3-chlorophthalic anhydride

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

3-chlorophthalic anhydride

EC Number:

204-179-8

EC Name:

3-chlorophthalic anhydride

Cas Number:

117-21-5

Molecular formula:

C8H3ClO3

IUPAC Name:

4-chloro-1,3-dihydro-2-benzofuran-1,3-dione

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Start and end of the test from each replicate test vessel

Test solutions

Vehicle:

no

Details on test solutions:

Water used for the acclimation of test organisms and for all toxicity testing was carbon filtered, deionized water collected at T.R. Wilbury Laboratories in Marblehead, Massachusetts. Water was adjusted to a hardness of 40 to 48 mg/L as CaCO3 and stored in polyethylene tanks where it was aerated and recirculated through particle filters, activated carbon, and an ultraviolet sterilizer. A sample of dilution water collected at the start of the definitive test had a hardness of 44 mg/L as CaCO3.

The test substance was added directly to dilution water in test vessels to formulate the test concentration (18.3 g of test substance in 15 L dilution water). The solutions were mixed overnight with electric mixers and the pH was adjusted to approximately 7.7 with 1.0N sodium hydroxide. Nominal concentrations of the active ingredient were 0 mg/L (control) and 1,200 mg/L.

Test organisms

Test organisms (species):

Pimephales promelas

Details on test organisms:

Juvenile fathead minnow, Pimephales promelas, were procured from a commercial supplier (Aquatic Biosystems, Fort Collins, Colorado, USA). Prior to testing, the fish were maintained under flow-through conditions in a 220 L polyethylene tank. During acclimation, fish were not treated for disease and they were free of apparent disease, injuries, and abnormalities at the beginning of the test. Fish were fed dry commercial fish food once per day except during the 48 hours preceding the test and they were not fed during the toxicity test. During the 14 day period before the start of the test, the acclimation temperature range was 22.0 to 22.7°C and the dissolved oxygen concentration was always at least 7.2 mg/L. Mortality during the last 48 hours of acclimation was less than 3%. At the conclusion of the test, the control fish had an average wet weight of 0.32 g (loading rate was 0.21 g/L) and they ranged in length from 2.9 to 3.8 cm.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test conditions

Hardness:

44 mg/L as CaCO3 (at study start)

Test temperature:

21.6 - 21.9°C

pH:

6.7 - 8.0

Dissolved oxygen:

7.3 - 9.6 mg/L

Nominal and measured concentrations:

0 mg/L (control) and 1,200 mg 3-CLPA/L

Details on test conditions:

A range-finding test with 3-CLPA was conducted under static conditions with a control and five concentrations of test substance: 0.10, 1.0, 10, 100, and 1,000 mg/L. After 96 hours there was 100% survival in the control and at all tested concentrations. No sublethal effects were observed at any tested concentration. A small amount of insoluble material settled out of the 1,000 mg/L test solutions and no other insoluble material was observed.

The definitive test was also conducted under static conditions. Twenty fathead minnows were randomly distributed among three replicates of the treatment and control. The test was performed in 20 liter glass aquaria that contained 15 liters of test solution (water depth was approximately 18 cm). Test vessels were randomly arranged in a water bath during the 96 hour test (random numbers tables were used to add test organisms to test vessels, to select the location of each vessel, as well as the assignment of test concentrations). A 16 hour light and 8 hour dark photoperiod was automatically maintained with cool white fluorescent lights that provided a light intensity of approximately 40 footcandles. A 15 minute transition period was provided between dark and light. Aeration was not required to maintain dissolved oxygen concentrations above acceptable levels. The number of surviving organisms, and the occurrence of sublethal effects and insoluble material were determined visually and recorded every 24 hours.

Dissolved oxygen, pH, conductivity, and temperature were measured and recorded daily in each test vessel. The temperature in a control vessel was recorded continuously during the test.

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 1 200 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 1 200 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Effect concentrations 3

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 1 200 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Effect concentrations 4

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 200 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Effect concentrations 5

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

1 200 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

No insoluble material was observed at any tested concentration during the definitive toxicity test. Nominal concentrations of 3-chlorophthalic anhydride were: 0 mg/L (control) and 1,200 mg/L. Mean measured concentrations were: ND (not detected at or above the limit of quantitation) and 1,200 mg/L. The mean measured concentration was 100% of the nominal concentration. One hundred percent survival occurred and no sublethal effects were observed in the control or test group.

Reported statistics and error estimates:

Results of the toxicity test could not be interpreted by standard statistical techniques because there was 100% survival at the single tested concentration. The no observed effect concentration is the tested concentration at which there was no toxicant-related mortality or sublethal effect.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Nominal Concentration of 3-CLPA (mg/L)	Measured Concentration of 3-chlorophthalic anhydride (mg/L)
	0 Hour	96 Hours	Mean	% Recovery
Test Media Samples
Control	ND	ND	ND	---
	ND	ND
	ND	ND
1,200	1,200	1,300	1,200	100
	1,200	1,300
	1,100	1,300
Matrix Spike Sample
1,200	1,100	1,300	1,200	100
	1,200	1,200
	1,200	1,200
Blank
0	ND	ND	ND	---
ND = none detected at or above the limit of quantitation.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 96-h LC50 was determined to be >1200 mg/L

Executive summary:

The short-term toxicity to freshwater fish was determined in a limit test according to OECD TG 203 and in compliance with GLP criteria. In this study, three fathead minnows (Pimephales promelas) were exposed to nominal test concentrations of 0 (control) and 1200 mg/L for 96 hours under static conditions. The nominal test concentration was analytically verified at the start and end of exposure and remained within ±20% of nominal. Therefore, the effect value is expressed as nominal. Fish mortality and general symptoms of toxicity were recorded after 24, 48, 72 and 96 hours exposure. In this study the OECD validity criteria are met. No mortality or general symptoms of mortality was observed during the test. Based on these findings the 96-h LC50 value was determined at >1200 mg/L.