Ammonium undecafluorohexanoate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 Feb - 04 Mar 2016

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Non-GLP study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

GLP compliance:

no

Remarks:

This study was not initially performed for REACh purposes but for other regulatory programs where the studies are not needed to be in compliance with GLP.

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Samples from all test solutions were analysed at test start and test end.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Required amount of the test sample and dilution water were mixed and stirred in test vessel to prepare the test solution.
- Eluate: no
- Differential loading: yes
- Controls: yes, test medium control

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: medaka
- Source: CERI Kurume (in-laboratory production)
- Length at study initiation (length definition, mean, range and SD): 2.0 ± 1.0 cm
- Age at study initiation: four months

ACCLIMATION
- Acclimation conditions (same as test or not): flow-through conditions at 24 ± 1 °C and 80% air saturation
- Type and amount of food during acclimation: Feed of Medaka for education (KYORlN); Amount corresponding to 3% of body weight was fed every day.
- Health during acclimation (any mortality observed): healthy status

FEEDING DURING TEST
- None

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

37 mg/L as CaCO3

Test temperature:

23.5 - 24.3 °C

pH:

7.4 - 7.8

Dissolved oxygen:

7.4 - 8.2 mg/L

Nominal and measured concentrations:

nominal: control, 911, 1280, 1790, 2500, 3500 mg/L
measured: n.d., 976, 1380, 1870, 2580, 3640 mg/L (geom. mean)

Details on test conditions:

TEST SYSTEM
- Test vessel: 3 L glass tank (diameter: 16 cm, depth: 16 cm), covered with a transparent plastic lid maintained in a water bath
- Aeration: gently aerated
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap water
- Total organic carbon: < 0.5 mg/L
- Particulate matter: < 1 mg/L
- Pesticides: not detected
- Chlorine: < 0.02 mg/L
- Alkalinity: 38 mg/L
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: Room light, 16-hour light/8-hour dark

EFFECT PARAMETERS MEASURED
- Mortality and visible abnormality were observed at 3, 24, 48, 72 and 96 h after the start of exposure.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.4
- Range finding study
- Test concentrations: 100, 316, 1000, 3160, 10000 mg/L
- Results used to determine the conditions for the definitive study: above 1000 mg/L all fish were dead

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1 850 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: 0%
- Abnormal responses: No abnormal response was obtained in the control. At the end of the study one fish was swimming at the surface and partial loss of equilibrium at 1280 mg/L.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The test solutions in all exposure levels were colorless and clear at the start of exposure, and a little bubbles were observed. The test solutions in exposure levels of 2500 and 3500 mg/L were colorless and clear at the time that all test organisms were confirmed to be dead, and a little bubbles were also observed. The test solutions in other exposure levels were slightly white suspended at the end of exposure. The test solution in the control was colorless and clear at the start and end of exposure.
- Effect concentrations exceeding solubility of substance in test medium: no

Reported statistics and error estimates:

Binominal test to determine the LC50.

Sublethal observations / clinical signs:

 Table 1: Cumulative mortality during exposure.

Nominal concentration [mg/L]	Cumulative mortality [%]
	3 h	24 h	48 h	72 h	96 h
Control	0	0	0	0	0
911	0	0	0	0	0
1280	0	0	0	0	0
1790	0	0	14	29	43
2500	14	100	100	100	100
3500	14	100	100	100	100

 

The measured concentrations of the test item in the test solutions were 945-3610 mg/L (102-106% of the nominal concentrations) at the start of exposure and 1010-3680 mg/L (104-111% of the nominal concentrations) at the end of exposure (including the time that all test organisms were confirmed to be dead), and kept within ± 20% of the nominal concentrations.

Validity criteria fulfilled:

yes

Reference 2

Endpoint:

fish embryo acute toxicity (FET)

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

12 - 17 Apr 2018

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 236 (Fish embryo acute toxicity (FET) test)

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test item was weighed and directly dissolved in dilution water.
- Eluate: no
- Controls: yes, test medium control

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Source: National Institute for Environmental Studies, Japan and Tsunashima Fishing Co.
- Collection of fertilized eggs: Freshly fertilized eggs were obtained from natural mating. Eggs were collected in 0.003% sodium hypochlorite solution as fungicide and rinsed three times with rearing water. Viable eggs were selected by microscopic observation.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

120 h

Test temperature:

28 ± 0.5 °C

Nominal and measured concentrations:

nominal: 100, 500, 1000, 1500, 2000 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 24-multiwell plate with cover
- Fill volume: 2 mL/well
- Renewal rate of test solution (frequency/flow rate): batchwise renewal at 48 h after start of exposure)
- No. of organisms per vessel: 20 eggs/test group (1 egg/well)
- No. of vessels per concentration (replicates): 1 vessel/test group (20 wells/test group)
- No. of vessels per control (replicates): 1 vessel/control (20 wells/control)

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated Yokohama city tap water processed with an activated charcoal filter and sodium thiosulfate

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light/8 h dark
- Light intensity: 800 lux or less

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Morphology of the embryo or larvae was assessed after 24, 48, 96 and 120 h. At the end of the test viable larvae were anesthetized with 1000 mg/L 2-phenoxyethanol and checked for morphological abnormalities. Mortality and hatching rate were assessed as well.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.3 - 5

Reference substance (positive control):

yes

Remarks:

sodium valproate (33.2 mg/L)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1 560 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI: 1380 - 1770 mg/L

Duration:

120 h

Dose descriptor:

LC50

Effect conc.:

1 220 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CI: 1080 - 1340 mg/L

Duration:

120 h

Dose descriptor:

NOEC

Effect conc.:

500 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

morphology

Reported statistics and error estimates:

The LC50 was derived by probit analysis. The NOEC was determined by Chi²-test (Yukms Statlight #5 software, Yukms Corp., Tokyo)

Sublethal observations / clinical signs:

Table 1: Mortality

Test group [mg/L]	Cumulative number of dead (mortality, %)
	24 hpf	48 hpf	96 hpf	120 hpf
Control	0 (0)	0 (0)	0 (0)	0 (0)
Positive control	0 (0)	0 (0)	0 (0)	0 (0)
100	0 (0)	0 (0)	0 (0)	0 (0)
500	0 (0)	0 (0)	0 (0)	0 (0)
1000	0 (0)	0 (0)	2 (10)	4 (20)
1500	0 (0)	0 (0)	7 (35)	16 (80)
2000	0 (0)	0 (0)	17 (85)	20 (100)

Hpf: hours post fertilization

Table 2: Hatching rate

Test concentration [mg/L]	Number of hatching	Hatching rate [%]
Control	20	100
Positive control	20	100
100	20	100
500	20	100
1000	18	90
1500	18	90
2000	18	90

Table 3: Frequency of morphological abnormalites

Test concentration [mg/L]	Number of morphological assessments	Number of morphological abnormalities *1	Frequency of morphological abnormality *2
Control	20	2	10
Positive control	20	20	100
100	20	1	5
500	20	4	20
1000	15	15	100
1500	4	4	100
2000	-	-	-

*1: Surviving larvae were assessed
*2: Non-inflated swim bladder was not considered as morphological abnormality

Table 4: Results of the morphological assessment

Organs	Major parameters observed as abnormality
	Positive control	Test substance
Body	Somite anomalies Notochord anomalies Fin anomalies	Somite anomalies Notochord anomalies Fin anomalies
Face/head	Lower jaw anomalies	-
Thoracic region	Pericardial edema Heart anomalies	Pericardial edema
Abdominal region	Abdominal edema	Abdominal organ anomalies Opaque intestinal tissue
Circlation	Blood circular anomalies Heartbeat anomalies	Blood circular anomalies Heartbeat anomalies

Description of key information

LC50 (96 h): 1850 mg/L (nominal; OECD 203)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

1 850 mg/L

Additional information

Two experimental studies are available investigating the short-term toxicity of ammonium undecafluorohexanoate (CAS 21615-47-4) to freshwater fish. The key study was performed according to OECD 203 (non-GLP) and the supporting study was performed according to OECD 236 (non-GLP).

Both studies were not performed in accordance with GLP since it was not initially performed for the purpose of a REACh registration but for other regulatory programs where the studies are not needed to be in compliance with GLP. Nevertheless, both studies were performed according to internationally accepted guidelines in a GLP accredited laboratory without any deviations to the guideline. Thus, the results are considered to be reliable and can be used for the hazard assessment. In the key study Oryzia latipes was exposed to nominal concentrations of 911, 1280, 1790, 2500, 3500 mg/L under static conditions for 96 h. The actual concentrations were analytically verified by HPLC/UV-VIS analysis and it was shown that the concentrations remained stable throughout the exposure. Geometric mean concentrations of 976, 1380, 1870, 2580, 3640 mg/L were determined. At the end of exposure a LC50 (96 h) of 1850 mg/L. The effect concentrations indicate that the substance will not cause mortality up to a limit concentration of 100 mg/L.

In the supporting study freshly fertilized eggs of Danio rerio were exposed to the test item up to 2000 mg/L in a semi-static testing regime. Morphological changes as well as mortality were recorded each 24 h up to 120 h of exposure. In accordance with the guideline the LC50 of 1560 mg/L after 96 h was used as key value. This effect concentration is in the same order of magnitude compared to the key study indicating low toxicity of the test item to fish.