD50 was determined to be > 6000 mg/kg bw, the highest dose level tested.

Under the conditions of the main study, the acute oral LD50 of the test material was > 4000 mg/kg bw, the highest dose level tested.

Acute oral: Read across to structurally similar substance monobutyltin trichloride (MBTC, CAS No.: 1118-46-3).

A study was conducted in male and female Wistar rats to investigate the oral LD50 for the test material.

The animals were administered the test material orally as a 10% emulsion in distilled water. Six doses were used: 0.5, 1.0, 1.4, 2.0, 2.8, and 4.0 g/kg body weight. After treatment the animals were observed for 12 days for clinical signs and mortality. Necropsy of all animals were performed at death or at the end of the observation period.

The method of Litchfield and Wilcoxon was used to calculate the LD50 which was reported as 2.2 g/kg body weight. The confidence limits were reported as 1.9 -2.7 g/kg body weight. Necropsy findings in animals that died were reported as bleeding and erosion in the gastrointestinal tract, bleeding in the pancreas, focal necrosis in the liver, kidney, and pancreas, and bloody infiltration of the mesenteric lymph nodes. The necropsy findings in the animals that survived the 12-day observation period were reported as unremarkable.

Under the conditions of the study the oral LD50 for the test material in rats was reported to be 2.2 g/kg bw, confidence limits (1.9-2.7 g/kg bw). However, the volumes of dosage administration were large (up to 4 mL/100 g). Hence, LD50 should be reported as > 2 g/kg bw (> 2000 mg/kg bw).

Inhalation

In accordance with REACH, Annex VIII, column 2 of information requirement 8.5, in addition to the oral route, for substances other than gases, the information required under 8.5.2 to 8.5.3 shall be provided for at least one other route. The choice for the second route will depend on the nature of the substance and the likely route of human exposure.

Information is provided for the dermal route which is deemed to be most appropriate as inhalation of the substance is unlikely. Testing by the inhalation route is therefore omitted.

Acute Dermal

Calandra (1976)

The acute dermal toxicity of the test material was investigated using young adult male New Zealand White rabbits. The study was awarded a reliability score of 2 in accordance with the criteria set forth by Klimisch et al. (1997).

Twenty-four hours prior to the dermal applications, the backs of the rabbits were shaved free of hair with electric clippers. A range of several dose levels was studied using 1 rabbit per level. The test site of each animal was covered by wrapping the trunk with impervious plastic sheeting which was securely taped in place. The test material remained in contact with the skin for 24 hours. At the end of this period, the plastic sheeting and all residual test material were removed. The test sites were examined for local skin reactions and the animals were returned to their cages. Observations for mortality, local skin reactions and behavioural abnormalities were continued for a total of 14 days following the skin applications. Initial, 7- and 14-day body weights were recorded. A necropsy examination was conducted on all animals.

No mortality occurred during the observation period. No pharmacotoxic symptoms were observed in the rabbits following dermal exposure to the test material. The test material was non-irritating to the skin of the albino rabbit.

Necropsy examination revealed a cyst on the liver and a haemorrhage in the lung in rabbit 2-M. These lesions appeared to be associated with naturally occurring disease. No other gross pathologic alterations were found.

Under the conditions of this study the acute dermal LD50 was greater than 2000 mg/kg.

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the substance does not require classification with respect to acute toxicity.