Diphenylacetonitrile

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Test item weighed individually for all concentrations & formulation was prepared in glass vial, then respective volumes of solution were transferred from vial to aquarium to obtain the required concentrations for the experiment.

All the test concentrations were analysed for test substance verification.

Test vessels were immediately analysed after sampling

Vehicle:

no

Details on test solutions:

The test solution was prepared by dissolving 500 mg of the test substance in 500 mL of potable water, which was prepared passing water through reverse osmosis system. The above stock solution was stirred for 72 hrs and filtered through vaccum filter. The stock solution was analytically detectedand the fincal expsure concentration was 0.58, 0.88, 1.33, 2.0 and 3.0 mg/l respectively.

- Controls: potable water
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Not applicable
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): Not applicable

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Source: Koyal aquarium, India
- Age at study initiation (mean and range, SD): Juveniles (originate fromsame source and population)
- Length at study initiation (length definition, mean, range and SD): 1.4 cm on average
- Weight at study initiation (mean and range, SD): 0.012 g on average


ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions (same as test or not): same as test
- Type and amount of food during acclimation: Commercial fish food.at adlibitum
- Feeding frequency during acclimation: Daily
- Health during acclimation (any mortality observed): healthy



FEEDING DURING TEST
NO feeding during test

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

158 mg CaCO3/L

Test temperature:

22.-22.7 °C

pH:

6.4-7.1

Dissolved oxygen:

6.8-8.5 mg/L

Salinity:

-

Conductivity:

-

Nominal and measured concentrations:

Nominal Concentrations: 0.58, 0.88, 1.33, 2.0 and 3.0 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquarium
- Material, size, headspace, fill volume: Glass aquarium 5 liters
- Aeration: no aeration duration test
- No. of organisms per vessel: 07
- No. of vessels per concentration (replicates): 01
- No. of vessels per control (replicates): 01
- No. of vessels per vehicle control (replicates): 01
- Biomass loading rate: 0.012 g fish/L


OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod: 16 h light- 8 h dark
- Light intensity: 730-784 Lux


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.5
- Range finding study : No, Main study was directly initated based on the readily availble data.
- Test concentrations: 0.58, 0.88, 1.33, 2.0 and 3.0 mg/l

Reference substance (positive control):

not required

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.437 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: behvaioural abonromalitie were observed in control group and exposure group
- Mortality of control: No
- Other adverse effects control: No

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Based on the mortality data, LC50 value was determined statistically by probit analysis.

Sublethal observations / clinical signs:

Conc. mg/L	2.5h	5h	24h	48h	72h	96h	Cumulative mortality
Control	0	0	0	0	0	0	0
0.58	0	0	0	0	0	0	0
0.88	0	0	0	0	1	0	1
1.33	0	0	1	0	1	0	2
2.0	0	0	3	0	1	2	6
3.0	0	0	3	4	-	-	7

Validity criteria fulfilled:

yes

Remarks:

No mortality in control were found throughout test duration. Dissolve oxygen concentration was maintained above 60% in all test vessels throughout the test. The test item was found to be between 80-120% up to 96h.

Conclusions:

The 96 h LC50 of test item to zebrafish (Danio rerio) was evaluated to be 1.43 mg/L.

Executive summary:

This study was conducted as per OECD 203 (2019) to assess the acute toxicity effects of test chemical on zebrafish (Danio rerio) following exposure up to 96 h under static condition. Juvenile fish of same age and normal in appearance were used in this (originate from same source and population). The average length and weight (10 fish) were observed 1.3 cm and 0.012g, respectively. Fish were fed (commercial fish food) daily during acclimatization. Photoperiod and light intensity were maintained such as16 h light- 8 h dark, 735 -800 Lux during experiment. Hardness of water was measured once during acclimatization and found to be 165 mg CaCO3/L, temperature, pH and dissolve oxygen were maintained between 22.-22.7 °C,6.4-7.1,6.8-8.5 mg/L, respectively, throughout the test. Fish were acclimatized for 7 days prior dosing. The test solution was prepared by dissolving 500 mg of the test substance in 500 mL of potable water, which was prepared passing water through reverse osmosis system. The above stock solution was stirred for 72 hrs and filtered through vaccum filter.  The stock solution was analytically detectedand the fincal expsure concentration was 0.58, 0.88, 1.33, 2.0 and 3.0 mg/l respectively. there wre no mortality in the control and lowest concentration. and at 0.88 mg/L, 1.33 mg/l, 2.0 mg/l, and 3.0 mg/l the mortality was observed to be 14.2%, 28.57%, 85.71% and 100%. The analytical monitoring of test subsatnce was performed, which was maintained to be with in the range of 80 -120 of nominal test concentrations. The test is valid as all the validity criteria are fulfilled: No mortality in control or vehicle control were found throughout the 96-h test duration; Dissolve oxygen conc. was maintained above 60% in all test vessels throughout the test; The recovery active ingredient content was found between 80-120% up to 96h. The 96 h LC₅₀of test item to zebrafish (Danio rerio) was 1.43 mg/. Thus, test chemical is considered as toxic and hazardous to aquatic fishes. Since the partition coefficient (log Kow) of the test chemical is ≤ 4 (i.e., reported as 2.79 to 3.2), test chemical can be considered as non-toxic to aquatic organisms and thus can be considered to be 'Not classified’ as per CLP classification criteria.

Description of key information

Short term toxicity to fish:

This study was conducted as per OECD 203 (2019) to assess the acute toxicity effects of test chemical on zebrafish (Danio rerio) following exposure up to 96 h under static condition. Juvenile fish of same age and normal in appearance were used in this (originate from same source and population). The average length and weight (10 fish) were observed 1.3 cm and 0.012g, respectively. Fish were fed (commercial fish food) daily during acclimatization. Photoperiod and light intensity were maintained such as16 h light- 8 h dark, 735 -800 Lux during experiment. Hardness of water was measured once during acclimatization and found to be 165 mg CaCO3/L, temperature, pH and dissolve oxygen were maintained between 22.-22.7 °C,6.4-7.1,6.8-8.5 mg/L, respectively, throughout the test. Fish were acclimatized for 7 days prior dosing. The test solution was prepared by dissolving 500 mg of the test substance in 500 mL of potable water, which was prepared passing water through reverse osmosis system. The above stock solution was stirred for 72 hrs and filtered through vaccum filter.  The stock solution was analytically detectedand the fincal expsure concentration was 0.58, 0.88, 1.33, 2.0 and 3.0 mg/l respectively. there wre no mortality in the control and lowest concentration. and at 0.88 mg/L, 1.33 mg/l, 2.0 mg/l, and 3.0 mg/l the mortality was observed to be 14.2%, 28.57%, 85.71% and 100%. The analytical monitoring of test subsatnce was performed, which was maintained to be with in the range of 80 -120 of nominal test concentrations. The test is valid as all the validity criteria are fulfilled: No mortality in control or vehicle control were found throughout the 96-h test duration; Dissolve oxygen conc. was maintained above 60% in all test vessels throughout the test; The recovery active ingredient content was found between 80-120% up to 96h. The 96 h LC₅₀of test item to zebrafish (Danio rerio) was 1.43 mg/L, Thus, test chemical is considered as toxic and hazardous to aquatic fishes. thus chemical can be classified into aquatic chronic category 2 as per CLP classification criteria

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

1.43 mg/L

Additional information

Experimental study of the test chemical and various supporting weight of evidence studies for its read across chemical were reviewed for the short term toxicity to fish end point which are summarized as below:

 

This study was conducted as per OECD 203 (2019) to assess the acute toxicity effects of test chemical on zebrafish (Danio rerio) following exposure up to 96 h under static condition. Juvenile fish of same age and normal in appearance were used in this (originate from same source and population). The average length and weight (10 fish) were observed 1.3 cm and 0.012g, respectively. Fish were fed (commercial fish food) daily during acclimatization. Photoperiod and light intensity were maintained such as16 h light- 8 h dark, 735 -800 Lux during experiment. Hardness of water was measured once during acclimatization and found to be 165 mg CaCO3/L, temperature, pH and dissolve oxygen were maintained between 22.-22.7 °C,6.4-7.1,6.8-8.5 mg/L, respectively, throughout the test. Fish were acclimatized for 7 days prior dosing. The test solution was prepared by dissolving 500 mg of the test substance in 500 mL of potable water, which was prepared passing water through reverse osmosis system. The above stock solution was stirred for 72 hrs and filtered through vaccum filter.  The stock solution was analytically detectedand the fincal expsure concentration was 0.58, 0.88, 1.33, 2.0 and 3.0 mg/l respectively. there wre no mortality in the control and lowest concentration. and at 0.88 mg/L, 1.33 mg/l, 2.0 mg/l, and 3.0 mg/l the mortality was observed to be 14.2%, 28.57%, 85.71% and 100%. The analytical monitoring of test subsatnce was performed, which was maintained to be with in the range of 80 -120 of nominal test concentrations. The test is valid as all the validity criteria are fulfilled: No mortality in control or vehicle control were found throughout the 96-h test duration; Dissolve oxygen conc. was maintained above 60% in all test vessels throughout the test; The recovery active ingredient content was found between 80-120% up to 96h. The 96 h LC₅₀of test item to zebrafish (Danio rerio) was 1.43 mg/L,

In an experimental study from study report (2017), an acute toxicity test was conducted for 96 hrs for assessing the effect of test chemical on Zebra fish (Danio rerio). The test was performed in accordance to OECD guideline No. 203 “Fish Acute Toxicity Test”. Zebra fish (Danio rerio) of average weight 0.473 g and average length of 1.76 cm was used as a test organism for the study. Test fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration. During the housing period, test fishes were fed once daily with standard brand fed. The test conditions during the housing of the test organisms were oxygen content of 7.8 mg/l, pH 7.65, water temperature 24.5°C and under a photoperiod of 16:8 hr light: dark conditions, respectively. The test solution was prepared by dissolving 2 g of the test substance in 2 liters of potable water (passed through reverse osmosis system) with 72 hrs continuous stirring. After that analytically detect that stock and set up experimental concentration according to that solubility value i.e, 7.08 mg/L. Hence, test concentrations decided are 1.39 mg/L, 2.09 mg/L, 3.14 mg/L, 4.72 mg/L and 7.08 mg/, respectively. Total 8 fishes were exposed to test chemical in a 5 lit bowl aquaria containing 4 liters of potable water. The test vessels were placed in a room at a temperature of 23°C, pH 7.03, hardness of water 152.5 mg of CaCO3 and under a photoperiod of 16:8 hr light: dark conditions, respectively. Aeration in test vessels was provided 1 day before the start of the experiment. The fishes were moving slowly in the test chemical conc. as compared to the control. No mortalities were observed in the control aquaria. The median lethal concentration (LC50 (96 h)) value was determined to be ranges between > 1.39 to < 2.09 mg/L.

 

In a supporting weight of evidence study from secondary source (2018),short term fish toxicity study was conducted for 72 hrs for assessing the effect of test chemical. Study was performed in a static system using Oncorhynchus mykiss (Rainbow trout) as a test organism. On the basis of effect of test chemical on the mortality of the test organism, the 72 hrs LC50 value was determined to be 2.4mg/l.

 

For the test chemical, short term fish toxicity study was conducted for 96 hrs for assessing the effect of test chemical (Handbook, 2008 and secondary source, 2004). Brachydanio rerio (Zebra fish) of length 2.5 to 3.5 cm was used as a test organism. Nominal test chemical conc. used for the study were 3.16, 10.0, 31.6 mg/l, respectively. Test was carried out using aquarium of 300mm x 135mm x 200mm dimensions as a test vessel. Constant aeration was provided with air during the study. Total 30 fishes (10 organisms/conc.) were exposed to the test chemical in a static system at a temperature of21.3±0.45°C, pH7.9±0.2, oxygen conc. 8.4 to 9.7 mg/l under a photoperiod of 16 hr: 8 hr light/dark. The required amount of test chemical was first dissolved in water and stirrer with a magnetic stirrer during 24 h. The suspension was then filtered in order to remove the undissolved particles. After a period of 48 hr, 9 fishes were died at a test chemical measured conc. of 5.6 mg/l (nominal conc. 31.6 mg/l) and at measured conc. of 3.9 mg/l (nominal conc. 10 mg/l), total 2 fishes were died after a period of 96 hr. On the basis of effect on mortality of the test organism Brachydanio rerio, the 96 hr LC0, LC50 and LC100 value was determined to be 0.91, 3.9 and 5.6 mg/l (measured conc.).

 

On the basis of the above results, it can be concluded that the test chemical can be considered as toxic and hazardous to aquatic fish.

Thus, test chemical is considered as toxic and hazardous to aquatic fishes. thus chemical can be classified into aquatic chronic category 2 as per CLP classification criteria