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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from April 28 to May 03, 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Qualifier:
according to
Guideline:
EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)
Qualifier:
according to
Guideline:
other: OECD Series on Testing and Assessment No 23 - Guidance Document On Aquatic Toxicity Testing Of Difficult Substances And Mixtures ENV/JM/MONO(2000)6
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Specific details on test material used for the study:
Test item identification: YELLOW 2747
Batch number: 78-242-15
Manufacturing date: February 01, 2017
Expiry date: February 01, 2018
Chemical name: 1H-Benz[de]isoquinoline-1,3(2H)-dione, 6-amino-,N,N’-bis[mixed 2-ethylhexyl and 3-[(2-ethylhexyl)oxy]propyl] derivs
C.A.S. number: 1971906-58-7
The test item was stored at room temperature in the dark.

Sampling and analysis

Analytical monitoring:
no

Test solutions

Vehicle:
yes
Remarks:
acetone
Details on test solutions:
The test concentration was chosen in accordance with the results of a no GLP pre-test (range-finding test), whose relevant Raw Data were archived under the code of the present study.
The main final test included five nominal test item concentration in a geometric series, namely 0.043, 0.094, 0.207, 0.455 and 1.0 mg/L.

Since the test item is insoluble in water, it was added to the reconstituted water by the means of a solvent vehicle (acetone). Therefore a solvent control (reconstituted water whit the same concentration of acetone used as carrier) was added in order to verify the absence of toxic effect due to the solvent; moreover a negative control (reconstituted water without test item) was prepared.

The test medium was prepared as follows:

the day of the test start, a 10000.0 mg/L Stock Solvent Solution (SS) was prepared by direct weighing 0.0502 g of test item into 5.0 mL of acetone.
The resulting solution was observed to be yellow, with no undissolved test item.
The test was performed in glass beakers filled with 40 mL of test medium. The test vessels were covered with glass plates to reduce the loss of water and to avoid dust entry into test solutions.
The test vessels were labelled with the internal study number and all necessary additional information to ensure unmistakable identification.

For the test concentration, for the negative control and for the solvent control, group of 20 daphnids were used divided into four replicates of five daphnids each. The daphnids were randomly distributed to the test vessels at test initiation. The daphnids loading rate was about 8 mL per one daphnid (5 daphnids for 40 mL, according to OECD guideline 202, 2004).

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
The study was performed on the freshwater crustacean Daphnia magna Straus. A strain of this species was supplied by MicroBioTests Inc., Belgium in December 2011 (batch number: DM290911).
No acclimation was needed prior to testing.
It was daily fed with a fixed amount of the laboratory cultured green algae Pseudokirchneriella subcapitata (formerly Selenastrum capricornutum) and with a suspension of the yeast Saccharomyces cerevisiae.

At the test start the organisms used in the test were less than 24 hours old and were not first brood progeny.



Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
The daphnids were observed by visual inspection for immobility after 24 and 48 hours of exposure (those organisms not able to swim within 15 seconds after gentle agitation of the test beaker were considered to be immobile).
The immobilization percentage was calculated and reported

Test conditions

Test temperature:
20.0°C
pH:
7.43 – 7.57
Dissolved oxygen:
6.10 – 6.64 mg O2/L
Nominal and measured concentrations:
The biological results were referred to nominal concentration of test item.
Details on test conditions:
Oxygen Concentration: 6.10 – 6.64 mg O2/L
Light intensity: 1356 – 1500 Lux
Light Regime: 16 hours light and 8 hours dark
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 1 mg/L
Nominal / measured:
nominal
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
0.6 mg/L
Nominal / measured:
nominal
Basis for effect:
mobility
Remarks on result:
other: LCL and UCL
Remarks:
0.361 – 1.575
Duration:
24 h
Dose descriptor:
NOEC
Effect conc.:
1 mg/L
Nominal / measured:
nominal
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
0.094 mg/L
Nominal / measured:
nominal
Basis for effect:
mobility
Results with reference substance (positive control):
In the negative control and in the solvent control 0% of immobilization was observed and no daphnid was trapped on the test water surface or showed other signs of disease or stress. These values comply with the validity criteria of the test, that provides a maximum immobilization or a maximum number of daphnids showing distress signs of 10% in the negative control or in the solvent control at the end of the test.

Any other information on results incl. tables

Immobilization effect of test item toDaphnia magna.

 

Nominal test item

concentration

(mg/L)

No. of
Daphnia
tested

No. of immobilized
Daphnia

% of immobilized
Daphnia

24 h

48 h

24 h

48 h

0.0 (negative control)

20

0

0

0

0

0.0 (solvent control)

20

0

0

0

0

0.043

20

0

0

0

0

0.094

20

0

4

0

20

0.207

20

0

7

0

35

0.455

20

0

9

0

45

1.0

20

0

11

0

55

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The acute toxicity of test item YELLOW 2747 was tested on Daphnia magna under static conditions.

The EC0, the EC100, the EC50 value and the NOEC at 24 and 48 hours were assessed.
Results, calculated in terms of nominal test item concentration, were as follows:

Time (h) EC0(mg/L) EC100(mg/L) EC50(mg/L) NOEC(mg/L)
24 1.0 > 1.0 > 1.0 1.0
48 0.043 > 1.0 0.6 0.094

LCL, Lower Confidence Level and UCL, Upper Confidence Level= 0.361 – 1.575
Executive summary:

Title:                    YELLOW 2747: Acute toxicity to Daphnia magna in a 48-hour immobilization test under static exposure.

Evaluations:          Immobilization after 24 and 48 hours of exposure, abnormal behaviour or appearance were compared with corresponding parameters in the negative control.

Test concentrations:              Negative control (reconstituted water without test item), solvent control (reconstituted water without test item but with the same concentration of solvent as in test item concentration tested) and 0.043, 0.094, 0.207, 0.455 and 1.0 mg test item/L.

Analytical check of tested concentrations: not performed

Exposure regime:   static

Test duration:        48 hours

Test environment:  controlled environment room.

Monitoring of test medium quality: pH and oxygen concentration was measured at the start and at the end of the test, in the negative control, solvent control and test item solution.

pH of test medium:               7.43 – 7.57

Oxygen Concentration:        6.10 – 6.64 mg O2/L

Light intensity:                       1356 – 1500 Lux

Light Regime:                        16 hours light and 8 hours dark

Test temperature:                 20.0°C

Results expression:               The biological results were referred to nominal concentration of test item.

Results, calculated in terms of nominal test item concentration, were as follows:

Time (h)       EC0(mg/L)       EC100(mg/L)       EC50(mg/L)       NOEC(mg/L)

24       1.0       > 1.0       > 1.0       1.0

48       0.043       > 1.0       0.6       0.094

LCL, Lower Confidence Level and UCL, Upper Confidence Level= 0.361 – 1.575