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Short-term toxicity to aquatic invertebrates

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Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
19.7.- 26.10.2000
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to guideline
Guideline:
other: method No. L 49: “Acute Toxicity for Daphnia, immobilization test. Chemical formulations and waste waters”
Deviations:
no
Principles of method if other than guideline:
Young daphnia born in 24 hours are segregated to the handling container from which they entered the experiment. The sensitivity of the test species is periodically monitored by testing with the reference substance, potassium dichromate.The sample is dissolved in the dilution water at a concentration of about 0.1 g / L and forms a dark opaque solution. Higher concentrations can not be used for the intense color and the swiming of Daphnia to the surface. The volume of the test mixture for one dilution is 50 ml.Preliminary test is carried on with 10 organisms, full test with 10 as well in two parallel rows in which the number of individuals for evaluating is aggregated. Immobilization is monitored at 24 and 48 hours. The test is repeated 2x. Solutions are not aerated. The temperature of the solution during the test is 21.0 ± 1.0 °C. The oxygen content and pH values are measured in each vessel at the beginning and end of the test.
GLP compliance:
no
Specific details on test material used for the study:
Ostazinová modř H-5R, mixture No. A0 671No. of sample: 698A/00 Curent name: Reactive Blue 234
Analytical monitoring:
no
Details on test solutions:
The stock solution of the test substance was prepared in the dilution water. 100 mg of the test substance was weighed into 1000 mL of the dilution water for the preliminary test; dissolved sample forms a dark opaque solution. Higher concentrations can not be used for the intense color and the swiming of Daphnia to the surface.The concentrations of solutions used in test were obtained by dilution of the stock solution with dilution water. The stock solution for the preliminary test: 0.1 g·L-1 The stock solution for the main test No.1, 2: 0.1038 g·L-1
Test organisms (species):
Daphnia magna
Details on test organisms:
Daphnia magna Straus were obtained by aperiodic parthenogenesis from own laboratory breeding. Daphnia were fed by a mixture of algae.The grow-up gravid females are separated into dilution water. Young daphnia born in 24 hours are segregated to the handling container from which they entered the experiment. The sensitivity of the test species and correctness of test performance is periodically verified by testing with the reference substance, potassium dichromate.TEST ORGANISM- Common name: Daphnia Magna- Strain: Straus- Source: own laboratory breeding- Age at study initiation: young daphnia born in 24 hours- Method of breeding: aperiodic parthenogenesis- Feeding during test: noACCLIMATIONThe grow-up gravid females are separated into dilution water. Young daphnia born in 24 hours are segregated to the handling container from which they entered the experiment. - Type and amount of food: a mixture of algae- Health during acclimation: healthy
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Test temperature:
21.0 ± 1.0 °CMain test No. 1: 20.8 - 21.1 °CMain test No. 1: 20.0 – 20.9 °C
pH:
Main test No. 1: pH of the control: 7.84 – 7.90pH of the tested concentrations: 7.90 – 8.24Main test No. 2: pH of the control: 7.86 – 7.91pH of the tested concentrations: 7.90 – 8.18
Dissolved oxygen:
Main test No. 1: Oxygen content in the control: 8.7 mg·L-1Oxygen content in the tested concentrations: 8.5 – 8.7 mg·L-1Main test No. 2: Oxygen content in the control: 8.7 – 8.8 mg·L-1Oxygen content in the tested concentrations: 8.6 – 8.8 mg·L-1
Nominal and measured concentrations:
Preliminary test: 80,70,60,50,40,30,20 mg/L mg/L + controlFull test 1 and 2: 103.8, 101.7, 99.6, 97.6 , 95.5, 93.4, 91.3, 89.3 mg/L + control
Details on test conditions:
TEST SYSTEM- Test vessel: glass beakers (volume of the test solution is 50ml in each test beaker)- Type (delete if not applicable): open- Aeration: no- No. of organisms per vessel: 10- No. of vessels per concentration (replicates): 2- No. of vessels per control (replicates): 2Preliminary Test: 10 animals Main test No.1, 2: 10 animals in two parallel rows (20 animals)Dilution water was prepared just before the test by admixing 2.5 mL of stored solutions A, B, C and D into 1 000 mL of deionized water.A. 117.6 g CaCl2.2H20 in 1 L of deionized waterB. 49.3 g MgSO4.7H2O in 1 L of deionized waterC. 25.9 g NaHCO3 in 1 L of deionized waterD. 2.3 g KCl in 1 L of deionized water
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
98.8 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mobility

The test quality criteria

These quality criteria given in the current guideline were fulfilled.

1.      Immobilisation in the controls must not exceed 10 % at the end of the test (there were no immobilisation)

2.      The dissolved oxygen concentrations (throughout the test) should remain above 3 mg·L-1

Validity criteria fulfilled:
yes
Executive summary:

The test substance, Reactive Blue 234, was tested in acute immobilisation test on Daphnia magna. The test was performed as static.

Test result: EC50 (48h) = 98.8 mg·L-1  (measured concentration - arithmetic mean)

Description of key information

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
98.8 mg/L

Additional information

An ErC50 value from acute toxicity test on invertebrates is 98.8 mg/l and the substance is not rapidly degradable. These experimental results would led to classification (Aquatic Chronic 3, H412) however the results on other taxonomic species - fish and algae - were above 100 mg/L and the result on daphnids is practically 100 mg/L as well, so it led to the decision not to classified the substance Reactive Blue 234 as toxic for aquatic environment.