Petroleum resins

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 Immobilisation Test”, adopted 30. May 2008

Deviations:

yes

Remarks:

See "Any other information" for details

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

No further details specified in the study report.

Analytical monitoring:

yes

Details on sampling:

The content of DOC in the test vessels were measured at the beginning and at the end of the test

Vehicle:

no

Details on test solutions:

The water-accommodated fraction (WAF) was prepared for the test. This was done by weighing the nominal loads on a glass coverslip, transferring it into a Schott-flask and adding the corresponding amount of dilution water and shaking vigorously for 23.25 hours. The resulting solutions were filtrated through 0.45 µm PTFE filters.

Test organisms (species):

Daphnia magna

Details on test organisms:

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, following SOP 115 002 01 („Zucht und Hälterung von Daphnia magna STRAUS“), Version 12 from 02. Feb. 2015.
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:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

Not 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:

9.2 - 8.6 mg/L

Salinity:

Not applicable

Conductivity:

Not specified

Nominal and measured concentrations:

Due to low solubility of test item tested concentrations were not clearly determinable. The measured DOC showed fluctuating values in all concentrations which were similar to those of the blank control. 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 set-tling-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 absor-bent 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 agitat-ed. Daphnia which are trapped at the surface of the test solution are also considered im-mobilised.
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 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

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

One valid experiment was performed.
The study was performed using 5 concentrations ranging from 4.6 to 100 mg/L. 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.
No significant toxicity was observed in the treatments. None of the animals was immobi-lised 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 tested concentrations were not clearly determinable. The measured DOC showed fluctuating values in all concentrations which were similar to those of the blank control. Therefore, the determination of the biological results was based on the nominal concentrations. This is in agreement with the OECD guidance document no. 23, where is stated: For tests with chemicals that cannot be quantified by analytical methods, the effect concentration can be expressed based on the nominal concentrations (loading rates).
Since no toxicity and no DOC content were observed, additional pre-tests were run to show, that another test design is not feasible.
No observations were made which might cause doubts concerning the validity of the study outcome. All validity criteria were met.
The result of the test is considered valid.

Results with reference substance (positive control):

Not examined

Reported statistics and error estimates:

The estimation of the biological results was accomplished using the software ToxRat® Professional, version 3.2.1

PRE-TESTS

Immobility

Nominal Concentration in mg/L	Immobility 48 hours
	Absolute				In %
Blank control	0	0	0	0	0
1	0	0	--	--	0
10	0	0	--	--	0
100	0	2	--	--	20

 

DOC content of test item at the beginning of the pre-test

Test item concentration	DOC content of test item (mg/L)
	PTFE filter (0.45 μm)	Unfiltered
1 mg/L	3.01	1.08
10 mg/L	3.09	2.43
100 mg/L	2.75	24.15

 

pH and O₂-value

Nominal Concentration in mg/L	pH		O2-Concentration in mg/L
	0 h	48 h	0 h	48 h
Blank control	7.7	7.8	9.5	8.5
1	7.6	7.7	8.9	8.1
10	7.3	7.4	8.9	8.2
100	7.2	7.2	8.9	8.1

 

Immobility

Nominal Concentration in g/L	Immobility 48 hours
	Absolute		In %
Blank control	0	0	0
1	3^*1	3^*1	60
1 Nylon	1^*1	1^*2	20
1 PTFE	1^*2	5^*2	60
10 Nylon	0	4^*2	40
10 PTFE	4^*2	5^*2	90
10	5^*1	4^*1	90

^*1= trapped at the surface partly clotted with the test item and dead

^*2= trapped at the surface but still alive

 

DOC content of test item at the beginning of the additional pre-test

Test item concentration	DOC content of test item (in mg/L)
	Nylon filter (0.45 μm)	PTFE filter (0.45 μm)	Unfiltered
1 g/L	2.45	3.80	67.42
10 g/L	5.51	2.70	107.80

 

pH and O₂-value

Nominal Concentration in g/L	pH		O2-Concentration in mg/L
	0 h	48 h	0 h	48 h
Blank control	7.8	7.8	8.7	8.8
1	7.7	7.7	8.4	8.0
1 Nylon	7.6	7.7	8.5	8.7
1 PTFE	7.7	7.7	8.4	8.5
10 Nylon	7.3	7.3	8.0	8.1
10 PTFE	7.5	7.5	8.3	8.2
10	7.5	7.5	8.4	8.1

 

MAIN STUDY

 

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	0	0	0	0	0
22	0	0	0	0	0	1	0	0	0	5
46	0	0	0	0	0	1	0	0	0	5
100	0	0	0	0	0	0	0	0	1	5

 

pH and O₂-value

Nominal Concentration in mg/L	pH		O2-Concentration in mg/L
	0 h	48 h	0 h	48 h
Blank control	7.8	7.8	9.2	8.6
4.6	7.8	7.9	8.9	8.7
10	7.7	7.9	9.0	8.6
22	7.7	7.9	9.0	8.6
46	7.7	7.9	9.4	8.6
100	7.7	7.9	9.0	8.9

 

 

Analytical Determinations

Measured Concentrations IC and TC

Nominal Concentration Test Item	Measured TC t = 0 h	Measured TC t = 48 h	Measured IC t = 0 h	Measured IC t = 48 h
mg/L	mg/L	mg/L	mg/L	mg/L
Blank control	10.55	12.93	10.45	11.88
4.6	10.93	13.24	10.47	11.65
10	10.74	13.32	10.44	11.90
22	10.85	13.18	10.47	12.18
46	11.32	13.78	10.73	12.24
100	10.65	13.67	10.46	12.27

LOQ (Limit of quantification) TC = 5.43 mg/L

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

 

Measured Concentrations DOC

Nominal Concentration Test Item	Measured DOC (TC-IC) t = 0 h	Measured DOC (TC-IC) t = 48 h	Measured DOC minus blank control t = 0 h	Measured DOC minus blank control t = 48 h
mg/L	mg/L	mg/L	mg/L	mg/L
Blank control	0.09	1.05	--	--
4.6	0.47	1.59	0.37	0.54
10	0.30	1.42	0.21	0.37
22	0.38	1.01	0.28	-0.04
46	0.59	1.54	0.50	0.49
100	0.20	1.40	0.10	0.35

 

Calculated Test Item Concentrations

Nominal Concentration Test Item	Calculated Concentration Test Item t = 0 h	Calculated Concentration Test Item t = 48 h	% of Nominal Concentration t = 0 h	% of Nominal Concentration t = 48 h
mg/L	mg/L	mg/L	mg/L	mg/L
Blank control	--	--	--	--
4.6	0.4	0.6	9	14
10	0.2	0.4	2	4
22	0.3	0.0	1	0
46	0.6	0.6	1	1
100	0.1	0.4	0	0

 

Validity criteria fulfilled:

yes

Conclusions:

Since no significant toxicity occurred, the biological results are given in a range and are presented below:
Biological Results Test Item
Parameter Value
24h EC50 > 100 mg/L
48h EC50 > 100 mg/L
48h NOEC ≥ 100 mg/L
48h LOEC > 100 mg/L

Executive summary:

Title of Study: Determination of short term toxicity of Petroleum Resins (Kendex 0897) against Daphnia magna Straus according to OECD 202 resp. EU C.2

 

Findings and Results:

One valid experiment was performed.

The study was performed using 5 concentrations ranging from 4.6 to 100 mg/L. 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.

No significant toxicity was observed in the treatments. None of the animals 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 tested concentrations were not clearly determinable. The measured DOC showed fluctuating values in all concentrations which were similar to those of the blank control. Therefore, the determination of the biological results was based on the nominal concentrations. This is in agreement with the OECD guidance document no. 23, where is stated: For tests with chemicals that cannot be quantified by analytical methods, the effect concentration can be expressed based on the nominal concentrations (loading rates).

The following results were determined for the test item Petroleum Resins (Kendex 0897) (species: Daphnia magna).

 

48h-NOEC ≥ 100 mg/L
48h-LOEC > 100 mg/L
24h-EC50 > 100 mg/L
48h-EC50 > 100 mg/L

Since no significant toxicity occurred, the biological results are given in a range.

Description of key information

The following results were determined for the test item Petroleum Resins (Kendex 0897) (species: Daphnia magna).

48h-NOEC ≥ 100 mg/L
48h-LOEC > 100 mg/L
24h-EC50 > 100 mg/L
48h-EC50 > 100 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

100 mg/L

Additional information

One valid experiment was performed.

The study was performed using 5 concentrations ranging from 4.6 to 100 mg/L. 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.

No significant toxicity was observed in the treatments. None of the animals 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 tested concentrations were not clearly determinable. The measured DOC showed fluctuating values in all concentrations which were similar to those of the blank control. Therefore, the determination of the biological results was based on the nominal concentrations. This is in agreement with the OECD guidance document no. 23, where is stated: For tests with chemicals that cannot be quantified by analytical methods, the effect concentration can be expressed based on the nominal concentrations (loading rates).

The following results were determined for the test item Petroleum Resins (Kendex 0897) (species: Daphnia magna).

 

48h-NOEC ≥ 100 mg/L
48h-LOEC > 100 mg/L
24h-EC50 > 100 mg/L
48h-EC50 > 100 mg/L

Since no significant toxicity occurred, the biological results are given in a range.