2-amino-4-nitrophenol

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

Justification for type of information:

Experimental test result performed according to the guideline.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Principles of method if other than guideline:

Study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test).

GLP compliance:

no

Analytical monitoring:

not required

Remarks:

substance is stable under recomended conditions

Vehicle:

no

Details on test solutions:

The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 62.5 mg, 125 mg, 250 mg, 500 mg & 1 g of the test substance in 10 liters of potable water (passed through reverse osmosis system) with continuous stirring for achieving test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L & 100 mg/L, respectively.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name:Zebra Fish
- Strain:Danio rerio
- Source: Orange City Aquarium, Sitabuldi, Nagpur, MS (India)
- Length at study initiation (length definition, mean, range and SD): average 2.42 cm
- Weight at study initiation (mean and range, SD): average 0.117 g
-Housing:The fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration.
-Duration of housing: 15 days
-Photoperiod: 12 hours light and 12 hours darkness
-Diet : Standard brand feed, once daily
-Acclimatization: seven days
-feeding frequency: Once in a day

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

51.2 mg of CaCO3

Test temperature:

24-26 °C

pH:

7.2

Nominal and measured concentrations:

6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L & 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Plastic aquaria
- Material, size, headspace, fill volume: 5 liters of plastic aquaria filled with 2 liter of water
- Aeration: Aeration in test vessels was provided 1 day before the start of experiment
- No. of organisms per vessel: 8 fishes

OTHER TEST CONDITIONS
- Photoperiod: 12 hours light and 12-hour darkness

Reference substance (positive control):

not specified

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Sublethal observations / clinical signs:

8.1. Mortality and symptoms

Nominal conc. (mg/l)	Number of fish	Mortality (cumulated) after
		24h	48h	72h	96h
0 (control)	8	-	-	-	-
6.25	8	-	-	-	-
12.5	8	-	-	-	-
25	8	-	-	-	-
50	8	1	-	-	-
100	8	8	-	-	-

Nominal conc. (mg/l)	Number of fish	Symptoms after
		24h	48h	72h	96h
0 (control)	8	NS8	NS8	NS8	NS8
6.25	8	NS8	NS8	NS8	NS8
12.5	8	NS8	NS8	NS8	NS8
25	8	NS8	NS8	NS8	NS8
50	8	NS7	NS7	NS7	NS7

Explanation of symptoms:

A: apathy

N: narcotic-like state

C: swimming near water surface

D: swimming at the bottom of bowl

M: Slow movements compared to control

NS: No abnormal symptoms

Number behind symbol for symptom= number of affected fish

8.2. pH and oxygen content:

Nominal conc. (mg/l)	pH readings after
	24h	48h	72h	96h
0 (control)	7.6	7.5	7.6	7.7
6.25	7.8	7.5	7.0	6.1
12.5	7.6	7.2	7.1	6.1
25	7.5	7.3	6.9	6.1
50	7.3	7.0	6.5	6.0

Nominal conc. (mg/l)	Oxygen content (mg/l) after
	24h	48h	72h	96h
0 (control)	6.79	6.72	6.69	6.68
6.25	6.76	6.73	6.10	6.09
12.5	6.47	6.21	6.01	5.93
25	6.14	6.01	5.99	5.71
50	6.05	6.01	5.77	5.67

8.3. Test temperature

Nominal conc. (mg/l)	Temperature (°C) after
	24h	48h	72h	96h
0 (control)	25	26	26	26
6.25	25	26	26	26
12.5	25	26	26	26
25	25	26	26	26
50	25	26	26	26

Validity criteria fulfilled:

yes

Conclusions:

Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for test material on Zebra Fish Danio rerio was determine to be >50 mg/l.

Executive summary:

Study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test). The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 62.5 mg, 125 mg, 250 mg, 500 mg & 1 g of the test substance in 10 liters of potable water passed through reverse osmosis system with continuous stirring for achieving test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L & 100 mg/L, respectively.  This test solution was then added to the remaining three liters of water for achieving test concentrations of 100 mg/L and Zebra FishDanio reriowere exposed to these concentration for 96 hours.Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure to test item to various nominal test concentrations, LC50 was determine to be >50 mg/l LC100 was observed at the concentration 100 mg/l . Based on the LC50, it can be consider that the chemical was toxic to fish and can be consider to be classified as aquatic chronic 3 as per the CLP classification criteria.

Description of key information

Short term toxicity to fish:

Study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test).The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 62.5 mg, 125 mg, 250 mg, 500 mg & 1 g of the test substance in 10 liters of potable water passed through reverse osmosis system with continuous stirring for achieving test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L & 100 mg/L, respectively.  This test solution was then added to the remaining three liters of water for achieving test concentrations of 100 mg/L and Zebra FishDanio reriowere exposed to these concentration for 96 hours.Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure to test item to various nominal test concentrations, LC50 was determine to be >50 mg/l LC100 was observed at the concentration 100 mg/l . Based on the LC50, it can be consider that the chemical was toxic to fish and can be consider to be classified as aquatic chronic 3 as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

50 mg/L

Additional information

Short term toxicity to fish:

The toxicity of test material for fish was evaluated using data from lab report along with a publication.Study was conducted to access the effect of test chemical on the growth of fish Danio rerio. Test conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test).The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 62.5 mg, 125 mg, 250 mg, 500 mg & 1 g of the test substance in 10 liters of potable water passed through reverse osmosis system with continuous stirring for achieving test concentrations of 6.25 mg/L, 12.5 mg/L, 25 mg/L, 50 mg/L & 100 mg/L, respectively.  This test solution was then added to the remaining three liters of water for achieving test concentrations of 100 mg/L and Zebra FishDanio reriowere exposed to these concentration for 96 hours.Bowl aquaria containing 2 liters of potable water (passed through reverse osmosis system) were loaded with 8 fishes. A static procedure was used for the study and it was conducted in compliance with the OECD guideline 203. After 96 hours of exposure to test item to various nominal test concentrations, LC50 was determine to be >50 mg/l LC100 was observed at the concentration 100 mg/l . Based on the LC50, it can be consider that the chemical was toxic to fish and can be consider to be classified as aquatic chronic 3 as per the CLP classification criteria.

In a peer reviewed journal ,short term toxicity to fish was evaluated for the test chemical for 48 h . Carp(Ciprinus carpio) and was provided by Lu-Kan Fish Culture Station, Taiwan Fisheries Research Institute, Taichung, Taiwan. Before the experiment, the test organisms were acclimatized in aquaria for 2 weeks under conditions similar to those under which the tests was performed. Four carp , 2 to 6 cm in size, were introduced to each 10-L beaker containing5 L of different concentrations of test chemical . Five different concentrations 0.1 , 0.5 , 1 , 2 and 4 mg/l were prepared. Sub-lethal concentration (LC50) values were calculated by the trimmed Spearman} act concentration EC50 of test material on fish when observed after 48 h was noted to be 0.453 mg/l. Based on the above effect concentration it can be concluded that the test material is highly toxic to fish and can be classified as aquatic acute 1.

As we have two studies which are classifying test material in two diffrent categories , hence we will consider effect concentration obtained from lab report as it has higher relaibility then that of data from publication .Based on the LC50, it can be consider that the chemical was toxic to fish and can be consider to be classified as aquatic chronic 3 as per the CLP classification criteria.