Oligo

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:

27 Nov - 01 Dec 2017

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Analytical method was not sensitive enough to detect test item at limit loading rate.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: control and 100 mg/L
- Sampling method: Samples were taken at test start and end.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The weighed test sample (0.28 g) and dilution water (2.8 L) were mixed to prepare the nominal concentration of 100 mg/L. The solution was stirred with a magnetic stirrer für 48 h to prepare the test solution.
- Eluate: no
- Differential loading: yes
- Controls: yes, test medium control
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): Precipitate was recorded at the exposure vessel at test start and test end. Particles were floating on top of the water layer.
- Appearance of test solutions: Test solution at start: white, at the end: colorless and clear

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: medaka
- Source: CERI Kurume (in-laboratory production)
- Length at study initiation (length definition, mean, range and SD): 2.0 ± 1.0 cm
- Weight at study initiation (mean and range, SD): 0.14 ± 0.02 g

ACCLIMATION
- Acclimation period: 12 d
- Acclimation conditions (same as test or not): same as test
- Type and amount of food during acclimation: Feed of Medaka fry for education (KYORIN); Amount corresponding to 3% of body weight
- Feeding frequency during acclimation: daily (fish were not fed 24 h before test initiation)
- Health during acclimation (any mortality observed): 0%; no external medication was provided

FEEDING DURING TEST
- None

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

23.2 - 23.9 °C

pH:

7.8 - 8.0

Dissolved oxygen:

8.3 - 8.7 mg/L

Nominal and measured concentrations:

nominal: control, 100 mg/L
measured: < LOD for control and limit concentration

Details on test conditions:

TEST SYSTEM
- Test vessel: 3 L glass tank filled with 2.8 L of test medium (diameter: 16 cm, depth: 16 cm) stored in a temperature controlled water bath
- Type: covered with a transparent plastic lid
- Aeration: yes, gentle aeration
- 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: Dechlorinated tap water, aerated sufficiently and temperature-controlled
- Total organic carbon: < 0.5 mg/L
- Particulate matter: < 1 mg/L
- Pesticides: < LOQ
- Chlorine: < LOQ
- Alkalinity: 36 mg/L
- Culture medium different from test medium: same as test

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light / 8 h darkness
- Light intensity: room light

EFFECT PARAMETERS MEASURED
- Mortality and visible abnormality were observed after 3, 24, 48, 72 and 96 h after the start of exposure.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable, limit test
- Range finding study
- Test concentrations: 100 mg/L
- Results used to determine the conditions for the definitive study: No mortality was observed after 96 h.

Reference substance (positive control):

yes

Remarks:

Copper (Il) sulfate pentahydrate was periodically tested.

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

WAF

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: 0%
- Abnormal responses: No abnormal response was observed in the exposure level and control during exposure.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The exposure solutions were colorless and clear with floating particles on the surface. The solution in the control was colorless and clear.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- LC50 (96 h): 0.33 mg/L (standard deviation: 0.14 - 0.90 mg/L)
- Other: Study was performed from 13-17 Nov 2017

Sublethal observations / clinical signs:

Table 1: Validity criteria for OECD 203.

Criterion from the guideline	Outcome	Validity criterion fulfilled
The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test	0%	yes
The dissolved oxygen concentration must have been at least 60 per cent of the air saturation value throughout the test	8.3 - 8.7 mg/L (saturation level: 8.26 - 8.58 mg/L)	yes

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Analytical method is not considered to be appropriate.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: control, 1000 mg/L
- Sampling method: Water samples were taken from the middle of the test chamber by pipetting approx. 20 mL into a screw-neck glass bottle at study start, and after 48 and 96 h.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test substance was weighed into a beaker. Water for dilution was added, the mixture was homogenized using an Ultra-Turrax and poured into the chamber under stirring with a glass rod. The chamber contents were then stirred for approximately 30 min with a KPG-stirrer and a glass rod.
- Eluate: no
- Differential loading: yes
- Controls: yes, test medium control
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): The tested concentration showed a turbidity. Additionally substance deposits at the water surface and on the bottom of the test chamber were observed.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Source: Charles River Aquatics, Someren, Netherlands (date of hatching: 15 Dec 1995; date of delivery: 13 Mar 1996)
- Length at study initiation (length definition, mean, range and SD): 2.7 - 3.0 cm (mean: 2.8 cm)

ACCLIMATION
- Acclimation period: 12 d
- Acclimation conditions (same as test or not): 22 ± 1 °C, oxygen content >= 80% of the saturation value, 12 h light daily
- Type and amount of food during acclimation: commercial fish food (Tetra Min, Tetra Werke, Melle)
- Feeding frequency during acclimation: twice daily ad libitum

FEEDING DURING TEST
- None

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

21.6 - 21.8 °C (test concentration)
21.2 - 21.9 °C (control)

pH:

7.8 - 8.1 (test concentration)
8.0 - 8.2 (control)

Dissolved oxygen:

6.9 - 9.5 mg/L (test concentration)
8.0 - 9.9 mg/L (control)

Nominal and measured concentrations:

nominal: control, 1000 mg/L
measured: < 1 (control), 342 mg/L (geometric mean; 1000 mg/L)

Details on test conditions:

TEST SYSTEM
- Test vessel: The test chambers, which were calibrated to 10 L, were made of glass (length 30 cm, width 22 cm, height 24 cm) and stood in a water bath with a plexiglas viewing panel.
- Aeration: no
- 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: Reconstituted water, composition according to ISO/DIS 7346/1 was used as water for dilution.
- Hardness: 2.3 - 2.5 mmol Ca2+ + Mg2+/L
- Culture medium different from test medium: same as test
- Intervals of water quality measurement: Water parameters were measured regularly.

OTHER TEST CONDITIONS
- Photoperiod: 12 h light / 12 h darkness
- Light intensity: 700 lux

EFFECT PARAMETERS MEASURED
- Mortality was recorded after 3, 6, 24, 48, 72 and 96 h

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Other biological observations: Recording of symptoms was disturbed because of the turbidity of the test water until 24 h after study start. Ouring this first 24 h the fish showed slight, reversible symptoms as swimming posture tail-heavy and projecting opercula.
- Mortality of control: 0%

Sublethal observations / clinical signs:

Table 1: Validity criteria for OECD 203.

Criterion from the guideline	Outcome	Validity criterion fulfilled
The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test	0%	yes
The dissolved oxygen concentration must have been at least 60 per cent of the air saturation value throughout the test	6.9 - 9.5 mg/L (test concentration) 8.0 - 9.9 mgL (control) 8.8 mg/L (=100% saturation)	yes

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.

Description of key information

LL50 (96 h) > 100 mg/L (nominal, OECD 203)

Additional information

Two experimental studies are available investigating short term toxicity of Fatty acids C16-18 (even numbered), oligoesters with sucrose to freshwater fish.
The key study was performed according to OECD 203 (GLP) with Oryzias latipes. A limit loading rate of 100 mg/L was tested and mortality of fish was determined after 96 h. The actual concentration in the test vessels was tried to analyze using HPLC. However, the analytical method was not sensitive enough (limit of detection: 0.505 mg/L) to detect the substance in test solutions. This result reflects the very low water solubility of the test item. After 96 h no mortality was recorded and the LL50 was derived to be > 100 mg/L (nominal).
In the supporting study Danio rerio was exposed to the test item for 96 h at a limit concentration of 1000 mg/L. The study was performed according to OECD 203 (GLP). The test item was directly added to the test vessels without any further processing step (e.g. filtration). HPLC analysis of the test item concentration in test medium resulted in a geometric mean measured concentration of 342 mg/L. This result is unexpectedly high and does not fit to the very low water solubility of the test item which was reported to be < 0.509 mg/L. Thus, the applied analytical method is not considered to be robust and the results are based on the nominal concentration. After 96 h no mortality was recorded and a LC50 of > 1000 mg/L was derived.