Fatty acids, C18 unsaturated, C12-14 (even numbered) alkyl esters

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:

weight of evidence

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)

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

yes (incl. QA statement)

Remarks:

Ministerium für Arbeit, Integration und Soziales des Landes Nordrhein-Westfalen, Germany

Analytical monitoring:

yes

Details on sampling:

- Concentrations: control, 100 mg/L. The concentrations of tetradecyl myristate in the test media were analysed at test start directly after distribution of the stock test item WAF into the 60 mL glass beakers (fresh test media at test start). Furthermore, a second sample of the stock WAF was taken at this time to investigate a possible 'wall effect' of the used glass beakers. Another sample was taken at the end of the test after 48 hours (aged test media at test end).
- Sampling method: As the volume of the analytical sub-sample required for sampling was large compared to the total test media volume, the test item concentration was determined from samples taken from additionally prepared glass beakers. Five millilitres were pipetted directly into 20 mL extraction (centrifuge) tubes and 15 mL of purified water added. The pipette tips were rinsed multiple times and the surface wiped with a lint free paper towel prior taking the analytical sub-sample.
- Sample storage conditions before analysis: The samples were measured immediately after sampling without further storage.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test substance pellets were crushed by mortar and pestle and the Water Accommodated Fraction (WAF) prepared with the test item powder. A test preparation with a loading of 100 mg/L dilution water was prepared according to the OECD guideline No. 23, and stirred slowly at 300 rpm to avoid bubble and foam formation using a stirring bar on a magnetic stirrer for 48 h at room temperature (about 20 °C). Finally, the test preparation was filtered using a 0.2 µm membrane filter (Sartolab 150V, vacuum capsule sterile - Sartorius) to separate insoluble parts from the aqueous phase in accordance with the OECD guideline No. 29. The aqueous phase (WAF) was drawn off and used for testing. The control consisted of dilution water only and was not stirred for 48 h.
- Eluate: no
- Differential loading: yes
- Controls: yes, dilution water control

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: Obtained from the German Federal Environment Agency, Institut für Wasser-, Boden- und Lufthygiene, Germany. Specimens used in the test were bred in the laboratory at Fraunhofer IME.
- Age at study initiation (mean and range, SD): < 24 h
- Method of breeding: Adult Daphnia, at least 3 weeks old, were separated from the stock population by sieving. Batches of 30 - 50 animals were held at room temperature in ca. 1.8 L of dilution water for one week. During this week, the daphnids were fed daily with an algal suspension (Desmodesmus subspicatus) and LiquizellR. Algae growing in the log-phase were centrifuged and the pellet was re-suspended in a few mL of medium. Thirty milliliters of this suspension was given to 1 L of medium. The water was changed once per week. Newborn Daphnia (between 4 - 23 h old) were removed by wide-bore pipette (to avoid damage) and isolated in fresh dilution water for at least 1 h prior to being added randomly to the test vessels containing the appropriate test or control media.
- Feeding during test: no

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

20.2 - 20.4 °C

pH:

7.91 - 8.49

Dissolved oxygen:

6.3 - 8.2 mg O2/L

Nominal and measured concentrations:

nominal: control, 100 mg/L
measured: < LOQ, 0.128 µg/L (geometric mean)

Details on test conditions:

TEST SYSTEM
- Test vessel
- Type: covered with a glass pane
- Material, size, headspace, fill volume: glass, 60 mL, headspace: 10 mL, fill volume: 50 mL
- Aeration: no
- 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: Purified drinking water was used according to the OECD guideline. The purification included filtration with activated charcoal, passage through a lime-stone column and aeration. To avoid copper contamination, plastic water pipes were used for the test facilities. The water was aerated to the point of oxygen saturation.
- Chlorine: < 0.02 mg/L
- Alkalinity: 2 mmol/L
- Conductivity: 244.5 µS/cm
- Nitrate: 1.7 mg/L
- Nitrite: 0.005 mg/L
- Ammonium: < 0.01 mg/L
- Phosphate: 0.73 mg/L
- Culture medium different from test medium: no
- Intervals of water quality measurement: regularly

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light / 8 h dark
- Light intensity: 518 - 536 lux

EFFECT PARAMETERS MEASURED: After 24 h and 48 h, the number of immobile animals in each beaker was counted. The animals were considered to be immobile if they were not able to swim within 15 sec of gentle agitation of the test vessels. Any abnormalities in appearance and behaviour were also recorded.

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 1, 10 and 100 mg/L (nominal WAF)
- Results used to determine the conditions for the definitive study: EL50 > 100 mg/L

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

>= 0.128 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOELR

Effect conc.:

> 0.128 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 0.128 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOELR

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mobility

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

Details on results:

- Other biological observations: No signs of disease or stress were observed. All surviving specimens gave the impression of healthy condition.
- Mortality of control: 0%

Results with reference substance (positive control):

- Results with reference substance valid? yes
- EC50/LC50: EC50 (24 h): 0.87 mg/L

Table1: Measured concentrations of tetradecyl myristate [µg/L].

Nominal loading	Test duration				Geometric mean
	Mean measured 0 h		Mean measured 48 h
[mg/L]	[µg/L]	% nominal	[µg/L]	% nominal	[µg/L]	[%]
100	0.172	1.72 E-4	0.096	9.59 E -5	0.128	1.28 E-4
Control	<LOQ	-	<LOQ	-	-	-

 

 Table 2: Immobility raw data

Geometric mean conc. [mg/L]	Control	100
0 h	5	5
	5	5
	5	5
	5	5
Number of replicates:	4	4
Total Introduced:	20.00	20.00
24 h	0	0
	0	0
	0	0
	0	0
Number of replicates:	4	4
Total Immobile:	0.00	0.00
48 h	0	0
	0	0
	0	0
	0	0
Number of replicates:	4	4
Total Immobile:	0.00	0.00

Validity criteria fulfilled:

yes

Conclusions:

Based on the geometric mean measured concentrations of the test item, the 48 hour EL50 for Daphnia magna was 0.128 µg/L (nominal > 100 mg/L). The NOEL after 48 hours was >/= 0.128 µg/L (nominal >/= 100 mg/L).

Executive summary:

In the acute immobilisation test with Daphnia magna, the effects of tetradecyl myristate (CAS No 3234-85-3), a constituent of the registration substance, were determined according to OECD 202. The study was conducted under static conditions over a period of 48 h with the WAF of a 100 mg/L solution (nominal concentration; measured concentration < LOQ, 0.128 µg/L).

20 daphnids were exposed to the test item solution and dilution water control.

The concentration of tetradecyl myristate was analytically by GC verified at the study start and termination (48 h).

Table1: Measured concentrations of tetradecyl myristate [µg/L].

Nominal loading	Test duration				Geometric mean
	Mean measured 0 h		Mean measured 48 h
[mg/L]	[µg/L]	% nominal	[µg/L]	% nominal	[µg/L]	[%]
100	0.172	1.72 E-4	0.096	9.59 E -5	0.128	1.28 E-4
Control	<LOQ	-	<LOQ	-	-	-

 

The EL50 and NOEL after 48 h were 0.128 µg/L (nominal > 100 mg/L) and >/= 0.128 µg/L (nominal >/= 100 mg/L), respectively.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

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

Adequacy of study:

weight of evidence

Study period:

30 Jan - 01 Feb 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
Please refer to cread across justification justification setcion 13.2

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
Please refer to cread across justification justification setcion 13.2

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Please refer to cread across justification justification setcion 13.2

3. ANALOGUE APPROACH JUSTIFICATION
Please refer to cread across justification justification setcion 13.2

4. DATA MATRIX
Please refer to cread across justification justification setcion 13.2

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF)

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF) + Emulsifier

Basis for effect:

mobility

Details on results:

- Mortality of control: 0%

Results with reference substance (positive control):

- EC50 (24 h): 0.98 mg/L which is in the range of 0.9 and 1.9 mg/L proposed for validity of the test.

Table 1: Immobilization of test species after 24 h and 48 h.

sample name	replicate	immobilised test species [%]
		24 h	48 h
control	1	0	0
	2	0	0
Solvent control	1	0	0
	2	0	0
WAF, filt. (100 mg/L)	1	0	0
	2	0	0
WAF (100 mg/L) + TAGAT	1	0	0
	2	0	0

Validity criteria fulfilled:

yes

Conclusions:

Based on the reported observation of the present short term toxicity test with the test item, fatty acids, C16-18, C12-18-alkyl esters, in Daphina magna the EL50 (nominal) with and without emusifier was greater than 100 mg/L.

Executive summary:

In the acute immobilisation test with Daphnia magna, the effects of fatty acids, C16 -18, C12 -18,-alkyl esters (CAS No 93912-87-1) were determined according to OECD 202. The study was conducted under static conditions over a period of 48 h with the WAF of a 100 mg/L solution.

20 daphnids were exposed to the test item solution (with and without emulsifier) and vehicle control.

The concentration of test item was analytically verified by GC.

Result:

Table 1: Immobilization of test species after 24 h and 48 h.

sample name	replicate	immobilised test species [%]
		24 h	48 h
control	1	0	0
	2	0	0
Solvent control	1	0	0
	2	0	0
WAF, filt. (100 mg/L)	1	0	0
	2	0	0
WAF (100 mg/L) + TAGAT	1	0	0
	2	0	0

Description of key information

No toxic effects up to the limit of water solubility (< 0.02 mg/L) for Daphnia magna (OECD 202).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

Two GLP Guideline studies conducted according to OECD 202 are available investigating the short-term effects of tetradecyl myristate (CAS 3234-85-3), a constituent of the registration substance, and fatty acids, C16 -18, C12 -18-alkyl esters (CAS No 95912 -87-1), a strucutral analogue, to freshwater invertebrates. Daphnia magna was exposed to a Water Accommodated Fraction (WAF) of nominal 100 mg/L for 48 h.

1. Result of study with tetradecyl myristate:

GC/EI-MS analysis resulted in a measured geometric mean concentration of 0.128 µg/L. No immobilization was recorded after 48 h resulting in an EL50 of > 0.128 µg/L (measured, geometric mean) and > 100 mg/L (nominal), respectively. Thus it can be concluded that no toxicological effects on aquatic invertebrates are expected up to the limit of water solubility for tetradecyl myristate.

2. Result with fatty acids, C16 -18, C12 -18-alkyl esters:

no effects on the test organisms were observed resulting in an EL50 > 100 mg/L (nominal).