Fatty acids, C12-14 (even numbered), methyl ester

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21 Oct - 26 Oct 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Circular on Test Methods of New Chemical Substances (Japan), Daphnia acute immobilisation test

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES
- Melting point: 5 °C
- Boiling point: 250 °C
- Vapour pressure: 1.3 hPa at 88 °C
- Water solubility: 0.4301 mg/L (calculated)
- log Kow: 6.0

Analytical monitoring:

yes

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: One day before the test initiation, 10000 mg/L stock solution was prepared by dissolving test substance in dimethylformamide (DMF) and dilution water. Stock solution was stored at room temperature during exposure period and it was confirmed that it stabled within 9 days. Test water was continuously prepared with propotional diluter and a magnetic stirrer.
- Controls: blank (dilution water) control and vehicle control
- Chemical name of vehicle: N,N-dimethylformamide (DMF)
- Concentration of vehicle in test medium (final test solution): 0.100 mL/L

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Medaka
- Source: breed in-house
- Age at study initiation: about four months
- Length at study initiation: 2.3 ± 1.2 cm
- Feeding during test: no

ACCLIMATION
- Acclimation period: 60 day
- Type and amount of food: general pellet (2C)
-Withdrawal of food: 1 day before exposure
- Health during acclimation (any mortality observed): no mortality was observed during 7 days before study initiation

Test type:

flow-through

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

120 h

Remarks on exposure duration:

because of low water solubility of the test item, exposure was continued until 120 hours

Hardness:

29.2 mg/L ( as CaCO3)

Test temperature:

23.2 - 23.8 °C

pH:

7.6 - 7.8

Dissolved oxygen:

8.3 - 8.5 mg/L (> 60 % of saturated oxygen)

Nominal and measured concentrations:

nominal concentration: 1 mg/L
measured concentration (arithmetic mean): 0.524 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 3.0 L glass vessel
- Type: closed
- Material, size, fill volume: glass, diameter 16 cm: depth 17 cm, 1.8 L
- Aeration: no
- Type of flow-through: proportional diluter
- Renewal rate of test solution (frequency/flow rate): Control: dilution water 120 mL/min, Vehicle control: dilution water 120 mL/min + DMF 12 μL/min, Test water: Dilution water 120 mL/min + stock solution 12 μL/min
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated aerated tap water
- Alkalinity: 20 mg/L
- Ca/mg ratio: 8.4/2.0

OTHER TEST CONDITIONS
- Adjustment of pH: not adjusted
- Photoperiod: 16 hours in light / 8 hours in dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
LC50 after 96 hours and 120 hours

TEST CONCENTRATIONS
- Range finding study: performed
- Test concentrations: 1 mg/L and control
- Results used to determine the conditions for the definitive study: test type: closed, type of flow-through: not semi-static but flow-through, concentration of vehicle

Reference substance (positive control):

yes

Remarks:

copper sulfate pentahydrate (analytical grade)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.52 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: not observed during exposure
- Observations on body length and weight: length: 2.7 ± 0.095 cm, weight: 0.16 ± 0.017 g
- Other biological observations: no abnormality was observed
- Mortality of control: 0%
- Other adverse effects control: not observed
- Abnormal responses: not observed
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: test water was clear at the stud initiation and after expoxure

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- LC50: 0.423 mg/L (reference range: 0.111 - 1.01 mg/L)

Sublethal observations / clinical signs:

Table 1. Measured concentration of test substance in test solution

Nominal Concentration (mg/L)	Measured Concentration (mg/L) (Percentage of solubility in dilution water)		Arithmetic Mean (mg/L)
	0 Hour new	48 Hours old
Control	n.d.	n.d.	-
Vehicle Control	n.d.	n.d.	-
1	0.495 (132)	0.554 (147)	0.524 (139)

n.d.: Not detected (< 0.0495 mg/L)

Solubility in dilution water: 0.376 mg/L

Table 2. Cumulative mortality

Measured Concentration * (mg/L)	Cumulative mortality (%)
	3-hour	24-hour	48-hour	72-hour	96-hour	120-hour
Control	0	0	0	0	0	0
Vehicle Control	0	0	0	0	0	0
0.524	0	0	0	0	0	0

*: Arithmetic mean of the measured concentration

Validity criteria fulfilled:

yes

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07 Jan - 11 Jan 1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions (no analytics)

Qualifier:

according to guideline

Guideline:

ISO 7346-1 (Determination of the Acute Lethal Toxicity of Substances to a Freshwater Fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)] - Part 2: Semi-static method)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Direct weight of test substance into test vessel, filled up to final volumes of 5 L with standard dilution water. Thereafter, stirring 5 min by ultrasound and 20 seconds by ultraturrax

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Feeding during test: no
- Pretreatment: no
- Health during acclimation (any mortality observed): <0.1% mortality prior to the test

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

approx. 250 mg CaCO3/L

Test temperature:

23.0 - 23.5 °C

pH:

8.0-8.3

Dissolved oxygen:

80-97% air saturation value

Nominal and measured concentrations:

Nominal: control, 1000, 3000 and 10000 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 10 L fish vessel, containing 5 L of test water
- Renewal rate of test solution: daily
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1


OTHER TEST CONDITIONS
- Photoperiod: 16 hours light, 8 hours darkness

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality at 0, 6, 24, 48, 72 and 96 hours

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1 700 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

3 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: none
- Test concentrations were considerably above the water solubility of the test substance (0.05 mg/L)

Sublethal observations / clinical signs:

Table 1: Mortality [%] of Danio rerio at 6, 24, 48, 72 and 96 h incubation time

Concentration	Incubation time (h)
in mg/L	0	6	24	48	72	96
0	0	0	0	0	0	0
1000	0	0	0	0	0	0
3000	0	70	100	100	100	100
10000	0	100	100	100	100	100

Validity criteria fulfilled:

yes

Description of key information

No effects up to the limit of water solubility (WS < 0.06 mg/L)

Additional information

No experimental data evaluating the acute toxicity of Fatty acids, C12-14 (even numbered), methyl esters (CAS No. 308065-15-8) on fish are available. Therefore, toxicity data from two structurally related category members, methyl laurate (CAS No. 111-82-0) and methyl myristate (CAS No. 124-10-7) are used as read-across in accordance with Regulation (EC) No. 1907/2006, Annex XI, 1.5. All three substances are fatty acids esterified with methanol, containing fatty acid C-chain lengths of C12-14 (CAS No. 308065-15-8), C12 (CAS No. 111-82-0) and C14 (CAS No. 124-10-7). Therefore, the two read-across substances represent the individual fatty acid components contained in the target substance. The log Pow (> 5) and water solubility values (< 0.06 mg/L) of methyl myristate and Fatty acids, C12-14 (even numbered), methyl esters indicate that these two substances are expected to show a comparable behavior in the water phase. Despite the fact that the water solubility value of methyl laurate is not comparable (7.7 mg/L), the target substance has a high content of C12 fatty acids (30-100%). Methyl laurate (C12 FA content > 80%) exhibits the highest toxicity to aquatic organisms (Daphnia and algae) throughout the whole SCAE Me category, with EC50 and NOEC values < 1 mg/L. Therefore, methyl laurate is a worst-case read across substance for Fatty acids, C12-14 (even numbered), methyl esters in accordance with Regulation (EC) No. 1907/2006, Annex XI, 1.5.

The test conducted with methyl laurate was performed according to OECD Guideline 203 under GLP conditions (Japanese Ministry of Environment, 2004). Oryzias latipes was exposed to the substance for 120 h within a flow-through water regime, at a nominal concentration of 1 mg/L (limit test). No mortality was observed in either treatment or control groups after the exposure period and therefore the LC50 (96 h) was determined to be > 0.52 mg/L (measured concentration). The test conducted with methyl myristate (Richterich, 2001) was performed according to ISO Guideline 7346-1. Danio rerio was exposed for 96 h to the test substance within a semi-static water regime, at concentrations ranging from 1000 to 10,000 mg/L (nominal). Mortality was observed at the highest concentrations tested (3000 and 10000 mg/L, nominal), resulting in a LC50 (96 h) of 1700 mg/L (nominal). Due to the poorly soluble nature of the substance in water (water solubility 0.05 mg/L) and considering that it was directly added to the test solution without any further filtration step, the observed effects are expected to be caused rather by physical interference with undissolved test material (e.g. gill clogging) than due to toxicity.

Based on the results obtained for the read-across substances (in accordance with Regulation (EC) No. 1907/2006, Annex XI, 1.5), acute toxicity of Fatty acids, C12-14 (even numbered), methyl esters to fish species up to the limit of its water solubility (< 0.06 mg/L) is not expected.