Benzene-1,2:4,5-tetracarboxylic dianhydride

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

Study period:

April 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

test item is unstable in water, therefore the hydrolysis product PMA was tested

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Description: white powder
- Batch number: 8M30N
- Storage conditions: room temperature in the dark until 04 April 2008, thereafter at approximately 4 °C, over silica gel, in the dark

Analytical monitoring:

yes

Details on sampling:

The concentration and stability of the test material in the test preparations were verified by chemical analysis at 0, 24 and 96 hours. Water samples were taken from the control and each replicate test vessel at 0 (fresh media), 24 and 96 hours (old media) for quantitative analysis. Duplicate samples and samples at 24 (fresh media), 48 and 72 hours (fresh and old media) were taken and stored at approximately -20°C for further analysis if necessary.

Vehicle:

no

Details on test solutions:

For the purpose of the definitive test the test material was dissolved directly in dechlorinated tap water. An amount of test material (4000 mg) was dissolved in dechlorinated tap water and the volume adjusted to 1 litre to give a 4000 mg/l stock solution. An aliquot (500 ml) of this stock solution was dispersed in a final volume of 20 litres of dechlorinated tap water, and stirred using a flat bladed mixer for approximately 1 minute, to give the 100 mg/l test concentration. This dilution was conducted in duplicate to give replicates R1 and R2. The stock solution was inverted several times to ensure adequate mixing and homogeneity.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

The test was carried out using juvenile rainbow trout (Oncorhynchus mykiss). Fish were obtained from Brow Well Fisheries Limited, Hebden, near Skipton, Yorkshire, UK and maintained in-house since 19 March 2008.

Fish were maintained in a glass fibre tank with a "single pass" water renewal system. Fish were acclimatised to test conditions from 9 April 2008 to 21 April 2008. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods.
The water temperature was controlled at approximately 14°C with a dissolved oxygen content of greater than or equal to 10.3 mg O2/1. These parameters were recorded daily. The stock fish were fed commercial trout pellets which was discontinued approximately 24 hours prior to the start of the definitive test. There was zero mortality in the 7 days prior to the start of the test and the fish had a mean standard length of 4.7 cm (sd = 0.2) and a mean weight of 1.36 g (sd = 0.22) at the end of the definitive test. Based on the mean weight value this gave a loading rate of 0.48 g body weight/litre.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

approximately 140 mg/1 as CaC03

Test temperature:

The water temperature was controlled at approximately 14°C.

pH:

5.7 to 7.8

Dissolved oxygen:

9.35 to 11.1 mg O2/l

Salinity:

freshwater used

Nominal and measured concentrations:

The test concentration to be used in the definitive test was determined by a preliminary range-finding test. Test fish were exposed to a series of nominal test concentrations of 1.0, 10 and 100 mg/l. Based on the results of the range-finding test a Limit test was conducted at a concentration of 100 mg/l to confirm that at the maximum concentration given in the OECD/EC Test Guidelines, no mortalities or sub-lethal effects of exposure were observed.
Analysis of the test preparations at 0, 24 and 96 hours showed measured test concentrations to range from 93% to 97% of nominal and so it was considered justifiable to estimate the LC50 values in terms of the nominal test concentrations only.

Details on test conditions:

The test concentration to be used in the definitive test was determined by a preliminary rangefinding test. In the range-finding test fish were exposed to a series of nominal test concentrations of 1.0, 10 and 100 mg/l. The test material was dissolved directly in dechlorinated tap water. An amount of test material ( 4000 mg) was dissolved in dechlorinated tap water and the volume adjusted to I litre to give a 4000 mg/l stock solution. Aliquots (5.0, 50 and 500 ml) of this stock solution were each separately dispersed in a final volume of 20 litres of dechlorinated tap water, and stirred using a flat bladed mixer for approximately 1 minute, to give the 1.0, 10 and 100 mg/l test concentrations respectively. The stock solution was inverted several times to ensure adequate mixing and homogeneity. In the range-finding test, 3 fish were added to each 20 litre test and control vessel and maintained at approximately 14°C in a temperature controlled room with a photoperiod of 16 hours light and 8 hours darkness with 20 minute dawn and dusk transition periods for a period of 96 hours under
static test conditions. The control group was maintained under identical conditions but not exposed to the test material. Each vessel was covered to reduce evaporation. After 3, 6, 24, 48, 72 and 96 hours any mortalities or sub-lethal effects of exposure were determined by visual inspection of the test fish.
Based on the results of the range-finding test a "Limit test" was conducted at a concentration of 100 mg/l to confirm that at the maximum concentration given in the OECD/EEC Test Guidelines, no mortalities or sub-lethal effects of exposure were observed.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

MORTALITY:
There were no mortalities in 14 fish exposed to a test concentration of 100 mg/1 for a period of 96 hours. The results of the definitive test showed the highest test concentration resulting in 0% mortality to be greater than or equal to 100 mg/l, the lowest test concentration resulting in 100% mortality to be greater than 100 mg/l and the No Observed Effect Concentration (NOEC) to be 100 mg/l. The No Observed Effect Concentration is based upon zero mortalities and the absence of any sublethal effects of exposure at this concentration. It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/1.

SUBLETHAL EFFECTS:
There were no sub-lethal effects of exposure observed in 14 fish exposed to a test concentration of 100 mg/L for a period of 96 hours.

TEST MATERIAL SOLUBILITY:
The test preparations were observed to be clear, colourless solutions throughout the duration of the test.

Results with reference substance (positive control):

not tested

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test material to the freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated and gave a 96-Hour LC50 of greater than 100 mg/l. Correspondingly the No Observed Effect Concentration was 100 mg/l.

Executive summary:

As the substance to be registered, PMDA, is unstable in water, the corresponding hydrolysis product Pyromellitic acid (PMA) was tested. The study was performed to assess the acute toxicity of the test material to rainbow trout (Oncorhynchus mykiss). The method followed that described in the OECD Guidelines No. 203, "Fish, Acute Toxicity Test". Following a preliminary range-finding test fish were exposed, in two groups of seven, to an aqueous solution of the test material, at a single concentration of 100 mg/l for a period of 96 hours at a temperature of approximately 14°C under semi-static test conditions. The number of mortalities and any sub-lethal effects of exposure in each test and control vessel were determined 3 and 6 hours after the start of exposure and then daily throughout the test until termination after 96 hours.

The 96 -Hour LC50 based on nominal test concentrations was greater than 100 mg/1 and correspondingly the No Observed Effect Concentration was 100 mg/l. It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/l.

Analysis of the test preparations at 0, 24 and 96 hours showed measured test concentrations to range from 93% to 97% of nominal and so the results are based on nominal test concentrations only.

Description of key information

As the substance to be registered, PMDA, is unstable in water (t1/2 approx. 20 min), the corresponding hydrolysis product Pyromellitic acid (PMA) was tested. The study was performed to assess the acute toxicity of the test material to rainbow trout (Oncorhynchus mykiss). The method followed that described in the OECD Guidelines No. 203, "Fish, Acute Toxicity Test". The 96 -Hour LC50 based on mean measured test concentrations was greater than 100 mg/l and correspondingly the No Observed Effect Concentration was 100 mg/l.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

Source: GLP-report