[(±)-(4-amino-2-hydroxy-4-oxobutyl)trimethylammonium] chloride

Endpoint summary

Description of key information

Key study: Acute oral toxicity. OECD guideline and EU method. GLP study.
The oral LD50 for the test substance was greater than 2000 mg/kg bw.
Key study: Acute dermal toxicity. OECD guideline and EU method. GLP study.
The dermal LD50 for the test substance was greater than 2000 mg/kg bw.

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:

From 27 November 2012 to 04 February 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline and EU method. GLP study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Germany
- Age at study initiation: 9-10 week old
- Weight at study initiation: 193-205 g
- Fasting period before study: On the day before the start of the experiment, i.e. about 19 hours before the administration of the test item, the food was withheld but the water was still available.
- Housing: The animals were kept in plastic cages covered with wire bar lids. The dimensions of the cages were as follows: 58 x 37 x 21 cm (length x width x height). In the sighting study, the animal was caged individually, whereas in the main study, there were four animals per cage.
- Diet (e.g. ad libitum): “Murigran” standard granulated laboratory fodder, ad libitum
- Water (e.g. ad libitum): drinking tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 – 22 °C
- Humidity (%): 38 – 70%
- Air changes (per hr): about 16 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light – 12 hours dark

IN-LIFE DATES: From: 30 November 2012 To: 14 December 2012

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 0.5 mL/100 g of animal body weight

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The evaluation of general condition of the animals, i.e. the observation of all animals for morbidity and mortality was conducted twice a day or once a day (on days off) during the 14-day period of the experiment. The detailed clinical observations were performed on the day of the test item administration (day 0), i.e. 10, 30 and 60 minutes after the administration and then at hourly intervals up to 5 hours. From the 1st to the 14th day of the observation period, the detailed clinical observations were made once a day. The animals’ body weight was determined individually directly before the administration of the test item (day 0) and then on the 7th and 14th day.
- Necropsy of survivors performed: yes
After the 14-day observation period, all animals which survived the experiment were killed by an intraperitoneal administration of morbital at a dose of 200 mg/kg b.w. and subjected to the post mortem examination.

Preliminary study:

Following a single administration of the test item at a dose of 2000 mg/kg b.w. to one animal used in the sighting study, some signs of toxicity were visible on the administration day. These included a slight decrease in locomotor activity, dejection and a bristled coat – from the 1st to the 5th hour after the administration. On 1st and 2nd day after the administration, the animal’s gait was wavering. The animal survived the 14-day observation period. During the entire experiment, the animal’s body weight increased. The animal necropsy did not reveal any pathological changes.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed.

Clinical signs:

other: Following a single administration of the test item at a dose of 2000 mg/kg b.w. to four animals used in the main study, no clinical signs were observed in these animals. All animals survived the 14-day observation period.

Gross pathology:

The animal necropsy did not reveal any pathological changes.

Table 1:    Summary of acute oral toxicity

Females
Dose	Mortality
2000 mg/kg bw	0/5

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The oral LD50 for the test substance was greater than 2000 mg/kg bw.

Executive summary:

The acute oral toxicity study – fixed dose procedure, according to the OECD Guideline No. 420 / EU Method B.1.bis, was performed in order to obtain information on health hazards likely to arise from a single oral exposure caused by the consumption of the test item. The experiment commenced with a sighting study in which the test item at a single dose of 2000 mg/kg b.w was administered to one animal. On the grounds of the sighting study results, four animals used in the main study were given the test item at a dose of 2000 mg/kg b.w. (the animal of the sighting study was included in the main study). After the administration of the test item, the animals were observed for 14 days. General and detailed clinical observations were conducted daily during the entire experiment. The animals’ body weight was determined individually on day 0 (directly before the administration of the test item) as well as on the 7th and 14th day.

After the 14-day observation period, all animals which survived the experiment were euthanized and subjected to a necropsy and a detailed gross examination. Following a single administration of the test item at a dose of 2000 mg/kg b.w. to one animal, some signs of toxicity were observed. The animal survived the 14-day observation period. Following a single administration of the test item at a dose of 2000 mg/kg b.w. to the next four animals used in the main study, no clinical signs were observed during the 14-day observation period. All animals survived the observation period. Body weight gains were stated in all animals. The animal necropsy did not reveal any pathological changes. The oral LD50 for the test substance was greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

Klimisch 1 and the study was carried out in accordance with internationally valid GLP principles.

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:

From 27 November 2012 to 04 February 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline and EU method. GLP study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Germany
- Age at study initiation: 8-week old males; 10-week old females
- Weight at study initiation: males 273-300 g; females 209-241 g
- Housing: The animals were kept in plastic cages with the following dimensions: 58 x 37 x 21 cm (length x width x height). The cages were covered with wire bar lids. After the test item application, the animals were caged individually. After the removal of the test item from the animals’ skin, there were five rats per cage. Each sex was kept separately.
- Diet (e.g. ad libitum): “Murigran” standard granulated laboratory fodder, ad libitum
- Water (e.g. ad libitum): drinking tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 – 22 °C
- Humidity (%): 38 – 67%
- Air changes (per hr): about 16 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light – 12 hours dark

IN-LIFE DATES: From: 28 November 2012 To: 12 December 2012

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: The area of skin treated with the test item was about 41 cm2 (males) and 31 cm2 (females).
- % coverage: about 10% of the total body skin area.
- Type of wrap if used: The powdered test item was applied to gauze patches. Then, the patches were laid on the prepared skin and covered with PVC foil; an elastic bandage was used to make a circular protecting band.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The band and the gauze patches were taken off and the residual test item was removed using water
- Time after start of exposure: After 24 hours.

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 animals per sex per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The evaluation of general condition of the animals, i.e. the observation of all animals for mortality and morbidity was conducted twice a day or once a day (on days off) during the 14-day observation period. The detailed clinical observations were made on the administration day (day 0) at hourly intervals up to 5 hours. From the 1st to the 14th day of the observation period, the detailed clinical observations were performed once a day. The animals’ body weight was determined individually directly before the test item administration (day 0) and then on the 7th and 14th day.
- Necropsy of survivors performed: yes
After the 14-day observation period, all animals were humanely killed by an intraperitoneal administration of morbital at a dose of 200 mg/kg and transferred to the gross examination.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed.

Clinical signs:

other: Following a single application of the test item, no general clinical signs were stated in the animals. Pathological changes on the skin in the site of the test item application were not observed.

Gross pathology:

The gross examination did not reveal any pathological changes in the examined animals.

Table 1:    Summary of acute dermal toxicity

	Males	Females
Dose	Mortality	Mortality
2000 mg/kg bw	0/5	0/5

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The dermal LD50 for the test substance was greater than 2000 mg/kg bw.

Executive summary:

The acute dermal toxicity study, according to the OECD Guideline No. 402 / EU Method B.3, was performed in order to obtain information on health hazards resulting from the possibility of a single exposure due to skin contact with the test item. The test item at a single dose of 2000 mg/kg b.w. was applied to the shaved dorsal skin of 5 males and 5 females for 24 hours. After the administration of the test item, the animals were observed for 14 days. The animals’ body weight was determined individually on day 0 (directly before the administration of the test item) as well as on the 7th and 14th day. After the 14-day observation period, all animals were humanely killed, dissected and subjected to a detailed gross necropsy. Following a single application of the test item, no general clinical signs were stated in the animals.Pathological changes on the animals’ skin in the site of the test item application. All animals survived the period of the experiment were not observed. During the 14-day period of the experiment, all animals’ body weight increased. The gross examination did not reveal any pathological changes in the examined animals. The dermal LD50 for the test substance was greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

Klimisch 1 and the study was carried out in accordance with internationally valid GLP principles.

Additional information

Key study: Acute oral toxicity. OECD guideline and EU method. GLP study.

The oral LD50 for the test substance was greater than 2000 mg/kg bw.

Key study: Acute dermal toxicity. OECD guideline and EU method. GLP study.

The dermal LD50 for the test substance was greater than 2000 mg/kg bw.

Justification for selection of acute toxicity – oral endpoint
Only one study available.

Justification for selection of acute toxicity – dermal endpoint
Only one study available.

Justification for classification or non-classification

Based on the available data, the substance is not classified for acute toxicity.

Oral LD50 > 2000 mg/kg bw: not classified.

Dermal LD50>2000 mg/kg bw: not classified.