Sodium [[α,α'-(ethylenediimino)bis[2-hydroxybenzene-1-acetato]](4-)]ferrate(1-)

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:

other: see 'Remark'

Remarks:

Basic information reported. In accordance to a test method reported by Bathe et.al., 1974. GLP compliance not applicable, since it was established one year later.

Qualifier:

no guideline followed

Principles of method if other than guideline:

In accordance with the method reported by Bathe et. al. (1974) which is a modification of the procedure proposed by the United States Department of the Interior Fish and Wildlife Service, reported in "Procedure for Evaluation of Acute Toxicity of Pesticides to Fish and Wildlife".

GLP compliance:

not specified

Remarks:

Not applicable. GLP principles are applied since 1978, not before.

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
- Name of test material: technical CGA 65047, identical to Fe(Na)EDDHA
- Analytical purity: 100 % (UVCB)

Analytical monitoring:

not specified

Details on sampling:

Oxygen dissolved in the water and pH were monitored at 24 h intervals throughout the 96 h testing.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
Various concentrations of the substance were prepared by adding the test material dissolved in aqua dest. (stock solution), (solubility of active ingr. in water: well miscible) at the appropriate amount ensuring that the total volume in each vessel remained the same.

Test organisms (species):

other: Rainbow trout, Carp, Catfish and Bluegill

Details on test organisms:

TEST ORGANISM
1. Rainbow trout (Salmo gairdneri), supplied by "Mändli", Liestal, Switzerland, fish breeding station.
Average Weight: 6.8 g
Average Length: 82 mm

2. Carp (Cyprinus carpio), supplied by "Riegger", Ettenheim, W.Germany, fish breeding station.
Average Weight: 7.5 g
Average Length: 80 mm

3. Catfish (Ictalurus nebulosus), supplied by "Weber", Wohlen Switzerland.
Average Weight: 3.9 g
Average Length: 65 mm

4. Bluegill (Lepomis macrochirus), supplied by "Exotarium Stubenvoll", Allschwil, Switzerland.
Average Weight: 4.3 g
Average Length: 60 mm

Prior to testing, the fish were adapted to laboratory conditions for a minimum of 10 d. They were kept in 250 L glass tanks containing normal drinking water which was aerated and filtered continuously through a filter bed of activated charcoal. The fish were fed daily. Prior to testing the food was withdrawn for 3 d.

Test type:

not specified

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

No post exposure observation period reported.

Hardness:

No details available.

Test temperature:

14 +/- 2 °C (Trout, Bluegill)
22 +/- 2 °C (Carp, Catfish)

pH:

Rainbow trout: 7.2 - 7.4
Carp: 6.3 - 7.4
Catfish: 6.8 - 7.6
Bluegill: 6.4 - 7.3

Dissolved oxygen:

Rainbow trout: not determined
Carp: 1.4 - 8.1
Catfish: 5.7 - 8.4
Bluegill: 4.8 - 9.2

Salinity:

Not applicable.

Nominal and measured concentrations:

Rainbow trout: 28 mg/kg, 100 mg/L
Carp: 49 mg/kg, 100 mg/L
Catfish: 49 mg/kg, 100 mg/L
Bluegill: 49 mg/kg, 100 mg/L

Details on test conditions:

TEST SYSTEM
The fish were tested in glass tanks, of 12 and 15 litre capacity. Testing was performed in reconstituted water prepared of deionised water to which salts were added in the following amount per litre: 30 mg calcium sulphate, 30 mg magnesium sulphate, 48 mg sodium bicarbonate, 3 mg potassium chloride. With exception of experiments on trout the water was not aerated during testing. Three to four fish were placed in each tank resulting in the following weights of fish per volume of water:
Rainbow trout: 1. 8 g/L
Carp: 2.0 g/L
Catfish: 1. 0 g/L
Bluegill: 1.4 g/L

Reference substance (positive control):

no

Remarks:

Even in OECD Guidelines not mandatory.

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)

Remarks on result:

other: for Rainbow trout, Carp, Catfish and Bluegill

Details on results:

Symptoms for each particular species:
Rainbow trout: slight discolorations, hypersensitivity, retarded movements and hyperventilation (all test concentrations)
Carp: slight hypersensitivity (within 48 h), slight diminished sensitivity (after 72 h), slight retarded movements and hyperventilation (all test concentrations)
Catfish: slight hypersensitivity at a test concentration of 49 ppm (within 48 h), slight diminished sensitivity (at 100 ppm and after 72 h for 49ppm), slight retarded movements
Bluegill: slight hypersensitivity at a test concentration of 100 ppm, slight diminished sensitivity, retarded movements and decreased respiration rate (at all test concentrations)

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

No statistics and error estimates.

Validity criteria fulfilled:

not specified

Remarks:

Basic scientific principles were met.

Conclusions:

In this acute toxicity test with Rainbow trout, Carp, Catfish and Bluegill the 96 h LC50 was greater than 100 mg/L. These results are based on nominal concentrations. The test material is practically devoid of acute toxicity to Trout, Carp, Catfish and Bluegill when tested and evaluated according to the procedure of Bathe et al.

Executive summary:

The acute toxic effects of Fe(Na)EDDHA to Rainbow trout, Carp, Catfish and Bluegill were determined (Sachssee and Barthe, 1977). The fish were tested in glass tanks of 12 and 15 litre capacity. Testing was performed in reconstituted water prepared of deionised water. Various concentrations of Fe(Na)EDDHA were prepared by adding the test material dissolved in aqua dest. (stock solution). (solubility of active ingr. in water: well miscible) at the appropriate amount ensuring that the total volume in each vessel remained the same.

In this acute toxicity test with Rainbow trout, Carp, Catfish and Bluegill the 96 h LC50 was greater than 100 mg/L. These results are based on nominal concentrations. Fe(Na)EDDHA is practically devoid of acute toxicity to trout, carp, catfish and bluegill when testet and evaluated according to the procedure of Bathe et al.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010-04-27 - 2010-06-24

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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)

Remarks:

State Office for the Environment, Water and Trade Control, Mainz, Germany

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
- Name of test material (as cited in study report): Acetic acid,oxo-,sodium salt (FeNaEDDHA)
-Synonym: Acetic acid,oxo-,sodium salt, reaction products with ethylenediamine and phenol,iron sodium salts
- Physical state: Solid/ red, dark
- Analytical purity: 100 % (UVCB)
- Storage condition of test material: At ambient temperature under exclusion of light

Analytical monitoring:

yes

Details on sampling:

Samples for analytical determination of the concentration were taken within 30 minutes before insertion of the test organisms (start of exposure) and at the end of the exposure after approx. 96 hours. Water samples were taken from the middle of the test vessel using a glass pipette and were transported to the analytical laboratory in glass ampoules.
The transport to the Analytical Laboratory was done on the day of sampling.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
The test substance (1.20168 g) was weighed into a beaker. The defined volume of test water (10 L) was filled into the test vessel and an aliquot was removed and mixed with the test substance in a beaker using an ultra turrax high shear mixer. The mixture was then poured into the test vessel and the entire volume of test solution was homogenized again with an ultra turrax.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Source: ZF BioLabs, Ronda da Valdecarrizo 41°B. 28760, Tres Cantos (Madrid), Spain
- Age at study initiation: Approx. 12 months
- Length at study initiation: 3.3 cm (3.2 – 3.4 cm)
- Weight at study initiation: 0.35 g (0.25 – 0.48 g)
- Feeding during test: ad libitum, additionally generally on workdays live brine shrimp (artemia nauplii).
- Food type: Commercial fish diet “Tetramin“

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

No post exposure observation period reported.

Hardness:

Approx. 1mmol/L = 100 mg/L CaCO3

Test temperature:

24 °C

pH:

7.5 - 8.5

Dissolved oxygen:

> 80 % saturation

Salinity:

Not applicable.

Nominal and measured concentrations:

120 mg/L (Nominal concentration based on test substance mass without a correction for composition)

Details on test conditions:

TEST SYSTEM
- Test vessel: Stainless steel aquaria (29 cm long, 21 cm wide, 22 cm high) partially covered with a black cover
- Volume of water: 10 L
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- Aeration: Slight

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Non chlorinated charcoal filtered tap water (Frankenthal, Germany) mixed with deionized water, aerated
- Conductivity: Approx. 250 µS/cm (at 25 °C)

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light, 8 hours dark
- Light intensity: The experiment was performed under reduced light intensity, since the test substance is potentially light sensitive.
- Feeding: None

EFFECT PARAMETERS MEASURED:
The fish were observed for mortality and toxic signs (changes appearance and abnormal behaviour in comparison to the control group) within 1 hour after start of exposure and 6, 24, 48, 72 and 96 hours after start of exposure. Fish were considered dead if there was no visible movement and no reaction after touching. Dead fish were removed from the test vessels.

Reference substance (positive control):

no

Remarks:

Not mandatory according to the applied test guideline.

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No toxic signs were observed during the entire exposure time.

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

Since the study is designed as a limit test, no statistical analysis was performed.

Sublethal observations / clinical signs:

Cumulative mortality (number of dead animals):

Nominal concentration [mg/L]	Number of fish	1 hour	6 hours	24 hours	48 hours	72 hours	96 hours
0	7	0	0	0	0	0	0
120	7	0	0	0	0	0	0

Toxic signs:

Nominal concentration [mg/L]	Number of fish	1 hour	6 hours	24 hours	48 hours	72 hours	96 hours
0	10	n	n	n	n	n	n
120	10	n	n	n	n	n	n

Explanation of abbreviations

n = no toxic signs observed                   

The following effect concentrations (mg/L) were obtained based on nominal concentrations:

	Observation time (hours after start of exposure)
	1 hour	6 hours	24 hours	48 hours	72 hours	96 hours
LC0	>120	>120	>120	>120	>120	>120
LC50 (95% confidence limits)	> 120 -	> 120 -	> 120 -	> 120 -	> 120 -	> 120 -
LC100	> 120	> 120	> 120	> 120	> 120	> 120
NOEC	>120	>120	>120	>120	>120	>120

- insufficient data for calculation

Validity criteria fulfilled:

yes

Remarks:

All study results are consistent with all validity criteria of the applied guideline.

Conclusions:

The study is regarded as a valid guideline study with certificated GLP compliance. According to the reported LC50 value (> 120 mg/L) the test substance is not classified as toxic to fish.

Executive summary:

The acute toxicity effects of the test substance to zebrafish (Danio rerio) over a 96 hour static exposure period were determined according to the principles of OECD-Guideline 203 and EU Method C.1 (Salinas, 2010). The experiment was conducted as limit test with an analytically confirmed test concentration of 120 mg/L. Therefore, no statistical analysis was performed. Since neither mortality nor toxic signs were observed, the LC50(96h) value is greater than tested concentration of 120 mg/L based on nominal concentrations. Based on the mean of analytically determined concentration, a value of greater or equal to 122 mg/L is reported. Since the analytically determined concentrations of the test substance in the test solutions were within +/- 20% of the nominal concentrations, the effect concentration is expressed relative to the nominal concentration. The corresponding NOEC was therefore determined to be greater than or equal to 120 mg/L.

Description of key information

LC50 (96 h): > 120 mg/L [OECD 203; zebrafish (Danio rerio); nominal; read-across substance Fe(Na)EDDHA]

LC50 (96 h): > 100 mg/L [no guideline followed; Rainbow trout, Carp, Catfish and Bluegill; nominal; read-across substance Fe(Na)EDDHA]

Additional information

The acute toxicity to freshwater or marine fish was not investigated so far for the substance o-o-EDDHA (CAS 16455-61-1). To fulfil this relevant endpoint for the intended registration under REACH, a read-across from the closely related substance Fe(Na)EDDHA (CAS 84539-55-9) was performed.

Two experimental reports are available for Fe(Na)EDDHA (CAS 84539-55-9) concerning short-term toxicity to freshwater fish. The zebrafish (Danio rerio) was used as test organism for an exposure period of 96 hour in an experiment conducted according to the principles of OECD Guideline 203 and EU Method C.1 (Salinas, 2010). A limit test was performed with an analytically confirmed test concentration of 120 mg/L. Therefore, no statistical analysis was performed. Since neither mortality nor toxic signs were observed, the LC50(96h) value is greater than the tested concentration of 120 mg/L based on nominal concentrations. Based on the mean of analytically determined concentration, a value of greater or equal to 122 mg/L is reported. Since the analytically determined concentrations of the test substance in the test solutions were within +/- 20 % of the nominal concentrations, the effect concentration is expressed relative to the nominal concentration. The corresponding NOEC was therefore determined to be greater than or equal to 120 mg/L. Furthermore, the acute toxic effects of Fe(Na)EDDHA to Rainbow trout, Carp, Catfish and Bluegill were determined by Sachssee and Barthe (1977). The fish were tested in glass tanks of 12 and 15 litre capacity. Testing was performed in reconstituted water prepared of deionised water. Various concentrations of Fe(Na)EDDHA were prepared by adding the test material dissolved in aqua dist. (Stock solution). Solubility of active ingr. in water: well miscible. At the appropriate amount ensuring that the total volume in each vessel remained the same. In this acute toxicity test with Rainbow trout, Carp, Catfish and Bluegill the 96 h-LC50 was greater than 100 mg/L. These results are based on nominal concentrations. Fe(Na)EDDHA is practically devoid of acute toxicity to Trout, Carp, Catfish and Bluegill when tested and evaluated according to the procedure of Barthe et al.