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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:

1996-01-12 (date of study plan) - 1996-03-06 (date of final report)

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

EC Directive 92/69/EEC, Annex, method C.1, Official Journal of the European Communities No. L383A, December 29 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Stability under test conditions: apparently stable (> 80 %)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: The method of the analytical determination of the concentrations is described in the related report "Determination of the test item in Combination with the Acute Toxicity Fish Test and the Pseudomonas Cell Multiplication Inhibition Test"

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
The solubility of the active ingredient of the test item in potable water is 14.3 mg/l (as stated by the sponsor). The stated nominal concentrations after dilution and filtration were based on this value.
Each pool contained 10l dilution water.
The control pool (pool No. 1) contained dilution water only.
0.5 ml of the test item was transferred into a 5000 ml volumetric flask and filled up to volume with dilution water. This solution was stirred overnight for the purpose of saturation. Afterwards the solution was filtrated into pool No. 2 containing 5000 ml dilution water. Resulting concentration: half saturated solution of the test item (7.2 mg/l).
0.5 ml of the test item were transferred into a 5000 ml volumetric flask and filled up to volume with dilution water. This solution was stirred overnight for the purpose of saturation. Subsequently 2500 ml of this solution was filtrated into pool No. 3 containing 7500 ml dilution water. Resulting concentration: 1/4 saturated solution of the test item (3.6 mg/l).
1250 ml of the 2nd saturated solution made before was filtrated into pool No. 4 containing 8750 ml dilution water. Resulting concentration: 1/8 saturated solution of the test item (1.8 mg/l).
625 ml of the 2nd saturated solution made before was filtrated into pool No. 5 containing 9375 ml dilution water. Resulting concentration: 1/16 saturated solution of the test item (0.9 mg/l).
312 ml of the 2nd saturated solution made before was filtrated into pool No. 6 containing 9688 ml dilution water. Resulting concentration: 1/32 saturated solution of the test item (0.45 mg/l).
The saturated solutions and the dilution water as well were prepared the day before (in the afternoon). They were stirred overnight in the same cabinet and at the same temperature as the fish test was performed.
No vehicle was used. The control for the vehicle was omitted.
The pools were permanently aerated.
To all appearances not dissolved test item was not discernible in the pools.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Strain: Brachydanio rerio, Teleostei, Cyprinidae
- Source: Südharz-Aquarium GmbH, Scharzfelder Str. 107 D-37431 Bad Lauterberg, Germany
- Length at study initiation (length definition, mean, range and SD): 2.5 - 3.5 cm

ACCLIMATION
The test fish were in good health and were acclimatized at least 12 -15 days of which at least 7 days they were kept in water of the quality used during the test.

FEEDING DURING TEST
fish were fed daily (Tetra Min Haupfutter) and discontinued at least 24 hours prior to the test period

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

none

Hardness:

200 mg/l CaCO3

Test temperature:

22.1-22.5°C

pH:

7.3 - 7.8

Dissolved oxygen:

8.2 - 8.

Conductivity:

Conductivity of deionized water: 0.4 -1.5 yS/cm

Nominal and measured concentrations:

nominal: 0, 0.45, 0.9, 1.8, 3.6, 7.2 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: 10 litre pools
- Aeration: The pools were permanently aerated, at least 80 % of the air saturation value.
- Renewal rate of test solution (frequency/flow rate): semi-statically; daily renewal of the test solutions
- Loading: maximum loading of 1.0 g fish/litre
- Groups: 10 fish per test concentration and per control
- Use of vehicles: No use of vehicles

TEST MEDIUM / WATER PARAMETERS
Conductivity of deionized water: 0.4 -1.5 µS/cm
Dilution water which was used in the test period was reconstituted water:
CaCl2: 2 mmol/l
MgSO4: 0.5 mmol/l
NaHCO3: 0.75 mmol/l
KCl: 0.075 mmol/l
The sum of calcium and magnesium ions in the dilution water was 2.5 mmol/l. The proportion of Ca : Mg ions was 4 : 1 and of Na : K ions 10 : 1. The acid capacity Ka4.3 of the solution was 0.8 mmol/l.
Derived from the CaCl2 addition, the hardness of water was equal to 2 mmol/l CaCO3 which corresponds to a hardness of 200 mg/l CaCO3 (valid: 10 - 250 mg/l).
pH values of test waters: neutral (7.3 - 7.8)
For each water renewal the reconstituted water was always left ready prepared overnight.

OTHER TEST CONDITIONS
- Photoperiod: 16 h light, 8 h dark
- Temperature: 20 - 24 ± 1 °C

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality, behaviour, length, weight, pH, dissolved oxygen

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.55 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Other biological observations: At 7.2 mg/l after 4 hours, 10 fish perished. At 3.6 mg/ 1 fish lies upside down at the bottom of the pool, with respiration movements. 6 fish show intensive respiration activity and uncontrolled movements. after 6 h, 1 fish shows intensive respiration
activity. At any other time of inspection no visible signs of abnormalities of surviving test fish were observed.
- Mortality of control: One fish of 150 died during acclimatization, no fish in control died.
- Other adverse effects control: The control fish showed neither mortality nor any visible sign of abnormality during the test time of 96 hours. Incidents in the course of the test which might have influenced the results could not be observed during the test period.

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test item towards fish was determined scientifically reasonable acc. EU method C.1, the given information indicates that testing was properly conducted, hence, the results can be considered sufficiently reliable. The LC50(96h) was determined to be 2.55 mg/l, so the substance does not need to be classified as acutely toxic as it is above the limit value for classification (1 mg/l) according to regulation 1272/2008 and amendments. However, as the substance is not readily biodegradable, it must be considered as Aquatic Chronic Cat. 2.

Executive summary:

The acute toxicity of Aminic salt of dialkyldithiophosphoric acid, formulation in mineral oil towards zebra fish (Brachydanio rerio) was determined semi-statically (daily renewal of the test solutions) according to EC Directive 92/69/EEC, Annex, method C.1 under GLP. The results are:

Table concentrations of the test item (nominal/measured)

	24h	48h	72h	96h
NOEC	1.8/0.09	1.8/--	1.8/--	1.8/0.13
LC0	1.8/0.09	1.8/--	1.8/--	1.8/0.13
LC50	2.55	2.55	2.55	2.55
LC100	3.6/1.3	3.6/--	3.6/--	3.6/--

Only moderate degradation was observed at relevant concentrations.

The substance is classified as Aquatic Chronic Cat. 2 according to regulation 1272/2008.

Description of key information

Acute Toxicity to fish: LC50(96h) = 2.55 mg/l (nominal) for Brachidanio rerio (semi-statically, EU method C.1, GLP)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

2.55 mg/L