L-Glutamic acid, N-coco acyl derivs., compds. with triethanolamine (1:1)

Endpoint summary

Description of key information

Acute Oral Key Study (Read-Across, Kiss, 2011)

Under the conditions of the study, no signs of acute toxicity were observed, and it can be concluded that the acute oral LD50 is greater than 2000 mg/kg bw.

Acute Oral Key Study (Read-Across, Masuyama, 2008)

Under the conditions of the test, the acute oral LD50 was > 5.0 g/kg body weight and < 10.0 g/kg body weight.

Acute Oral Supporting Study (Muroi, 1971)

Under the conditions of the study the LD50 value of the test material was estimated to be 6.50 g/kg bw (95 % Confidence limit: 5.66 – 7.48 g/kg).

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:

05 November 1986 to 28 November 1986

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

Reason / purpose for cross-reference:

other: Read-across target

Qualifier:

no guideline followed

Principles of method if other than guideline:

The test material was administered once orally to 20 male mice in dosages of 5.0 g/kg, 10.0 g/kg and 20.0 g/kg. A volume of 0.67 mL of the sample to be administered per 10 g of body weight was orally administered to the animals into their stomachs using gastric catheters.
Clinical signs, mortality and body weight were assessed during the study and necropsies performed on all animals.

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

no

Species:

mouse

Strain:

CD-1

Remarks:

ICR (Crl: CD-1)

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 4 weeks oldon arrival, 5 weeks old at the time the test material was administered.
- Weight at study initiation: 25.2 to 29.3 g
- Fasting period before study: The animals were fasted for approximately 8 hours before administration. Feeding resumed 2 hours after administration.
- Housing: Polyethylene cages with 5 animals in each cage.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23.3 ± 3 °C
- Humidity (%): 55 ± 5 %
- Air changes (per hr): 12 cycles per hour (all-fresh air system)
- Photoperiod (hrs dark / hrs light): 12 hours light:dark with light intensity of between 200 and 500 lx.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: The test material was dissolved in distilled water, and solutions of 7.5 %, 15.0 %, and 30.0 % were prepared immediately before administration.

MAXIMUM DOSE VOLUME APPLIED: 0.67 mL per 10 g of body weight was administered, representing that dosages of 5.0 g/kg, 10.0 g/kg, and 30.0 g/kg, of the test material were administered, respectively.

Doses:

5.0 g/kg, 10.0 g/kg, and 30.0 g/kg.

No. of animals per sex per dose:

20 males

Control animals:

yes

Remarks:

The vehicle (distilled water)

Details on study design:

- Duration of observation period following administration: 8 days
- Frequency of observations and weighing: From the day of administration (Day 1) until Day 8, observations were made once a day as to whether there were any deaths of the animals. On Day 1, observation of acute toxicity symptoms was continuously conducted, starting immediately after administration for approximately 2 hours. Observation of clinical signs was also conducted on Day 1 before administration and once a day between Day 2 and Day 8.
The body weight of all surviving animals was measured on Days 1, 2, 6, and 8.
Dates were counted based on Day 1 being the day that administration was carried out.
- Necropsy of survivors performed: Yes
- Other examinations performed: Clinical signs, body weight,organ weights.

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 - <= 10 000 mg/kg bw

Based on:

test mat.

Mortality:

During the study period, it was confirmed that upon orally administering the test material once to mice, there was 1 death on the 2nd day after administration (Day 3) in the group that was administered 5.0 g/kg. In addition, in the groups that were administered 10.0 g/kg and 20.0 g/kg, symptoms thought to be resulting from administration were observed immediately after administration, and all of the animals died.

Clinical signs:

other: Upon orally administering the test material once to mice, a decrease of locomotor activity and diarrohea were observed immediately after administration in the group that was administered 5.0 g/kg, and the death of 1 animal was confirmed on Day 3. The othe

Gross pathology:

Upon conducting necropsy examinations of the animals that died immediately after administration, haemorrhaging was found in the stomach or the intestinal tract of all animals, and there was also accumulation of water fluid in the intestinal tract. Upon conducting necropsy examinations of the surviving animals on the 7th day after administration, there were no abnormal findings for any of the animals.

Macroscopic findings

Dose (g/kg)	Sex	Animal No.	Type of death	Findings
0	Male	1	S	-
		2	S	-
		3	S	-
		4	S	-
		5	S	-
5.0	Male	1	S	-
		2	S	-
		3	D2	C
		4	S	-
		5	S	-
10.0	Male	1	D1	A,B
		2	D1	A,B
		3	D1	A,B
		4	D1	A,B
		5	D1	A,B
20.0	Male	1	D1	A,B
		2	D1	A,B
		3	D1	A,B
		4	D1	A,B
		5	D1	A,B

-: No abnormal findings,

A: Water fluid and haemorrhage in intestine

B: Dehydration (abdominal cavity)

C: No significant gross pathological findings were recorded because of the severe post-mortem change

Type of death:

S: Scheduled

D1: Death (Day 1)

D2: Death (Day 3)

Interpretation of results:

other: Not classified according to EU criteria.

Conclusions:

Under the conditions of the test, the acute oral LD50 was > 5.0 g/kg body weight and < 10.0 g/kg body weight.

Executive summary:

The test material was administered once orally to mice in dosages of 5.0 g/kg, 10.0 g/kg and 20.0 g/kg.

Immediately after administration, a decrease in locomotor activity and diarrhoea were observed in the group that was administered 5.0 g/kg. One animal in this group died the second day after administration but the other 4 animals recovered. In the groups that were administered 10.0 g/kg and 20.0 g/kg, a decrease in locomotor activity, diarrhoea, and discharge of the substance administered from the naris were observed. In the group that was administered 20.0 g/kg, cyanosis, opisthotonus, and convulsions were also observed. Afterwards, all animals in the groups that were administered 10.0 g/kg and 20.0 g/kg died.

Based on the above, it was suggested, LD50 for this compound is greater than 5.0 g/kg body weight, less than 10.0 g/kg body weight.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

For the justification for read-across, please refer to the read-across assessment framework report that is attached to Section 13.

Reason / purpose for cross-reference:

read-across source

Sex:

male

Dose descriptor:

LD50

Effect level:

> 5 000 - <= 10 000 mg/kg bw

Based on:

test mat.

Interpretation of results:

other: Not classified according to EU criteria.

Conclusions:

Under the conditions of the test, the acute oral LD50 was > 5.0 g/kg body weight and < 10.0 g/kg body weight.

Executive summary:

The test material was administered once orally to mice in dosages of 5.0 g/kg, 10.0 g/kg and 20.0 g/kg.

Immediately after administration, a decrease in locomotor activity and diarrhoea were observed in the group that was administered 5.0 g/kg. One animal in this group died the second day after administration but the other 4 animals recovered. In the groups that were administered 10.0 g/kg and 20.0 g/kg, a decrease in locomotor activity, diarrhoea, and discharge of the substance administered from the naris were observed. In the group that was administered 20.0 g/kg, cyanosis, opisthotonus, and convulsions were also observed. Afterwards, all animals in the groups that were administered 10.0 g/kg and 20.0 g/kg died.

Based on the above, it was suggested, LD50 for this compound is greater than 5.0 g/kg body weight, less than 10.0 g/kg body weight.

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Not stated

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

The acute oral toxicity of the test material was investigated in a study using male mice. Mice were administered with a single dose of test material by oral gavage. The animals were observed for mortality for a 14 day period.

GLP compliance:

no

Remarks:

This study pre-dates the inception of GLP

Test type:

standard acute method

Limit test:

no

Species:

mouse

Strain:

other: Dd

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 4 weeks
- Weight at study initiation: 16.0 to 18.0 g
- Fasting period before study: The animals were fasted for approximately 8 hours before administration.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 1 °C
- Humidity (%): 50 ± 10 %

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Distilled water

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 0.5 mL/ 10 g bodyweight administered orally using a feeding tube.

DOSAGE PREPARATION: The test material was weighed immediately before use and dissolved in the distilled water to make the test solution of specified concentrations.

Doses:

0.0, 4.0, 4.8, 5.8, 6.9, 8.3, 10.0, 12.0 and 14.3 g/kg

No. of animals per sex per dose:

10 males per dose

Control animals:

yes

Remarks:

Distilled water was administered to animals of the control group in the same manner.

Details on study design:

- Duration of observation period following administration: 14 days (336 hours)
- Frequency of observations: Mortality and viability was observed at the specified periods: Immediately after administration (0 hour), 2, 16, 24, 48, 72, 96, 120, 144, 168 and 336 hours after administration.

Statistics:

Litchfield-Wilcoxon method was used for calculation of LD50 value.

Sex:

male

Dose descriptor:

LD50

Effect level:

6 500 mg/kg bw

Based on:

test mat.

95% CL:

>= 5.66 - <= 7.48

Mortality:

All animals in 0 and 4 g/kg dose groups survived the duration of the observation period. 1, 2 and 6 animals died in the 4.8, 5.8 and 6.9 g/kg dose groups respectively during the 14 day observation period. In the 10.0, 12.0 and 14.3 g/kg dose groups no animals survived the 14 day observation period.

Table 1: Mortality and viability results within the study

Dose level (g/kg)	Number of survival animals at each observation period (h)											Mortality number at 336 h	Mortality rate at 336 h (%)
	0	2	16	24	48	72	96	120	144	168	336
0.0	10	10	10	10	10	10	10	10	10	10	10	0	0
4.0	10	10	10	10	10	10	10	10	10	10	10	0	0
4.8	10	9	9	9	9	9	9	9	9	9	9	1	10
5.8	10	10	10	8	8	8	8	8	8	8	8	2	20
6.9	10	7	7	4	4	4	4	4	4	4	4	6	60
8.3	10	2	1	0	0	0	0	0	0	0	0	10	100
10.0	10	0	0	0	0	0	0	0	0	0	0	10	100
12.0	10	0	0	0	0	0	0	0	0	0	0	10	100
14.3	10	0	0	0	0	0	0	0	0	0	0	10	100

Interpretation of results:

study cannot be used for classification

Conclusions:

Under the conditions of the study the LD50 value of the test material was estimated to be 6.50 g/kg bw (95 % Confidence limit: 5.66 – 7.48 g/kg).

Executive summary:

The acute oral toxicity of the test material was investigated in a study using male mice.

Mice were administered with a single dose of test material by oral gavage at 0.0, 4.0, 4.8, 5.8, 6.9, 8.3, 10.0, 12.0 and 14.3 g/kg. The animals were observed for mortality for a 14 day period at the following time points: Immediately after administration (0 hour), 2, 16, 24, 48, 72, 96, 120, 144, 168 and 336 hours after administration.

All animals in the 0 and 4 g/kg dose groups survived the duration of the observation period. 1, 2 and 6 animals died in the 4.8, 5.8 and 6.9 g/kg dose groups respectively during the 14 day observation period. In the 10.0, 12.0 and 14.3 g/kg dose groups no animals survived the 14 day observation period.

Under the conditions of the study the LD50 value of the test material was estimated to be 6.50 g/kg bw (95 % confidence limit: 5.66 – 7.48 g/Kg).

Reference 4

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 September 2011 to 12 October 2011

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Reason / purpose for cross-reference:

other: Read-across target

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

Qualifier:

according to guideline

Guideline:

other: JMAFF 12 Nousan 8147 (2000)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

other: CRL:(WI)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 9 weeks old.
- Weight at study initiation: 238 – 265 g.
- Fasting period before study: Yes, animals were fasted the day before dosing, food being returned 3 hours after treatment.
- Housing: Animals were housed in groups of 3 in Type II polypropylene/polycarbonate cages.
- Diet: Complete feed for rats and mice, ad libitum.
- Water: Municipal tap water, ad libitum.
- Acclimation period: At least 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): 15-20 air exchanges/hour.
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 06:00 to 18:00 hours.

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on oral exposure:

DOSAGE PREPARATION: Test material was freshly formulated at a concentration of 200 mg/mL in the vehicle on the day of administration. The formulation container was stirred continuously up to finishing the treatment.

CLASS METHOD
- Rationale for the selection of the starting dose: The initial dose level was selected by the study director to be that which was most likely to produce mortality in some of the dosed animals.

ADMINISTRATION: Group 1 was dosed first at 2000 mg/kg bw . The results were then confirmed by dosing Group 2 in the same manner.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Six females per dose, tested in two groups of 3 animals.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days.
- Frequency of observations and weighing:
> Clinical observations were performed at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and then daily until day 14. Observations included assessment of the skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
> Bodyweights were measured on Days -1, 0, 7 and 14 prior to necropsy.
- Necropsy of survivors performed: yes
- Other examinations performed: After examination of the external appearance, the cranial, thoracic and the abdominal cavities were opened and the organs and the tissues were observed. Macroscopic abnormalities were recorded.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality was observed in either Group 1 or 2.

Clinical signs:

other: Liquid faeces was observed in all animals, treated at a dose level of 2000 mg/kg bw, on the day of dosing. All animals fully recovered and were symptom free from 6 hours after the treatment until the end of the observation period.

Gross pathology:

There was no evidence of treatment related findings at necropsy. Pelvic dilatation of the right kidney was incidentally observed in one animal.

Table1: Clinical Observations

Group	Animal No.	Observations	Observation Days							Frequency
			0						1 - 14
			30 min	1 h	2 hrs	3 hrs	4 hrs	6 hrs
1	8861	Symptom Free	+	+	+	-	+	+	+	19/20
		Faeces liquid	-	-	-	1	-	-	-	1/20
	8862	Symptom Free	+	+	+	-	+	+	+	19/20
		Faeces liquid	-	-	-	1	-	-	-	1/20
	8863	Symptom Free	+	+	+	-	-	+	+	18/20
		Faeces liquid	-	-	-	1	1	-	-	2/20
2	8864	Symptom Free	+	+	+	-	+	+	+	19/20
		Faeces liquid	-	-	-	1	-	-	-	1/20
	8865	Symptom Free	+	+	+	-	+	+	+	19/20
		Faeces liquid	-	-	-	1	-	-	-	1/20
	8866	Symptom Free	+	+	+	-	+	+	+	19/20
		Faeces liquid	-	-	-	1	-	-	-	1/20

Frequency of observations = number of occurrence of observations / total number of observations.

 

Table 2: Bodyweights (g)

Group No.	Animal No.	Body weight (g) on Days				Weight Gain (g)
		-1	0	7	14	-1 - 0	0 - 7	7 -14	-1 - 14
1	8861	281	265	296	301	-16	31	5	20
	8862	273	256	288	296	-17	32	8	23
	8863	269	251	290	304	-18	39	14	35
2	8864	266	249	281	290	-17	32	9	24
	8865	257	239	267	277	-18	28	10	20
	8866	255	238	257	274	-17	19	17	19
Mean:		266.8	249.7	279.8	290.3	-17.2	30.2	10.5	23.5
Standard Deviation:		9.8	10.3	15.0	12.5	0.8	6.6	4.3	6.0

Interpretation of results:

other: not classified according to EU criteria

Conclusions:

Under the conditions of the study, no signs of acute toxicity were observed, and it can be concluded that the acute oral LD50 is greater than 2000 mg/kg bw.

Executive summary:

The acute oral toxicity of the test material was assessed in study performed under GLP conditions and in line with OECD 423 and EU Method B.1 tris according to the acute toxic class method. Female CRL:(WI) rats were treated with the test material at a dose level of 2000 mg/kg bw in two groups. Initially, three females (Group 1) were treated at a dose level of 2000 mg/kg bw. As no mortality was observed, a confirmatory group (Group 2) was treated at the same dose level.

Under the conditions of the study no mortality was observed in either group. Animals showed normal weight gain. Liquid faeces were observed in all animals on the day of dosing; all animals had fully recovered, and were symptom free, from 6 hours after the treatment until the end of the observation period. There was no evidence of treatment related findings at necropsy. Pelvic dilatation of the right kidney was incidentally observed in one animal. It can therefore be concluded that the acute oral LD50 of the test material is in excess of 2000 mg/kg bw.

Reference 5

Endpoint:

acute toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

For the justification for read-across, please refer to the read-across assessment framework report that is attached to Section 13.

Reason / purpose for cross-reference:

read-across source

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Interpretation of results:

other: not classified according to EU criteria

Conclusions:

Under the conditions of the study, no signs of acute toxicity were observed, and it can be concluded that the acute oral LD50 is greater than 2000 mg/kg bw.

Executive summary:

The acute oral toxicity of the test material was assessed in study performed under GLP conditions and in line with OECD 423 and EU Method B.1 tris according to the acute toxic class method. Female CRL:(WI) rats were treated with the test material at a dose level of 2000 mg/kg bw in two groups. Initially, three females (Group 1) were treated at a dose level of 2000 mg/kg bw. As no mortality was observed, a confirmatory group (Group 2) was treated at the same dose level.

Under the conditions of the study no mortality was observed in either group. Animals showed normal weight gain. Liquid faeces were observed in all animals on the day of dosing; all animals had fully recovered, and were symptom free, from 6 hours after the treatment until the end of the observation period. There was no evidence of treatment related findings at necropsy. Pelvic dilatation of the right kidney was incidentally observed in one animal. It can therefore be concluded that the acute oral LD50 of the test material is in excess of 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

> 2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute Oral Key Study (Read-Across, Kiss, 2011)

The acute oral toxicity of the test material was assessed in study performed under GLP conditions and in line with OECD 423 and EU Method B.1 tris according to the acute toxic class method. Female CRL:(WI) rats were treated with the test material at a dose level of 2000 mg/kg bw in two groups. Initially, three females (Group 1) were treated at a dose level of 2000 mg/kg bw. As no mortality was observed, a confirmatory group (Group 2) was treated at the same dose level.

Under the conditions of the study no mortality was observed in either group. Animals showed normal weight gain. Liquid faeces were observed in all animals on the day of dosing; all animals had fully recovered, and were symptom free, from 6 hours after the treatment until the end of the observation period. There was no evidence of treatment related findings at necropsy. Pelvic dilatation of the right kidney was incidentally observed in one animal. It can therefore be concluded that the acute oral LD50 of the test material is in excess of 2000 mg/kg bw.

Acute Oral Key Study (Read-Across, Masuyama, 2008)

The acute oral toxicity of the read-across material was assessed. The test material was administered once orally to mice in dosages of 5.0 g/kg, 10.0 g/kg and 20.0 g/kg.

Immediately after administration, a decrease in locomotor activity and diarrhoea were observed in the group that was administered 5.0 g/kg. One animal in this group died the second day after administration but the other 4 animals recovered. In the groups that were administered 10.0 g/kg and 20.0 g/kg, a decrease in locomotor activity, diarrhoea, and discharge of the substance administered from the naris were observed. In the group that was administered 20.0 g/kg, cyanosis, opisthotonus, and convulsions were also observed. Afterwards, all animals in the groups that were administered 10.0 g/kg and 20.0 g/kg died.

Based on the above, it was suggested, LD50 for this compound is greater than 5.0 g/kg body weight, less than 10.0 g/kg body weight.

Acute Oral Supporting Study (Muroi, 1971)

The acute oral toxicity of the test material was investigated in a study using male mice. The study was awarded a reliability score of 4 in accordance with the criteria set forth by Klimisch et al. (1997).

Mice were administered with a single dose of test material by oral gavage at 0.0, 4.0, 4.8, 5.8, 6.9, 8.3, 10.0, 12.0 and 14.3 g/kg. The animals were observed for mortality for a 14 day period at the following time points: Immediately after administration (0 hour), 2, 16, 24, 48, 72, 96, 120, 144, 168 and 336 hours after administration.

All animals in the 0 and 4 g/kg dose groups survived the duration of the observation period. 1, 2 and 6 animals died in the 4.8, 5.8 and 6.9 g/kg dose groups respectively during the 14 day observation period. In the 10.0, 12.0 and 14.3 g/kg dose groups no animals survived the 14 day observation period.

Under the conditions of the study the LD50 value of the test material was estimated to be 6.50 g/Kg bw (95 % confidence limits: 5.66 – 7.48 g/Kg).

Acute Inhalation Toxicity

Endpoint has been waived as testing by the inhalation route is not appropriate as exposure via inhalation is unlikely as the substance is a sticky, paste-like solid.

Acute Dermal Toxicity

Endpoint has been waived. The substance does not meet the criteria for classification as acute toxicity or STOT-SE by the oral route. In addition no systemic effects were observed in in vivo studies with dermal exposure, namely the in vivo skin irritation study and skin sensititsations studies. It can reasonably be predicted that the substance is not harmful by acute dermal exposure.

Justification for classification or non-classification

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.