1,2-dihydroxyanthraquinone

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 2017-09-13 to 2017-10-20

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

3 April 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

1,2-dihydroxyanthraquinone

EC Number:

200-782-5

EC Name:

1,2-dihydroxyanthraquinone

Cas Number:

72-48-0

Molecular formula:

C14H8O4

IUPAC Name:

1,2-dihydroxy-9,10-dihydroanthracene-9,10-dione

Test material form:

solid: particulate/powder

Details on test material:

- Appearance: solid yellow powder

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 10.0, 4.55, 2.07, 0.939, 0.427 mg/L and control
- Sampling method: samples taken from all test item concentrations and control at test start and after 24 hours from fresh and aged solutions and after 48 hours from aged solutions. For each sampling also a retain sample was taken.
- Sample storage conditions before analysis: the test medium samples (10 mL) were mixed with 50 μL formic acid and stored deep-frozen (≤ - 18 °C)

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Stock solution: weight necessary amount of test item in volumetric flask and added test medium up to the benchmark, then homogenised by shaking and one minute of sonication.
- Lower test solutions: dilution of stock solution with test medium
- Test concentration: 10.0, 4.55, 2.07, 0.939, 0.427 mg/L and control.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna Straus
- Strain/clone: Clone V
- Source: bred in the laboratory and were originally purchased from the Federal Environment Agency in Berlin/Germany.
- Age: Freshly hatched daphnids less than 24 hours old
- Feeding during test: no
- Breed method: bred as single culture (1 daphnid per 100 mL) in Elendt M4 medium at 20 ± 2 °C in a climatic chamber with 16 hours of illumination and 8 hours of darkness. medium changed three times per week. Fed with single cell green algae at least three times a week
- Condition: pH value of the aerated water: within a range of 6.0 – 9.0; dissolved oxygen: above 60 % saturation; total hardness: 140 - 250 mg/L (as CaCO3), corresponding to 7.8 - 14°dH.
- Young daphnids separation: using pipette

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

13°dH (232 mg/L CaCO3)

Test temperature:

20.0 ± 0.3 °C

pH:

7.92 ± 0.13

Dissolved oxygen:

9.1 ± 0.1 mg/L

Nominal and measured concentrations:

Nominal: 10.0, 4.55, 2.07, 0.939, 0.427 mg/L
Measured: 7.40, 3.50, 1.86, 0.911, 0.346 mg/L

Details on test conditions:

TEST SYSTEM
- Feeding: none
- Test vessels: four 100 mL glass beakers per concentration, each filled with ~ 50 mL, one additional replicate for physico–chemical measurements without test organisms.
- Type: covered with a glass plate
- Loading: ~ 10 mL of test solution for each animal
- Aeration: none
- Number of animals: 20 per concentration in 4 replicates of 5 daphids
- Renewal: 24 h
- Exposure to light: none

TEST MEDIUM / WATER PARAMETERS
- Test medium: Elendt M4 medium
- pH-value: 7.91
- Dissolved oxygen concentration: 9.3 mg/L
- Total hardness: 13°dH (232 mg/L as CaCO3).

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Biological assessment: immobilised daphnids counted after 24 h and 48 h
- Physico-chemical assessment: test temperature, pH-value, oxygen concentration measured at t = 0 h fresh and t = 24 h from fresh test solutions and after t = 24 h and t = 48 h from aged test solutions in one separate replicate per test item concentration without test organisms

RANGE-FINDING STUDY
- Test concentrations: 100, 10.0 and 1.00 mg/L and control
- Results used to determine the conditions for the definitive study: After 48 hours of exposure no immobilisation was observed in the control and at test item concentration 1.00 mg/L. 100 % of the daphnids were immobile at 10.0 and 100 mg/L.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1.72 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.939 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1.58 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.911 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Study validity: control immobilisation after 48 hours was 0 %. Dissolved oxygen concentration at the end of the test was ≥ 8.9 mg/L. The validity criteria were met.
- Biological Assessment: see any other information on results

Results with reference substance (positive control):

Tested at 1.00 and 2.00 mg/L with 20 test organisms per test concentration.
The results indicate an EC50 (24 h) of the reference item potassium dichromate between 1.00 mg/L and 2.00 mg/L.
Since the results are in accordance with the requirements of the OECD guideline 202 and fall within the historical data generated with the reference item at the testing facility, the daphnids were suitable for the determination of the toxicological effects of the test item.

Reported statistics and error estimates:

The values for EC50 were determined by Weibull analysis using linear max. likelihood regression.

Any other information on results incl. tables

Biological results after 24h

	Test concentration (mg/L) - Nominal
	control	0.427	0.939	2.07	4.55	10
group 1	0	0	0	0	5	5
group 2	0	0	0	2	5	5
group 3	1	0	0	0	5	5
group 4	0	0	0	3	5	5

Biological results after 48h:

	Test concentration (mg/L) - Nominal
	control	0.427	0.939	2.07	4.55	10
group 1	0	0	0	5	5	5
group 2	0	0	0	4	5	5
group 3	0	0	0	3	5	5
group 4	0	0	0	4	5	5

Analytical results

Test item nominal concentration (mg/L)	Sampling	Measured concentration (mg/L)	% of nominal	Geometric mean concentration (mg/L)
Control	0 fresh	n.d.	-	-
	24 aged	n.d.
	24 fresh	n.d.
	48 aged	n.d.
0.427	0 fresh	0.441	103	81
	24 aged	0.182	43
	24 fresh	0.47	110
	48 aged	0.372	87
0.939	0 fresh	1.13	120	97
	24 aged	0.755	80
	24 fresh	1.13	120
	48 aged	0.714	76
2.07	0 fresh	1.61	78	90
	24 aged	1.72	83
	24 fresh	2.3	111
	48 aged	1.86	90
4.55	0 fresh	4.21	93	77
	24 aged	2.38	52
	24 fresh	4.29	94
	48 aged	3.51	77
10	0 fresh	9.13	91	74
	24 aged	4.74	47
	24 fresh	9.56	96
	48 aged	7.27	73

-=not calculated; n.d.=not detectable; LOQ= 0.04 mg test item/L

EC50 and NOEC-values of daphnids exposed to the test item:

	Nominal [mg/L]		Actual [mg/L]
	24 h	48 h	24 h	48 h
NOEC	0.939	0.939	0.911	0.911
EC50	2.54	1.72	2.17	1.58
95% confidence limit of EC50	2.11-3.48	1.49-1.96	1.88-3.45	1.37-1.79

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The EC50 (48 h) for immobilisation was determined to be 1.72 mg/L (nominal) and 1.58 mg/L (actual). The corresponding NOEC (48 h) was 0.939 mg/L (nominal) and 0.911 mg/L (actual).

Executive summary:

The immobilisation effect of test item was determined on the water flea Daphnia magna under worst-case exposure conditions according to OECD 202. Following a semi-static non-GLP range-finding test with nominal concentrations of 100, 10.0 and 1.00 mg/L and control, a semi-static main test with nominal concentrations of 10.0, 4.55, 2.07, 0.939 and 0.427 mg/L and control was performed. Since he substance is known to undergo photomodification, the test was performed in the dark.

The values for EC50 were determined by Weibull analysis using linear max. likelihood regression. The NOEC was established based on the highest concentration at which the immobilisation is not higher than the allowed control immobilisation (≤10 % immobilisation). Water quality parameters of pH-value and dissolved oxygen concentration measured at 0 and 48 h were determined to be within the acceptable limits. The validity criteria of the test guideline were fulfilled. The measured content of test item was between 80 and 120 % of nominal at test start.

According to the results of the test, the EC50 (48 h) was determined to be 1.72 mg/L (nominal) and 1.58 mg/L (actual). The corresponding NOEC (48 h) was 0.939 mg/L (nominal) and 0.911 mg/L (actual).