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Ecotoxicological information

Short-term toxicity to fish

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Link to relevant study record(s)

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
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
no
Specific details on test material used for the study:
- Test Item Name 3',6'-bis(diethylamino)spiro[isobenzofuran- 1(3H),9'-[9H]xanthene]-3-one
- CAS No. 509-34-2
- Batch No. KCP/FS/42/17
- Physical Appearance pinkish red powder
- Date of manufacturing 2017-01-03
- Date of expiry 2017-12-02
- Purity not available in coa
- Stability Stable in recommended storage conditions
- Storage Conditions 1. Ambient Temp (23 to 27 °C)
Analytical monitoring:
not specified
Vehicle:
yes
Details on test solutions:
Preparation of the test Solutions : The test substance was soluble in water. Therefore, the test solution was prepared by dissolving 1 g 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 6.25mg/L, 12.5mg/L, 25mg/L, 50mg/L,100mg/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: Koyal aqurium,Nagpur.
- Age at study initiation (mean and range, SD):-
- 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: Withdrawal of food 1 day before and during exposure
- 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:
7.80 mg/L
Salinity:
Not specified
Conductivity:
Not specified
Nominal and measured concentrations:
6.25mg/L,12.5mg/L, 25mg/L, 50mg/L and 100mg/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
Key result
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:
LC0
Effect conc.:
25 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:

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

-

-

-

-

-

-

-

-

-

-

-

-

6.25

4

4

-

-

-

-

-

-

-

-

-

-

-

-

12.5

4

4

-

-

-

-

 

-

-

-

-

-

-

-

25

4

4

-

-

-

-

-

-

-

-

-

-

-

-

50

4

4

-

-

-

-

-

-

-

-

1

1

2

2

100

4

4

-

-

-

-

4

4

4

4

4

4

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

6.25

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

12.5

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

25

4

4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

NS4

50

4

4

NS4

NS4

NS4

NS4

NS4

NS4

M4

M4

M3

M3

M2

M2

100

4

4

NS4

NS4

M4

M4

-

-

-

-

-

-

-

-

 

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

6.25

4

4

7.41

7.42

8.12

8.12

8.21

8.22

8.36

8.36

12.5

4

4

7.50

7.50

8.15

8.16

8.29

8.30

8.39

8.38

25

4

4

7.55

7.56

8.24

8.24

8.31

8.31

8.40

8.41

50

4

4

7.61

7.61

8.29

8.30

8.35

8.36

8.45

8.42

100

4

4

-

-

-

-

-

-

-

-

 

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

6.25

4

4

7.09

7.08

7.34

7.34

7.21

7.21

7.00

7.01

12.5

4

4

6.88

6.89

7.32

7.32

7.14

7.15

7.02

7.03

25

4

4

6.82

6.83

7.31

7.31

7.17

7.17

7.01

7.00

50

4

4

6.98

6.95

7.29

7.29

7.21

7.22

7.03

7.05

100

4

4

-

-

-

-

-

-

-

-

 

Validity criteria fulfilled:
yes
Conclusions:
Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for CAS No.509-34-2 on Zebra Fish (Danio rerio) was found to be 50 mg/L.
Executive summary:

A 96 h study was conducted for CAS No. 509-34-2 on Zebra fish(Danio rerio), at a test concentrations of 6.25mg/L,12.5mg/L, 25mg/L, 50mg/L and 100mg/L. The fishes were moving slowly as compared to control. No mortalities were found in the control aquaria. The lethal concentrations LC50 was found to be 50 mg/L.

LC0 (96 hours) (highest loading at which no mortality was observed) = 25 mg/L

LC50 (96 hours) Experimental = 50 mg/L

LC100 (96 hours) (lowest loading at which 100% mortality was observed) = 100 mg/L

This value indicates that the substance 3',6'-bis(diethylamino)-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one is toxic to fish and can be classified as Aquatic chronic category 3.

Description of key information

Based on nominal concentrations, experimental median lethal Concentrations [LC-50 (96 h)] for CAS No.509-34-2 on Zebra Fish (Danio rerio) was found to be 50 mg/L. This value indicates that the substance 3',6'-bis(diethylamino)-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one is toxic to fish and can be classified as Aquatic chronic category 3.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
50 mg/L

Additional information

Various studies, considering its adequacy as one key and other supporting, were reviewed for the endpoint short term toxicity to fish of the substance 3',6'-bis(diethylamino)-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one and same are summarized as below:

In a key study, 96 h study was conducted by SSS for CAS No. 509-34-2 on Zebra fish (Danio rerio), at a test concentrations of 6.25mg/L,12.5mg/L, 25mg/L, 50mg/L and 100mg/L. The fishes were moving slowly as compared to control. No mortalities were found in the control aquaria. The lethal concentrations LC50 was found to be 50 mg/L.

LC0 (96 hours) (highest loading at which no mortality was observed) = 25 mg/L

LC50 (96 hours) Experimental = 50 mg/L

LC100 (96 hours) (lowest loading at which 100% mortality was observed) = 100 mg/L

This value indicates that the substance 3',6'-bis(diethylamino)-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one is toxic to fish and can be classified as Aquatic chronic category 3.

In supporting study for read across Rhodamine B (Cas no. 81-88-9) from Experimental data (J. Toxicol. Sci. 7: 193; 1982) indicated that fish toxicity study was conducted for 48 hrs for evaluating the lethal concentration of test substance Rhodamine B on fish Oryzias latipes on the basis of mortality effect. During experiment the lethal concentration (LC50) for test substance Rhodamine B was found to be 17 mg/l for 24 hrs. and 12 mg/l for 48 hrs. This LC50 value indicate that the test substance Rhodamine B is toxic to fish and classified in aquatic chronic 3 category as per the CLP criteria.

For the same read across substance, i.e Rhodamine B (Cas no. 81-88-9); Fish toxicity study (Japan chemicals collborative knowledge database (J-check); 2017) was conducted for 48 hrs for evaluating the lethal concentration of test substance Rhodamine B on fish Oryzias latipes on the basis of mortality effect. During experiment the lethal concentration (LC50) for test substance Rhodamine B was determined to be 33.9 mg/l. This LC50 value indicate that the test substance Rhodamine B is toxic to fish and classified in aquatic chronic 3 category as per the CLP criteria.

Both studies for read across chemical give assistance to experimental result of target chemical.

Thus, based on experimental study conducted by SSS with EC50= 50 mg/l, it can be concluded that the target chemical 3',6'-Bis(N,N-diethylamino)-3H-spiro[isobenzofuran-1,9'-xanthen]-3-one (CAS no. 509-34-2) is toxic to fish and thus can be classified in Aquatic Chronic 3 category as per the CLP criteria.