Glycerides, rape-oil mono-, di- and tri-, hydrogenated

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 Aug - 29 Nov 2012

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP - Guideline study. According to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Qualifier:

according to guideline

Guideline:

EU Method C.20 (Daphnia magna Reproduction Test)

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 850.1300 (Daphnid Chronic Toxicity Test)

Qualifier:

according to guideline

Guideline:

other: ISO 10706 (2000)

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 (information from sponsor). Therefore the test solution(s) was prepared following general guidance provided in OECD 23 in order to achieve water accommodated fraction (WAF) of the test substance. Generally, each test solution was prepared by directly adding test substance to test medium. A defined volume of test media was placed into a beaker (3-L nominal volume) and a glass sampling tube was 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 on the water surface outside of the glass tubes. 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 (test solution) was drawn off for testing via the glass sampling tube. The exposure was started after separation of the undissolved material. Fresh test solution was prepared for each test solution renewal day (days 0, 3, 5, 7, 10, 12, 14, 17, 19) during the experiment. Undissolved test substance was visible outside the glass sampling tube at the water surface in the treatment group after each solution preparation. The test solutions removed from the sampling tube were visibly colorless and clear with no visible undissolved substance. The test solution pH was not adjusted prior to the start of exposure.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Source: The clone of Daphnia magna STRAUS 1820 used was supplied by the Institut National de Recherche Chimique Appliquée, France, in 1978. From this date on this clone was cultured and bred continuously in the Ecotoxicology Laboratory of Experimental Toxicology and Ecology, BASF SE, Ludwigshafen Germany.
- Culture conditions: Daphnia brood stock are kept in mass cultures consisting of approx. 20 – 30 individuals for a maximum of 4 weeks. All individuals in the mass culture originate from a single female. 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. Detailed records are kept (in test facility archives) to monitor the health of Daphnia brood stock cultures including observations of young production, mortality, ephippia, and measurement of water chemistry parameters. Only young from healthy cultures without signs of stress are used for testing.
- Feeding during test: yes
- Food type: live green algae Desmodesmus subspicatus
- Amount: ≤109 µL of the algae concentrate
- Frequency: daily

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, temperature (20 ± 1 °C), and diet.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Hardness:

2.46 - 2.54 mmol/L

Test temperature:

20 - 21 °C

pH:

7.7 - 8.4

Dissolved oxygen:

7.6 - 9.1 mg O2/L

Nominal and measured concentrations:

Nominal: control and 10 mg/L (loading rate, WAF)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 100 mL glass beakers filled with 50 mL test solution
- Aeration: none
- Renewal rate of test solution (frequency/flow rate): 3 times per week
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10
- Biomass loading rate: 0.02 animals/mL


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light : 8 hours darkness
- Light intensity: About 620 - 685 lux at a wave length of 400 - 750 nm

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Parent mortality, abnormal effects, and numbers of live and dead offspring were assessed daily throughout the experiment. Reproductive success was measured by counting and discarding the offspring produced by each parent. Body length was measured after the 21-day exposure period.

Reference substance (positive control):

yes

Remarks:

Sodium chloride

Key result

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOELR

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

other: EL0

Effect conc.:

>= 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mortality

Details on results:

- The test substance had no observable adverse chronic effect on Daphnia magna at a loading rate of 10 mg/L in test media and under test conditions.

Results with reference substance (positive control):

EC50(48h) = 5.13 mg/L

Reported statistics and error estimates:

Student´s t-test was used to evaluate any differences in reproduction and growth as length between the test concentration and control. Wilcoxon’s test (one sided analysis) was used to evaluate any differences in the time to the first brood.

Table: Reproduction and Mortality Summary after 21 days

Test Groups [mg/L]	Reproduction		Mortality
	Mean Living Young per surviving adult	% effect(b)	Parent animals	% effect(b)
0 (control)	167 (35.3%)(a)	-	0	-
10	196	-	0	-

(a) Coefficient of variation for control fecundity

(b) Effect relative to control. Only calculated for statistically significant effects

Table: Biological observations among surviving parent animals in each concentration after a 21 day exposure

Test Groups [mg/L]	Growth (mean length, mm)	Mean immobile young (per surviving adult)	Mean Days to First Brood	Mean aborted eggs (per surviving adult)
0 (control)	4.6	0.6	8.4	0.9
10	4.7*	0.2	8.1	1.0

* p≤ 0.01

Table: Summarized effect parameters for the main endpoints, reproduction (living young) and % mortality of the parent animals after 21 days based on loading rates

Endpoint	21 day effect concentrations [mg/L], loading
	NOELR	LOELR	EL10*	EL20*	EL50*
Reproduction	≥ 10	> 10	> 10	> 10	> 10
Mortality	≥ 10	> 10	> 10	> 10	> 10

* ECx values are estimated since the study is designed as a limit test

Validity criteria fulfilled:

yes

Conclusions:

The substance is not harmful to aquatic invertebrates at chronic exposure within its limit of solubility in water.

Reference 2

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

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

Adequacy of study:

key study

Study period:

23 Aug - 29 Nov 2012

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP - Guideline study. According to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Justification for type of information:

Toxicity data from a structurally related source substance (Glycerides, C14-C18 and C 16-C18 unsatd., mono-, di-, and tri- (even numbered), CAS 91052-28-7)are used as read-across in accordance with Regulation (EC) No. 1907/2006, Annex XI, 1.5. Both are UVCB substances, formed from the esterification of fatty acids and glycerol, having a high content of C18 fatty acids and to a certain extent C14 and C16 fatty acids. Both are composed of mono- to tri esters (Target substance: monoester: 61.9%, diester<: 29.7%, triester: 3.2%; Source substance: monoester: 20-95%, diester: 5-60%, triester: 0.5-15%). Furthermore, their water solubilities are comparable (< 1 mg/L). Therefore, these two substances are expected to show similar environmental behaviour and comparable degrees of toxicity to aquatic organisms. Based on the above information, the use of data fromGlycerides, C14-C18 and C 16-C18 unsatd., mono-, di-, and tri- (even numbered)as read-across for Glycerides, C14-18 mono- and di- is considered justified.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

reproduction

Description of key information

No effects up to the limit of water solubility (WS < 1 mg/L)

Additional information

No study is available investigating the long-term toxicity of Glyceride, rape oil mono-, di- and tri-, hydrogenated (CAS 91744-94-4) to aquatic invertebrates. Therefore, toxicity data from a structurally related source substance (Glycerides, C14-C18 and C 16-C18 unsatd., mono-, di-, and tri- (even numbered), CAS 91052-28-7)are used as read-across in accordance with Regulation (EC) No. 1907/2006, Annex XI, 1.5. Both are UVCB substances, formed from the esterification of fatty acids and glycerol, having a high content of C18 fatty acids and to a certain extent C14 and C16 fatty acids. Both are composed of mono- to tri esters (Target substance: monoester: 61.9%, diester<: 29.7%, triester: 3.2%; Source substance: monoester: 20-95%, diester: 5-60%, triester: 0.5-15%). Furthermore, their water solubilities are comparable (< 1 mg/L). Therefore, these two substances are expected to show similar environmental behaviour and comparable degrees of toxicity to aquatic organisms. Based on the above information, the use of data fromGlycerides, C14-C18 and C 16-C18 unsatd., mono-, di-, and tri- (even numbered)as read-across for Glycerides, C14-18 mono- and di- is considered justified.

 The chronic toxicity of Glycerides,C14-18 and C16-18 unsaturated, mono-, di- and tri- (CAS No. 91052-28-7)to aquatic invertebrates has been evaluated by Salinas (2013). This test was conducted according to OECD 211, under GLP conditions. Daphnia magna was exposed to the substance at a single loading rate of 10 mg/L (limit test) for 21 days within a semi-static water regime. At the end of the exposure period, no effects on survival, reproduction or any other adverse effects were reported. Therefore the NOELR was determined to be ≥ 10 mg/L (nominal, loading rate) indicating that the substance is not harmful to aquatic invertebrates at long-term exposure within its limit of solubility in water. Based on the suitability of the read across approach this is also considered to be true for the target substance.