Oligomerisation products of plasma treated triglycerides, C16-18 (even numbered) and C18 (unsaturated)

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:

2-6 September 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

Apr. 2004

Deviations:

yes

Remarks:

Uncritical deviation: no acclimatation period. Switching from M4-medium (husbandry) to Dilution water (test) has been shown not to cause any detrimental effects for test daphnia, therefore no acclimation period of 48 h was performed.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

May 2008

Deviations:

yes

Remarks:

idem as above

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

vegetable oil mono- and polyunsaturated C16-C22 triglycerides

Molecular formula:

C51H92O6 to C69H128O6

IUPAC Name:

vegetable oil mono- and polyunsaturated C16-C22 triglycerides

Constituent 2

Reference substance name:

dimers of vegetable oil mono- and polyunsaturated C16-C22 triglycerides

Molecular formula:

C102H186O12 to C138H256O12

IUPAC Name:

dimers of vegetable oil mono- and polyunsaturated C16-C22 triglycerides

Constituent 3

Reference substance name:

trimers of vegetable oil mono- and polyunsaturated C16-C22 triglycerides

Molecular formula:

C153H280O18 to C207H388O18

IUPAC Name:

trimers of vegetable oil mono- and polyunsaturated C16-C22 triglycerides

Constituent 4

Reference substance name:

tetramers and higher oligomers of vegetable oil mono- and polyunsaturated C16-C22

Molecular formula:

C204H374O24 and higher oligomers

IUPAC Name:

tetramers and higher oligomers of vegetable oil mono- and polyunsaturated C16-C22

Test material form:

liquid: viscous

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampling method: the content of the test item was estimated in the test solutions at the start and the end (48 h) of the test

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The water-accommodated fraction (WAF) was prepared for the test. This was done by mixing the nominal load of 100 mg/L resp. 108.7 µL/L test item (based on a density of 0.92 g/mL stated in the SDS) with the corresponding amount of dilution water and stirring moderately for 24 hours at 120 rpm on a magnetic stirrer. The lower phase was used unfiltered as test solution
- Evidence of undissolved material: No turbidity of the test solution was visible.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Species: Daphnia magna Straus
- Strain: Berlin
- Age at study initiation: 0-24 hours
- Sex: female
- Source: Umweltbundesamt Berlin
- Feeding during test: no
- Food type: Desmodesmus subspicatus - during breeding only

ACCLIMATION
- Acclimation period: none: Switching from M4-medium (husbandry) to Dilution water (test) has been shown not to cause any detrimental effects for test daphnia, therefore no acclimation period of 48 h was performed.

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES, INCLUDING CULTURING CONDITIONS: Daphnia magna was 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 keeping 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

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

Observations after 24 and 48 h.

Post exposure observation period:

not applicable

Test conditions

Hardness:

no data

Test temperature:

18.9 - 20.5°C

pH:

7.5 - 7.6

Dissolved oxygen:

8.3 - 8.4 mg/L

Salinity:

not applicable

Conductivity:

no data

Nominal and measured concentrations:

Loading rate: 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beakers
- Type: open
- Fill volume: 50 mL beakers containing 20 +/- 5 mL test solution
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:For preparation of the dilution water demineralised water was used, with the following specifications: 293.80 mg/L CaCl2*2H2O, 123.30 mg/L MgSO4*7H2O, 64.80 mg/L NaHCO3 and 5.80 mg/L KC. Deviations from the nominal weighted loads were less than 5 %. Exact values are rec-orded in the raw data. After preparation, the dilution water was aerated and the pH was measured. The pH was 7.6.
- Total organic carbon: 0.97 - 1.11 mg/L

EFFECT PARAMETERS MEASURED : immobility every 24 h

RANGE-FINDING STUDY : yes
- Test concentrations: 1, 10 and 100 mg/L in a non-GLP pre-test
- Results used to determine the conditions for the definitive study: yes, no toxicity observed at highest tested dose.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

other: immobility

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

other: immobility

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Relevant effect levels: The value was within the requested range for the 24h-EC50 between 0.6 - 2.1 mg/L of potassium dichromate K2Cr2O7 as specified in the OECD guideline.
- ECx: The 24h-EC50 value was determined as 1.98 mg/L.

Reported statistics and error estimates:

Since no immobility has occurred, no statistical evaluation was carried out.

Any other information on results incl. tables

Non-GLP pre-test

Nominal Concentration in mg/L	Immobility 48 hours
	absolute		in %
Blank control	0	0	0
1	0	0	0
10	0	0	0
100	0	0	0

Main test

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
100	0	0	0	0	0	0	0	0	0	0

Analytical determinations

The concentrations measured in the test solutions are in the same very low range as the blank value. It is detectable but not quantifiable. During the preparation of the WAF, the test solutions were in direct contact with the test item while stirring. This means that the soluble components must have dissolved. Therefore, the results are related to the loading rate; see Guidance Doc. No.23 (§176).

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	10.41	11.77	9.44	10.66
100	10.30	12.34	10.10	10.70

LOQ (Limit of quantification) TC = 822.33 µg/L

LOQ (Limit of quantification) IC = 447.52 µg/L

Calculated concentrations TOC

Nominal Concentration Test Item	Calculated concentration TOC (TC-IC)  t = 0 h	Calculated concentration TOC (TC-IC)  t =48 h	Calculated concentration TOC minus blank control t = 0 h	Calculated concentration TOC minus blank control t = 48 h
mg/L	mg/L	mg/L	mg/L	mg/L
Blank control	0.97	1.11	--	--
100	0.20	1.65	-0.77	0.54

Calculated test item concentrations

Nominal Concentration Test Item	Calculated Concentration t = 0 h	Calculated Concentration t = 48 h	% of Nominal concentration t = 0 h	% of Nominal concentration t = 48 h
mg/L	mg/L	mg/L	%	%
Blank control	--	--	--	--
100	-1.0	0.7	-1	1

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

A 48h exposure of the test item at 100 mg/L WAF did not induce any immobility of Daphnia magna. Results indicated an NOELR and EL50 >= 100 mg/L.

Executive summary:

A study performed as limit study in a Water Accommodated Fraction (WAF) test setup exposing the crustacean, Daphnia magna, for 48 h to load of 100 mg/L (loading rate) of the test item. Test was performed according to the OECD guideline 202 and it fulfilled the validity criteria. The test solution was prepared following the water-accommodated fractions (WAF) method at the load of 100 mg/L. Measured concentrations were determined using measurement of total organic carbon (TOC). The concentrations measured in the test solutions are in the same very low range as the blank value. It is detectable but not quantifiable. Therefore, the results are related to the loading rate; see Guidance Doc. No.23 ( § 153). No immobilisation was observed after 48 h, determining a NOELR, LOELR and EL50 greater than 100 mg/L.