Reaction mass of aluminium tris(5,7,12,14-tetrahydro-7,14-dioxoquino[2,3-b]acridine-2-sulphonate) and 5,12-dihydro-quino[2,3-b]acridine-7,14-dione

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

According to OECD 202 and GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Aluminium tris(5,7,12,14-tetrahydro-7,14-dioxoquino[2,3-b]acridine-2-sulphonate)

EC Number:

278-207-2

EC Name:

Aluminium tris(5,7,12,14-tetrahydro-7,14-dioxoquino[2,3-b]acridine-2-sulphonate)

Cas Number:

75431-69-5

Molecular formula:

C20H12N2O5S.1/3Al

IUPAC Name:

aluminum tris(7,14-dioxo-5,7,12,14-tetrahydroquino[2,3-b]acridine-2-sulfonate)

Constituent 2

Reference substance name:

5,12-dihydroquino[2,3-b]acridine-7,14-dione

EC Number:

213-879-2

EC Name:

5,12-dihydroquino[2,3-b]acridine-7,14-dione

Cas Number:

1047-16-1

Molecular formula:

C20H12N2O2

IUPAC Name:

5,12-dihydroquino[2,3-b]acridine-7,14-dione

Constituent 3

Reference substance name:

Dialuminium tris(5,7,12,14-tetrahydro-7,14-dioxoquino[2,3-b]acridine-2,9-disulphonate)

EC Number:

243-319-2

EC Name:

Dialuminium tris(5,7,12,14-tetrahydro-7,14-dioxoquino[2,3-b]acridine-2,9-disulphonate)

Cas Number:

19795-24-5

Molecular formula:

C20H12N2O8S2.2/3Al

IUPAC Name:

dialuminium tris(5,7,12,14-tetrahydro-7,14-dioxoquino[2,3-b]acridine-2,9-disulphonate)

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

sample preparation

each sample in a volume of 10 mL was taken and analyzed in a TOC analyzer.

nominal concentration [mg C/L] determined TOC in replicates [mg C/L]

1 2 3 4 5 Average Recovery [%] Standard Deviation

control 0.772 0.796 - - - 0.784 - 0.017
1.0 1.086 1.081 1.076 1.042 1.161 1.089 108.92 0.044
10.0 9.508 9.339 9.691 9.659 9.591 9.558 95.58 0.141

Test solutions

Vehicle:

no

Details on test solutions:

Preparation of the test solution
100 mg/L test item was weighed out in a glass flask with ISO test water yielding a heterogeneous mixture with undissolved particles. The mixture was left for mechanical shaking (48 hours, 90 rpm, in the dark). The still heterogeneous mixture was filtered twice (conditioned paper filter and membrane filter 0.22 µm, Millipore). The filtrate was visually clear. After 24 hours of exposure the test item solution was renewed.


Test concentration
The effects of the test item was tested in a limit test.

Control
Dilution water without test item exposed under the same conditions as the test groups.


In a preliminary non-GLP study under semi-static conditions three test item concentrations were used for exposure: filtrates of loading 100 mg/L, 10 mg/L and 1 mg/L were used for exposure. respectively. The solutions were prepared in analogy to the concentration of the definite test.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Test system
Daphnia magna STRAUS, up to 24 hours old

Reason for the selection of the test system
Daphnia magna is the preferred species in accordance with the test guideline and is bred at the test facility.

Origin
standard laboratory culture, Institute of Industrial Organic Chemistry, Branch Pszczyna

Breeder
Institute of Industrial Organic Chemistry, Branch Pszczyna, Department of Ecotoxicology, Doswiadczalna 27, 43-200 Pszczyna, Poland

Culture
In glass vessels (150 mL capacity) with 100 mL ISO test water, at room temperature, 16 hours illumination, one parent per vessel

Culture medium
ISO test water as recommended by OECD 202 was used. The ISO test water was prepared based on deionised water by adding appropriate amounts of stock solutions; ingredients are reagent grade chemicals.The composition of the culture medium is presented
in Table 2.

Culture feeding
The culture daphnids are fed ad libitum with a mixture of two species in a 2:1 ratio originating from different cultures, Pseudokirchneriella subcapitata and Desmodesmus subspicatus. The algae are cultured in the Laboratory of Aquatic Toxicity. Group B vitamins and micronutrients necessary for proper growth were supplied with the lyophilized suspension of Spirulina sp.

Origin of the food algae
Sammlung von Algenkulturen (SAG), Pflanzenphysiologisches Institut der Universität Göttingen, Nikolausberger Weg 18, 37073 Göttingen, Germany

 

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Test temperature:

19 - 21°C, constant within ± 1°C

pH:

Water Quality Parameters at the Start of the Exposure (0 hours)


Nominal loading
of the test item
[mg/L] pH-value Dissolved O2 concentration[ mg/L]

100
(filtered solution) 7.69 8.9
Control 7.61 9.5


Water Quality Parameters after 24 h and 48 h of exposure

Nominal loading
of the test item
[mg/L] pH-values Dissolved O2 concentration [mg/L]

after 24 h after 48 h after 24 h after 48 h
spent fresh spent spent fresh spent

100
(filtered solution) 7.69 7.62 7.59 7.85 9.1 9.9 9.8
Control 7.62 7.58 7.55 7.83 8.7 9.8 9.8

values in fresh media were measured in samples before split up into replicates
values in spent media were measured in samples of pooled replicates

Dissolved oxygen:

Water Quality Parameters at the Start of the Exposure (0 hours)


Nominal loading
of the test item
[mg/L] pH-value Dissolved O2 concentration[ mg/L]

100
(filtered solution) 7.69 8.9
Control 7.61 9.5


Water Quality Parameters after 24 h and 48 h of exposure

Nominal loading
of the test item
[mg/L] pH-values Dissolved O2 concentration [mg/L]

after 24 h after 48 h after 24 h after 48 h
spent fresh spent spent fresh spent

100
(filtered solution) 7.69 7.62 7.59 7.85 9.1 9.9 9.8
Control 7.62 7.58 7.55 7.83 8.7 9.8 9.8

values in fresh media were measured in samples before split up into replicates
values in spent media were measured in samples of pooled replicates

Details on test conditions:

Test method
semi-static procedure

Test duration
48 hours

Test vessels
Glass beakers of 150 mL capacity, loosely covered with transparent lids

Test volume
20 mL, either test item concentration or the control

Dilution water ISO Test water, according to OECD 202, Annex 3

Number of daphnids
20 daphnids, divided into 4 replicates, each with 5 daphnids and replicates used for the test solution and the control.

Age of the daphnids at the start of the exposure
less than 24 hours old daphnids from a healthy stock were used for the study. No first brood progeny was used for the test

 
Application
the daphnids were exposed in glass beakers of 150 mL capacity, containing either 100 mL of eiterh test item concentration or the control. Each treatment was split up into four replicates. Five daphnids were introduced into each replicate; hence there wer 20 mL of a given treatment per daphnid.

Test temperature (target)
19 - 21°C, constant within ± 1°C, the temperature was continuously recorded using a sensor submerged in an additional test vessel containing 100 mL of the test medium.


Photoperiod (target)
16/8 hours light/dark cycle, fluorescent lightning

Feeding
The daphnids were not fed during the study.

RANGE-FINDING STUDY

In a preliminary non-GLP study under semi-static conditions three test item concentrations and a control were used for exposure: filtrates of loading 100 mg/L, 10 mg/L and 1 mg/L were used for exposure. respectively. No effects were observed.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Effect concentrations 2

Duration:

24 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Effect concentrations 3

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Effect concentrations 4

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Effect concentrations 5

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Effect concentrations 6

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Details on results:

See below

Results with reference substance (positive control):

A reference test was conducted as an acute immobilisation test under static conditions with a test duration of 48 hours in order to prove the validity of the test system and test conditions at the test facility.

Reference item Potassium dichromate p.a.
Purity 100.0 %

Test concentrations 0.32 - 3.2 mg/L (nominal)

24 h                     48 h

EC50 1.01 0.80
(0.87 - 1.18)      (0.70 - 0.91)

Reported statistics and error estimates:

Methods of evaluation

According to OECD Guidance Document No 23 the effects of the saturated solution were deduced directly from the observed immobilisation rates. If an effect cannot be detected in a test with a saturated solution, the result have to be reported as no toxic effects at saturation.
Therefore, the effect levels (NOEL, LOEL, EL50) were given based on the nominal loading of the test item.

EC-values and statistical
An EC50-value was calculated only for the reference item

Any other information on results incl. tables

composition of ISO test water, according to OECD 202 (Annex 3)

stock solution		Volume of stock solution added to each 1 L of ISO test water [mL]	Concentration in ISO test water [mg/L]
ingredient	Amount of substance added to 1 L [g]
CaCl2x 2 H2O	11.76	25	294
MgSO4x 7 H2O	4.93	25	123.3
NaHCO3	2.59	25	64.7
KCl	0.23	25	5.7

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In the saturated solution with a nominal loading of 100 mg/L of the test item no effects were observed on Daphnia magna.

Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the saturated and filtered solution with a nominal loading of 100 mg/L of the test item were determined according to OECD 202 (2004).The limit test was conducted under semi-static conditions over a period of 48 hours. The test medium was renewed after an exposure time of 24 hours.

The test item is hardly soluble in water. Therefore, a saturated solution (100 mg/L) was prepared with dilution water (ISO test water, acc. to OECD 202, Annex 3) by shaking with approximately 90 rpm for 48 hours at room temperature followed by filtration with a membrane filter (0.22 µm, Millipore) to remove undissolved test item and to achieve the saturated solution. The saturated solution was visually clear throughout the exposure period of 48 hours.

Twenty daphnids were exposed to the saturated solution and the control.

Total organic carbon (TOC) analysis was performed in samples of the filtrate of a loading of 100 mg/L and the control at exposure initiation, at renewal and at exposure termination.

The results were corrected by the value determined for the control sample in a series and presented as the concentrations of TOC. each sample in a volume of 10 mL was taken and analyzed in a TOC analyzer. The linearity of the response of the analytical method, its precision, recovery and limit of quantification (LoQ) were assessed in the process of the analytical method validation. Solutions of potassium hydrogen phthalate were used as standard of carbon. This analytic approach did not account for any degradation or metabolisation effects which may have occured during exposure. Therefore, the endpoint values were determined based on the nominal test item concentration.

Water quality parameters (i.e. pH-value and dissolved oxygen concentration), measured at the start (0 hours), after 24 hours and at the end of the exposure (48 hours), were within the acceptable limits. The validity criteria of the test guideline were fulfilled.

In the saturated solution with a nominal test item loading of 100 mg/L no effects were observed. Therefore, the test item has not to be classified according to Regulation (EC) no 1272/2008.