Alkali metal tetrakis (perfluoro-alcoholato)metallate

Endpoint summary

Description of key information

The LD50 of the test item AM(pfa)4 is greater than 50 mg/kg and lower than 300 mg/kg body weight after single oral administration to Wistar rats.

Based on Annex 2d Test Procedure with a Starting Dose of 2000 mg/kg body weight of OECD Guideline 423 it can be concluded that the test item AM(pfa)4 is classified GHS Category 3 with a LD50 cut off value 200 mg/kg body weight, after single oral administration to Wistar rats.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

September 2020

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)

Version / remarks:

17.12.2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Specific details on test material used for the study:

Test Item: AM(pfa)4
Appearance: white powder

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Female rats were used as recommended by the OECD TG 423 guideline. Test Animal Species: Wistar rats, sourced from Institute of Experimental Pharmacology and Toxicology, Center of Experimental Medicine of the Slovak Academy of Sciences, Dobrá Voda, Slovak Republic
Number and Sex of Animals used in study: 12 females
Age at First Dose: 8 - 12 weeks; female animals were non-pregnant and nulliparous; the health condition of animals was examined by a veterinarian before initiation of the study and animals were acclimated to the condition identical to the condition during the experiment at least 5 days prior to the start of treatment. The acclimation was according to standard operation procedures.
Housing Condition: The animals were housed in plastic cages suspended on stainless steel racks, up to 3 animals per cage, in a room equipped with central air-conditioning. The room temperature was maintained within the range of 22 ±2 °C. The relative humidity was 55 ±10 %. The light regime was set to a 12-hour light / 12-hour dark cycle. The sanitation was performed according to standard operation procedures.
Diet: A standard laboratory food KKZ-P/M (UEFT CEM SAS) was available ad libitum. The animals received tap water for human consumption. Supply of drinking water was unlimited. The quality of drinking water is periodical monitored (including microbiological control) and recorded.
Bedding: AlpenSpan Eco, Johann Pabst Holzindustrie GmbH, Zeltweg, Austria
Animals Identification: Each animal was marked with an ID number. Each cage was affixed with a cage card containing pertinent animal and study information. The animals in cages were marked by a line on the tail with an ink marker.

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Aqua Pro Injectione Braun, B. Braun Melsungen AG, Germany, Lot Number 19144011

Details on oral exposure:

The required amount of the test item (according to the body weight and dose) was mixed with appropriate vehicle (Aqua pro injectione) shortly before administration. Dose was recalculated to concentration 100% of AM(pfa)4 (purity of test item was ˃ 96 %, typically 98.5 %). The test item was administered in a single dose by gavage using an oral gavage. Animals were fasted prior to dosing (food but not water was withheld over-night). Following the period of fasting, the animals were weighed and the test item was administered. After the test item administration, food was withheld for further 3 ‒ 4 hours.
The animals were observed individually immediately after the administration of the test item and then 0.5, 1, 2 and 4 hours later. Each animal was inspected daily for the next 14 days. Observations included: changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems, and somatomotor activity and behavioural patterns. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Individual weights of animals were determined shortly before the test item administration and weekly thereafter. Weight changes after first and second week after administration were calculated and recorded.
All test animals were subjected to gross necropsy. All gross pathological changes were recorded for each animal. Examinations included: external body surface and orifices, the appearance of tissues and organs in the thoracic cavity (trachea, esophagus, heart, aorta, lungs with main stem bronchi, thymus, tracheobronchial lymph node) and in the abdominal cavity (liver, spleen, adrenal glands, kidneys, ovaries, uterus including cervix, urinary bladder, small intestine, large intestine, pancreas, stomach, mesenteric lymph nodes).

Doses:

2000 mg/kg bw, 300 mg/kg bw, 50 mg/kg bw

No. of animals per sex per dose:

3 females per dose

Control animals:

no

Details on study design:

The starting dose was to be selected from the fixed dose levels of 5, 50, 300, and 2000 mg/kg body weight. Available information indicated that the test item was likely to be non-toxic regarding acute toxicity; therefore, a dose of 2000 mg of AM(pfa)4 per kg body weight to be used as a starting dose was selected. In the first step, one group of 3 females was dosed. The test item in limit dose caused mortality of all three animals within 4 hours after administration. In a second step, 3 females were treated at dose of 300 mg/kg body weight. The test item at this dose level caused mortality of all three animals within 4 hours after administration. In a third step, 3 females were treated at a dose of 50 mg/kg body weight. Test item-related mortality was observed only in Animal No. 9 within 24 hours and therefore, in a fourth step, 48 hours later another 3 females were treated at the same dose level and no mortality was found.

Sex:

female

Dose descriptor:

LD50

Effect level:

200 mg/kg bw

Based on:

test mat.

Mortality:

Mortality of 3/3 females at limit dose of 2000 mg/kg body weight and 3/3 females at the dose of 300 mg/kg body weight was observed. Five out of six female animals survived at the dose of 50 mg/kg body weight, and one animal died within 24 hours after administration. Summary results of clinical observations are presented in Tables 1 - 3.

Clinical signs:

other: Summary results of clinical observations are presented in Tables 1 - 3.

Gross pathology:

All animals were necropsied. No visible pathological findings were observed in animals dosed with 2000 and 300 mg/kg body weight. Similar findings were observed in Animal No 9, which died within 24 hours after administration of test substance. Necropsy of survived Animals 14 days after administration of test substance did not reveal any macroscopic findings (see Table 5).

Table 1. Clinical observation - 2000 mg/kg body weight, Animal No 1 - 3.

Observation	Time After Administration
	Hour					Day
	I	0.5	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
Skin and Hair**	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Eyes	-	-	-	-	2 lacrim	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Mucosa	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Respiratory System*	1,3	1,2,3	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Circulatory
System	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
CNS	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Somatomotoric Activity	1,3	1,2,3	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Tremor	1,3	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Spasms	1,3	1,2	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Salivation	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Diarrhoea	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Lethargy	1,2,3	1,2	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Sleep	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Coma	-	3	2	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Death	-	3	1	-	2	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Sacrificed	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Others	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

- No observed signs, I- immediately. * - dyspnoe, ** - piloerection

 

Sex	Dose	ID	Administration Result	Clinical Observation
☿	2000 mg/kg	1	death	- lethargy and sleep/sedative behaviour, tremors and spasms immediately after administration of test item, coma after 1 hour and 2 hours later: death of animal
		2	death	- lethargy and sleep/sedative behaviour, tremors and spasms immediately after administration of test item, coma after 2 hour and 4 hours later: death of animal, presence of lacrimation
		3	death	- lethargy and sleep/sedative behaviour immediately after administration of test item, ½ an hour later: death of animal

 

Table 2. Clinical observation - 300 mg/kg body weight, Animal No 4 - 6.

Observation	Time After Administration
	Hour					Day
	I	0.5	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
Skin and Hair**	-	4,5,6	4,5,6	4,5,6	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Eyes	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Mucosa	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Respiratory System*	-	-	5	4,5,6	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Circulatory
System	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
CNS	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Somatomotoric Activity	4,5,6	4,5,6	4,5,6	4,5,6	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Tremor	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Spasms	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Salivation	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Diarrhoea	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Lethargy	4,5,6	4,5,6	4,5,6	4,5,6	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Sleep	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Coma	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Death	-	-	-	-	4,5,6	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Sacrificed	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Others	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

- No observed signs, I - immediately. * - dyspnoe, ** - piloerection

 

Sex	Dose	ID	Administration Result	Clinical Observation
☿	300 mg/kg	4	death	- lethargy and somatomotor activity disturbances immediately after administration of test item, piloerection½an hour later, 1 hour later respiratory depression and 4 hours later death of animal
		5	death	- lethargy and somatomotor activity disturbances immediately after administration of test item, piloerection ½ an hour later, 2 hours later respiratory depression and 4 hours after administration of test item death of animal
		6	death	- lethargy and somatomotor activity disturbances immediately after administration of test item, piloerection ½ an hour later, 2 hours later respiratory depression and 4 hours after administration of test item death of animal

 

Table 3. Clinical observation - 50 mg/kg body weight, Animal No 7 - 12.

Observation	Time After Administration
	Hour					Day
	I	0.5	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
Skin and Hair**	-	10,11,12	10,11,12	10,11,12	10,11,12	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Eyes	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Mucosa	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Respiratory System*	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Circulatory
System	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
CNS	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Somatomotoric Activity	7,9	7,9	7,9	7,9
11	7,9	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Tremor	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Spasms	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Salivation	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Diarrhoea	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Lethargy	7,9	7,9	7,9	7,9	7,9	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Sleep	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Coma	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Death	-	-	-	-	-	9	-	-	-	-	-	-	-	-	-	-	-	-	-
Sacrificed	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
Others	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

- No observed signs, I - immediately. * - dyspnoe, ** - piloerection

 

Sex	Dose	ID	Administration Result	Clinical Observation
☿	50 mg/kg	7	alive	- lethargy and somatomotor activity disturbances immediately after administration of test item, which persisted for 4 hours. No other adverse reactions during 14-days observation period were observed.
		8	alive	no signs of intoxication, change of health, nor any other adverse reactions during 24 hours and 14-days observation period
		9	death	- lethargy and somatomotor activity disturbances immediately after administration of test item, which persisted for 4 hours. - 1 day after administration death of animal
		10	alive	- piloerection ½ an hour after administration and persisted for 4 hours. No other adverse reactions during 14-days observation period were observed.
		11	alive	- piloerection ½ an hour after administration and persisted for 4 hours, somatomotor activity disturbances observed 2 hours after. No other adverse reactions during 14-days observation period were observed.
		12	alive	- piloerection ½ an hour after administration and persisted for 4 hours. No other adverse reactions during 14-days observation period were observed.

 

Table 4. Body Weight

Sex	Dose	ID	Body Weight (g)			Body Weight Difference (g)
			Initial	Week 1	Week 2	Week 1 - Initial	Week 2 - Initial	Week 2 - Week 1
☿	2000 mg/kg	1	165	-	-	-	-	-
		2	168	-	-	-	-	-
		3	160	-	-	-	-	-
☿	300 mg/kg	4	188	-	-	-	-	-
		5	178	-	-	-	-	-
		6	195	-	-	-	-	-
☿	50 mg/kg	7	177	182	198	5	21	16
		8	203	218	237	15	34	19
		9	175	-	-	-	-	-
		10	165	203	217	38	52	14
		11	181	199	223	18	42	24
		12	171	202	219	31	48	17

 

Table 5. Necropsy Results                                          

Sex	Dose	ID	Result
☿	2000 mg/kg	1	no pathological findings
		2	no pathological findings
		3	no pathological findings
☿	300 mg/kg	4	no pathological findings
		5	no pathological findings
		6	no pathological findings
☿	50 mg/kg	7	no pathological findings
		8	no pathological findings
		9	no pathological findings
		10	no pathological findings
		11	no pathological findings
		12	no pathological findings

 

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

The LD50 of the test item AM(pfa)4 is greater than 50 mg/kg and lower than 300 mg/kg body weight after single oral administration to Wistar rats.
Based on Annex 2d Test Procedure with a Starting Dose of 2000 mg/kg body weight of OECD Guideline 423 it can be concluded that the test item AM(pfa)4 is classified GHS Category 3 with a LD50 cut off value 200 mg/kg body weight, after single oral administration to Wistar rats.

Executive summary:

The purpose of the study was to evaluate the potential toxic effect of the test item AM(pfa)4 when administered as a single oral dose to Wistar rats. The procedure according to OECD Guideline 423 Acute Toxic Class (ATC) method was used.

A limit dose of 2000 mg/kg body weight was used as a starting dose. Available information indicated that the test item is likely to be non-toxic regarding acute toxicity; therefore, a limit dose of 2000 mg/kg body weight was used as a starting dose. In the first step, one group of 3 females was dosed. The test item in limit dose caused mortality of all animals within 4 hours after administration. In a second step, 3 females were treated with dose of 300 mg/kg body weight. The test item in the dose of 300 mg/kg caused mortality of all animals within 4 hours after administration. In a third step, 3 females were treated with dose of 50 mg/kg body weight. The test item in dose 50 mg/kg caused death of Animal No 9 within the 24 hours. Animals No 7 and 8 survived and therefore another 3 females were treated 48 hours later at the same dose level and Animals No 10 - 12 survived.

The test item AM(pfa)4 administered to 3 females Wistar rats at a limit dose of 2000 mg/kg caused strong reaction immediately after administration of test item (tremor, spasm, breathing difficulties, coma). Within four hours all animals died. During necropsy no obvious pathological changes in gastrointestinal tract or other organs were observed.

The dose of 300 mg/kg of body weight caused immediate somatomotoric depression and lethargy with piloerection and breathing difficulties 1 hour after administration of test item. Within four hours all animals died. During necropsy no obvious pathological changes in gastrointestinal tract or other organs were seen.

The dose of 50 mg/kg caused death within 24h after administration of animal No 9, which preceded by lethargy and somatomotor disturbances. No other death at this dose was observed. During necropsy of animal No 9 no visible pathological changes were found. Necropsy of survived Animals 14 days after administration of test substance did not reveal any macroscopic findings at this dose level.

The LD50 of the test item AM(pfa)4 is greater than 50 mg/kg and lower than 300 mg/kg body weight after single oral administration to Wistar rats.

Based on Annex 2d Test Procedure with a Starting Dose of 2000 mg/kg body weight of OECD Guideline 423 it can be concluded that the test item AM(pfa)4 is classified GHS Category 3 with a LD50 cut off value 200 mg/kg body weight, after single oral administration to Wistar rats.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

200 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

Justification for classification or non-classification

Based on the results of an acute toxicity study (toxic class method) the test item AM(pfa)4 is classified for acute oral toxicity to CLP (Regulation EC No 1272/2008) Category 3 with a LD50 cut off value 200 mg/kg body weight.