2,4,6-trichloroaniline

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from peer reviewed journal

Justification for type of information:

Data is from peer reviewed journal

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Principles of method if other than guideline:

Acute juvenile immobilization and embronic hatching inhibition assay was performed to study the adverse effects of 2, 4, 6 trichloroaniline.

GLP compliance:

not specified

Specific details on test material used for the study:

Details on test material
- Name of test material (as cited in study report): 2,4,6-trichloroaniline
- Molecular formula (if other than submission substance): C6-H4-Cl3-N
- Molecular weight (if other than submission substance): 196.464 g/mol
- Smiles notation (if other than submission substance): c1(c(cc(Cl)cc1Cl)Cl)N
- InChl (if other than submission substance): 1S/C6H4Cl3N/c7-3-1-4(8)6(10)5(9)2-3/h1-2H,10H2
- Substance type: organic
- Physical state: Solid

Analytical monitoring:

not specified

Details on sampling:

Details on sampling
- Concentrations: Concentrations not specified; atleast five test concentrations were used
- Sampling method: the test compound was dissolved in N,N- Dimethylformamide (DMF) at a concentration of 0.1% v/v in test water

Vehicle:

yes

Details on test solutions:

Details on test solutions
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): N,N- Dimethylformamide (DMF)
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 0.1% v/v

Test organisms (species):

Daphnia magna

Details on test organisms:

Details on test organisms
TEST ORGANISM
- Common name: Water flea
- Strain/clone: Daphnia magna
- Justification for species other than prescribed by test guideline:
- Source: National Institute for Environmental Studies, Japan
- Age of parental stock (mean and range, SD): Young female juveniles aged less than 24 hrs
- Feeding during test: Unicellular green alga Chlorella vulgaris at levels of 0.2 mg organic carbon /Daphnia /day
- Food type: Green alga
- Amount: 0.2 mg organic carbon /Daphnia /day
- Frequency: Daily

ACCLIMATION
- Acclimation period: Since 1995 till the conduct of the study
- Acclimation conditions (same as test or not): 20OC under a 16/8-h light/dark cycle (less than 600 lux) since 1995
- Type and amount of food: Unicellular green alga Chlorella vulgaris
- Feeding frequency: Daily
- Health during acclimation (any mortality observed): Daphnia showed no signs of stress such as high mortality, presence of males and ephippia, delay in the production of first brood, discoloured animals, etc.

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: the culturing period for one generation was 2-4 weeks before testing.

Test type:

static

Water media type:

freshwater

Total exposure duration:

48 h

Test temperature:

Test temp: 20°C

pH:

pH: 8.0- 9.1

Details on test conditions:

Details on test conditions
TEST SYSTEM
- Test vessel: No data available
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: Teflon sheet
- Aeration: No data available
- No. of organisms per vessel: 20 animals

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt’s M4 medium containing 293.8 mg CaCl2. 2H2O, 123.3 mg MgSO4. 7H2O, 5.8 mg KCl, 64.8 mg NaHCO3, 10mg Na2SiO3. 9H2O, 0.274 mg NaNO3, 0.143 mg KH2PO4, 0.184 mg K2HPO4, 2.86 mg H3BO3, 0.996 mg FeSO4. 7H2O, 0.361 mg MnCl2. 4H2O, 0.306 mg LiCl, 0.071 mg RBCl, 0.152 mg SrCl2. 6H2O, 0.016 g NaBr, 0.063 mg Na2MoO4. 2H2O, 0.0168 mg NaBr, 0.063 mg Na2MoO4.2H2O, 0.0168 mg CuCl2.2H2O, 0.013 mg ZnCl2, 0.010 mg CoCl2. 6H2O, 3.25 µg KI, 2.19 g Na2SeO3, 0.575 g NH4VO3, 2.50 mg Na2EDTA. 2H2O, 75µg thiamine HCl, 1.0 g cyanocobalamine (vitamin B12) and 0.75 g biotin

- Spacing factor for test concentrations: Atleast five concentrations were arranged in a geometric series with a ration lower than 2.0
- Range finding study: yes, no detailed data available
- Test concentrations: Not specified

Reference substance (positive control):

yes

Remarks:

Potassium Dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

2.3 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Development of juvenile

Basis for effect:

other: test material provided

Remarks on result:

other: 95% confidence interval was 1.5- 3.0 mg/L

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

0.83 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Embryo hatching inhibition

Remarks on result:

other: 95% confidence interval was 0.65-1.1 mg/L

Reported statistics and error estimates:

EC50 value along with the 95% confidence interval limit was calculated either by binomial, moving average or probit analysis.

Validity criteria fulfilled:

not specified

Conclusions:

After the exposure of chemical with daphnia magna effect were observed on the juvenile immobilization and embronic hatching inhibition. The EC50 was determine to be 2.3 mg/l based on the immobilisation and 0.83 on the basis of embronic hatching inhibition for 3 days.

Executive summary:

Acute juvenile immobilization and embronic hatching inhibition assay was performed to study the adverse effects of 2, 4, 6 trichloroaniline on daphnia. Test performed in accordance with OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Atleast five test concentrations were used. N,N- Dimethylformamide (DMF) were used as a solvent. Before test daphnia acclimated and daphnia showed no signs of stress such as high mortality, presence of males and ephippia, delay in the production of first brood, discoloured animals, etc.Young female juveniles were exposed to the test solution and the immobilized juveniles were observed after 24 and 48 hrs and the embronic hatching inhibition after 72 hrs. After the exposure of chemical with daphnia magna effect were observed on the juvenile immobilization and embronic hatching inhibition. The EC50 was determine to be 2.3 mg/l based on the immobilisation and 0.83 on the basis of embronic hatching inhibition for 3 days. Based on the EC50 (2.3 mg/l) chemical concluded to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.

Description of key information

Acute juvenile immobilization and embronic hatching inhibition assay was performed to study the adverse effects of 2, 4, 6 trichloroaniline on daphnia. Test performed in accordance with OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Atleast five test concentrations were used. N,N- Dimethylformamide (DMF) were used as a solvent. Before test daphnia acclimated and daphnia showed no signs of stress such as high mortality, presence of males and ephippia, delay in the production of first brood, discoloured animals, etc.Young female juveniles were exposed to the test solution and the immobilized juveniles were observed after 24 and 48 hrs and the embronic hatching inhibition after 72 hrs. After the exposure of chemical with daphnia magna effect were observed on the juvenile immobilization and embronic hatching inhibition. The EC50 was determine to be 2.3 mg/l based on the immobilisation and 0.83 on the basis of embronic hatching inhibition for 3 days. Based on the EC50 (2.3 mg/l) chemical concluded to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

2.3 mg/L

Additional information

Based on the experimental data from various database for the target chemical study have been reviewed to determine the mode of action of2,4,6-trichloroaniline (CAS no. 634-93-5)on the mobility rate and behavior of aquatic invertebrates. The studies are as mentioned below:

 

In the first key study from peer reviewed journal 2001, Acute juvenile immobilization and embronic hatching inhibition assay was performed to study the adverse effects of 2, 4, 6 trichloroaniline on daphnia. Test performed in accordance with OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). At least five test concentrations were used. N,N- Dimethylformamide (DMF) were used as a solvent. Before test daphnia acclimated and daphnia showed no signs of stress such as high mortality, presence of males and ephippia, delay in the production of first brood, discoloured animals, etc. Young female juveniles were exposed to the test solution and the immobilized juveniles were observed after 24 and 48 hrs and the embronic hatching inhibition after 72 hrs. After the exposure of chemical with daphnia magna effect were observed on the juvenile immobilization and embronic hatching inhibition. The EC50 was determine to be 2.3 mg/l based on the immobilisation and 0.83 on the basis of embronic hatching inhibition for 3 days. Based on the EC50 (2.3 mg/l) chemical concluded to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.

 

First key study was supported by the second study from authoritative database 2018. Study was conducted to determine the effect of test chemical on the growth of aquatic invertebrates for 48 hrs. Test performed according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). Based on the immobility of daphnia magna due to the chemical effect, the EC50 was determine to be 4.3 mg/l. Thus considering the CLP Criteria for aquatic classification of the substance, it is concluded that 2,4,6-trichloroaniline exhibits short term toxicity to aquatic invertebrate and is therefore classified as Aquatic Chronic category 2.

 

Similarly above results and classification was supported by another studies from secondary sources 2018. Study was conducted to determine the effect of test chemical on the growth of aquatic invertebrates. After the exposure of test chemical effect were observed and measured at the time when 50 % population affected. Based on the Distance moved, change in direct movement, the EC50 was determine to be 6 mg/l.

 

Based on the overall studies from peer reviewed journal, authoritative databases and secondary sources 2018, it was concluded that the chemical 2,4,6-trichloroaniline was toxic to the aquatic invertebrates and consider to be classified as aquatic chronic 2 as per the CLP classification criteria.