Amines, C12-16 alkyl dimethyl, reaction products with ethyl chloroacetate, 2-(2-aminoethylamino)ethanol and 2-propenoic acid

Endpoint summary

Description of key information

Acute oral toxicity: LD50 >2000 mg/kg bw,
Acute dermal toxicity: LD50 > 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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well-documented publication that meets basic scientific principles

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

not specified

GLP compliance:

not specified

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: Charles River Wiga, Sulzfeld
- Age at study initiation: 4 - 7 weeks old
- Weight at study initiation: male 100 - 125 g, female 90 - 115 g
- Fasting period before study: overnight
- Housing: single
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 23°C
- Humidity (%): 40 - 70 %
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 hours / 12 hours

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg bw

Doses:

4 dose levels: 4000, 5040, 6350 and 8002 mg/kg bw

No. of animals per sex per dose:

5 male and 5 female per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observations: 15 min, 30 min, 1 hour, 2 hours, 4 hours on day of application, once per day thereafter
weighing on day -1, 0, 7 and 14
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: no

Preliminary study:

Range finding study with 3 doses: 1000, 2500, 5000 mg/kg bw,
Result: 50 % mortality at 5000 mg/kg

Sex:

male/female

Dose descriptor:

LD50

Effect level:

5 106 mg/kg bw

Based on:

test mat.

95% CL:

4 306 - 5 822

Sex:

male/female

Dose descriptor:

LD50

Effect level:

2 553 mg/kg bw

Based on:

act. ingr.

Mortality:

Dose 4000 mg/kg bw: mortality 2/10
Dose 5040 mg/kg bw: mortality 5/10
Dose 6350 mg/kg bw. mortality 7/10
Dose 8002 mg/kg bw: mortality 10/10

Clinical signs:

other: rough fur, eyes closed, sedation, diarrhoea, ataxia, ventral position, gasping, respiration increased, exophthalmos

Gross pathology:

The necropsy 14 days after application showed no substance related morphological visible pathologic organ findings in the survived animals.

Interpretation of results:

practically nontoxic

Remarks:

Migrated information Criteria used for interpretation of results: expert judgment

Conclusions:

LD50 > 2000 mg/kg, the test item is not acute toxic following single oral application.

Executive summary:

In an acute oral toxicity study similar to OECD guideline 401, each 5 male/ 5 female, fasted, 4 -7 weeks old Sprague-Dawley rats were given a single oral dose of the test substance by gavage at doses of 4000, 5040, 6350 and 8002 mg/kg bw and observed for 14 days. Clinical signs and death was observed in all dose groups; mortality was 2/10, 5/10, 7/10 and 10/10 in the doses 4000, 5040, 6350 and 8002 mg/kg bw, respectively. The LD₅₀ was calculated to be 5106 mg/kg bw for the test item and 2553 mg/kg bw for the active ingredient. All surviving animals gained the expected weight at the end of the study period. No pathological changes were observed in these animals at necropsy.

 

Oral LD₅₀ (rat) > 2000  mg/kg bw

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 553 mg/kg bw

Quality of whole database:

Acceptable, well-documented study report

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline 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

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Sex:

male/female

Details on test animals or test system and environmental conditions:

Strain / Stock CD / Crl: CD(SD)
Supplier: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
Body weight (at dosing):Males: 213 - 223 g; Females: 206 - 234 g
Age (at dosing): Males: approx. 8 weeks; Females: approx. 9 weeks
Identification of animals: By coloured marks and cage label

Commercial diet, ssniff® R/M-H V1534 (ssniff Spezialdiäten GmbH, 59494 Soest, Germany) served as food. Feeding was discontinued approx. 16 hours before administration; only tap water was then available ad libitum.
Periodic analysis of the food for contaminants based on EPA/USA is conducted at least twice a year by LUFA-ITL. Certificates of analysis of the composition and for contaminants were provided by the manufacturer and are included in the raw data.
Housing
Granulated textured wood (Granulat A2, J. Brandenburg, 49424 Goldenstedt, Germany) was used as bedding material for the cages. The cages were changed and cleaned twice a week.
Periodic analysis of the bedding material for contaminants based on EPA/USA is conducted at least once a year by LUFA-ITL (see Appendix 2 'Limitation for Contaminants in the Bedding Material').
During the 14-day observation period the animals were kept singly in MAKROLON cages (type III plus) at a room temperature of 22°C ± 3°C (maximum range) and a relative humidity of 55% ± 15% (maximum range). Deviations from the maximum range caused for example during cleaning procedures are dealt with in SOPs.
The rooms were lit (150 lux at approx. 1.50 m room height) and darkened for periods of 12 hours each.

Drinking water
Drinking water in bottles was offered ad libitum.
Drinking water is examined according to the 'Deutsche Trinkwasserverordnung 2011 [German Regulations on drinking water 2011] by the Hamburger Wasserwerke, 20539 Hamburg, Germany, at least four times a year (see Appendix 2 'Limitation for Contaminants in the Drinking Water').
In addition, drinking water samples taken at LPT are analysed by LUFA-ITL once a year for means of bacteriological investigations according to the 'Deutsche Trinkwasserverordnung 2011, Anlage 1' [German Regulations on drinking water 2011, Addendum 1].

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

The test item was applied once for 24 hours on the shaved intact dorsal skin of rats (5 cm x 6 cm, approx. 1/10 of body surface). This treatment was followed by an observation period of 2 weeks.

Procedure

The intact dorsal skin of the animals was shaved free of hair with a shaver approximately 18 hours before administration of the test item. The site was situated on the animal´s back between the fore and hind extremities and had an area of at least 5 cm x 6 cm (approx. 1/10 of body surface).
The test patch was occlusive. The test item was held in contact with the skin with 8 layers of gauze. The gauze was covered with a plastic sheet and secured with adhesive plaster strips on the application site for 24 hours.

Duration of exposure:

Two weeks

Doses:

Dose level: 2000 mg/kg b.w. The dose level refers to the active ingredient.
Administration volume: 3.73 mL/kg b.w.

No. of animals per sex per dose:

5 animals/sex

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 5, 15, 30 min after administration, as well as 3, 6 and 24 hours after administration
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology: changes of skin and fur, eyes and mucous membranes, respiratory and circulatory function, autonomic and central nervous system and somatomotor activity as well as behaviour pattern, were observed at least once a day until all symptoms subsided, thereafter each working day. Attention was also paid to possible tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Observations on mortality were made at least once daily to minimize loss of animals during the study. Individual body weights were recorded before administration of the test item and thereafter in weekly intervals up to the end of the study. Changes in weight were calculated and recorded.
The skin was observed for the development of erythema and oedema.

Preliminary study:

not performed

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

act. ingr.

Mortality:

No deaths

Clinical signs:

other: None

Gross pathology:

No signs of abnormalities were noted at necropsy.

Summarized results

Symptoms/Criteria	Test substance 2000mg/kg b.w. (n = 5)
	males	females
Clinical signs	none	none
Skin reactions	none	none
Mortality within 6 h within 27 h within 7 d within 14 d	0 0 0 0	0 0 0 0
Mean body weight (in g) start	217.6	218.2
After 7 days	275.8 (+ 26.7)	238.6 (+ 9.3)
After 14 days	331.8 (+ 52.5)	263.8 (+ 20.9)
Inhibition of body weight gain	none	None
Necropsy findings	none	none

 

In brackets: body weight gain in %, compared to the start value

The dose level refers to the active ingredient

Conclusions:

The dermal LD50 in rats is greater than 2000 mg/kg bw (related to the active ingredient).

Executive summary:

The dermal LD50 of the test substance was investigated in male and female rats according to OECD guideline 402 in a limit test. A value of > 2000 mg/kg was obtained relating to the solid content of the test substance. No deaths, clinical signs or necroscopy findings were observed.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

mg/kg bw

Quality of whole database:

Acceptable, well-documented study report

Additional information

Acute oral toxicity

In an acute oral toxicity study similar to OECD guideline 401, each 5 male/ 5 female, fasted, 4 -7 weeks old Sprague-Dawley rats were given a single oral dose of the test substanceby gavage at doses of 4000, 5040, 6350 and 8002 mg/kg bw and observed for 14 days. Clinical signs and death was observed in all dose groups; mortality was 2/10, 5/10, 7/10 and 10/10 in the doses 4000, 5040, 6350 and 8002 mg/kg bw, respectively. The LD₅₀ was calculated to be 5106 mg/kg bw for the test item and 2553 mg/kg bw for the active ingredient.All surviving animals gained the expected weight at the end of the study period. No pathological changes were observed in these animals at necropsy.

 

Oral LD₅₀(rat) > 2000  mg/kg bw

Acute Inhalation

Given that inhalation is not a relevant route of exposure, testing by the inhalation route is not necessary according to REACH Regulation Annex VIII 8.5.2 Column 2. Inhalation is not a relevant route of exposure to test substance. This applies to both workers and the general population and is due to the physicochemical properties of the substance and the nature of the products where it is used. The test substance is a waxy solid paste. Generation of inhalable particles such as dust or aerosols is therefore not to be expected. Vaporisation needs not to be considered due to the substance’s very low vapour pressure of < 1 Pa at 20 °C. The generation of aerosols is excluded by technical means or product design. The substance is not used in spray applications. The most likely route of human exposure for workers and consumers is the dermal route. Results of laboratory animal studies show a low acute toxicity of closely related substances after oral and dermal exposure. Therefore the acute intrinsic toxic activity of the test substance is considered to be low. The occurrence of a systemic toxicity relevant to humans after inhalation is unlikely and therefore the conduct of an acute inhalation toxicity study is unjustified.

According to REACH Annex XI, the conduct of an acute inhalation toxicity study is scientifically unjustified.

Results of laboratory animal studies show a very low acute toxicity after oral or dermal exposure. Therefore the acute intrinsic toxic activity of the Formamidopropylbetaine is considered to be very low. The occurrence of a systemic toxicity relevant to humans after inhalation is unlikely and therefore the conduct of an acute inhalation toxicity study is unjustified.

Furthermore, the substance is exclusively manufactured in liquid form (aqueous solution) and has a low vapor pressure, so the potential for the unintended generation of inhalable forms such as vapors is low. The generation of aerosols is excluded by technical means or product design. The most likely route of human exposure for workers and consumers is the dermal route.

Acute dermal toxicity

The study was carried out based on the guidelines OECD No 402 "Acute Dermal Toxicity".

The test substance was administered to five rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice. No mortality and no systemic toxicity occurred. No signs of local skin reactions were noted among the animals. The body weight gain during the observation period was within the range expected for rats used in this type of study. No abnormalities were found at macroscopic post mortem examination of the animals. The dermal LD50 value in rats was established to exceed 2000 mg/kg body weight for the test item.

Justification for selection of acute toxicity – oral endpoint
Data from an acceptable, well-documented study report with reliability 2.

Justification for selection of acute toxicity – inhalation endpoint
Inhalation is not a relevant route of exposure and testing by the inhalation route is not required.

Justification for selection of acute toxicity – dermal endpoint
Data from an acceptable, well-documented study report with reliability 2.

Justification for classification or non-classification

Acute oral toxicity

Based on relevant, reliable and adequate data the test substance does not need to be classified and labelled according to the CLP Regulation (EC) No 1272/2008 and Directive 67/548/EEC with respect to acute oral toxicity.

 

Acute dermal toxicity

Based on relevant, reliable and adequate data the test substance has to be classified and labelled according to the CLP Regulation (EC) No 1272/2008 and Directive 67/548/EEC with respect to acute dermal toxicity, Category 4