Hexadecyl palmitate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

Refer to analogue justification provided in IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other:

Remarks:

Source: CAS 3234-85-3, Evonik, 2012, D. rerio, 96 h

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.189 µg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other:

Remarks:

Source: CAS 3234-85-3, Evonik, 2012, D. rerio, 96 h

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other:

Remarks:

Source: CAS 3234-85-3, Evonik, 2012, D. rerio, 96 h

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 0.189 µg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other:

Remarks:

Source: CAS 3234-85-3, Evonik, 2012, D. rerio, 96 h

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF)

Basis for effect:

mortality (fish)

Remarks on result:

other:

Remarks:

Source: CAS 36078-10-1, Dako, 2013, D. rerio, 96 h

Duration:

96 h

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF)

Basis for effect:

mortality (fish)

Remarks on result:

other:

Remarks:

Source: CAS 36078-10-1, Dako, 2013, D. rerio, 96 h

Conclusions:

No effects on D. rerio (OECD 203) up to the limit of water solubility (< 0.02 mg/L and < 0.05 mg/L, respectively).

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 Sep - 14 Sep 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

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
- Sampling method: Samplings out of the test aquaria were done by the staff of the analytical laboratory using a 'research 5000' pipette.
The aqueous samples of 5.0 mL were pipetted directly into the extraction (centrifuge) tubes; the pipette tips were rinsed multiple and the surface was wiped with a lint free paper towel prior taking the analytical sub-sample.
- Sample storage conditions before analysis: The samples were processed and measured directly after sampling without any time delay.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Due to the very low water solubility of the test item, a Water Accommodated Fraction (WAF) of the test item was applied. The test substance pellets were crushed by mortar and pestle and the WAF prepared with the test item powder. Therefore, 1050 mg test item powder was weighed in a glass bottle. The bottle was filled with 10.5 L water. The solution was stirred slowly to avoid bubble and foam formation using a star shape magnetic stirring bar for about 48 h at room temperature (about 20 °C). After stirring the test solution was filtered by using a 0.2 µm filter (Sartolab 150v, Vacuum capsule sterile, Charge 21/000143, Sartorius). 10 L of the filtrate was used for the study resulting in a nominal concentration of 100 mg/L.
- Eluate: no
- Differential loading: yes
- Controls: yes, test medium control

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Source: Test facility bred.
- Length at study initiation: 2.2 - 2.6 cm (SD: 0.1 cm)
- Weight at study initiation: 0.11 g (mean; SD: 0.02 g)
- Method of breeding: The fish were held in water of the same quality as used in the test (purified drinking water) until the start of exposure. Fish were fed ad libitum throughout the holding period with live brine shrimp (Artemia spp.) nauplii and ground flake food TetraMin (Tetra Werke, Melle, Germany) once daily. No feeding of the fish occurred 24 h before test start.
- Feeding during test: no

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

1 mmol/L

Test temperature:

23.1 - 23.2 °C

pH:

7.9 - 8.6

Dissolved oxygen:

74 - 100%

Nominal and measured concentrations:

nominal: control, 100 mg/L
measured: < LOQ, 0.189 µg/L (0 h), < LOQ (96 h)

Details on test conditions:

TEST SYSTEM
- Test vessel
- Material, size, headspace, fill volume: full glass aquaria, 15 L, headspace: 5 L, fill volume: 10 L
- Aeration: slightly aerated via glass capillary
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.07 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Purified drinking water was used according to the OECD-Guideline. The purification includes filtration with activated charcoal, passage through a lime-stone column and aeration until oxygen saturation. To avoid copper contamination, plastic water pipes are used for the test facilities.
- Metals: < detection limit
- Chlorine: < 0.02 mg/L
- Alkalinity: 2 mmol/L
- Conductivity: 244.5 µS/cm
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 12 h light / 12 h dark

EFFECT PARAMETERS MEASURED: Mortality and clinical signs (sub-lethal and abnormal behavior) were recorded at 3 h, 24 h, 48 h, 72 h and 96 h after fish addition.

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: No mortality, clinical signs or any distress were observed throughout the range-finding test.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.189 µg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 0.189 µg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: No clinical signs nor mortality could be were observed throughout the study.
- Mortality of control: 0%

Sublethal observations / clinical signs:

The effects of the test substance on fish were assessed in the acute limit study with zebrafish (Danio rerio), which were exposed to a single treatment of nominal 100 mg test item/L. The chemical analysis of tetradecyl myristate in the fresh and aged test media revealed measured concentrations of ≤0.01% of nominal 100 mg/L and <LOQ at the start and the end of the test respectively. The LOQ was 0.1 µg/L.

Table 1: Analytical concentrations in the stock solution at the beginning of the study and at the end of exposure.

Nominal conc. 100 mg/L	Samp- ling time	Conc. factor, FC	Quan. data, CGC/MS [ng/mL]	Analyzed a.s. conc., CW [µg/L]	Percent of nominal
Stock, 100 mg/L *)	t0 fresh test media	25.0	4.08	0.163	1.6E-4
Control		25.0	n.d. **)	< LOQ	-
Test media, 100 mg/L		25.0	4.73	0.189	1.9E-4
Control, sample 1	96 h, aged test media	25.0	n.d.	< LOQ	-
Control, sample 2		25.0	n.d.	< LOQ	-
Control, sample 3		25.0	n.d.	< LOQ	-
Test media, sample 1		25.0	n.d.	< LOQ	-
Test media, sample 2		25.0	n.d.	< LOQ	-
Test media, sample 3		25.0	n.d.	< LOQ	-

*)       Analytical sub-sample was taken before the stock WAF was distributed into the test aquaria

**)     n.d. = not detected

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03 Dec - 07 Dec 2012

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

GLP compliance:

yes (incl. QA statement)

Remarks:

LUBW Landesanstalt für Umwelt, Messungen und Naturschutz, Baden-Württemberg, Germany

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Samples were taken from the control and the exposure vessel (100 mg/L).
- Sampling method: Sampling at start (0 h) and end (96 h), 4.8 mL were taken from the test and the control aquarium for chemical analysis.
- Sample storage conditions before analysis: Sample storage in 4 mL brown glass vials (filled up to the brim to avoid oxidative processes), maximum storage period of 8 d at 5 °C ± 3 °C before analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Due to sparingly soluble properties of the test item, the study was performed with the water accommodated fraction (WAF) prepared with synthetic fish water according to OECD 203. The test solutions were withdrawn from the middle of the suspension og the beakers after 1 h settling using a glass tube and transferred into the test vessels (aquarium).
Two beakers were prepared like this and stirred for a period of 48 h at 12-22 °C in diffuse light. For stirring an overhead high-grade stainless steel radial impeller was used (about 500 rpm). The impeller was placed about 5 cm above the bottom of the beakers. The depth of the vortex was about 10% of the total height of the liquid. The test item is very mobile and was therefore added by pipetting. According to the density indicated by the sponsor of 0.8529 g/cm3 the amount to be added was converted into volume.
The filtration of the WAF to obtain the water soluble fraction (WSF) was not taken into consideration to avoid unwanted impacts of the filtration process and to have a worst case scenario.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): Slightly whitish oily spots were still floating on the surface of the solutions. The test solutions were withdrawn from the middle of the suspension on the beakers using a glass tube and transferred into the test vessels (aquarium).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Source: Aquarium Dietzenbach Fischzucht, Dietzenbach, Germany, delivery date: 20 Nov 2012
- Length at study initiation (length definition, mean, range and SD): 2 ± 1 cm
- Feeding during test: none

ACCLIMATION
- Acclimation period: 8 days before the test started.
- Acclimation conditions (same as test or not): same as test
- Feeding frequency: 2 x every working day.
- Health during acclimation (any mortality observed): No mortality, the fish behaved normally.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

22.3 - 23.9 °C

pH:

8.0 - 8.1

Dissolved oxygen:

Control: 7.9 - 8.6 mg O2/L
100 mg/L: 7.9 - 8.7 mg O2/L

Nominal and measured concentrations:

Nominal: control, 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Size: 8 L aquarium
- Aeration: yes
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: according to guideline
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 14 h light / 10 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality every 24 h (and 6 h after study initiation); behavioural abnormalities

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF)

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction (WAF)

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: no
- Mortality of control: no

Sublethal observations / clinical signs:

No fish died or showed any abnormal behaviour during the 96 h exposure period.

Description of key information

No acute effects up to the limit of water solubility (OECD 203, D. rerio); read-across

Additional information

There is no study available assessing the short-term toxicity of the target substance hexadecyl palmitate (CAS 540-10-3) to fish. Therefore, read-across to two structurally related source substances was conducted in accordance with Regulation (EC) No 1907/2006 Annex XI, 1.5, namely to lauryl oleate (dodecyl oleate, CAS 36078-10-1) and to tetradecanoic acid, tetradecyl ester (CAS 3234-85-3). Both substances are characterized by similar fatty acid chain lengths as well as alcohol components compared to the target substance. The target substance hexadecyl palmitate (CAS 540-10-3) is an ester derived from a fatty acid and fatty alcohol with carbon atom chain lengths of 16 each, i.e. from palmitic acid and hexadecanol. The source substance dodecyl oleate (CAS 36078-10-1) is an ester of a C18 fatty acid and a C12 fatty alcohol. Tetradecyl myristate (CAS 3234-85-3) is an ester of both a C14 fatty acid and fatty alcohol. Therefore, both source substances are considered suitable representatives for the assessment of the acute toxicity of the target substance to fish. A detailed read-across justification is provided in in the analogue justification in IUCLID section 13.

The study with the source substance dodecyl oleate (CAS 36078-10-1) was performed according to OECD guideline 203 and GLP (Dako, 2013). In a static limit test, D. rerio was exposed to a water accommodated fraction (WAF) with a nominal loading concentration of 100 mg/L for 96 h. No mortality was observed at test end and a LL50 (96 h) of > 100 mg/L was derived based on the nominal concentrations. The chemical analysis resulted in concentrations below the analytical detection limit of 0.005 mg/L reflecting the very low water solubility of the test item.

The study with the source substance tetradecyl myristate (CAS 3234-85-3) was performed according to OECD guideline 203 and GLP (Evonik, 2012). In a static limit test, D. rerio was exposed to a WAF with a nominal loading concentration of 100 mg/L for 96 h. GC/EI-MS analysis resulted in a measured initial concentration of 0.189 µg/L which dropped below the detection limit of 0.1 µg/L at the end of exposure after 96 h. At the end of the test, no mortality was observed, resulting in a LL50 (96 h) of > 0.189 µg/L (measured initial concentration) and > 100 mg/L (nominal), respectively.

Based on the structural and chemical similarity of the target and source substances, the target substance is expected to exhibit a similar ecotoxicological profile as the two source substances. Therefore, it can be concluded that hexadecyl palmitate (CAS 540-10-3) will not exhibit short-term toxic effects to fish up to the limit of water solubility (< 0.846 µg/L).