Fatty acids, sunflower-oil, conjugated, maleated,reation products with diethanolamine, maleated tall-oil fatty acids and triethanolamine

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-12-03 - 2017-12-08

Reliability:

1 (reliable without restriction)

Data source

Reference

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, sunflower-oil, conjugated, maleated,reation products with diethanolamine, maleated tall-oil fatty acids and triethanolamine

EC Number:

800-003-4

Cas Number:

1415316-96-9

IUPAC Name:

Fatty acids, sunflower-oil, conjugated, maleated,reation products with diethanolamine, maleated tall-oil fatty acids and triethanolamine

Test material form:

liquid

Details on test material:

52% active content (solid phase) in water

Specific details on test material used for the study:

Name of test substance: Efka FA 4671
Test substance No.: 16/0408-2
Batch identification: 0016381869
CAS No.: 1415316-96-9
Purity: 100 % UVCB (Substances of Unknown or Variable composition,
Complex reaction products or Biological materials), see Certificate of Analysis
Content: Solid content 52.0 % (Certificate of Analysis)
Identity: Confirmed (Certificate of Analysis)
Homogeneity: Given visual
Storage stability: Expiry date: 24 Nov 2019
Chemical Name: Fatty acids sunflower oil, conjugated, maleated, reaction products with diethanolamine, maleated tall-oil fatty acids and triethanolamine
Physical state/
Appearance: Liquid, yellowish to brownish, clear
Water solubility: Miscible with water at room temperature (T=23°C) and at T=20°C ± 0.5°C in any ratio
The test item is a mixture (UVCB) and undissolved components of the test item were visible at 220 mg/L in test media (observation of the ecotoxicology laboratory).
The test item is a mixture (UVCB) and as such every constituent contributes to a different degree to overall solubility, depending on its own individual solubility and its mass fraction in the test item. Thus, the water solubility depends on the proportions of
test item and water used in the experiment.

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Details on test solutions:

Test Substance Preparation:
The test substance is a multi-component mixture (UVCB), which is designated as miscible with water. Thetest substance was homogenized by shaking thecontainer before use. The test solution was prepared separately (differential loading) by directly adding test substance to test water. The necessary amount of test substance was weighed into a 20 L glass flask and filled to the mark with test water. The solution was then stirred in a closed glass flask for approximately 10 min using a magnetic stirrer. After stirring, the solution was left undisturbed for 24 hours to allow separation of the undissolved material. Undissolved material was observed after 24 hours as fluff and filaments on the bottom and the walls of the glass flask. The aqueous fraction was slightly turbid with faint visible fluff or fibrous material (slight positive Tyndall effect). Approx. 10L of the aqueous fraction was drawn off for testing with a peristaltic pump (500 series, Watson Marlow GmbH, Germany) into a 10 L glass flask.
Throughout the 24 hours intervals the aqueous phase of the tested concentration was colorless but slightly turbid. Undissolved test substance was slightly visible as fluff or fibrous material. Fresh test solutions were prepared daily.

Concentration Control/ Analysis: Specific analyses of the test substance preparations were not conducted since the test substance is an UVCB.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Test species: Zebrafish (Danio rerio)
Animal supplier: The fish were hatched at the testing facility.
Body length(a): 2.9 cm (range 2.5 cm –3.2 cm)
Body weight(a): 0.17 g (range 0.11 g –0.30 g)
Age of the animals: Approx. 9 months
Hatching date: 24-26 Feb 2017
All animals used for the study were derived from the same batch of fish. The test fish were acclimatized to testing conditions prior to test initiation. During acclimatization, the general health status of the fish was observed for signs of sickness, injury, or other abnormalities. All fish were < 3.0 g and the total length of the longest was not more than twice the shortest . The mean length was within the OECD guideline [Ref. 2] recommended range for zebrafish (2.0 ± 1.0 cm), although some individual fish used in this study slightly exceeded the length range. However, the loading of fish mass per volume of test water is much more important in determining fish sensitivity and this aspect was clearly fulfilled (< 1.0 g fish/L). As long as the maximum loading rate in the guideline is not exceeded (1.0 g fish/L) in the test, the use of some slightly larger fish has no influence on the test results.

HOUSING AND DIET
The fish were acclimatized to the testing conditions including test water, temperature and light regime for at least 14 days. The general conditions during acclimatization are as follows:
Housing: Prior to testing, the batch of fish were housed in a glass tank (approx. 45 L) receiving a continuous supply of fresh test water.
Photoperiod: 16 hours light, 8 hours dark
Water quality: The water used for housing and test purposes is nonchlorinated charcoal filtered drinking water (Frankenthal, Germany) mixed with deionized water and aerated to achieve the following water characteristics. The mixed test water is sanitized by UV treatment prior to entering the aquaria.
Total hardness: Approx. 1 mmol/L = 100 mg/L CaCO3
Conductivity: Approx. 250 μS/cm (at 25°C)
Ca content: Approx. 40 mg/L
Mg content: Approx. 5 mg/L
Acid capacity: Approx. 2.5 mmol/L
pH-value: Generally 7.5 – 8.5
Oxygen content: >80% saturation
TOC: Generally <2mg/L
Temperature: 23 ±1°C
Diet: Commercial fish diet ”Tetramin“ (manufacturer Tetra-Werke, Melle, Germany) daily ad libitum, additionally generally on workdays live juvenile brine shrimp (Artemia nauplii). Withdrawal of feed: No feeding from approximately 24 hours before start
of exposure Medical treatment: None during acclimatization Mortality during the last week before start of exposure: 0%

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test conditions

Test temperature:

22 °C

pH:

No adjustment of the pH-value

Dissolved oxygen:

>60% ASV (air saturation value)

Details on test conditions:

The test substance is a multi-component mixture (UVCB), which is designated as miscible with water. The test substance was homogenized by shaking the container before use. The test solution was prepared separately (differential loading) by directly adding test substance to test medium.
The necessary amount of test substance was weighed into a 20 L glass flask and filled to the mark with test water. The solution was then stirred in a closed glass flask for approximately 10 min using a magnetic stirrer. After stirring, the solution was left undisturbed for 24 hours to allow separation of the undissolved material. Undissolved material was observed after 24 hours as fluffy material and filaments on the bottom and the walls of the glass flask. The aqueous fraction was slightly turbid with faint visible fluffy or fibrous material (slight positive Tyndall effect).
Approx. 10 L of the aqueous fraction was drawn off for testing with a peristaltic pump (500 series, Watson Marlow GmbH, Germany) into a 10 L glass flask. Throughout the 24 hours intervals the aqueous phase of the tested concentration was colorless
but slightly turbid. Undissolved test substance was visible as slight fluffy or fibrous material. Fresh test solutions were prepared daily.

Test temperature: 22 °C
Test water: Non chlorinated charcoal-filtered drinking water from the municipal water works of the city of Frankenthal mixed with deionized water, hardness approx. 100 mg/L as CaCO3, pH generally 7.5 - 8.5
Test vessels: Approx. 10 Liter glass flask covered with a glass plug
Test volume: Approx. 10L
Loading: 7 Fish/ test vessel, max. 1 g fish/L
Randomization: The test organisms were introduced into the test vessels according to a randomization plan prepared by using a program of the laboratory data evaluation group of the testing facility.
Feeding: None
Photoperiod 16h light, 8h dark
Light intensity: 149–494 Lux (the light intensity is determined in regular intervals at the surface of the aquaria)
Oxygen content: >60% ASV (air saturation value)
Aeration: None
pH-value: No adjustment of the pH-value
Route of exposure: Static-renewal, exposure via test water.
Reason: In order to insure constant exposure conditions this
study was conducted as a static-renewal exposure. The renewal period was 24 hours.

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

192.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

Concentration control analysis was not performed because a sufficiently sensitive method for analyses in the required concentration range was not available. Since the test substance is a multi-component mixture (UVCB), the test solution is considered a water accommodated fraction (WAF). The term “loading rate” is advocated to express exposure to a WAF and is considered analogous to the nominal concentration. According to OECD 23 [Ref. 7], for tests with chemicals that cannot be quantified by analytical methods at the concentrations causing effects, the effect concentration can be expressed based on the nominal concentrations or loading rate (for mixtures).

There was no mortality in any replicate over the exposure period.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Executive summary:

Zebrafish (Danio rerio) were exposed for 96 h under semi-static conditions towards the test item according to OECD 203 criteria. The EC50 was determined to be 192 mg/L test item corresponding to 100 mg/L acitve substance. All validity criteria were fulfilled.