Trifluoroacetic acid

Long-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

fish early-life stage toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2017-05-23 to 2019-02-20

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

For the Reaction mass of potassium trifluoroacetate and potassium trifluoromethanesulphinate a new chemical substance notification in China has been prepared under the Regulation ‘Measures on the Environmental Management of New Chemical Substances' (Decree No. 7 of the Ministry of Environmental Protection of the P.R. China, also known as ‘China REACH’). Under this regulation a long-term toxicity test in fish is part of the data requirements for substances that are produced or imported in volumes >100 t/y. For this reason a long-term toxicity test in fish was performed at Bioassay and Safety Assessment Laboratory, APM (Shanghai, China) under GLP conditions. The results of the study were included in the dossier and serve as the valid testing for this endpoint. More information about the read-across justification is included in the Reporting format as attached to the respective IUCLID entry (section 13).

Qualifier:

according to guideline

Guideline:

OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)

Version / remarks:

2013

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Certificate from 25 September 2017

Analytical monitoring:

yes

Remarks:

Ionic Chromatography (IC)

Details on sampling:

- Concentrations: Sampling in the control vessel and the vessel exposed to the limit concentration (10 mg/L test substance nominal).
- Sampling method: 10 mL of test solution was taken from the test vessel which was selected randomly from each group, at the beginning of the test, twice each week thereafter and at the end of the test.
- Sample storage conditions before analysis: The samples were analyzed directly without storage.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A stock solution of 1 g/L was prepared by dissolving 2.0004 g test substance in 2 L test medium. The secondary stock solution of 100 mg/L was prepared by dilution of the stock solution and the test solution of 10 mg/L was prepared by dilution of the secondary stock solution with test water. The secondary stock solution was renewed once in two days (0-10 days) and every day after that.
- Controls: dilution medium

Test organisms (species):

other: Gobiocypris rarus

Details on test organisms:

TEST ORGANISM
- Common name: Chinese Rare Minnow
- Strain: Gobiocypris rarus
- Source: Shanghai Academy of Public Measurement (ID: GR-SAPM-171026)

METHOD FOR PREPARATION AND COLLECTION OF FERTILIZED EGGS
- Numbers of parental fish (i.e. of females used to provide required number of eggs): Embryos collected for use in the test were from three individual spawns.
- Method of collection of fertilised eggs: The embryos were removed from the spawning substrates and examined under dissection microscope to select healthy, viable specimens embryos.
- Subsequent handling of eggs: The embryos were immersed in the test solutions before cleavage of the blastodisc started. The embryos used in the test were distributed at random and unproperly treated embryos were discarded. The test started within 8 hours post-fertilization of the eggs.

POST-HATCH FEEDING
- Start date: after hatching
- Type/source of feed: newly bred fish were fed with live food of brine shrimp (Ocean Star International, Inc., USA)
- Amount given: ad libitum; the food ration was adjusted to the number of living fish per test vessel.
- Frequency of feeding: twice a day and two feeds were separated by 6 hours. Uneaten food and fecal material were removed from the test vessels each day at 30 minutes after the second feeding by carefully cleaning the bottom of each tank using a suction siphon.

Test type:

flow-through

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

35 d

Post exposure observation period:

not applicable

Hardness:

Test initiation: 95 mg CaCO3/L (control), 102 mg CaCO3/L (test solution)
Test termination: 98mg CaCO3/L (control), 104 mg CaCO3/L (test solution)

Test temperature:

24.3-26.1°C

pH:

6.59 - 8.03

Dissolved oxygen:

60.3 - 93.7% ASV

Salinity:

not applicable

Nominal and measured concentrations:

- Nominal concentrations: 10 mg test item/L, plus a control
- Analysed concentrations: 10.3 mg/L (arithmetic mean measured concentration).
The results of the measured concentrations are provided in Table 1 "Any other information on results incl. tables"

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass test chambers containing 100 mL test solution at day 0-10; glass test chambers containing 250 mL test solution at day 11-35.
- Type of flow-through: peristaltic
- Renewal rate of test solution (frequency/flow rate): The test solutions flowed through each test chamber at a flow-rateof 0.347 mL/min and 0.5 litres of test solution was renewed per 24 hours from day 0 to day 10; the flow rate was increased to 1.04 mL/min and 1.5 litres of test solution flowed through each test chamber per 24 hours from days 11 to day 35.
- No. of fertilized eggs/embryos per vessel: 20
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: 0.359 g fish/L per 24h (at the end of the test in control) and about 2.15 g fish/L of solution at any time.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The test water used was tap water, dechlorinated by activated carbon and sterilized by UV lamp. The water was aerated continuously over 24h.
- Total organic carbon: 1.10 mg/L
- Particulate matter: 3.8 mg/L
- Total hardness: 108 mg/L
- Total chlorine: 0.24 mg/L
- Oxygen concentration: >90% ASV
- Nitrite: 2.0 mg/L
- Nitrate: 3.6 mg/L
- Metals:
· Cobalt (Co): 0.0945 µg/L
· Copper (Cu): 0.776 µg/L
· Lead (Pb): 0.0430 µg/L
· Silver (Ag): ND
· Cadmium (Cd): 0.0155 µg/L
· Mercury (Hg): ND
· Chromium (Cr): 0.286 µg/L
· Arsenic (As): 0.533 µg/L
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Temperature: 21-25°C maintained within a range of ± 2°C
- Photoperiod: 12/12 hours light/dark cycle

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
The fish were inspected and recorded once a day for hatching, mortality and other observed effects (i.e. abnormal appearance and abnormal behaviour). Dead embryos and fish were removed when observed. All survivors were treated by anaesthesia with 80mg/L MS-222 at the end of the test. The wet weight (blotted dry) and lengths are measured individually by test vessel, dry weight (24 hours at 60℃) were weighed in groups by test vessel. Fish were not weighed during the test.
Total hardness was measured in the control and the highest concentration at the beginning and the end of the test. During the test, measurements of pH and dissolved oxygen were carried out three times every week, the temperature in the control and the treatment group was measured daily.

DATA EVALUATION
To determine the lowest observed effect concentration (LOEC) and the no observed effect concentration (NOEC), the variances among the means of the specific growth rate in all groups was analyzed by one-way ANOVA and the mean of treatment groups was compared with the control group using Williams Multiple Sequential t-test. The data treatment was carried out with software TOXRAT (v3.0.beta). The results were expressed as the nominal concentration.

RANGE-FINDING STUDY
- No range-finding study was performed.

Reference substance (positive control):

no

Duration:

35 d

Dose descriptor:

NOEC

Effect conc.:

>= 10 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: Hatching success, mortality (post-hatch success), numbers of healthy fish, length of the surviving fish, dry weight of the surviving fish

Duration:

35 d

Dose descriptor:

LOEC

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: Hatching success, mortality (post-hatch success), numbers of healthy fish, length of the surviving fish, dry weight of the surviving fish

Details on results:

- The measured concentrations of test substance were within ± 20% (-14.6%-11.9%) of the nominal concentration (see Table 1, "Any other information on results incl. tables"). Therefore, the results are reported in nominal concentration.
- Mortality/survival: Larval mortality on Day 35 in the control and treatment groups were both 0%.
- Days to hatch and hatching succes: All of the fish were hatched on Day 5 and the hatching success rate in the control group and treatment group were both 100% (see Table 2, "Any other information on results incl. tables").
- Observations on body length and weight of young: Growth measurements for total length, wet weight and dry weight at the end of the 35-day period are summarized in Tables 3 and 4 ("Any other information on results incl. tables"). Length of the fish hatching in the control group for each replicate were 1.2 ± 0.1 cm; 1.1 ± 0.1 cm; 1.1 ± 0.1 cm and 1.1 ± 0.1 cm（Mean ± SD) and fresh weight were 0.0288 ± 0.007 g ;0.0253 ± 0.006 g; 0.0264 ± 0.007 g and 0.0272 ± 0.004 g (Mean ± SD) at the end of the test. Based on the fresh weight of the survived fishes at the end of the test, the treatment group was not significantly different from that of the control group ( p = 0.175, p >0.05).
Based on the dry weight of the survived fishes at the end of the test, the treatment group was not significantly different from that of the control group ( p = 0.598, p >0.05). Based on the length of the survived fish at the end of the test, the treatment group was not significantly different from that of the control group (p = 0356, p >0.05). Under the test conditions, the EC50 of the growth inhibition for Gobiocypris rarus exposed to the test substance for 35 days was >10 mg/L; the NOEC was ≥10 mg/L and the LOEC was >10 mg/L.
- Number of healthy fish at end of test: All surviving fish appeared healthy at the end of exposure, i.e., behavioural or morphological abnormalities were not observed. Therefore, a separate statistical evaluation was not performed on the number of healthy fish.
- Validity Criteria:
· The hatching success of the fertilised eggs in the control was 100%
· The overall survival of the post-hatch fish in the control was 100%
· The dissolved oxygen concentrationh was greater than 60.0% (60.3% - 93.7%) of the air saturation value throughout the test
· The water temperature of 25°C did not differ by more than ± 1.5°C (24.3°C - 26.1°C) during the test.
So the validity criteria of the OECD Test Guideline were fulfilled and the test is considered valid.

Table 1: Measured concentration of the test substance in the test solutions

Duration	Blank control	Treatment group (nominal concentration 10 mg/L)
0d	ND	11.2
4d	ND	10.4
6d	ND	11.5
8d	ND	9.74
11d	ND	10.5
13d	ND	10.2
15d	ND	9.93
18d	ND	9.51
20d	ND	9.92
22d	ND	11.3
25d	ND	10.5
27d	ND	10.1
29d	ND	10.4
32d	ND	10.2
35d	ND	8.78
Arithmetic mean measured concentration	-	10.3
Variation (%)	-	-12.2 – 15.0

ND = < LOD (< 2.12 mg/L)

‘-’ = not available

Table 2: Egg survival and hatching success

Group		Duration (d)
		0	1	2	3	4	5
Control group	1	0	0	0	13	20	20
	2	0	0	0	13	20	20
	3	0	0	0	14	20	20
	4	0	0	0	17	20	20
Treatment group (Arithmetic mean measured concentration, 10.3mg/L)	1	0	0	0	10	19	20
	2	0	0	0	15	20	20
	3	0	0	0	8	20	20
	4	0	0	0	10	20	20

Table 3: Body weight and the length of the fish at the end of the test period

Replicate Group		Body weight (g)				length (cm)
		1	2	3	4	1	2	3	4
Blank control	1	0.0257	0.0166	0.0144	0.0303	1.2	0.9	0.9	1.2
	2	0.0416	0.0318	0.0194	0.0330	1.4	1.2	1.0	1.2
	3	0.0389	0.0230	0.0245	0.0213	1.3	1.0	1.2	1.1
	4	0.0266	0.0294	0.0302	0.0285	1.1	1.3	1.2	1.1
	5	0.0209	0.0303	0.0216	0.0326	1.1	1.2	1.0	1.3
	6	0.0407	0.0362	0.0191	0.0260	1.2	1.4	1.1	1.2
	7	0.0346	0.0221	0.0403	0.0326	1.3	1.1	1.3	1.2
	8	0.0290	0.0379	0.0274	0.0269	1.2	1.3	1.1	1.1
	9	0.0262	0.0417	0.0304	0.0270	1.2	1.4	1.1	1.2
	10	0.0251	0.0293	0.0248	0.0255	1.2	1.1	1.2	1.1
	11	0.0375	0.0233	0.0278	0.0276	1.4	1.1	1.2	1.2
	12	0.0312	0.0170	0.0322	0.0257	1.3	1.1	1.2	1.1
	13	0.0262	0.0160	0.0269	0.0208	1.1	0.9	1.1	1.1
	14	0.0250	0.0223	0.0233	0.0308	1.1	1.2	1.0	1.1
	15	0.0305	0.0253	0.0222	0.0260	1.3	1.1	1.0	1.1
	16	0.0248	0.0309	0.0273	0.0293	1.3	1.3	1.2	1.2
	17	0.0237	0.0255	0.0274	0.0268	1.2	1.2	1.2	1.1
	18	0.0241	0.0281	0.0195	0.0224	1.2	1.1	1.0	1.1
	19	0.0198	0.0144	0.0284	0.0276	0.9	0.8	1.1	1.1
	20	0.0245	0.0264	0.0198	0.0230	1.1	0.9	1.1	1.0
	Mean	0.0288	0.0264	0.0253	0.0272	1.2	1.1	1.1	1.1
	SD	0.007	0.007	0.006	0.004	0.11	0.17	0.10	0.07
	CV	8.98				6.77
Treatment group (Arithmetic mean meas concentration, 10.3 mg/L)	1	0.0288	0.0305	0.0372	0.0267	1.3	1.2	1.3	1.0
	2	0.0274	0.0282	0.0246	0.0262	1.2	1.1	1.1	1.1
	3	0.0168	0.0371	0.0174	0.0340	1.0	1.2	1.0	1.2
	4	0.0326	0.0279	0.0194	0.0221	1.4	1.1	1.1	1.0
	5	0.0288	0.0211	0.0220	0.0312	1.2	1.0	1.1	1.1
	6	0.0228	0.0224	0.0299	0.0297	1.1	1.1	1.2	1.2
	7	0.0187	0.0303	0.0177	0.0219	1.0	1.2	1.1	1.1
	8	0.0172	0.0218	0.0254	0.0194	1.0	1.0	1.1	1.0
	9	0.0159	0.0234	0.0135	0.0456	0.9	1.1	0.9	1.4
	10	0.0246	0.0200	0.0250	0.0273	1.1	1.0	1.1	1.2
	11	0.0278	0.0245	0.0229	0.0235	1.1	1.1	1.1	1.0
	12	0.0246	0.0285	0.0256	0.0274	1.2	1.1	1.2	1.1
	13	0.0192	0.0233	0.0188	0.0171	1.0	1.1	1.1	0.9
	14	0.0290	0.0297	0.0171	0.0283	1.2	1.2	1.0	1.1
	15	0.0169	0.0356	0.0309	0.0371	0.9	1.2	1.2	1.3
	16	0.0307	0.0266	0.0254	0.0245	1.3	1.1	1.1	1.1
	17	0.0239	0.0257	0.0191	0.0259	1.0	1.1	1.0	1.1
	18	0.0224	0.0313	0.0196	0.0216	1.0	1.1	1.1	1.1
	19	0.0186	0.0214	0.0228	0.0278	1.0	1.1	1.2	1.2
	20	0.033	0.0200	0.0355	0.0196	1.1	1.1	1.2	1.1

 

Table 4: Dry body weight of the fish at the end of the test period

Replicate	Dry weight (g)
	Control group	Treatment group (Arithmetic mean measured concentration,10.3 mg/L)
1	0.113	0.0965
2	0.113	0.111
3	0.0988	0.101
4	0.107	0.117
Mean	0.108	0.106

 

Validity criteria fulfilled:

yes

Remarks:

see "Details on results"

Conclusions:

A GLP-compliant Fish Early-Life Stage Toxicity Test was conducted with the Reaction mass of TFAK/TFSK according to OECD 210 with Gobiocypris rarus under flow-through conditions as part of a new chemical substance notification in China.
Based on nominal concentrations, the NOEC was ≥10 mg/L and the LOEC was >10 mg/L (corresponding to ≥ 3.8 mg TFA/L).

Executive summary:

A GLP-compliant Fish Early-Life Stage Toxicity Test was conducted with the Reaction mass of potassium trifluoroacetate (TFAK) and potassium trifluoromethanesulphinate (TFSK) according to OECD 210 with Gobiocypris rarus under flow-through conditions as part of a new chemical substance notification in China. Based on the results of the acute fish test showing no acute toxicity at the limit concentration, a limit test at a nominal concentration of 10 mg/L was performed. At the start of the test, 80 fertilised eggs were randomly distributed and equally divided between four replicates in the control and treatment groups. The test duration was 35 days. The fish were inspected and recorded once daily for hatching, mortality and other observed effects. At the end of the test, the surviving fish were weighed and individual lengths were measured.

The hatching success of fertilised eggs in the control was 100% and the overall survival of the post-hatch fish in the control was 100%. The dissolved oxygen concentration was greater than 60% (60.3%-93.7%) of the air saturation value throughout the test. The water temperature of 25°C did not differ by more than ± 1.5°C (24.3 -26.1°C) during the test. Therefore the validity criteria were fulfilled and the test is valid.

The concentrations of the test substance in the test solutions were analyzed during Ion Chromatography and the results demonstrated that the exposure has been satisfactorily maintained within 20% of the nominal concentration. The arithmetic mean measured concentration of the test substance was 10.3 mg/L.

Under the test conditions, all embryos were hatched within 5 days and there was no mortality in the treatment group. Therefore the No Observed Effect Concentration (NOEC) was ≥10 mg/L, the Lowest Observed Effect Concentration was >10 mg/L and the EC50 was >10 mg/L. Based on the measurements of the fresh and dry weight at the end of the test, as well as the length of the surviving fish, the NOEC was ≥10 mg/L and the LOEC was >10 mg/L (corresponding to ≥3.8 mg TFA/L).

Description of key information

A GLP-compliant fish, early-life stage toxicity test was conducted with the Reaction mass of potassium trifluoroacetate and potassium trifluoromethanesulphinate according to OECD 210 with Gobiocypris rarus under flow-through conditions as part of a new chemical substance notification in China. Based on nominal concentrations, the NOEC was >10 mg/L and the LOEC was >10 mg/L (corresponding to >3.8 mg TFA/L).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

NOEC

Effect concentration:

>= 3.8 mg/L

Additional information

A GLP-compliant Fish Early-Life Stage Toxicity Test was conducted with the Reaction mass of potassium trifluoroacetate and potassium trifluoromethanesulphinate according to OECD 210 with Gobiocypris rarus under flow-through conditions as part of a new chemical substance notification in China. Based on the results of the acute fish test showing no acute toxicity at the limit concentration of 122 mg/L, a limit test at a nominal concentration of 10 mg/L was performed. At the start of the test, 80 fertilised eggs were randomly distributed and equally divided between four replicates in the control and treatment groups. The test duration was 35 days.

The hatching success of fertilised eggs and the overal survival of fish larvae in the blank control was 100% and the dissolved oxygen concentration was greater than 60.0% of the air saturation value throughout the test. The concentrations of the test substance in the test solutions were analyzed during Ion Chromatography and the results demonstrated that the exposure has been satisfactorily maintained within 20% of the nominal concentration. The arithmetic mean measured concentration of the test substance was 10.3 mg/L.

Under the test conditions, all embryos were hatched within 5 days and there was no mortality in the treatment group. Therefore the No Observed Effect Concentration (NOEC) was ≥10 mg/L, the Lowest Observed Effect Concentration was >10 mg/L and the EC50 was >10 mg/L (corresponding to >3.8 mg TFA/L). Based on the measurements of the fresh and dry weight at the end of the test, as well as the length of the surviving fish, the NOEC was ≥10 mg/L and the LOEC was >10 mg/L (corresponding to >3.8 mg TFA/L).

.