Glycerides, tall-oil mono-

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:

20. Mar. 2018 to 26. Mar. 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

OECD Guideline for Testing of Chemicals No. 202, adopted 13. Apr. 2004: ”Daphnia sp., Acute Immobilisation Test“

Deviations:

yes

Remarks:

See "Any other information" for details

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

Council Regulation (EC) No. 440/2008, Method C.2. “Daphnia sp. Acute Immobilisa-tion Test”, adopted 30. May 2008

Deviations:

yes

Remarks:

See "Any other information" for details

GLP compliance:

yes

Specific details on test material used for the study:

No further details specified in the study report.

Analytical monitoring:

yes

Details on sampling:

Not specified.

Vehicle:

no

Details on test solutions:

The water-accommodated fraction (WAF) was prepared for the test. This was done by weighing the nominal loads of 4.6 / 10 / 22 / 46 / 100 mg/L, adding the corresponding amount of dilution water and shaking vigorously for 23.25 hours. The resulting solution was filtrated through 0.45 μm PTFE filters (max. 20 mL per filter) and used for the test.

Test organisms (species):

Daphnia magna

Details on test organisms:

Specification
Species: Daphnia magna
Authority: STRAUS
Strain: Berlin
Sex: female
Age: between 0 and 24 hours
Origin: Umweltbundesamt Berlin
In-house breeding: since 27. September 2007
Selection of the test system was made following the proposal of the guidelines.

Animal Husbandry
Daphnia magna is bred in the LAUS GmbH throughout the year. The animals are kept for the use in toxicity tests. They multiply by parthenogenesis, thus being genetically identical. The husbandry is performed similar to the method described in the OECD guideline.
Vessels: preserving glasses, nominal volume 2 L
Medium: M4-Medium (recipe of ELENDT)
Food: green algae (Desmodesmus subspicatus)
Medium renewal: twice a week
Photo period: 16/8 hours, using neon tubes
Temperature: 20 ± 2 °C

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

Not post exposure observation period specified

Hardness:

Hardness in mmol/L: 2.502
Hardness in mg CaCO3/L: 250

Test temperature:

19.8 - 22.4°C

pH:

7.7 - 7.9

Dissolved oxygen:

8.6 - 9.2 mg/L

Salinity:

Not applicable - freshwater study

Conductivity:

Not specified

Nominal and measured concentrations:

Due to low solubility of test item only in the highest tested concentration (prepared WAF) test item was clearly determinable and was between 4 to 6% of the nominal concentration at the beginning and at the end of the test. The lower concentrations showed fluctuating DOC values. Since toxicity was observed in the four highest concentrations, test item must have been present. Therefore, the determination of the biological results was based on the nominal concentrations.

Details on test conditions:

Selection of Daphnia
22 hours 50 minutes before the start of the test, the adult animals were separated from the young. 45 minutes before test start, the adults were caught with the help of a glass tube, and the newborn daphnia, aged between 0 and 22 hours 5 minutes, were sieved from the medium and immediately placed into a beaker containing dilution water. After the settling-in period, animals which showed no apparent damage were used for the test.
Switching from M4-medium (husbandry) to dilution water (test) has been shown not to cause any detrimental effects for test daphnia.

Study Performance
Using a glass tube, the daphnia were caught and lifted from the beaker. They were put on a small sieve, and the medium surrounding the animals was sucked off using absorbent paper. Immediately after that, the animals were put into the respective test solution.
The test vessels were left to stand for 48 hours. After 24 and 48 hours, the immobilised daphnia were counted. Daphnia are considered immobilised when they do not perform any movements or are only able to move their antennae when the beaker is gently agitated. Daphnia which are trapped at the surface of the test solution are also considered immobi-lised.
The pH, the concentration of dissolved oxygen and the content of DOC in the test vessels were measured at the beginning and at the end of the test.

Experimental Conditions
Date of performance: 21. Mar. - 23. Mar. 2018
Treatments: 4.6 / 10 / 22 / 46 / 100 mg/L (nominal)
The concentrations to be tested were based on the result of a non-GLP pre-test.
Temperature: 19.8 - 22.4°C
Duration: 48 hours
Observation times: 24 and 48 hours
Medium renewal: none
Test vessels: glass beakers, nominal volume 50 mL, tall shape
Replicates: (Treatments) 4 vessels, each containing 20 ± 5 mL test solution and 5 daphnia
Replicates: (Blank control) 4 vessels, each containing 20 ± 5 mL dilution water and 5 daphnia

Reference substance (positive control):

yes

Remarks:

Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9)

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

4.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

30.01 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Four concentrations showed toxicity between 10 and 90% immobilisation. None of the animals was immobilised in the blank control.

Results with reference substance (positive control):

The 24h-EC50 value was determined as 1.9 mg/L. The value was within the requested range for the 24h-EC50 between 0.6 - 2.1 mg/L of po-tassium dichromate K2Cr2O7 as specified in the OECD guideline. Since the fits showed sufficient statistical correspondence of the data with the dose-response-equation, this ref-erence study can be considered valid.

Immobility

Nominal Concentration in mg/L	Immobility 24 hours					Immobility 48 hours
	Absolute				In %	Absolute				In %
Blank control	0	0	0	0	0	0	0	0	0	0
4.6	0	0	0	0	0	0	0	0	0	0
10	0	0	0	0	0	1	0	0	1	10
22	0	0	0	0	0	3	1	3	2	45
46	0	0	0	0	0	4	2	3	4	65
100	0	0	0	0	0	5	4	5	4	90

 

pH and O₂values

Nominal Concentration in mg/L	pH		O2Concentration in mg/L
	0 h	48 h	0 h	48 h
Blank control	7.8	7.8	9.2	8.6
4.6	7.6	7.9	8.9	8.6
10	7.7	7.9	9.0	8.6
22	7.7	7.9	9.0	8.7
46	7.7	7.9	9.0	8.6
100	7.7	7.9	9.0	8.6

 

Measured Concentration IC and TC

Nominal Concentration mg/L	Measured TC t = 0 h mg/L	Measured TC t = 48 h mg/L	Measured IC t = 0 h mg/L	Measured IC t = 48 h mg/L
Blank control	10.66	12.88	10.56	12.02
4.6	11.22	13.76	10.49	12.22
10	10.95	13.99	10.29	12.54
22	12.27	14.65	10.50	12.25
46	11.66	14.38	10.32	12.36
100	14.50	15.28	10.42	11.66

LOQ (Limit if quantification): TC = 5.43 mg/L

LOQ (Limit of quantification) IC = 2.03 mg/L

 

Measured Concentration DOC

Nominal Concentration Test Item mg/L	Measured DOC (TC-IC) t = 0 h mg/L	Measured DOC (TC-IC) t = 48 h mg/L	Measured DOC minus blank control t = 0 h mg/L	Measured DOC minus blank control t = 48 h mg/L
Blank control	0.09	0.86	--	--
4.6	0.73	1.54	0.64	0.68
10	0.65	1.45	0.56	0.59
22	1.77	2.40	1.68	1.54
46	1.35	2.02	1.26	1.16
100	4.07	3.61	3.98	2.75

 

Calculated Test Item Concentrations

Nominal Concentration Test Item mg/L	Calculated Concentration t = 0 h mg/L	Calculated Concentration t = 48 h mg/L	% of Nominal Concentration t = 0 h mg/L	% of Nominal Concentration t = 48 h mg/L
Blank control	--	--	--	--
4.6	0.9	1.0	19	21
10	0.8	0.8	8	8
22	2.4	2.2	11	10
46	1.8	1.6	4	4
100	5.6	3.9	6	4

 

Validity criteria fulfilled:

yes

Conclusions:

The following results were determined for the test item LUMULSE GMT-K (species: Daphnia magna).
48h-NOEC = 4.6 mg/L
48h-LOEC = 10 mg/L
24h-EC50 > 100 mg/L
48h-EC50 = 30.01 mg/L

Executive summary:

One valid experiment was performed.

The study was performed using 5 concentrations ranging from 4.6 to 100 mg/L nominal concentration. For each test concentration, 20 daphnia were exposed to the test item for 48 hours in a static test system. After 24 and 48 hours, the immobilised daphnia were counted.

Four concentrations showed toxicity between 10 and 90% immobilisation. None of the an-imals was immobilised in the blank control.

Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9) was used as positive control in a current reference study to assure that the test conditions are reliable.

 

Due to low solubility of test item only in the highest tested concentration (prepared WAF) test item was clearly determinable and was between 4 to 6% of the nominal concentration at the beginning and at the end of the test. The lower concentrations showed fluctuating DOC values and were not clearly assignable to the corresponding nominal concentration. Therefore, back-calculation from DOC content to test item concentration could not be con-ducted. However, since toxicity was observed in the four highest concentrations, test item must have been present. According to the OECD guidance document no. 23, chapter 5, test substance that cannot be quantified by analytical methods at the effect concentrations, the biological results can be expressed based on the nominal concentrations. Therefore, the determination of the biological results was based on the nominal concentrations.

 

The following results were determined for the test item LUMULSE GMT-K (species: Daphnia magna).

48h-NOEC = 4.6 mg/L

48h-LOEC = 10 mg/L

24h-EC50 > 100 mg/L

48h-EC50 = 30.01 mg/L

Description of key information

The following results were determined for the test item LUMULSE GMT-K (species: Daphnia magna).

48h-NOEC = 4.6 mg/L

48h-EC50 = 30.01 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

30.01 mg/L

Additional information

One valid experiment was performed.

The study was performed using 5 concentrations ranging from 4.6 to 100 mg/L nominal concentration. For each test concentration, 20 daphnia were exposed to the test item for 48 hours in a static test system. After 24 and 48 hours, the immobilised daphnia were counted.

Four concentrations showed toxicity between 10 and 90% immobilisation. None of the an-imals was immobilised in the blank control.

 

Due to low solubility of test item only in the highest tested concentration (prepared WAF) test item was clearly determinable and was between 4 to 6% of the nominal concentration at the beginning and at the end of the test. The lower concentrations showed fluctuating DOC values and were not clearly assignable to the corresponding nominal concentration. Therefore, back-calculation from DOC content to test item concentration could not be con-ducted. However, since toxicity was observed in the four highest concentrations, test item must have been present. According to the OECD guidance document no. 23, chapter 5, test substance that cannot be quantified by analytical methods at the effect concentrations, the biological results can be expressed based on the nominal concentrations. Therefore, the determination of the biological results was based on the nominal concentrations.

 

The following results were determined for the test item LUMULSE GMT-K (species: Daphnia magna).

48h-NOEC = 4.6 mg/L

48h-LOEC = 10 mg/L

24h-EC50 > 100 mg/L

48h-EC50 = 30.01 mg/L