N-methyl-N-[C18-(unsaturated)alkanoyl]glycine

Endpoint summary

Description of key information

Oral (similar OECD 401, rat): LD50 > 5000 mg/kg bw

Inhalation (similar OECD 403, rat): LC50 = 1.37 mg/L

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:

key study

Study period:

11 Dec - 23 Dec 1980

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)

Deviations:

yes

Remarks:

lack of test material details

GLP compliance:

not specified

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
- Age at study initiation: 7-8 weeks
- Weight at study initiation: 154 ± 6.8 g (males), 163 ± 10.5 g (females)
- Fasting period before study: animals were fasted overnight prior to administration.
- Diet: rat food (NAFAG, Gossau SG), ad libitum
- Housing: animals were housed in groups of 5 in Macrolon III cages
- Water: ad libitum
- Acclimation period: at least 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 55 ± 10
- Photoperiod (hrs dark / hrs light): 10/10

Route of administration:

oral: gavage

Vehicle:

other: 2% carboxymethylcellulose + 0.1% Tween 80

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 20 mL/kg bw

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: weekly
- Necropsy of survivors performed: yes

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study period.

Clinical signs:

other: Animals showed slight signs of dyspnoea from 1 h to 6 days after treatment. Animals had slight exophthalmos until 5 h after administration. Animals showed slight to moderate ruffled fur until Day 4. Slight to moderate diarrhoea was observed until Day 2 a

Gross pathology:

Necropsy revealed no substance-related findings.

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Basic data given. Lack of test material and study details.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

yes

Remarks:

lack on details on test material and study design

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

not specified

Sex:

not specified

Route of administration:

other: p.o. (presumably per os but no further information)

Sex:

not specified

Dose descriptor:

LD50

Effect level:

9 200 mg/kg bw

Based on:

test mat.

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

> 5 000 mg/kg bw

Quality of whole database:

The available studies are adequate and reliable (Klimisch score 2) and are thus sufficient to fulfil the standard information requirements set out in Annex VII, Section 8.5, of Regulation (EC) No. 1907/2006 (REACH).

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

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 403 (Acute Inhalation Toxicity)

Deviations:

yes

Remarks:

lack of details on test method

GLP compliance:

no

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: WIGA, Sulzfeld, Germany
- Weight at study initiation: 187 g (males), 204 g (females)
- Diet: Herilan MRH (Eggersmann KG, Rinteln/Weser, Germany), ad libitum
- Water: tap water, ad libitum

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose/head only

Vehicle:

not specified

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: dynamic head-nose apparatus (BUNDSCHUH, Griesheim, Germany) and constant infusion apperatus (UNITA I, BRAUN, Melsungen, Germany)
- Rate of air: 1000 L/h

TEST ATMOSPHERE
- Brief description of analytical method used: for the quantitative analysis an indirect indicator method was used. Therefore, the substance was mixed with 0.06 and 0.19% Oil Res 0 C.J. 26125. The samples were measured spectroscopically at 525 nm and the concentration of the test substance was evaluated using a calibration line.
- Samples taken from breathing zone: yes

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

0.3, 0.6, 2.2 and 3.7 mg/L (analytical concentration)
0.95, 1.67, 11.4 and 22.7 mg/L (nominal concentration)

No. of animals per sex per dose:

10

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: animals were observed daily for mortality and individual body weights were determined before start of the study and thereafter weekly
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Statistics:

The statistical analysis was performed according to the "Probitanalyse" (D.J. Finney, 1971, Syndics of the Cambridge University Press, London, UK).

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

1.37 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Sex:

female

Dose descriptor:

LC50

Effect level:

1.8 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Sex:

male

Dose descriptor:

LC50

Effect level:

ca. 1.05 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

0.6 mg/L: 3 males and 1 female animal died
2.2 mg/L: 10 males and 4 females died
3.7 mg/L: 8 males and 10 females died

Clinical signs:

other: 0.3 mg/L: flight attempts and gasping breathing 0.6 mg/L: additional to the signs of the low dose group and noisy breathing and slight staggering. One female showed lateral position. 2.2 mg/L: additional to the signs of the 0.6 mg/L dose group and bloody

Body weight:

2.2 mg/L (females): weight loss (mean 8 g) from Day 7-14
3.7 mg/L (males): weight loss (mean 4 g) from the Day of exposure to Day 7

Gross pathology:

Animals that died during the study period:
0.6 mg/L: heart: acute dilatation, acute hyperemia; lung: oedema with emphysema at the edges, pulmonary lobes were wet and fleshy and excess of blood were observed
2.2 mg/L: acute dilatation, acute hyperemia; lung: severe exhalation especially at the peripheral areas, severe oedema formation, single areas wet and fleshy. One animal with slight hydrothorax
3.7 mg/L: acute dilatation, acute hyperemia; lung: laminar bleedings, slight exhalation

Surviving animals:
Organs showed no signs of toxicity.

Table 1. Table for acute inhalation toxicity.

Target concentration [mg/L air]	Toxicological results*	Duration of clinical signs	Mortality (%)
Males
0.3	0/10/10	Day 2-7	0
0.6	3/10/10	Day 2-7	30
2.2	10/10/10	Day 2-7	100
3.7	8/10/10	Day 2-7	80
Females
0.3	0/10/10	Day 2-7	0
0.6	1/10/10	Day 2-7	10
2.2	4/10/10	Day 2-7	40
3.7	10/10/10	Day 2-7	100
LC50 = 1.37 (1.01-1.85) mg/L air

* first number = number of dead animals

 second number = number of animals with clinical signs

 third number = number of animals used

Interpretation of results:

other: Acute tox. Inhalation 4, H332. Classification according to Regulation (EC) No. 1272/2008 (CLP/EU GHS).

Reference 2

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

supporting study

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 403 (Acute Inhalation Toxicity)

Deviations:

yes

Remarks:

lack on details on test method

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: Sprague-Dawley, Mura: SPRA (SPF 68 Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:MUS RATTUS, Brunnthal, Germany
- Weight at study initiation: 185±15 g
- Diet: Herilan MRH (Eggersmann KG, Rinteln/Weser, Germany), ad libitum
- Water: tap water, ad libitum

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose/head only

Vehicle:

other: Ethanol 10%

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: constant infusion apperatus (UNITA I, BRAUN, Melsungen, Germany) and two-substance diffusor (Rhema)
- Method of holding animals in test chamber:animals were placed in tubes and stick out with the nose in to the inhalation area
- Source and rate of air: compressed air, 1500 L/h

TEST ATMOSPHERE
- Brief description of analytical method used: for the quantitative analysis an indirect indicator method was used. Therefore, the substance was mixed with 0.5% Oil Res 0 C.J. 26125. The samples were measured spectroscopically at 525 nm and the concentration of the test substance was evaluated using a calibration line.
- Samples taken from breathing zone: yes

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

1 h

Concentrations:

0.17, 1.35, 1.84, 1.85, 3.28 and 4.96 mg/L (analytical concentration)
1.13, 3.42, 6.84, 6.82, 24.55 and 25.6 mg/L (nominal concentration)

No. of animals per sex per dose:

10

Control animals:

other: a control group for body weight gain observation (no further information)

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: animals were observed daily for mortality and individual body weights were determined before start of the study and thereafter weekly
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight

Statistics:

The statistical analysis was performed according to the "Probitanalyse" (D.J. Finney, 1971, Syndics of the Cambridge University Press, London, UK).

Sex:

male/female

Dose descriptor:

LC50

Effect level:

2.3 mg/L air

95% CL:

1.9 - 2.8

Exp. duration:

1 h

Mortality:

0.17 mg/L: no animals died
1.35 mg/L: 1 male and 3 female animals died
1.84 mg/L: 6 males died
1.85 mg/L: 7 male and 3 female animals died
3.28 mg/L: 8 male and 3 female animals died
4.96 mg/L: 10 male and 9 female animals died

Clinical signs:

other: 0.17 mg/L: no clinical signs of toxicity observed. The animals of all other groups had eye and nose exudate, eyelid closure, dyspnoea, staggering, cowering position, scrubby substance-clotted fur. Animals that died during the study died within the first

Body weight:

3.28 mg/L (males) and 1.84 mg/L (females): reduced body weight gain during the first days of the study. After 14 days, the body weight was comparable to the control.

Gross pathology:

Animals that died during the study period:
heart: acute dilatation, acute hyperemia: lung: exhalation, excess of blood and severe oedemas

Surviving animals:
Organs showed no signs of toxicity.

Table 1. Table for acute inhalation toxicity.

Target concentration [mg/L air]	Toxicological results*	Duration of clinical signs	Mortality (%)
Males
0.17	0/0/10	-	0
1.35	1/10/10	Day 3	10
1.84	6/10/10	Day 7	60
1.85	7/10/10	Day 14	70
3.28	8/10/10	Day 14	80
4.96	10/10/10	Day 7	100
Females
0.17	0/0/10	-	0
1.35	3/10/10	Day 3	30
1.84	0/10/10	Day 7	0
1.85	3/10/10	Day 14	30
3.28	3/10/10	Day 14	30
4.96	9/10/10	Day 7	90
LC50 = 2.3 (1.9 - 2.8) mg/L air

* first number = number of dead animals

 second number = number of animals with clinical signs

 third number = number of animals used

Interpretation of results:

other: Acute tox. Inhalation 4, H332. Classification according to Regulation (EC) No. 1272/2008 (CLP/EU GHS).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

1 370 mg/m³ air

Quality of whole database:

The available studies are adequate and reliable (Klimisch score 2) and are thus sufficient to fulfil the standard information requirements set out in Annex VIII, Section 8.5, of Regulation (EC) No. 1907/2006 (REACH).

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute oral

Two studies investigating the acute toxicity via the oral route of N-methyl-N-(C18-(unsaturated)alkanoyl)glycine (EC No. 701-177-3) are available.

One study for acute oral toxicity was performed in Sprague-Dawley rats similar to OECD guideline 401 (Ciba Geigy, 1980). A group of 10 rats (5 males and 5 females) was dosed with 5000 mg/kg bw of the test substance by gavage. The animals were observed for a period of 14 days following administration. During the study period, no mortality occurred in any animal. Observed clinical signs included slight dyspnoea, slight exophthalmos and slight to moderate ruffled fur and slight to moderate diarrhoea and a slightly curved body position. All animals recovered within 7 days. No effects on body weight were noted and necropsy revealed no substance-related findings. Thus, the oral LD50 for male and female rats was considered to be greater than 5000 mg/kg bw.

In a further study with limited data given, the registered substance was tested for acute oral toxicity similar to OECD guideline 401 (BASF, 1979). Rats were given the test material per oral administration. No details about the study are given. The authors evaluated an oral LD50 for rats to be greater than 9200 mg/kg bw.

 

Acute inhalation

The acute inhalation toxicity was evaluated in two studies similar to OECD guideline 403 (BASF, 1979a and b). In the key study, groups of 10 Sprague-Dawley rats were exposed nose/head only to 0.3, 0.6, 2.2 and 3.7 mg/L air for 4 h. The animals were observed for a period of 14 days following administration. Mortality occurred in male animals in the 0.6, 2.2 and 3.7 mg/L air groups (30, 100 and 80% mortality). In female animals, mortality was observed in the 0.6, 2.2 and 3.7 mg/L air groups (10, 40 and 100% mortality), as well. Clinical signs of toxicity were observed in all surviving animals including flight attempts, gasping and noisy breathing, slight staggering, bloody nose area, low motility, hair loss in the head area and salivation in varying degrees. Surviving animals were free of symptoms from Day 2 - 8 after treatment and the organs showed no signs of toxicity at necropsy. Body weight loss was observed in females of the 2.2 mg/L and in males of the high-dose group. In animals that died during the study period, clinical signs of toxicity were acute dilatation, acute hyperemia, oedema and hyperemia of the lungs, severe exhalation especially at the peripheral areas, severe oedema formation and laminar bleedings of the lungs (high-dose group). Thus, the LC50 for female animals is calculated to be 1.8 mg/L air and 1.05 mg/L air for males. The combined LC50 value for males and females is considered to be 1.37 mg/L air after 4 h exposure to the test material.

In the second study, similarly performed as described above, 10 Sprague-Dawley rats were exposed nose / head only to 0.17, 1.35, 1.84, 1.85, 3.28 and 4.96 mg/L air (analytical concentration) for 1 h. Within the first 3 days after exposure, mortality occurred in male animals in the 1.35, 1.84, 1.85, 3.28 and 4.96 mg/L air groups (10, 60, 70, 80 and 100% mortality) and in female animals in the 1.35, 1.85, 3.28 and 4.96 mg/L air groups (30, 30, 30 and 90% mortality). Except in the lowest dose group, the animals of all other groups had eye and nose exudate, eyelid closure, dyspnoea, staggering, cowering position and scrubby substance-clotted fur. Male animals of the 3.28 mg/L and females of the 1.84 mg/L dose groups showed reduced body weight gain at the beginning of the study period being comparable to the control within 14 days. At gross pathology in animals that died during the study period, acute dilatation of the heart and acute hyperemia of the lung were observed. In surviving animals, no signs of toxicity were observed at gross pathology.

Thus, the LC50 for male and female animals is calculated to be 2.3 mg/L air after 1 h exposure.

Justification for classification or non-classification

The available data on acute oral toxicity with N-methyl-N-(C18-(unsaturated)alkanoyl)glycine (EC No. 701-177-3) do not meet the criteria for classification according to Regulation (EC) No 1272/2008 (CLP) and are therefore conclusive but not sufficient for classification. The data on acute inhalation toxicity meet the classification criteria for Acute Tox. 4 (H332) according to Regulation (EC) No. 1272/2008 (CLP).