4-Decenal, 9-hydroxy-5,9-dimethyl-

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 July to 14 August 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Not applicable

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Storage condition of test material: Protect from light, refrigerator.

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 2.2, 4.6, 10, 22, 46, 100 mg/L
- Sampling method: For the determination of the actual test item concentrations, duplicate samples were taken from each treatment at the start and end of each test medium renewal period. For sampling from the aged test media, the contents of the respective replicates were combined prior to sampling
- Sample storage conditions before analysis: All samples were stored deep-frozen (at about -20 °C) immediately after sampling.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
The test medium of the highest nominal concentration of 100 mg/L was prepared by completely dissolving 100.3 and 100.2 mg of test item in 1000 mL of test water using intense stirring for 15 minutes at room temperature in the dark, on Day 0 and 1, respectively. The test media of the lower test item concentrations were prepared in a series of further dilution steps. The test media were prepared just before introduction of the daphnids (i.e., start of the test and prior to the test medium renewal).

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia
- Strain: Daphnia magna Straus
- Age at study initiation: 6-24 hours old
- Source: A clone of this species (defined by the supplier as clone 5) was originally supplied by the University of Sheffield / UK in 1992. Since that time, the clone has been bred at Harlan Laboratories in reconstituted water under temperature and light conditions identical to those of the tests. The parental daphnids were maintained in test water for at least 48 hours prior to the start of the test.
- Feeding during test: No
- Food type: an algal suspension of the green algae Desmodesmus subspicatus CHODAT, Strain No. 86.81 SAG, supplied by the Collection of Algal Cultures (SAG, Institute for Plant Physiology, University of Göttingen, 37073 Göttingen / Germany) and cultivated at Harlan Laboratories under standardized conditions or a mixture of this algal suspension and a commercial fish diet (Tetra Min® Hauptfutter, supplied by TETRA-Werke, 49304 Melle / Germany).
- Frequency: three times a week

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

150 mg/L as CaCO3

Test temperature:

21 °C

pH:

7.6-7.8

Dissolved oxygen:

8.3 mg/L

Conductivity:

< 5μS/cm

Nominal and measured concentrations:

Nominal Concentration: 2.2, 4.6, 10, 22, 46 and 100 mg/L
Measured Concentration: -, - , -, 23.5, 50.3 and 109.4 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 250-mL glass beakers
- Type (delete if not applicable): closed
- Volume of solution: 125 mL
- Aeration: Not aerated during the test
- No. of organisms per vessel: 20
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Ca/mg ratio: 4/1
- Culture medium different from test medium: No
- Intervals of water quality measurement: at the start and end of each test medium renewal period

OTHER TEST CONDITIONS
- Adjustment of pH: Not done
- Photoperiod: Test was performed as far as possible in the dark to avoid a photolytic degradation of the test item.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Mobility, pH, Dissolved Oxygen Concentrations and Temperature.

VEHICLE CONTROL PERFORMED: no

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

68 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

74 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

46 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

50.3 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

109.4 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

59 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

71 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

22 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

23.5 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

109.4 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

22 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

23.5 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

In the analysed test medium samples from the start and end of the test medium renewal periods, the measured test item concentrations ranged between 97 and 113% of the nominal values. Consequently, the test water renewal ensured nearly constant concentration of the test item during the test. The reported biological results are related to both the nominal and following mean measured test item concentrations.

Table 6.1.3/3: The concentrations of the test item analysed

Nominal test concentration [mg/L]	Mean measured concentration of the test item (geometric mean)
	[mg/L]	% of nominal
2.2	---	---
4.6	---	---
10	---	---
22	23.5	107
46	50.3	109
100	109.4	109

 

--- were not takeninto account for the calculation since these concentrations were below the 48-hour NOEC determined in this test and, thus, were not relevant for the interpretation of the biological results.

During the first 24 hours of the test, no immobilized test organisms were determined in the control and up to and including the test item concentration of 46 mg/L. At the highest test concentration of 100 mg/L, all test organisms were found to be immobile after 24 hours.

The 24-hour EC₅₀ of the test item was calculated to be 68 mg/L with 95% confidence limits of 46 and 100 mg/L (based on nominal concentrations) and 74 mg/L with 95% confidence limits of 50 and 109 mg/L (based on mean measured concentrations). The 24-hour EC₀ was 46 mg/L (nominal), 50.3 mg/L (mean measured). The 24-hour EC₁₀₀ was 100 mg/L (nominal), 109.4 mg/L (mean measured).

After 48 hours of exposure, no immobilized test organisms were determined in the control and up to and including the nominal test item concentration of 22 mg/L. At the concentration of 46 mg/L, 10% of the daphnids were found to be immobile. At the highest test concentration of 100 mg/L, all test organisms were found to be immobile.

The 48-hour EC₅₀ was calculated to be 59 mg/L with 95% confidence limits of 37 and 94 mg/L (nominal) and 71 mg/L with 95% confidence limits of 57 and 89 mg/L (mean measured). The 48-hour EC₀ and NOEC of test item were both 22 mg/L (nominal), 23.5 mg/L (mean measured), since no toxic effect was observed up to and including this test concentration. The 48-hour EC₁₀₀ was 100 mg/L (nominal), 109.4 mg/L (mean measured).

No remarkable observations were made concerning the appearance of the test media. All test media were clear solutions throughout the entire test duration.

At the beginning and end of the test medium renewal periods, the pH values of the test media were between 7.6 and 7.8. The dissolved oxygen concentrations in the test media and controls were 8.3 mg/L, and water temperature was maintained at 21 °C during the test.

The test is considered to be valid, as in the control no daphnids showed immobilization or other signs of disease or stress (e.g., discoloration or unusual behaviour such as trapping at the surface water). Furthermore, the dissolved oxygen concentration at the end of the test was ≥3 mg/L in the control and test vessels.

Table 6.1.3/4: Effect of test substance on the Mobility of Daphnia magna

Nominal test item concentration [mg/L]	No. of daphnids tested	Immobilized daphnids after 24 hours		Immobilized daphnids after 48 hours
		No.	[%]	No.	[%]
Control	20	0	0	0	0
2.2	20	0	0	0	0
4.6	20	0	0	0	0
10	20	0	0	0	0
22	20	0	0	0	0
46	20	0	0	2	10
100	20	20	100	20	100

With the exception of the immobilization, no other adverse effects were observed in the test animals.

 

Table 6.1.3/5: Appearance of the Test Media during the Test Period

 

Abbreviations:

0: no remarkable observations, clear test medium

1: homogeneous dispersion in the water, turbidity observable

2: noticeable turbidity caused by the test item

3: noticeable coloration caused by the test item

4: inhomogeneous dispersion of the test item

5: precipitation of the test item

6: test item at the surface

7: test item lying at the bottom of the flask

Nominal test item concentration [mg/L]	Exposure time
	0 h	24 h		48 h
	new	old	new	old
2.2	0	0	0	0
4.6	0	0	0	0
10	0	0	0	0
22	0	0	0	0
46	0	0	0	0
100	0	0	0	--

--: not determined since all daphnids were immobile

 

Table 6.1.3/6: pH Values in the Treatments

Nominal test item concentration [mg/L]	Exposure time
	0 h	24 h		48 h
	new	old	new	old
Control	7.6	7.7	7.7	7.7
2.2	7.6	7.7	7.8	7.7
4.6	7.6	7.7	7.8	7.7
10	7.6	7.7	7.8	7.7
22	7.6	7.7	7.8	7.7
46	7.6	7.7	7.8	7.7
100	7.6	7.7	7.8	--

--: not determined since all daphnids were immobile

 

Table 6.1.3/7: Dissolved Oxygen Concentrations [mg/L] in the Treatments

Nominal test item concentration [mg/L]	Exposure time
	0 h	24 h		48 h
	new	old	new	old
Control	8.3	8.3	8.3	8.3
2.2	8.3	8.3	8.3	8.3
4.6	8.3	8.3	8.3	8.3
10	8.3	8.3	8.3	8.3
22	8.3	8.3	8.3	8.3
46	8.3	8.3	8.3	8.3
100	8.3	8.3	8.3	--

--: not determined since all daphnids were immobile

 

Table 6.1.3/8: Temperature [°C] in the Treatments

Nominal test item concentration [mg/L]	Exposure time
	0 h	24 h		48 h
	new	old	new	old
Control	21	21	21	21
2.2	21	21	21	21
4.6	21	21	21	21
10	21	21	21	21
22	21	21	21	21
46	21	21	21	21
100	21	21	21	--

--: not determined since all daphnids were immobile

Validity criteria fulfilled:

yes

Conclusions:

The test item had acute toxic effects on Daphnia magna. The 48-hour EC50 was determined to be 59 mg/L with 95%-confidence limits of 37 and 94 mg/L (nominal) and 71 mg/L with 95% confidence limits of 57 and 89 mg/L (mean measured).

Based on Nominal; Based on Mean Measured
24-hour EC50: 68 mg/L; 74 mg/L
95% confidence limits: (46 and 100 mg/L); (50 and 109 mg/L)
24-hour EC0: 46 mg/L; 50.3 mg/L
24-hour EC100: 100 mg/L; 109 mg/L
48-hour EC50: 59 mg/L; 71 mg/L
95% confidence limits: (37 and 94 mg/L); (57 and 89 mg/L)
48-hour EC0 and 48-hour NOEC: 22 mg/L; 23.5 mg/L
48-hour EC100: 100 mg/L; 109.4 mg/L

Executive summary:

The acute toxicity of the test item to Daphnia magna was determined in a 48-hour semi-static test according to the EU Method C.2 (Acute Toxicity for Daphnia), and the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test), as well as according to the OPPTS No. 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Fresh water Daphnids). The nominal test item concentrations tested were 2.2, 4.6, 10, 22, 46 and 100 mg/L. Additionally, a control group was tested in parallel.

Analysis of test samples was performed using GC-FID. In the analysed test medium samples from the start and end of the test medium renewal periods, the measured test item concentrations ranged between 97 and 113% of the nominal values. Consequently, the test water renewal ensured nearly constant concentration of the test item during the test. The reported biological results are related to both the nominal and following mean measured test item concentrations:

Nominal test concentration (mg/L)	Mean measured concentration of the test item (geometric mean)
	[mg/L]	% of nominal
2.2	---	---
4.6	---	---
10	---	---
22	23.5	107
46	50.3	109
100	109.4	109

---: were not takeninto account for the calculation since these concentrations were below the 48-hour NOEC determined in this test and, thus, were not relevant for the interpretation of the biological results.

During the first 24 hours of the test, no immobilized test organisms were determined in the control and up to and including the test item concentration of 46 mg/L. At the highest test concentration of 100 mg/L, all test organisms were found to be immobile after 24 hours.

The 24-hour EC₅₀ of the test item was calculated to be 68 mg/L with 95% confidence limits of 46 and 100 mg/L. The 24-hour EC₀ was 46 mg/L. The 24-hour EC₁₀₀ was 100 mg/L.

After 48 hours of exposure, no immobilized test organisms were determined in the control and up to and including the test item concentration of 22 mg/L. At the concentration of 46 mg/L, 10% of the daphnids were found to be immobile. At the highest test concentration of 100 mg/L, all test organisms were found to be immobile.

The 48-hour EC₅₀ was calculated to be 59 mg/L with 95% confidence limits of 37 and 94 mg/L. The 48-hour EC₀ and NOEC of test substance were both 22 mg/L, since no toxic effect was observed up to and including this test concentration.

The 48-hour EC₁₀₀ was 100 mg/L The biological test results (based on nominal and mean measured concentrations) were as follows:

	Based on Nominal	Based on Mean Measured
24-hour EC50:	68 mg/L	74 mg/L
95% confidence limits:	(46 and 100 mg/L)	(50 and 109 mg/L)
24-hour EC0:	46 mg/L	50.3 mg/L
24-hour EC100:	100 mg/L	109.4 mg/L
48-hour EC50:	59 mg/L	71 mg/L
95% confidence limits:	(37 and 94 mg/L)	(57 and 89 mg/L)
48-hour EC0 and 48-hour NOEC:	22 mg/L	23.5 mg/L
48-hour EC100:	100 mg/L	109.4 mg/L

 

The test is considered to be valid, as in the control no daphnids showed immobilization or other signs of disease or stress (e.g., discoloration or unusual behaviour such as trapping at the surface water). Furthermore, the dissolved oxygen concentration at the end of the test was ≥3 mg/L in the control and test vessels.

Description of key information

Under the experimental conditions and based on measured concentrations, the 48-hour ErC50 was estimated to be of 71 mg/L. Moreover, the highest measured concentration without observed effects was estimated to be at least 23.5 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

71 mg/L

Additional information

For that endpoint, a study on the registered substance is available.

The acute toxicity of the test item to Daphnia magna was determined in a 48-hour semi-statictest according to the EU Commission Directive 92/69/EEC, Part C.2, theCommission Regulation (EC) No. 440/2008, Part C.2 and the OECD Guideline for Testing ofChemicals, No.202 (2004), as well as according to the OPPTS Guideline No. 850.1010 (PublicDraft, April 1996).Thenominal test item concentrations tested were2.2, 4.6, 10, 22, 46 and 100 mg/L. Additionally,a control group was tested in parallel.

The test is considered to be valid, as in the control no daphnids showed immobilization or other signs of disease or stress (e.g., discoloration or unusual behavior such as trapping at the surfacewater). Furthermore, the dissolved oxygen concentration at the end of the test was ≥3 mg/L inthe control and test vessels.

Under the experimental conditions and based on measured concentrations, the 48 -hour ErC50 was estimated to be of 71 mg/L (nominal: 59 mg/L). Moreover, the highest measured concentration without observed effects was estimated to be at least 23.5 mg/L (nominal:22 mg/L).