1-Propanaminium, 3-amino-N-(carboxymethyl)-N,N-dimethyl-, N-C8-10 (even numbered) acyl derivs., hydroxides, inner salts

Endpoint summary

Description of key information

 
    
    

Oral:
- LD50 > 2000 mg/kg bw (based on test material) / LD50 > 830 mg a.i./kg bw; OECD TG 423, rat (female) oral: gavage (RL1, GLP); read-across: Formamidopropylbetaine
- LD50 = 2335 mg a.i./kg bw; Similar to OECD TG 401, standard acute method, rat oral: gavage (RL1; pre-GLP); read-across: C8-18 and C18 unsatd. AAPB






  
  

Dermal:
- LD50 > 2000 mg a.i./kg bw; OECD TG 402, rat, Type of coverage: Occlusive (RL1, GLP); read-across: Formamidopropylbetaine
- LD50 > 2000 mg/kg bw (based on test material) / LD50 > 620 mg a.i./kg bw; OECD TG 402, rat, Type of coverage: Occlusive (RL1, GLP); read-across: C8-18 and C18 unsatd. AAPB
Inhalation
- No relevant route of exposure






 

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

according to guideline

Guideline:

other: US Guideline: Appraisal of the Safety of Chemicals in Food, Drugs and Cosmetics, FDA, 1959

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

no

Remarks:

Study performed before implementation of GLP

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS:
- Age: no data
- Sex: males and females
- Weight at study initiation: females: 150 - 170 g, males: 175 - 210 g
- Housing: individual housing in a battery of cages
- Diet: ad libitum, standardised laboratory diest Ssniff/Intermast
- Water: ad libitum
- Fasting period before study: 16 hours

ENVIRONMENTAL CONDITIONS:
- Roomtemperature: 22 +/- 1° C
- Relative humidity: 45 - 55 %
- Illumination: 12 hours daily

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

applied volumes: 0.75 - 2.1 ml/animal (depending on individual body weight and designated dose level)
Test substance was applied in the original state (aqueous solution, a.i. according to producer information 30 %) as delivered by the sponsor.

Doses:

5.0, 6.3, 7.94, 10.0 ml/kg bw, test substance applied as delivered by the sponsor (30 % a.i.)

No. of animals per sex per dose:

5 females
5 males

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 14 d
- Body weights were recorded before treatment (day 0) and after 14 days (animals which survived)
- Clinical observation: In each animal a number of clinical-toxicological signs were evaluated. Any change from the normal condition was noted and degree of severity of any clinical symptoms was assessed. The animals were examined at the following post-treatment intervals: 20 min, 1 h, 3 h, 24 h, 7 d, 14 d.
- Necropsy of the survivors performed: yes
- Other examinations performed: necropsy on died animals

Statistics:

Method of determining LD 50: according to Litchfield & Wilcoxon in combination with Gauß Integral

Key result

Sex:

male/female

Dose descriptor:

LD50

Remarks:

after 14 d

Effect level:

2 335 mg/kg bw

Based on:

act. ingr.

95% CL:

1 944 - 2 571

Sex:

male/female

Dose descriptor:

LD50

Remarks:

after 14 d

Effect level:

7.45 mL/kg bw

Based on:

test mat.

95% CL:

6.48 - 8.57

Sex:

male/female

Dose descriptor:

LD50

Remarks:

after 24 h

Effect level:

8.1 mL/kg bw

Based on:

test mat.

95% CL:

6.82 - 9.64

Sex:

male/female

Dose descriptor:

LD50

Remarks:

after 24 h

Effect level:

2 430 mg/kg bw

Based on:

act. ingr.

95% CL:

2 046 - 2 892

Mortality:

Number of deaths at each dose:
- 5 ml/kg bw: 2/10 (1 animal died within 24 h, 1 animal died within 48 h)
- 6.30 ml/kg bw: 2/10 (within 24 h)
- 7.94 ml/kg bw: 6/10 (5 animals died within 24 h, 1 animal died within 48 h)
- 10 ml/kg bw: 8/10 (7 animals died within 24 h, 1 animal died within 4 days)

Clinical signs:

>= 5 ml/kg bw: decreased motor activity, coordination disturbances, abnormal body posture, piloerection, diarrhoea, skin/mucosa cyanosis,
decreased body temperature with dose-dependant increase of effects. Clinical signs were observed at 20 minutes, 1 h and 3 h after application. Except of slight diarrhoea in one animal in dose groups 6.3, 7.94 and 10 ml/kg bw each, all symptoms were reversible after 24 hours. 7 days after application, all surviving animals were free of clinical symptoms.
>= 7.94 ml/kg bw: prone position

Body weight:

Weight gains were normal in all animals.

Gross pathology:

- Decedents: reddened gastric and intestinal mucosa
- Animals sacrificed at study termination 14 days p.a.: light reddened intestinal mucosa

LD50 values reported by the study authors refer to the test substance as specified above (30 % a.i.): 7.45 (6.48 - 8.57) ml/kg after 14 days and 8.10 (6.81 - 9.64) ml/kg after 24 hours

Density of the substance: roughly 1 g/cm3.
LD50 (14 day) referring to 100 % active substance: 2235 mg/kg bwLD50 (24 hours) referring to 100 % active substance: 2430 mg/kg bw 

Interpretation of results:

GHS criteria not met

Conclusions:

On the basis of the results obtained after a single oral administration, the oral LD50 was determined to be 7.45 ml/kg bodyweight based on product, corresponding to 2335 mg/kg bw active ingredient.

Executive summary:

In an acute oral toxicity study according to US Guideline Appraisal of the Safety of Chemicals in Food, Drugs and Cosmetics, FDA, 1959, which is comparable to the OECD guideline 401 (1981), 5 male and 5 female Wistar rats were given a single oral dose of Coco AAPB (30 % a.i.) as delivered by the sponsor at doses of 5.0, 6.3, 7.94, and 10.0 ml/kg bw. Animals were then observed for 14 days.

At 5.0, 6.3, 7.94, and 10.0 ml/kg bw 2/10, 2/10, 6/10, and 8/10 animals died, respectively. Most animals died within 24 hours p.a.. Weight gains were normal in all animals. Clinical signs at >= 5 mL/kg bw were decreased motor activity, coordination disturbances, abnormal body posture, piloerection, diarrhoea, skin/mucosa cyanosis and decreased body temperature with dose response realtionship. At >= 7.94 mL/kg bw animals showed prone position. Clinical signs were observed at 20 minutes, 1 h and 3 h after application. Except of slight diarrhoea in one animal in dose groups 6.3, 7.94 and 10 mL/kg bw, each, all symptoms were reversible after 24 hours. 7 days after application, all surviving animals were free of clinical symptoms. Gross pathology examination of animals found dead revealed reddened gastric and intestinal mucosa. Animals sacrificed at study termination 14 days p.a. had light reddened intestinal mucosa.

Oral LD50 Combined =  7.45 mL/kg bw after 14 d

Oral LD50 Combined =  8.1 mL/kg bw after 24 h

LD50 determined refers to the test substance as delivered by the sponsor. There is no information on content of active ingredient of the test substance in the study report. However, according to producer information tested Coco AAPB has 30% active ingredient. The density is roughly 1 g/mL. Therefore the calculated oral LD50 combined referring to 100 % active substance = 2335 mg/kg bw after 14 d

Coco AAPB is of low toxicity based on the LD50 in males and females.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

06. Jun. 2005 - 22. Jun. 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Species: rat
Breed: White Wistar
Breeder: Harlan Winkelmann, Borchen, Germany
Date of receipt: 2005-05-31
Number / Sex: 12 female
Body weights: 159.9 - 180.7 g at day of application
Age: about 8 weeks at day of application
Identification: dye marks and cage numbers
Diet: Altromin International, Lage, Germany, Type 1324 - 1188,
Water: Drinking water consisted of normal tap water from municipal sources (Stadtische Werke Krefeld AG, Krefeld, Germany). The water was monthly examined for pollutants which might interfere with the study. The data are retained in the archive.
Bedding material: Lignocel BK 10/20, Rettenmaier & Sohne GmbH & Co., Batch 02101 40915
Experimental animals:
Young healthy rats acclimatized for at least 5 days to laboratory conditions were used for this test. During the acclimatization period animals were observed for their health condition to assure that they were in best condition for this investigation. Only female rats were used.
Husbandry:
Three rats were housed in one Makrolon cage type VI each. A non-barrier system with air condition was used. The air conditioning had following nominal values: Temperature: 22°C ± 3°C, Humidity: 30 - 70 %. Climate control was run automatically. The lighting was in a 12-hour light/dark-cycle. Temperature and humidity were recorded continuously using a thermohygrometer, Lambrecht GmbH, Gottingen, Germany. The data were archived. There has been a deviation from nominal humidity values. On 2005-06-17 and 2005-06-18 the humidity was slightly higher (up to 75 %). However, this deviation is not supposed to have any effect on the experimental result.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

The starting dose of 300 mg/kg body weight was applied to six female animals as 10 % (w/w) solution in tap water. Therefore 1.5 g of the test substance were added to 13.5 g tap water and mixed on a magnetic stirrer. The density of the 10 % solution was 1.025 g/mL and of the undiluted test substance 1.22 g/mL. The density of the undiluted test substance was higher than 1.05 g/mL and therefore was considered in calculation of the application volume. The dose of 2000 mg/kg body weight was applied to six female animals with the undiluted test substance.
Dose volume: 300 mg/kg group: 0.48 - 0.51 mL; 2000 mg/kg: 0.27 - 0.30 mg/kg.

Doses:

300 mg/kg, 2000 mg/kg

No. of animals per sex per dose:

6 animals per dose (female only)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical examination was performed immediately, 0.5, 1.5 and 3.5 hours after test substance administration and thereafter daily; weighing at day 0, day 7, day 14
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 1 000 mg/kg bw

Based on:

other: solid matter (incl. NaCl)

Sex:

female

Dose descriptor:

LD50

Effect level:

> 830 mg/kg bw

Based on:

act. ingr.

Remarks:

test substance Formamidopropyldimethylbetain 50% (50% based on solid matter, active matter: 41.5 %)

Mortality:

No mortality occured.

Clinical signs:

No clinical signs observed

Body weight:

The body weight development of the animals was positive and within normal ranges.

Gross pathology:

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

Body weights

Animal ID	Sex	Dose [mg/kg]	Day 0 [g]	Mean±SD	Day 7 [g]	Mean±SD	Day 14 [g]	Mean±SD	Average weight gain [%]
425	F	300	168.4	165.6 ±3.8	190.7	186.0 ±9.6	197.6	198.0 ±7.6	+ 19.6
425 RV	F	300	166.8		183.8		197.7
425 RH	F	300	161.6		171.0		185.7
426	F	300	169.2		183.3		201.1
426 RV	F	300	159.9		187.2		196.8
426 RH	F	300	167.4		200.0		209.3
427	F	2000	173.7	171.2 ±5.8	196.0	191.7 ±8.6	208.7	204.7 ±6.1	+ 19.6
427 RV	F	2000	165.4		180.0		197.3
427 RH	F	2000	166.4		185.3		198.0
428	F	2000	180.7		204.3		212.9
428 RV	F	2000	167.9		189.8		205
428 RH	f	2000	173.0		194.7		205.9

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50-value of the test substance Formamidopropyldimethylbetain (41.5% a.i.) is greater than 2000 mg/kg body weight, corresponding to >830 mg/kg bw in terms of active substance.

Executive summary:

The test substance Formamidopropyldimethylbetain (50% based on solid matter, active matter: 41.5%) is a clear yellowish liquid. For labelling and classification purposes the test substance was tested regarding its acute toxic potential following oral application. Slight acute oral toxicity of the test substance was expected. Therefore the starting dose of 300 mg/kg body weight was chosen in the Acute-Toxic-Class-Method procedure, according to the OECD Guideline for the Testing of Chemicals No. 423 "Acute Oral Toxicity - Acute Toxic Class Method". A total of twelve female rats of the strain White Wistar were used in the study. The test substance was applied by gavage undiluted and as 10% (w/w) solution in tap water. No toxic symptoms after administration of 300 mg/kg body weight were observed in six animals. In the next step the test substance was applied to six rats at a dose level of 2000 mg/kg body weight. No toxic symptoms were observed in this group, as well. No death occurred in all groups of the dose levels 300 mg/kg and 2000 mg/kg body weight. Body weight development of all animals was positive 7 days and 14 days post application. The necropsy 14 days after oral application showed no substance related morphological visible pathologic organ findings. Thus, the LD50-value for the test substance is higher than 2000 mg/kg bw, corresponding to > 830 mg/kg bw in terms of active substance. Therefore, the test substance Formamidopropyldimethylbetain 50% does not need to be classified according to the Globally Harmonised System.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

All available data are reliable and of high quality (guideline studies, GLP).

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1987-07-27 to 1987-08-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same 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

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: CD (Crl:COBS CD(SD)BR)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 7 - 10 weeks
- Weight at study initiation: 200 to 232 g
- Fasting period before study: no
- Housing: individually housing in metal cages with wire mesh floors
- Diet: ad libitum, standard laboratory rodent diet "Labsure LAD 1"
- Water: ad libitum
- Acclimation period: 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 23
- Humidity (%): 66
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

One day prior to treatment hair was removed from the dorsolumbar region of each rat with electric clippers exposing an area equivalent to 10% of the total body surface. No shaving or chemical depilation was used.

The test substance was applied by spreading it evenly over the prepared skin. Total volume applied was 1.92 ml/kg bw. Test substance was applied in the original state (aqueous solution, a.i. 31 %) as delivered by the sponsor. The treated area was then promptly covered with gauze which was held in place with an impermeable dressing encircled firmly around the trunk.

At the end of the 24-hours exposure period, the dressings were carefully removed and the treated area of skin decontaminated by washing in warm (30°-40°C) water and blotting dry with absorbent paper.

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw (based on product),

No. of animals per sex per dose:

5 males
5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 d
- Body weights were recorded on Day 1 (day of dosing), 8 and 15
- Clinical observation: Animals were observed soon after dosing and at frequent intervals for the remainder of Day 1. On subsequent days the animals were observed once in the morning and again at the end of the experimental day. This latter observation was at approximately 16.30 hours on week days or 11.30 hours on Saturday and Sunday. Clinical signs were recorded at each observation. The nature, severity, approximate time of
onset and duration of each toxic sign.
- Necropsy of the survivors performed: yes
- Other examinations performed: skin reactions - The treated areas of skin were examined daily for signs of dermal irritation and assessed according to the following arbitrary scoring system.
Erythema and eschar formation:
No erythema = score 0
Slight erythema = score 1
Well-defined erythema = score 2
Moderate to severe erythema = score 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) = score 4
Oedema formation:
No oedema = score 0
Slight oedema = score 1
Well-defined oedema (area well-defined by definite raising) = score 2
Moderate oedema (raised approximately 1 millimetre) = score 3
Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure) = score 4
A separate record was kept of dermal changes other than erythema and oedema.

Sex:

male/female

Dose descriptor:

LD0

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Sex:

male/female

Dose descriptor:

LD0

Effect level:

> 620 mg/kg bw

Based on:

act. ingr.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: mortality 0/10

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 620 mg/kg bw

Based on:

act. ingr.

Remarks on result:

other: mortality 0/10

Mortality:

0/10

Clinical signs:

- Application site: Sites of application of the test substance showed slight or well-defined erythema in all animals on Day 2 and/or 3. On Day 4 and Day 5, slight erythema were seen in three male and four female rats, sloughing in two male rats and one female rat and hyperkeratinisation in three female rats. All skin reactions were completely reversible by Day 6 in all animals.
Slough or hyperkeratinisation affected the treated skin of 4, 5, 6, 8, 9, 10 rats on Days 4 and 5 only.

- systemic: There were no clinical signs of systemic reaction to treatment.

Body weight:

Slightly low bodyweight gains were recorded for three females on Day 8.
All other rats achieved anticipated bodyweight gains throughout the study.

Gross pathology:

Terminal autopsy findings were normal.

Interpretation of results:

GHS criteria not met

Conclusions:

The acute lethal dermal dose to rats was found to be > 2000 mg/kg bodyweight (based on product).

Executive summary:

In an acute dermal toxicity study according to EU Method B.3 and OECD Guideline 402, 5 male and 5 female CD rats (Crl:COBS CD(SD)BR) were dermally exposed to Coco AAPB (a.i 31 %) as delivered by the sponsor for 24 hours to 10% of total body surface at a dose of 2000 mg/kg bw (limit test). Test sites were covered with an occlusive dressing. After 24 hours, the test sites were rinsed with warm water. Animals then were observed for 14 days after dosing.

There were no clinical signs of systemic reaction to treatment. Sites of application of the test substance showed slight or well-defined erythema in all animals on Day 2 and/or 3. On Day 4 and Day 5, slight erythema were seen in three male and four female rats, sloughing in two male rats and one female rat and hyperkeratinisation in three female rats. All skin reactions were completely reversible by Day 6 in all animals. Slightly low bodyweight gains were recorded for three females on Day 8. All other rats achieved anticipated bodyweight gains throughout the study. Terminal autopsy findings were normal.

Dermal LD₀ Combined: 2000 mg/kg bw

Dermal LD₅₀ Combined: > 2000 mg/kg bw

LD₀ and LD₅₀ determined refer to the test substance as delivered by the sponsor. Amount of active ingredient in test substance is 31 %. Therefore the calculated oral LD₀and LD₅₀ combined referring to 100 % active substance is 620 and > 620 mg/kg bw, respectively.

Coco AAPB (a.i. 31 %) is of low toxicity based on the LD₅₀ in males and females.

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

28. Dec. 2005 - 11. Jan. 2006

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

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Species : Rat, Wistar strain, Crl:WI (outbred, SPF-Quality). Recognized by international guidelines as the recommended test system (e.g. OECD, EC).
Source : Charles River Deutschland, Sulzfeld, Germany.
Number of animals : 5 males and 5 females (females were nulliparous and nonpregnant).
Age and body weight : Young adult animals (approx. 9 weeks old) were selected. Body weight variation did not exceed +/- 20% of the sex mean.
Identification : Earmark
Health inspection : A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health. Special attention was paid to the skin to be treated, which was intact and free from any abnormality.
Conditions : Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour, a temperature of 21.0 ± 3.0°C (actual range: 20.5 - 22.4°C), a relative humidity of 30-70% (actual range: 45 - 70%) and 12 hours artificial fluorescent light and 12 hours darkness per day.
Accommodation : Individually housed in labeled Macrolon cages (Mill type, height 18 cm.) containing sterilized sawdust as bedding material (Woody-Clean type 3/4; Tecnilab-BMI BV, Someren , The Netherlands) and paper as cage-enrichment (Enviro-dri, Tecnilab-BMI BV, Someren, The Netherlands ). Acclimatization period was at least 5 days before start of treatment under laboratory conditions. During the acclimatization period the animals were group housed in Macrolon cages (MIV type).
Diet : Free access to standard pelleted laboratory animal diet (from Altromin (code VRF 1), Lage, Germany).
Water : Free access to tap water.
Results of analysis for each batch of diet (nutrients) and results of quarterly analysis of diet (contaminants), sawdust, paper and water were assessed and did not reveal any findings that were considered to have affected the study integrity. All certificates and results of analysis are retained in the NOTOX archives.

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

Method : Dermal application.
Clipping : One day before exposure (day -1) an area of approximately 5x7 cm on the back of the animal was clipped.
Application : The test substance was applied in an area of approx. 10% of the total body surface, i.e. approx. 25 cm2 for males and 18 cm² for
females. The test substance was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1 D)·, successively covered with aluminum foil and Coban elastic bandage*. A piece of Micropore tape* was additionally used for fixation of the bandages in females only.
*. Manufacturers: Laboratoires Stella s.a., Liege, Belgium (surgical gauze) and 3M, St. Paul, Minnesota, U.S.A. (Coban & Micropore).
Frequency : Single dosage, on day 1.
Dose level (volume) : 2000 mg/kg (3.8 mllkg) body weight. Purity of the test substance was taken into account. Dose volume calculated as dose level: specific gravity.
Application period : 24 hours, after which dressings were removed and the skin cleaned of residual test substance using tap water.

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

Mortality/Viability : Twice daily.
Body weights : Days 1 (pre-administration), 8 and 15.
Clinical signs : At periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15. The time of onset, degree and duration were recorded and the symptoms graded according to fixed scales: Maximum grade 4: grading slight (1) to very severe (4), Maximum grade 3: grading slight (1) to severe (3), Maximum grade 1: presence is scored (1).
Necropsy : At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

act. ingr.

Remarks on result:

no indication of skin irritation up to the relevant limit dose level

Mortality:

No mortality occurred,

Clinical signs:

Chromodacryorrhoea was noted among the animals on days 1 and 2. Scales were seen in the treated skin-area of some animals between days 3 and 12.

Body weight:

The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity,

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals,

BODY WEIGHTS (GRAM)

Sex/dose level	animal	Day 1	Day 8	Day 15
Males 2000 mg/kg bw	1	290	318	348
	2	286	307	332
	3	292	316	341
	4	326	352	400
	5	319	337	375
	Mean	303	326	359
	St Dev	18	18	28
	N	5	5	5
Females 2000 mg/kg bw	6	211	229	243
	7	195	206	209
	8	206	228	244
	9	217	233	241
	10	220	241	251
	Mean	210	227	238
	St Dev	10	13	16
	N	5	5	5

Interpretation of results:

GHS criteria not met

Conclusions:

The dermal LD50 value of FORMAMIDOPROPYLBETAINE in Wistar rats was established to exceed 2000 mg/kg body weight (corrected for purity of the test substance).

Executive summary:

Assessment of acute dermal toxicity with FORMAMIDOPROPYLBETAINE in the rat.

The study was carried out based on the guidelines described in: OECD No 402 (1987) "Acute Dermal Toxicity" EC, Council Directive 67/548/EEC, Annex V, B.3 (1992) "Acute Toxicity (Dermal)" EPA, OPPTS 870.1200 (1998), "Acute Dermal Toxicity" JMAFF Guidelines (2000), including the most recent revisions.

FORMAMIDOPROPYLBETAINE was administered to five Wistar 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 (day 15). No mortality occurred, Chromodacryorrhoea was noted among the animals on days 1 and 2, Scales were seen in the treated skin-area of some animals between days 3 and 12. 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 of FORMAMIDOPROPYLBETAINE in Wistar rats was established to exceed 2000 mg/kg body weight (corrected for purity of the test substance).

Based on these results FORMAMIDOPROPYLBETAINE does not have to be classified and has no obligatory labelling requirement for dermal toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (New York and Geneva, 2003) and EC criteria for classification and labelling requirements for dangerous substance and preparations (Council Directive 67/548/EEC).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

All available data are reliable and of high quality (guideline studies, GLP).

Additional information

No experimental data are available for the target substance C8-10 Alkylamidopropyl betaine. However, acute oral and dermal toxicity studies are available for the closely related source substances C8-18 and C18 unsatd. AAPB and Formamidopropylbetaine. A justification for read-across is given below.

 

Acute oral toxicity

In an acute oral toxicity study according to US Guideline Appraisal of the Safety of Chemicals in Food, Drugs and Cosmetics, FDA, 1959, which is comparable to the OECD guideline 401 (1981), 5 male and 5 female Wistar rats were given a single oral dose of C8-18 and C18 unsatd. AAPB (30 % a.i.) as delivered by the sponsor at doses of 5.0, 6.3, 7.94, and 10.0 ml/kg bw. Animals were then observed for 14 days.

At 5.0, 6.3, 7.94, and 10.0 mL/kg bw 2/10, 2/10, 6/10, and 8/10 animals died, respectively. Most animals died within 24 hours p.a.. Weight gains were normal in all animals. Clinical signs at >= 5 mL/kg bw were decreased motor activity, coordination disturbances, abnormal body posture, piloerection, diarrhoea, skin/mucosa cyanosis and decreased body temperature with dose response realtionship. At >= 7.94 mL/kg bw animals showed prone position. Clinical signs were observed at 20 minutes, 1 h and 3 h after application. Except of slight diarrhoea in one animal in dose groups 6.3, 7.94 and 10 mL/kg bw, each, all symptoms were reversible after 24 hours. 7 days after application, all surviving animals were free of clinical symptoms. Gross pathology examination of animals found dead revealed reddened gastric and intestinal mucosa. Animals sacrificed at study termination 14 days p.a. had light reddened intestinal mucosa.

Oral LD50 Combined =  7.45 mL/kg bw after 14 d

Oral LD50 Combined =  8.1 mL/kg bw after 24 h

LD50 determined refers to the test substance as delivered by the sponsor. There is no information on content of active ingredient of the test substance in the study report. However, according to producer information tested C8-18 and C18 unsatd. AAPB has 30% active ingredient. The density is roughly 1 g/mL. Therefore the calculated oral LD50 combined referring to 100% active substance = 2335 mg/kg bw after 14 d

C8-18 and C18 unsatd. AAPB is of low toxicity based on the LD50 in males and females.

 

In an acute oral toxicity study according to according to the OECD Guideline for the Testing of Chemicals No. 423 "Acute Oral Toxicity - Acute Toxic Class Method” 6 female Wistar rats were given a single dose of Formamidopropylbetaine (50% based on solid matter, active matter: 41.5%).

The test substance was applied by gavage undiluted and as 10% (w/w) solution in tap water. No toxic symptoms after administration of 300 mg/kg body weight were observed in six animals. In the next step the test substance was applied to six rats at a dose level of 2000 mg/kg body weight. No toxic symptoms were observed in this group, as well. No death occurred in all groups of the dose levels 300 mg/kg and 2000 mg/kg body weight. Body weight development of all animals was positive 7 days and 14 days post application. The necropsy 14 days after oral application showed no substance related morphological visible pathologic organ findings. Thus, the LD50-value for the test substance is higher than 2000 mg/kg bw, corresponding to > 830 mg/kg bw in terms of active substance.

 

Based on these results, the oral LD50 of the target substance C8-10 Alkylamidopropyl betaine is expected to be >2000 mg/kg bw.

 

Acute dermal toxicity

In an acute dermal toxicity study according to EU Method B.3 and OECD Guideline 402, 5 male and 5 female CD rats (Crl:COBS CD(SD)BR) were dermally exposed to C8-18 and C18 unsatd. AAPB (a.i 31 %) as delivered by the sponsor for 24 hours to 10% of total body surface at a dose of 2000 mg/kg bw (limit test). Test sites were covered with an occlusive dressing. After 24 hours, the test sites were rinsed with warm water. Animals then were observed for 14 days after dosing.

There were no clinical signs of systemic reaction to treatment. Sites of application of the test substance showed slight or well-defined erythema in all animals on Day 2 and/or 3. On Day 4 and Day 5, slight erythema were seen in three male and four female rats, sloughing in two male rats and one female rat and hyperkeratinisation in three female rats. All skin reactions were completely reversible by Day 6 in all animals. Slightly low bodyweight gains were recorded for three females on Day 8. All other rats achieved anticipated bodyweight gains throughout the study. Terminal autopsy findings were normal.

Dermal LD0 Combined: 2000 mg/kg bw

Dermal LD50 Combined: > 2000 mg/kg bw

LD0 and LD50 determined refer to the test substance as delivered by the sponsor. Amount of active ingredient in test substance is 31 %. Therefore the calculated oral LD0and LD50 combined referring to 100 % active substance is 620 and > 620 mg/kg bw, respectively.

C8-18 and C18 unsatd. AAPB (a.i. 31 %) is of low toxicity based on the LD50 in males and females.

 

In an acute dermal toxicity study according to OECD Guideline 402 (1987) "Acute Dermal Toxicity" and EC, Council Directive 67/548/EEC, Annex V, B.3 (1992) Formamidopropylbetaine was administered to five Wistar 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 (day 15). No mortality occurred, Chromodacryorrhoea was noted among the animals on days 1 and 2, Scales were seen in the treated skin-area of some animals between days 3 and 12. 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 of Formamidopropylbetaine in Wistar rats was established to exceed 2000 mg/kg body weight (corrected for purity of the test substance).

 

Based on these results, the dermal LD50 of the target substance C8-10 Alkylamidopropyl betaine is expected to be >2000 mg/kg bw.

 

Acute inhalation toxicity

Acute toxicity studies by inhalation route are unjustified. Due to its very low vapour pressure, an exposure to C8-10 Alkylamidopropyl betaine vapour is negligible. Generation of aerosols may be theoretically possible, however the estimated exposure is very low. Furthermore, systemic toxicity relevant to humans did not appear neither in acute nor in repeated dose toxicity studies by other exposure routes. Considering the exposure probability and the available information on the intrinsic toxic activity of the substance, an inhalative risk to humans is unlikely and therefore, conduct of an inhalative toxicity study is unjustified.

 

Summary

There is no evidence on relevant intrinsic acute toxic activity of C8-10 Alkylamidopropyl betaine constituting a hazard to human health. The acute toxicity of closely related source substance in rats was demonstrated to be low, with a dermal LD50 > 2000 mg/kg bw, and an oral LD50 value of 2335 mg/kg bw.

There are no data gaps for the endpoint acute toxicity. No human data are available. However, there is no reason to believe that these results from rat would not be applicable to humans.

 

 

Justification for read-across

For details on substance identity and detailed toxicological profiles, please refer also to the general justification for read-across given at the beginning of the CSR and attached as pdf document to IUCLID section 13.

 

This read-across approach is justified based on structural similarities. The target and source substances contain the same functional groups. Thus a common mode of action can be assumed.

The only deviation within this group of substances is a variety in their carbon chain length, which obviously does not have a relevant impact on acute toxicity as demonstrated by the available data on the source substances.

a. Structural similarity and functional groups

The target substance C8 -10 Alkylamidopropyl betaine is a UVCB substance manufactured from fatty acids (C8 and C10) and N, N-dimethylpropylenediamine (DMAPA) and further reacted with sodium monochloroacetate.

The source substance C8-18 and C18 unsatd. AAPB is a UVCB substance manufactured from natural fatty acids or oils and N, N-dimethylpropylenediamine (DMAPA) and further reacted with sodium monochloroacetate. As their origin is from natural sources, the used fatty acids may have a mixed slightly varying composition with an even numbered chain length from C8 to C18. Unsaturated C18 amounts may be included.

The source substance Formamidopropylbetaine is a monoconstituent substance manufactured from formic acid and N, N-dimethylpropylenediamine (DMAPA) and further reacted with sodium monochloroacetate.

 

b. Differences

Differences in chemical and other intrinsic properties of the target and source substances could potentially arise from the following facts:

-Different amounts of different carbon chain lengths (carbon chain length distribution):

Higher amounts of higher chain lengths and corresponding lower amounts of lower chain length lead to a rising average lipophilicity as can be seen from the increasing log Kow from Formamidopropylbetaine (log Kow: -3.3), C8-10 Alkylamidopropyl betaine (log Kow: 2.2), C12 AAPB (log Kow: 3.54), C8-18 AAPB (log Kow: 4.23).

However, based on the available toxicological data it is demonstrated, that this read-across approach is nevertheless reliable.

- Different amounts of unsaturated fatty ester moieties:

The source substance C8-18 and C18 unsatd. AAPB contains considerable amounts of unsaturated C18 chains, which represents a worst case with respect to some toxicological endpoints, mainly local effects (e.g. irritation, sensitisation). But in general, variability in the fatty acid moiety is not expected to be relevant to the intrinsic systemic toxicity of the substances. 

The provided structural similarities and impurity profiles support the proposed read-across hypothesis with high confidence.

 

Comparison of acute toxicity data

Endpoints	Source substance	Target substance	Source substance
	Formamidopropylbetaine	C8-10 Alkalymidopropyl betaine	C8-18 and C18 unsatd. AAPB
Acute toxicity, oral	WoE_RA_LTOE 16396 Acute toxicity: oral   OECD TG 423, rat (female) oral: gavage   LD50 > 2000 mg/kg bw (based on test material) LD50 > 830 mg a.i./kg bw   Reliability: 1 (reliable without restrictions), GLP	No data, read-across	WoE_RA_Acute toxicity: oral: 61789-40-0_8.5.1_Th_Goldschmidt_AG_1977   Similar to OECD TG 401, standard acute method, rat oral: gavage LD50 = 7.45 mL/kg bw (based on test material; after 14 d) LD50 = 2335 mg a.i./kg bw (after 14 d) LD50 = 8.1 mL/kg bw (based on test material; after 24 h) LD50 = 2430 mg a.i./kg bw (after 24 h) Reliability: 1 (reliable without restrictions), pre-GLP
Acute toxicity, dermal	WoE_RA_NOTOX 454027 Acute toxicity: dermal   OECD TG 402, rat, Type of coverage: Occlusive   LD50 > 2000 mg a.i./kg bw   Reliability: 1 (reliable without restrictions), GLP	No data, read-across	WoE_RA_Acute toxicity: dermal: 61789-40-0_8.5.3_KAO Corporation_1987_OECD 402   OECD TG 402, rat, Type of coverage: Occlusive LD0 > 2000 mg/kg bw (based on test material) LD0 > 620 mg a.i./kg bw LD50 > 2000 mg/kg bw (based on test material) LD50 > 620 mg a.i./kg bw   Reliability: 1 (reliable without restrictions), GLP
Supporting information from repeated dose toxicity studies	WoE_RA_Repeated dose toxicity: oral 90-day OECD 408 NOTOX 497622   OECD TG 408, subchronic, rat, oral: gavage   NOAEL = 1000 mg/kg bw/d No toxicologically relevant effects were observed up to the highest dose level tested.   Reliability: 1 (reliable without restrictions), GLP	WoE_ Repeated dose toxicity: 73772-45-9 / 73772-46-0_8.6.1_Evonik_2016_OECD407   OECD TG 407, subacute, rat, oral: gavage   NOAEL = 500 mg/kg bw/d No toxicologically relevant effects were observed up to the highest dose level tested   Reliability: 1 (reliable without restrictions), GLP	WoE_RA_feeding_Repeated dose toxicity: 61789-40-0_8.6.2_90days_Unilever_A03_FT890785   OECD TG 408, subchronic, rat, oral: feed   NOEL effects relevant to humans: 247 mg a.i./kg bw/d (highest tested dose, 1 % in feed, 731 mg/kg bw/d based on product (a.i. 33.8 %)) LOEL: 97 mg a.i./kg bw/day) (0.4% in feed, 288 mg/kg bw/d based on product (a.i. 33.8 %))     Reliability: 1 (reliable without restrictions), GLP

There is no evidence of relevant intrinsic acute toxicity of the source substances:

-      The oral LD50 of C8-18 and C18 unsatd. AAPB was 2335 mg a.i/kg bw in rat.

-      The oral LD50 of Formamidopropylbetaine was > 2000 mg/kg bw (based on test material), corresponding to LD50 > 830 mg a.i./kg bw

-      Although C8-18 and C18 unsatd. AAPB has only been tested at 620 mg a.i./kg bw (corresponding to 2000 mg/kg bw in terms of test material) via the dermal route, the consideration of all available data on acute dermal toxicity, acute oral toxicity, toxicokinetics and dermal penetration leads to the conclusion, that the dermal LD50 will be > 2000 mg a.i./kg bw.

-      This is supported by the dermal LD50 of Formamidopropylbetaine of > 2000 mg a.i./kg bw

The source substances Formamidopropylbetaine and C8-18 and C18 unsatd. AAPB represent both extremes with shorter and longer C-chains compared to the target substance C8-10 Alkylamidopropyl betaine. Interpolation is judged to be appropriate to fulfil the information requirements for this endpoint.

Further support for this read-across is given by the lack of systemic toxicity in the repeated dose toxicity studies conducted with all three substances.

Quality of the experimental data of the analogues:

The available data are adequate and sufficiently reliable to justify the read-across approach.

The acute oral toxicity of C8-18 and C18 unsatd. AAPB was tested in a study similar to OECD Guideline 401 (RL1, study performed before implementation of GLP). The acute oral toxicity of Formamidopropylbetaine was tested in a study according to OECD Guideline 423 (RL1, GLP).

The acute dermal toxicity of C8-18 and C18 unsatd. AAPB as well as Formamidopropylbetaine was tested in a study according to OECD Guideline 402 (RL1, GLP).

The test materials used in the respective studies represent the source substance as described in the hypothesis in terms of substance identity and minor constituents.

Overall, the study results are adequate for the purpose of classification and labelling and risk assessment.

 

Conclusion

Based on structural similarities of the target and source substances as presented above and in more detail in the general justification for read across, it can be concluded that the available data from the source substances C8-18 and C18 unsatd. AAPB and Formamidopropylbetaine are also valid for the target substance C8-10 Alkylamidopropyl betaine.

There is no evidence of relevant intrinsic acute toxicity of C8-10 Alkylamidopropyl betaine, thus, classification and labelling with regard to acute toxicity is not warranted.

Justification for classification or non-classification

Based on the available data, C8-10 Alkylamidopropyl betaine does not need to be classified for acute toxicity according to regulation (EC) 1272/2008. Thus, no labelling is required.