2-nitro-4-(trifluoromethyl)benzonitrile

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

05 – 16 Mar 2019

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

GLP compliance:

yes

Remarks:

The test was performed in compliance with the requirements of the Principles of Good Laboratory Practices (GB/T 22278-2008).

Test material

Test material information

Constituent 1

Reference substance name:

2-nitro-4-(trifluoromethyl)benzonitrile

EC Number:

212-298-1

EC Name:

2-nitro-4-(trifluoromethyl)benzonitrile

Cas Number:

778-94-9

Molecular formula:

C8H3F3N2O2

IUPAC Name:

2-nitro-4-(trifluoromethyl)benzonitrile

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Control, 1.32, 1.98, 2.97, 4.45, and 6.67 mg/L
- Sampling method: Water samples were taken at 0, 24, 48, 72, and 96 h

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 0.2020 g was weighed, and test water used to dilute to 2 L, then stirred continuously for 30 minutes at 500 rpm at room temperature (23-25 °C), yielding the stock solution with a 100 mg/L concentration.
- Other relevant information: The test substance was fully dissolved.

Test organisms

Test organisms (species):

other: Gobiocypris rarus

Details on test organisms:

TEST ORGANISM
- Common name: rare minnow
- Strain: Batch number: F20181121G
- Source: Institute of Hydrobiology of the Chinese Academy of Sciences
- Length at study initiation (length definition, mean, range and SD): 3.37 cm (SD 4.04%)
- Weight at study initiation (mean and range, SD): 0.257 g (SD 8.38%)

ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions (same as test or not): The test fish underwent preliminary rearing in aerated, circulating water for a minimum of 12 days or more, during which period fish with abnormal growth or dead fish were removed. Prior to testing, they were acclimated in the test environment for 7 days.
- Type and amount of food during acclimation: They were fed newly hatched Artemia salina
- Feeding frequency during acclimation: Twice daily, terminating 24 h prior to the start of the test.

FEEDING DURING TEST
- Food type: No feeding

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Hardness:

148 mg (CaCO3)/L

Test temperature:

22.6 - 23.5 °C

pH:

7.82 - 8.29

Dissolved oxygen:

68.4 - 104.6%

Nominal and measured concentrations:

Nominal concentrations: 0.00 (Control), 1.32, 1.98, 2.97, 4.45, and 6.67 mg/L
Mean measured concentrations: < LOD/LOQ, 1.28, 1.95, 2.91, 4.30, and 6.62 mg/L

Details on test conditions:

TEST SYSTEM
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- Biomass loading rate: 0.257 g (wet weight)/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water aerated for 24 h or more
- Metals: Arsenic 0.5 μg/L; Cadmium <0.5 μg/L; Lead <2.5 μg/L; Cobalt < 0.02 mg/L; Nickel < 0.05 mg/L; Mercury 0.10 μg/L; Copper 5.2 μg/L; Chromium6+ 0.004 mg/L; Iron <0.01 mg/L; Zinc < 0.09 mg/L; Silver < 0.03 mg/L
- Culture medium different from test medium: No

OTHER TEST CONDITIONS
- Photoperiod: 12 h light each day

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
Every day, test temperature, pH, and dissolved oxygen water quality parameters were measured and recorded.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.5
- Range finding study: yes
- Test concentrations: Control, 0.100, 1.00, and 10.0 mg/L
- Results used to determine the conditions for the definitive study: Maximum concentration causing zero mortality in the test fish (LC0) and minimum concentration causing 100% mortality in the test fish (LC100) were recorded.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.47 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

1.28 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Other biological observations: At concentrations 2.97 mg/L and higher fish lay flat or side-on. After 96 h of exposure, the test fish in the 2.97 mg/L, 4.45 mg/L, and 6.67 mg/L treatment groups were all dead.
- Mortality of control: Swimming behaviour of the test fish in the blank control group was normal and there were no deaths.

Results with reference substance (positive control):

- Results with reference substance were valid.
- Mortality: Up to a potassium dichromate concentration of 200 mg/L no mortality was observed.
- LC50: The potassium dichromate 24 h-LC50 in the test fish was between 200 mg/L - 400 mg/L.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Table 1: Measurement Results for Stability of the Test Substance in Test Medium

Prepared concentration (mg/L)	Measured concentration (mg/L)
	0 h	24 h	48 h	72 h	96 h	Mean
0	ND(1)	ND	ND	ND	ND	-
1.32	1.33	1.31	1.28	1.26	1.23	1.28
1.98	2.00	1.97	1.96	1.94	1.88	1.95
2.97	3.00	2.96	2.91	2.88	2.80	2.91
4.45	4.41	4.33	4.28	4.16	-	4.30
6.67	6.71	6.59	6.55	-	-	6.62

(1) ND: lower than the limit of detection

- : 100% mortality in test fish not detected.

 

 

Table 2: Test Fish Fatalities 

Prepared concentration (mg/L)	Measured concentration (mg/L)		Number of fish (n)	Number of dead fish (n)
				3h	6h	24 h	48 h	72 h	96 h
0	ND		10	0	0	0	0	0	0
1.32	1.28		10	0	0	0	0	0	3
1.98	1.95		10	0	0	1	1	8	9
2.97	2.91		10	0	0	1	9	10	10
4.45	4.30		10	0	0	5	10	10	10
6.67	6.62		10	0	2	10	10	10	10
LC50 (mg/L) (based on actual measured concentration)						4.01	2.38	1.72	1.47
95% confidence interval (mg/L) (based on actual measured concentration)						3.38-4.75	2.13-2.65	1.55-1.90	1.28-1.70

 

 Table 3: Validity criteria for OECD 203 (2019)

Criterion from the guideline	Outcome	Validity criterion fulfilled
The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test	Mortality rate in the blank control group was 0%.	yes
The dissolved oxygen concentration must have been at least 60% of the air saturation value throughout the test	Dissolved oxygen: greater than 68.4% of the air saturation value.	yes
There must be evidence that the concentration of the substance being tested has been satisfactorily maintained, and preferably it should be at least 80% of the nominal concentration throughout the test. If the deviation from the nominal concentration is greater than 20%, results should be based on the measured concentration.	Change in concentration was less than 7% of the initial concentration.	yes

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

See Table 3 in "Any other information on results incl. tables".