Glycerides, C18-36

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

8 Nov-10 Nov 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study with acceptable restrictions (precipitation occurred in those test vessels where the substance was directly added, probably creating the observed effects by physical interference)

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

Additionally consideration of the "VCI-Konzept-Entwurfes für die (akute) aquatische Prüfung von schwerlöslichen Stoffen vom 31.1. 1994"

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Both test runs (a) directly weighed in and (b) separated from undissolved test substance
- Sampling method: Samples were taken at 0 and 48 hours

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Due to the low solubility of the test substance two different test solutions were prepared according to the VCI-concept for poorly soluble substances:
a) 100 mg/L directly weighed in solution: The test substance was directly weighed into the vessel and ultrasonicated for 5 min
b) Test solution with 100 mg/L and subsequently separation of undissolved test substance: 100 mg test substance were added to 1 L M4 medium and stirred for 24h. Afterwards the undissolved test substance was separated by filtration with a fibre glass filter (pretreated with 1 molar NaOH)

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain: STRAUS
- Source: BGA Berlin (May 1994), afterwards own breeding
- Age at study initiation: 6 - 24h
- Feeding during test: none

HOUSING
- Type and amount of food: Scenedesmus subspicatus

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

21.8 °C

pH:

7.8-7.9

Dissolved oxygen:

At least 90% oxygen saturation

Nominal and measured concentrations:

Nominal concentration: 100 mg/L
Measured concentrations:
Test solution a) directly weighed in without separation: 0h = 105.36 mg/L; 48h = 44.30 mg/L
b) separation of undissolved matter: 0h = 0.29 mg/L; 48 h = 0.15 mg/L. The recovery of the test substance at the beginning and after 48 hours was higher than the water solubility of the test substance determined with the column elution method, what means that the separation was not complete.

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 100 mL glass beaker, covered with a glass disc and filled with 50 mL
- Aeration: no
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- Biomass loading rate: 5 mL per daphnia


TEST MEDIUM / WATER PARAMETERS
- Test medium: M4-medium according to guideline


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilisation at 24 and 48 hours


TEST CONCENTRATIONS
- Spacing factor for test concentrations: Limit test - only 100 mg/L

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: solution a) test substance directly weighed in without separation of undissolved matter: No effect up to the limit of the water solubility of the test substance was observed.

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.01 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: solution b) separation of undissolved matter: No effect up to the limit of the water solubility of test substance (< 0.01 mg/L).

Details on results:

- Mortality of control: none
- Any observations that might cause a difference between measured and nominal values: Yes, in solution a) Black particles were observed in the suspension. Partly they adhered on the daphnids. A turbidity of the test suspension was observed.
- Effect concentrations exceeding solubility of substance in test medium: yes, for solution a) without separation of undissolved test substance

No effect was observed up to the limit of the water solubility of the test substance under conditions tested. Furthermore no effects were observed in an oversaturated solution with undissolved test substance.

Table 1: Immobilisation (%) of daphnids in solution a) (without separation of undissolved test substance)

Concentration (mg/L)	Incubation time (h)
	24	48
0/1	0	0
0/2	0	0
100/1	0*	10*
100/2	10*	10*

*Black particles were observed in the suspension. Partly, they adhered on the daphnids. A turbidity of the test suspension was observed.

 

Table 2: Immobilisation (%) of daphnids in solution b) (with separation of undissolved test substance)

Concentration (mg/L)	Incubation time (h)
	24	48
0/1	0	0
0/2	0	0
100/1	0	0
100/2	0	0

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 Aug - 23 Aug 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

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:

other: OECD Guidance Document No. 23 on Aquatic Toxicity of Difficult Substances and Mixtures, OECD 2000

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht, Mainz, Germany

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test substance is a mixture that is poorly soluble in water. Therefore the test solution(s) were prepared following general guidance provided in OECD 23 in order to achieve a water accommodated fraction (WAF) of the test substance. The test solution was prepared by directly adding test substance to test medium. A defined volume of test media was placed into a beaker (2L nominal volume) and a glass sampling tube is suspended in the beaker with the top of the tube sufficiently above the surface of the test medium to avoid overflow from the surrounding surface layer. The test substance was pipetted carefully (with a positive displacement pipette), on the water surface outside of the glass tube. The beakers were covered with Parafilm and shaken gently (approx. 50 RPM, to avoid emulsion formation) for about one day. After shaking, the required volume of aqueous fraction was drawn off for testing via the glass sampling tube. The exposure was started after separation of the undissolved material.
Undissolved test substance was visible outside the glass sampling tube at the water surface in the treatment group. The test solution removed from the sampling tube was visibly colorless and clear with no visible undissolved substance.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Source: originally obtained from the Institut National de Recherche Chimique Appliquée, France, in 1978. From this date on this clone was cultured and bred continuously in the Laboratory of Ecotoxicology of the Experimental Toxicology and Ecology of the BASF SE, Ludwigshafen, Germany
- Age at study initiation: <24 h old (at least 3rd brood progeny)
- Method of breeding: Daphnia brood stock are kept in mass cultures consisting of approx. 20 – 30 parthenogenetically reproducing females for a
maximum of 4 weeks and fed live unicellular algae (Desmodesmus subspicatus), cultured continuously at the test facility. All individuals in each mass culture originate from a single female and are thus genetic clones. After approximately 14 days the adults have produced at least 3 broods and the
young can be used in tests. Offspring are removed from the mass cultures at least once daily during the normal work week to ensure that young daphnia are <24 h old (first instar) at test initiation.
- Feeding during test: none

ACCLIMATION
- Acclimation conditions (same as test or not): The Daphnia are cultured under identical conditions as those in the final test, including test media (Elendt M4), water quality, and temperature (20 ± 1 °C)

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

2.51 mmol/L (medium)

Test temperature:

19.5 - 20.6 °C

pH:

7.9 - 8.0

Dissolved oxygen:

8.1 - 8.6 mg O2/L

Nominal and measured concentrations:

Nominal: control, 100 mg/L (WAF, limit test)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): Closed
- Material, size, headspace, fill volume: Test tubes (glass) with flat bottom (nominal volume 20 mL) filled with 10 mL test solution
- Aeration: none
- 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: A synthetic fresh water (Elendt M4) is used as media for culture and test purposes according to OECD guideline 202.
- Total organic carbon: 1 mg/L
- Ca/Mg ratio: about 4 : 1
- Conductivity: 636µS/cm
- Culture medium different from test medium: no
- Intervals of water quality measurement: daily

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light : 8 hours darkness
- Light intensity: 147 – 666 lux at a wave length of 400 - 750 nm

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Immobilisation of Daphnia is determined after 24 and 48 hours of exposure.

TEST CONCENTRATIONS
- Results used to determine the conditions for the definitive study: In a preliminary range-finding test (conducted under non-GLP conditions) a concentration of 100 mg/L caused no immobilisation or any other symptoms after 48 hours. According to the OECD guideline, the highest suggested test concentration is 100 mg/L for a limit test.

Reference substance (positive control):

yes

Remarks:

Sodium chloride

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EL0

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EL100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Details on results:

- No immobilization was reported at a nominal loading rate of 100 mg/L.
- No immobilization observed in the control.

Results with reference substance (positive control):

EC50 (48 h) = 5.13 mg/L.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

8 Nov-10 Nov 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study with acceptable restrictions (precipitation occurred in those test vessels where the substance was directly added, probably creating the observed effects by physical interference)

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

Additionally consideration of the "VCI-Konzept-Entwurfes für die (akute) aquatische Prüfung von schwerlöslichen Stoffen vom 31.1. 1994"

GLP compliance:

yes

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: solution a) test substance directly weighed in without separation of undissolved matter: No effect up to the limit of the water solubility of the test substance was observed.

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.01 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: solution b) separation of undissolved matter: No effect up to the limit of the water solubility of test substance (< 0.01 mg/L).

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

20 Aug - 23 Aug 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

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:

other: OECD Guidance Document No. 23 on Aquatic Toxicity of Difficult Substances and Mixtures, OECD 2000

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

ISO 6341 (Water quality - Determination of the Inhibition of the Mobility of Daphnia magna Straus (Cladocera, Crustacea))

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht, Mainz, Germany

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EL0

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EL100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Description of key information

No effects up to the limit of water solubility; read-across.

Additional information

Since no studies investigating the short-term toxicity of Dub TGI 24 to aquatic invertebrates are available, in accordance to Regulation (EC) No 1907/2006 Annex XI, 1.5 a read-across to the structurally related source substances, Glycerides, C16-18 and C18-unsatd. (CAS No. 67701-30-8) and Glycerides,C14-18 and C16-18 unsaturated, mono-, di- and tri- (CAS No. 91052-28-7) was conducted. The source substances are representative to evaluate the short-term toxicity of the target substance to aquatic invertebrates. The target substance is characterized as a di and triesters with glycerol and 2-decyltetradecanoic acid. The source substances are structurally very similar to the target substance.  Glycerides, C16-18 and C18-unsatd. (CAS No. 67701-30-8) is an UVCB substance consisting of C16 – C18 (even) and C18 unsaturated fatty acids with glycerol mainly as triglycerides. Glycerides,C14-18 and C16-18 unsaturated, mono-, di- and tri- (CAS No. 91052-28-7) is characterized as mono, di and triester of glycerol with fatty acids of C14-18 and C16-18 unsaturated.  

This read-across is justified in detail in the overall summary (IUCLID Section 6.1) and within the analogue justification in IUCLID Section 13. In this case of read-across, the best suited (highest degree of structural similarity, nearest physico-chemical properties) read-across substance was used for the assessment.

The first study with the read-across substance Glycerides, C16-18 and C18-unsatd. (CAS No. 67701-30-8) was performed as limit test according to EU-Method C.2, under GLP conditions. The test was performed with additional consideration of the VCI concept for poorly soluble substances. Thus, two different test solutions were prepared with 100 mg/L: One solution where the test substance was directly weighed into the test vessel and undissolved substance was not removed and a second solution with separation of undissolved substance through filtration. No effect was observed up to the limit of the water solubility of the test substance under conditions tested with an EC50 of > 0.01 mg/L (meas. initial) for the second solution with separation of undissolved substance. Furthermore no effects were observed in an oversaturated solution with undissolved test substance (EC50 (48h) > 100 mg/L).

The second study with the read-across substance Glycerides,C14-18 and C16-18 unsaturated, mono-, di- and tri- (CAS No. 91052-28-7) was performed according to OECD 202, under GLP conditions. Daphnia magna was exposed to the substance at a single loading rate of 100 mg/L (limit test) for 48 hours within a static water regime. At the end of the exposure period, no immobilization was observed. Therefore, the EL50 (48 h) was determined to be > 100 mg/L (nominal, loading rate).

Based on the available results from structurally related read-across substances (in accordance to Regulation (EC) No 1907/2006 Annex XI, 1.5) which are characterized by a similar ecotoxicological profile and comparable structure, it can be concluded that Dub TGI 24 will not exhibit short-term effects to aquatic invertebrates up to the limit of water solubility.