Fatty acids lanolin, di-, tri- and tetraesters with pentaerythritol and rape fatty acid

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:

29 Jan - 21 Feb 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Principles of method if other than guideline:

Since the substance is a UVCB the WAF (water accommodated fraction) was prepared in accordance with the method of Schäfers, C. et al. (2009, Environmental properties of long-chain alcohols, part 2: Structure-activity relationship for chronic aquatic toxicity of long-chain alcohols. Ecotoxicology and Environmental Safety, 72 (4), pp 996 – 1005).

GLP compliance:

yes (incl. QA statement)

Remarks:

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

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A concentrated stock solution of the test item was prepared by weighing an appropriate amount of the test item onto a small Teflon plate. The Teflon plate was placed inside a sterile brown glass Schott bottle with drain port. Dilution water was then added in an appropriate volume and the solution stirred at 300 rpm for 24 hours at room temperature (21 ± 2 °C). The WAF solution was then left to settle for 24 hours. The WAF solution formed the test item loading of 1 mg/L.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Age at start: aged less than 24 hours
- Source: German Federal Environment Agency, Institute for Water, Soil, and Air Hygiene (Berlin). Specimens used in the test were bred in the laboratory at Fraunhofer IME.
- Holding: 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 containing Artemio Fluid JBL). Algae growing in the log-phase were centrifuged and the pellet was re-suspended in a few mL of medium. Thirty millilitres of this suspension was given to 1 L of medium. The medium 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
- Food type: Concentrated suspensions of living cells of the unicellular alga Desmodesmus subspicatus
- Amount: The concentration of carbon (mg carbon/L) in the algal culture was measured by optical density at 585 nm and compared to a calibrated nomograph to calculate the volume required for feeding of adult daphnids.
- Frequency: Daily after the removal of juveniles and at exchange of test solutions

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Hardness:

1.0 mmol/L (dilution water)

Test temperature:

18.5 – 19.9 °C (with less than 2 °C variation)

pH:

7.70 – 9.62

Dissolved oxygen:

6.4 – 9.8 mg O2/L

Nominal and measured concentrations:

1 mg/L (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL glass flasks
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 100 mL glass flasks stoppered with silicone plugs and filled with pproximately 100 mL of each test medium
- Aeration: none
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Purified local tap water is used. The tap water is sourced from the Schmallenberg district water production plants, mostly fed by small springs and percolation. The purification process occurs on site at Fraunhofer IME and includes filtration with activated charcoal, passage through a lime-stone column, and aeration to the point of oxygen saturation. To avoid copper contamination, plastic water pipes are used in the test facilities.
- Chlorine: <0.02 mg/L
- Alkalinity: 1.3 – 1.6 mmol/L
- Conductivity: 203.7 – 216.8 µS/cm
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Photoperiod: Light/dark cycle of 16/8 hours
- Light intensity: 609 – 988 lux
- Sterile conditions: All test media was prepared under sterile conditions, used within 1-2 hours of final preparation, and replaced daily.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): The number of juvenile D. magna per replicate was recorded daily from day 6 onwards. Mortality, abnormalities in appearance, behaviour, condition (including male sex) or presence of winter eggs (ephippium) were also recorded. At study termination, body length (excluding the anal spine) of the adults in all treatments without significant mortality was measured by digital photography

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test mat. WAF

Basis for effect:

immobilisation

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test mat. WAF

Basis for effect:

other: mean age of first brood

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test mat. WAF

Basis for effect:

reproduction

Details on results:

- Statistically significant difference in length of treatment and control. The observed difference between the mean length of both groups was 6.04% (p = 0.039; 3.978 mm in the control vs 3.738 mm in the treatment) and should be thus considered of minor biological importance. Due to the small variance (s²=0.078) calculated for each group, the small difference turned out to be significantly different.

Reported statistics and error estimates:

The subject of the statistical evaluation was the replicate (n=10). The data were tested for variance homogeneity and normal distribution. The NOEC and LOEC were calculated by ANOVA followed by parametric pairwise comparisons of the limit treatment to the control using a Students T-Test or Fishers Exact Binomial Test with Bonferroni Correction. The NOEC was reported as more than the highest concentration tested when the statistical test failed to find a difference between the limit treatment and the control. Probit analysis modified for continuous data for the calculation of EC50 values can be applied when at least the two highest treatment levels show effects and inhibition of 100% is theoretically possible. As the response (i.e. number of treatments) was less than three the data was deemed inappropriate for the determination of EC50 values. All statistical evaluations were conducted using the computer software ToxRat® Professional v. 2.10 (ToxRat® Professional 2.10. ToxRat® Solutions GmbH. Dr. Monika M. Ratte, Naheweg 15, 52477 Alsdorf, Germany).

Table: Survival, growth and reproduction averages for all endpoints of Daphnia magna at the end of the 21 d exposure period to the test item compared to the control treatment.

Nominal treatment loading	Parental immobility	Growth (length on day 21)	Age at first brood	Cumulative offspring per female
(mg/L)	%	Mean ± SD (mm)	Mean ± SD (days)	Mean ± SD (n)
Control	0	3.98 ± 0.26	10.2 ± 0.95	64.7 ± 12.44
1	10	3.74 ± 0.3	10.8 ± 1.06	66.1 ± 9.08
NOEL	≥ 1 mg/L	n.d.	≥ 1 mg/L	≥ 1 mg/L

 n.d.: not able to determine as there were effects at the response level

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

Justification for type of information:

Please refer to the analogue justification attached to chapter 13.

Reason / purpose for cross-reference:

read-across source

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

reproduction

Remarks on result:

other: source: CAS 85711-45-1, Oleon, 2013, D. magna, 21 d

Conclusions:

The substance will not exhibit long-term effects to aquatic invertebrates up to the limit of water solubility.

Additional information

Since no studies investigating the long-term toxicity of Fatty acids lanolin, di, tri- and tetraesters with pentaerythritol and rape fatty acid to aquatic invertebrates are available, in accordance with Regulation (EC) No 1907/2006 Annex XI, 1.5, a read-across to the structurally related source substance Fatty acids, C16-18 and C18-unsatd., esters with pentaerythritol (CAS 85711-45-1) was conducted.

The source substance is representative to evaluate the short-term toxicity of the target substance to aquatic invertebrates. This read-across is justified in detail within the analogue justification in IUCLID Section 13.

The target substance Fatty acids lanolin, di, tri- and tetraesters with pentaerythritol and rape fatty acid is characterized as a UVCB substance with pentaerythritol as alcohol component. Esterification of pentaerythritol gives mainly tri- and tetraesters of long chain fatty acids C16-C20.

The source substance Fatty acids, C16-18 and C18-unsatd., esters with pentaerythritol (CAS 85711-45-1) is characterized as a UVCB substance with pentaerythritol as alcohol component. Esterification of pentaerythritol gives a mixed degree of esters of mainly C16-C18 fatty acids.

The study with the source substance Fatty acids, C16-18 and C18-unsatd., esters with pentaerythritol (CAS 85711-45-1) was performed in a semi-static test design according to OECD 211 (GLP).Daphnia magnawere exposed to a single nominal loading rate of 1 mg/L (prepared as WAF). No immobility and no sub-lethal effects were observed after an exposure period of 21 days. Hence a NOELR of ≥ 1 mg/L was determined.

Based on the results for the structurally related read-across substance (in accordance with Regulation (EC) No 1907/2006 Annex XI, 1.5) which is characterized by a similar ecotoxicological profile and comparable structure, it can be concluded that no long-term toxicity of the target substance Fatty acids lanolin, di, tri- and tetraesters with pentaerythritol and rape fatty acid to aquatic invertebrates is expected up to the limit of the water solubility (< 0.15 mg/L).