Hexachlorocyclopentadiene

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

Data on light was not recorded

GLP compliance:

no

Specific details on test material used for the study:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: test compound dissolved in acetone

Analytical monitoring:

not specified

Details on sampling:

- Concentrations: 0.32 mg/L; 0.24 mg/L; 0.18 mg/L; 0.14 mg/L; 0.10 mg/L; 0.087 mg/L; 0.065 mg/L.

Vehicle:

yes

Remarks:

acetone

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test compound for each concentration, dissolved in acetone, was added directly into each jar and mixed by stirring.
- Controls: Negative controls (same dilution water and conditions but with no test substance or acetone) and positive controls (containing greatest amount of acetone introduced into any one test vessel).
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Acetone

Test organisms (species):

Lepomis macrochirus

Details on test organisms:

TEST ORGANISM
- Common name: Bluegill
- Source: commercial fish hatchery in Nebraska
- Length at study initiation (length definition, mean, range and SD): 26 ± 4.9 mm
- Weight at study initiation (mean and range, SD): 0.45 ± 0.17 g
- Maintenance of the brood fish: fish were held in 1700L concrete raceways, coated with an epoxy resin paint to prevent leaching of extraneous materials into the water. Well water flowed through these raceways at a minimum of 4 L / minute, providing an adequate water turn-over rate for maintaining the species.

ACCLIMATION
- Acclimation period: 48 hours
- Acclimation conditions (same as test or not): test conditions of temperature and water quality
- Feeding frequency during acclimation: fish were not fed during acclimation period
- Health during acclimation (any mortality observed): no mortality observed and fish appeared to be in excellent condition at the initiation of the tests

QUARANTINE
- Duration: 30 days
- Health/mortality: less than 2%

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

35 mg/L CaCO3

Test temperature:

21 ± 1 °C

pH:

7.1

Dissolved oxygen:

> 60% saturation

Details on test conditions:

TEST SYSTEM
- Test vessel: jars
- Type (delete if not applicable): open / closed
- Material, size, headspace, fill volume: 19.6 L glass jars containing 15 L test solution
- Aeration: Test solutions were not aerated during the tests.
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Intervals of water quality measurement: Dissolved oxygen concentration, pH and temperature of test solutions were checked at 0, 24, 48 and 96 hours of exposure in the highest, middle and lowest test concentrations, at a minimum.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : percentage mortality at 24, 48, and 96 hours

TEST CONCENTRATIONS
- Range finding study: The preliminary test defined a narrower range of concentrations to be used in the definitive test which would then assess the dose response relationship and permit calculation of LC50 and its 95% confidence interval from the mortality data produced.
- Test concentrations: 0.32 mg/L; 0.24 mg/L; 0.18 mg/L; 0.14 mg/L; 0.10 mg/L; 0.087 mg/L; 0.065 mg/L.

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

0.17 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

0.15 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.13 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.065 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Other biological observations: The mortality syndrome among fish from those concentrations in which mortality was observed was similar. Fish generally became dark and lethargic, lost equilibrium, and expired.
- Mortality of control: no mortality
- Other adverse effects control: no effects

Reported statistics and error estimates:

LC50 (24hr) = 0.14 - 0.21 mg/L (95% confidence interval)
LC50 (48hr) = 0.12 - 0.18 mg/L (95% confidence interval)
LC50 (96hr) = 0.11 - 0.17 mg/L (95% confidence interval)

Sublethal observations / clinical signs:

Table showing concentrations tested and corresponding observed percentage mortalities for bluegill (Lepomis macrochirus)

Nominal concentration (mg/L)	% mortality observed
	24 h	48 h	96 h
0.32	100	100	100
0.24	80 (a,b)	100	100
0.18	30 (a,b)	30 (a,b)	40 (b)
0.14	10 (a,b)	30 (a,b)	40 (b)
0.10	10	10	20 (b)
0.087	0	0	10
0.065	0	0	0
control (acetone)	0	0	0
control	0	0	0

(a) = Fish displayed dark colouration

(b) = Fish were lethargic

(c) = Fish displayed a partial loss of equilibrium

Validity criteria fulfilled:

yes

Conclusions:

The 96 hr LC50 of hexachlorocyclopentadiene for Lepomis macrochirus was 0.13 mg/L.

Executive summary:

The short term toxicity of hexachlorocyclopentadiene to fish was determined using a procedure equivalent to the OECD Guideline for Testing of Chemicals 203 with acceptable restrictions. The study was carried out on Bluegill (Lepomis macrochirus). The study determined the percent mortality of the test species to the registered substance over a range of concentrations between 0.065 and 0.32 mg/L. It was determined that the 96 hour LC50 of the registered substance was 0.13 mg/L.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

Data on light was not recorded

GLP compliance:

no

Specific details on test material used for the study:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: test compound dissolved in acetone

Analytical monitoring:

not specified

Details on sampling:

- Concentrations: 0.32 mg/L; 0.24 mg/L; 0.18 mg/L; 0.16 mg/L; 0.14 mg/L; 0.12 mg/L; 0.087 mg/L.

Vehicle:

yes

Remarks:

acetone

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test compound for each concentration, dissolved in acetone, was added directly into each jar and mixed by stirring.
- Controls: Negative controls (same dilution water and conditions but with no test substance or acetone) and positive controls (containing greatest amount of acetone introduced into any one test vessel).
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Acetone

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead minnow
- Source: commercial fish hatchery in Arkansas
- Length at study initiation (length definition, mean, range and SD): 38 ± 5.2 mm
- Weight at study initiation (mean and range, SD): 0.72 ± 0.3 g
- Maintenance of the brood fish: fish were held in 1700L concrete raceways, coated with an epoxy resin paint to prevent leaching of extraneous materials into the water. Well water with a hardness flowed through these raceways at a minimum of 4 L / minute, providing an adequate water turn-over rate for maintaining the species.

ACCLIMATION
- Acclimation period: 48 hours
- Acclimation conditions (same as test or not): test conditions of temperature and water quality
- Feeding frequency during acclimation: fish were not fed during acclimation period
- Health during acclimation (any mortality observed): no mortality observed and fish appeared to be in excellent condition at the initiation of the tests

QUARANTINE
- Duration: 30 days
- Health/mortality: less than 2%

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

35 mg/L CaCO3

Test temperature:

21 ± 1 °C

pH:

7.1

Dissolved oxygen:

> 60% saturation

Details on test conditions:

TEST SYSTEM
- Test vessel: jars
- Material, size, headspace, fill volume: 19.6 L glass jars containing 15 L test solution
- Aeration: Test solutions were not aerated during the tests
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Intervals of water quality measurement: Dissolved oxygen concentration, pH and temperature of test solutions were checked at 0, 24, 48 and 96 hours of exposure in the highest, middle and lowest test concentrations, at a minimum.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : percentage mortality at 24, 48, and 96 hours

TEST CONCENTRATIONS
- Range finding study: The preliminary test defined a narrower range of concentrations to be used in the definitive test which would then assess the dose response relationship and permit calculation of LC50 and its 95% confidence interval from the mortality data produced.
- Test concentrations: 0.32 mg/L; 0.24 mg/L; 0.18 mg/L; 0.16 mg/L; 0.14 mg/L; 0.12 mg/L; 0.087 mg/L.

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

0.24 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

0.21 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.087 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Other biological observations: The mortality syndrome among fish from those concentrations in which mortality was observed was similar. Fish generally became dark and lethargic, lost equilibrium, and expired.
- Mortality of control: no mortality
- Other adverse effects control: no effects

Reported statistics and error estimates:

LC50 (24hr) = 0.17 - 0.32 mg/L (95% confidence interval)
LC50 (48hr) = 0.18 - 0.25 mg/L (95% confidence interval)
LC50 (96hr) = 0.16 - 0.22 mg/L (95% confidence interval)

Sublethal observations / clinical signs:

Table showing concentrations tested and corresponding observed percentage mortalities for fathead minnow (Pimephales promelas).

Nominal concentration (mg/L)	% mortality observed
	24 h	48 h	96 h
0.32	100	100	100
0.24	50 (a,b,c)	80 (b,c)	90 (a,b)
0.18	0	20 (a)	20 (a)
0.16	0	0	20 (a)
0.14	0	0	0
0.12	0	0	10
0.087	0	0	0
control (acetone)	0	0	0
control	0	0	0

(a) = Fish displayed dark colouration

(b) = Fish were lethargic

(c) = Fish displayed a partial loss of equilibrium

Validity criteria fulfilled:

yes

Conclusions:

The 96 hr LC50 of hexachlorocyclopentadiene for Pimephales promelas was 0.18 mg/L.

Executive summary:

The short term toxicity of hexachlorocyclopentadiene to fish (Pimephales promelas) was determined using a procedure equivalent to the OECD Guideline for Testing of Chemicals 203, with acceptable restrictions. The study was carried out on Fathead minnow (Pimephales promelas). The study determined the percentage mortality of the test species to the registered substance over a range of concentrations between 0.087 and 0.32 mg/L. It was determined that the 96 hour LC50 of the registered substance was 0.13 mg/L.

Description of key information

Two non-GLP short-term toxicity studies were performed according to a method similar to the OECD Testing Guideline 203 on Lepomis macrochirus and Pimephales promelas respectively. The lowest 96-hour LC50 (0.13 mg/l) for Lepomis macrochirus was selected as the key representative value as it represents a worst-case scenario.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.13 mg/L

Additional information

The short-term toxicity of the registered substance to marine fish was not investigated as part of this REACH Registration. It was not considered as relevant taking into account the uses of the substance.