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Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
21-03-2017 - 25-03-2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Experimental test result performed using standard test guidelines
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
no
Specific details on test material used for the study:
Test Item Name 4-(phenylazo)benzene-1,3-diamine
CAS No. 495-54-5
Batch No. KCP/FS/43/17
Physical Appearance orange powder
Date of manufacturing 08-01-17
Date of expiry 08-12-17
Purity not available in coa
Stability Stable in recommended storage conditions
Storage Conditions 1. Ambient Temp (23 to 27 °C)
Precautions Wear respiratory protection. Avoid contact with skin and eyes. Avoid dust formation. Avoid breathing dust, fume, vapours, mist, spray or gas. Ensure adequate ventilation. Evacuate personnel to safe areas. Use only in a chemical fume hood.
Solubility in Water 348.393 mg/L
Analytical monitoring:
not specified
Vehicle:
yes
Remarks:
water
Details on test solutions:
Preparation of the test Solutions : The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 1g of the test substance in 1 liters of potable water (passed through reverse osmosis system) with 2 hours continuous stirring for achieving test concentrations of 0.0625mg/L,0.125mg/L,0.25mg/L,0.5mg/L,1mg/L ,respectively.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name:Zebra Fish
- Strain:-
- Source:Koyal aquarium, Nagpur
- Age at study initiation (mean and range, SD):3-4 months
- Length at study initiation (length definition, mean, range and SD):average 1.79 cm
- Weight at study initiation (mean and range, SD):0.1204g (average)
- Method of breeding: -
- Feeding during test-
- Food type:Standard brand feed,
- Amount:-
- Frequency: once daily
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Hardness:
148.5 mg of CaCO3
Test temperature:
23°C
pH:
7.03
Dissolved oxygen:
7.8 mg/l
Nominal and measured concentrations:
0.0625mg/L,0.125mg/L,0.25mg/L,0.5mg/L,1mg/L
Details on test conditions:
Photoperiod: 16 hours light and 8 hours dark
Volume of water: 2 liters
No. of animals per vessel: 8 fishes
Test temperature: 23 °C
Adaptation: Test conditions corresponded to housing conditions
Test dates: Test started at : 13-03-2017
Test Exposure date: 21-03-2017
Date of experimental termination : 25-03-2017
Withdrawal of food: 1 day before and during exposure
Aeration: Aeration in test vessels was provided 1 day before the start of experiment
Reference substance (positive control):
not specified
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
no mortality was observed

Mortality and symptoms:

Nominal

Concentration

(mg/l)

Number of

fish

Mortality (cumulated) after

3 h

6 h

24 h

48 h

72 h

96 h

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

-

-

-

-

-

-

-

-

-

-

-

-

0.0625

4

4

-

-

-

-

-

-

-

-

-

-

-

-

0.125

4

4

-

-

-

-

 

-

-

-

-

-

-

-

0.25

4

4

-

-

-

-

-

-

-

-

-

-

-

-

0.5

4

4

-

-

-

-

-

-

-

-

-

-

-

-

1

4

4

-

-

-

-

-

-

-

-

-

-

-

-

 

Visible Symptoms:

Nominal

Concentration

(mg/l)

Number of

fish

Symptom (cumulated) after

3 h

6 h

24 h

48 h

72 h

96 h

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

0.0625

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

0.125

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

0.25

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

0.5

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

1

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

 

Explanation of symptoms:

A: apathy

N: Narcotic-like state

C: Swimming near water surface

M: Slow movements compared to control

NS: No abnormal symptoms

S: Surfacing behaviour

L: Loss of equilibrium

SM: Shimming movement

H1: Hyperactivity

H2: Hypo activity

B: Bending behavior

F: Fin rot

IS: Invert Swimming

SB: Swimming behaviour

RF: Respiratory Function

P: Pigmentation

 

 

pH and oxygen content:

Nominal

Concentration

(mg/l)

Number of

fish

pH readings after

24 h

48 h

72 h

96 h

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

8.04

8.04

8.12

8.15

8.23

8.25

8.32

8.35

0.0625

4

4

7.17

7.15

8.00

8.01

8.13

8.13

8.30

8.31

0.125

4

4

7.19

7.20

8.05

8.05

8.17

8.18

8.35

8.36

0.25

4

4

7.23

7.24

8.09

8.09

8.21

8.20

8.39

8.39

0.5

4

4

7.25

7.26

8.12

8.13

8.24

8.24

8.40

8.41

1

4

4

7.31

7.35

8.17

8.19

8.26

8.28

8.42

8.45

 

Nominal

Concentration

(mg/l)

Number of

fish

Oxygen content (mg/L) after

24 h

48 h

72 h

96 h

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

7.22

7.21

7.01

7.01

6.90

6.90

6.80

6.82

0.0625

4

4

7.12

7.11

7.13

7.13

7.00

7.00

7.20

7.21

0.125

4

4

6.93

6.88

7.09

7.09

6.99

6.99

7.19

7.15

0.25

4

4

6.91

6.92

7.07

7.07

6.90

6.91

7.21

7.23

0.5

4

4

6.89

6.89

7.05

7.05

7.01

7.01

7.00

7.01

1

4

4

6.99

6.99

7.01

7.01

7.02

7.02

7.03

7.01

 

Test temperature:

Nominal

Concentration

(mg/l)

Number of

fish

Temperature (°C) after

24 h

48 h

72 h

96 h

V1

V2

V1

V2

V1

V2

V1

V2

Control

4

4

22.9

22.9

22.5

22.5

23.2

23.2

22.9

22.8

0.0625

4

4

23.1

23.1

22.5

22.5

23.2

23.2

22.9

22.9

0.125

4

4

23.2

23.2

22.5

22.5

23.2

23.2

22.9

22.9

0.25

4

4

23.2

23.2

22.5

22.5

23.2

23.2

22.9

22.9

0.5

4

4

23.2

23.2

22.5

22.5

23.2

23.2

22.9

22.9

1

4

4

23.1

23.1

22.5

22.5

23.2

23.2

22.9

22.9

Validity criteria fulfilled:
yes
Conclusions:
A 96 h study was conducted for the substance 4-(phenylazo)benzene-1,3-diamine CAS No. 495-54-5 on Zebra fish(Danio rerio), at a test concentrations of 0.0625mg/L, 0.125mg/L, 0.25mg/L, 0.5mg/L, 1mg/L . The fishes were moving slowly as compared to control. No mortalities were found in the control aquaria. The lethal concentrations LC50 for 4-(phenylazo)benzene-1,3-diamine was found to be >1 mg/L.
Executive summary:

Fish Acute Toxicity test according to OECD Guideline 203 was conducted for 4-(phenylazo)benzene-1,3 -diamine The nominal concentration selected for the experiment were 0.0625mg/L,0.125mg/L,0.25mg/L,0.5mg/L,1mg/Land test fish were exposed to these concentration for 96 hours. The lethal concentrations LC50 was found to be >1 mg/L.

LC0 (96 hours) (highest loading at which no mortality was observed) = 1 mg/L

LC50 (96 hours) Experimental = >1 mg/L

LC100 (96 hours) (lowest loading at which 100% mortality was observed) = No mortality was observed

Description of key information

A 96 h study was conducted for the substance 4-(phenylazo)benzene-1,3-diamine CAS No. 495-54-5 on Zebra fish(Danio rerio), at a test concentrations of 0.0625mg/L, 0.125mg/L, 0.25mg/L, 0.5mg/L, 1mg/L . The fishes were moving slowly as compared to control. No mortalities were found in the control aquaria. The lethal concentrations LC50 for 4-(phenylazo)benzene-1,3-diamine was found to be >1 mg/L.

Key value for chemical safety assessment

LC50 for freshwater fish:
1 mg/L

Additional information

Different experimental studies of the substance 4-(phenylazo)benzene-1,3-diamine were reviewed for Short term toxicity to fish endpoint and are summarised as follows:

Fish Acute Toxicity test according to OECD Guideline 203 was conducted for 4-(phenylazo)benzene-1,3 -diamine The nominal concentration selected for the experiment were 0.0625mg/L,0.125mg/L,0.25mg/L,0.5mg/L,1mg/Land test fish were exposed to these concentration for 96 hours. The lethal concentrations LC50 was found to be >1 mg/L.

LC0 (96 hours) (highest loading at which no mortality was observed) = 1 mg/L

LC50 (96 hours) Experimental = >1 mg/L

LC100 (96 hours) (lowest loading at which 100% mortality was observed) = No mortality was observed.

From peer reviewed journal (Yasuhide Tonogai et al;The Journal of Toxicological Sciences,1982) Short term toxicity to fish was performed for the test material 2,4 Diaminoazobenzene using test organism Oryzias latipes. The test material was dissolved in 1 mL of ethanol and then diluted with water at a concentration of 0.5 mg/L (500 µg/L)and 0.3 mg/L (300 µg/L) for the 24hrs and 48 hrs study period respectively. The LC50 for the test material 2,4 Diaminoazobenzene is 0.5 mg/L for 24 hrs duration and 0.3 mg/L for 48 hrs duration.

Considering the above data, the LC50 value of the substance for fish is observed to be 0.3 - >1 mg/L. This result is not conclusive for classiying a chemical, but considering the harmonised classification of the substance and for the purpose of classification and chemical safety assessment the substance 4-(phenylazo)benzene-1,3 -diamine is considered to classified as Aquatic acute 1 and aquatic chronic 1 category.