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Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
96 hours
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP compliant study conducted according to OECD guideline 203 (1992).

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report date:
2010

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
Fumaric acid
EC Number:
203-743-0
EC Name:
Fumaric acid
Cas Number:
110-17-8
IUPAC Name:
but-2-enedioic acid
Details on test material:
- Name of test material (as cited in study report): Fumaric acid
- Molecular formula (if other than submission substance): C4H4O4
- Smiles notation (if other than submission substance): C(=C/C(=O)O)\C(=O)O
- InChl (if other than submission substance): InChI=1/C4H4O4/c5-3(6)1-2-4(7)8/h1-2H,(H,5,6)(H,7,8)/b2-1+
- Physical state: white powder
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not applicable

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
Analytical samples were taken from freshly prepared and 24 hour aged samples for each of the four 24 hpur exposure periods. In addition a sample was taken after 4 hours during the initial 24 hour exposure period.

Test solutions

Vehicle:
no
Details on test solutions:
To increase the stability of the test substance, the fish media (local tap water and deionised water, 1:1) were autoclaved which resulted in partly precipitation of the ions accountable for the water hardness. This precipitate was removed through filtration with folded filters (Schleicher Schuell GmbH, D-37582, Germany), therefore the autoclaved medium showed decreased water hardness (which was in the range given by the guidelines) compared to the holding water.

A nominal test substance concentration of 100 mg/L was prepared. At each point of time (start, 24, 48, and 72 hours) nominal 500 mg test substance were diluted in autoclaved dilution water by manual homogenisation, stirring for 10 minutes or ultrasonication for 5 minutes. Subsequently the test substance solution was filled up to a total volume of 5 litres with autoclaved dilution water. As the autoclaved dilution water exhibits decreased oxygen content, the test media were aerated for about 15 minutes before introduction of the fish into the exposure tanks.

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
The fish were obtained from Fa. Tomy`s Zoo GmbH, Joseph Streblgasse 6, 2345 Brunn am Gebirge. The fish were acclimated in the laboratory for for six weeks prior to use.
Feed: TetraMin; Tetra Werke, Dr.rer.nat. Ulrich Baensch GmbH, D-49304 Melle.
Exposure: no feeding (last feeding about 24 hours before start of the exposure).
As the allocation of the fish to their groups was performed using a randomisation procedure the size and the weight of the control fish is assumed to be representative for the entity of all fish used.
Length - 2.69 (SD: 0.24) (mean of control group at the end of study).
Weight - 0.41 (SD: 0.11) (mean of control group at the end of study).

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Post exposure observation period:
Not applicable

Test conditions

Hardness:
Between 140 and 150 mg CaCO3/L.
Test temperature:
The temperature ranged between 19 and 21 °C.
pH:
pH: 7.7 to 8.1 in the control medium and between 4.3 and 5.0 in the test substance medium. Due to the acidic nature of "FUMARIC ACID", addition of the test substance resulted in a marked pH change which had at the concentration tested no influence on survival of the fish. The pH of the control as well as of the test substance group was sufficiently stable throughout the test period.
Dissolved oxygen:
At least 88 % of the saturation value at the corresponding temperature throughout the test period (range 88 - 100%)
Salinity:
Not applicable.
Nominal and measured concentrations:
Control and 100 mg/L.
Details on test conditions:
Test vessels were full glass vessels (29 cm x 20 cm x 24 cm), filled with 5 L of control and test medium and covered with transparent plastic
lids. Each test vessel had seven fish, with a single replicate per concentration. Maximum loading of 1.0 g fish/litre. A 16 hour light and 8 hour dark cycle was in operation during the study.
Reference substance (positive control):
no

Results and discussion

Effect concentrations
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
There was no mortality during the exposure period in either the control or the 100 mg/L treatment.
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
Not applicable.

Any other information on results incl. tables

Sublethal observations / clinical signs:

The analytical results are as follows:

Nominal concentration (mg/L)

Timepoint (h)

Measured concentration (mg/L)

Replicate 1

Replicate 2

Mean

100

0

99.82

99.22

99.52

100

4

101.37

100.97

101.17

100

24 (old)

101.54

101.34

101.44

100

24 (new)

105.68

105.68

105.68

100

48 (old)

99.65

102.66

101.16

100

48 (new)

96.95

98.26

97.61

100

72 (old)

97.71

96.52

97.12

100

72 (new)

91.23

91.13

91.18

100

96 (old)

99.59

96.19

98.36

Overall mean

-

-

-

99.03

The limit of quantification was 0.58 mg/L. All of the control measurements were less than the LOQ.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The 96 hour LC 50 for malic acid is conisdered to be > 100 mg/l
The 96-hour LC50 determined was >100 mg fumaric acid/L (nominal concentration).
The NOEC (no-observed effect concentration) was 100 mg fumaric acid/L.
Executive summary:

Juvenile D. rerio were exposed to a single fumaric acid concentration of 100 mg/L in a semi-static 96 -hour acute toxicity test according to OECD 203 guidelines. The test design included an untreated control comprising of dilution water only. The concentration of fumaric acid was verified analytically and the mean measured concentration was 99.03% of nominal. Due to the acidic nature of the mateiral addition of the test substance resulted in a marked pH change (pH 4.3 - 5.0) which had at the concentration tested no influence on survival of the fish.

The 96-hour LC50 determined was >100 mg fumaric acid/L (nominal concentration). The NOEC (no-observed effect concentration) was 100 mg fumaric acid/L.

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