Fatty acids, C18-unsatd., compds. with triethanolamine

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-12-08 to 2017-08-28

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD Guideline; EU Method; GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: OECD Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures"

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C18 unsatd., reaction products with triethanolamine

EC Number:

938-513-5

Cas Number:

1187872-27-0

IUPAC Name:

Fatty acids, C18 unsatd., reaction products with triethanolamine

Test material form:

liquid: viscous

Specific details on test material used for the study:

Name: Emulsogen TO
CAS No.: 10277-04-0
Certificate of Analysis Date: September 01, 2016
Expiry Date: September 01, 2018
Aggregate State at Room Temperature: Liquid
Colour: brown
Storage Conditions at Test Facility: At 20 ± 5 °C, in the dark

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Duplicate samples from the freshly prepared test medium of the only test concentration and the control were taken at the start of the test.
For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentration during the test period, duplicate samples from the test medium of the only test concentration and the control were collected at the end of the test (after 48 hours). Additional samples of the control were taken at test start and test end without any sample treatment.

Test solutions

Vehicle:

no

Details on test solutions:

Dosage of Test Item: The test item is not well soluble in test medium. To avoid physical effects of undissolved test item on the daphnids, no concentrations above the solubility limit of the test item in test water was tested.
Therefore, a supersaturated stock solution of the test item of nominal 100 mg test item/L was prepared by suspending 100.0 mg test item in 1000 ml test water. The test solution was carefully stirred for 48 hours in the dark to dissolve as much of the test item as possible. After stirring followed a 3 hours settling period to allow phase separation, the aqueous phase, i.e. the water accommodated fraction, was carefully extracted from below the water surface and used as the test medium.
The test media was prepared just before introduction of the daphnids (= start of the test).

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Daphnia magna (Straus), clone 5
Age at Test Start: From 3.25 to 21 hours old
Sex: Female
Origin: The daphnids introduced in the test were taken from ibacon's in house laboratory culture.
Breeding Conditions: The daphnids were bred in the laboratories of ibacon under similar temperature and light conditions as used in the test. The cultivation of the parental daphnids was performed in Elendt M4 medium (see 6.5). The test organisms were not first brood progeny. The daphnids in the stock culture were fed at least on all working days with green algae (Desmodesmus subspicatus) freshly grown in the laboratories of ibacon.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

Water Hardness: 2.5 mmol/L (= 250 mg/L) as Calcium Carbonate

Test temperature:

20 °C at test start;
20 °C at test end

pH:

8.2 at test start;
8.0 at test end; and thus the pH-value did not vary by more than 1.5 units

Dissolved oxygen:

8.8 to 9 mg/L at test start;
8.7 to 8.8 mg/L at test end

Nominal and measured concentrations:

Due to the low water solubility limit of the test item a water accommodated fraction (WAF) of a supersaturated stock solution of nominal 100 mg test item/L was tested. Thus, no concentrations above the solubility limit of the test item in the used test water was tested. Additionally, a control was tested in parallel.

Details on test conditions:

Test Environment: Controlled environment room
Measurement of pH, Dissolved
Oxygen and Water Temperature:
The water temperature, pH-values and dissolved oxygen concentrations were determined at test start and test end in each treatment group.
Water Temperature: 20 °C at test start;
20 °C at test end
pH-Values: 8.2 at test start;
8.0 at test end; and thus the pH-value did not vary by more than 1.5 units
Dissolved Oxygen Concentration: 8.8 to 9.0 mg/L at test start;
8.7 to 8.8 mg/L at test end
Light Regime: 16 h light : 8 h dark
Light Intensity: The light intensity was 420 to 590 lux (measured once during the test).

Reference substance (positive control):

yes

Remarks:

For the evaluation of the quality of the Daphnia clone and the consistency of the experimental conditions, the reference item potassium dichromate is tested at least twice a year to demonstrate satisfactory test conditions.

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

After 48 hours of exposure no immobilisation of the test animals was observed in the control and the only test item concentration with a loading rate of nominal 100 mg test item/L.

Results with reference substance (positive control):

In the most recent test with the reference item potassium dichromate performed in September 2016 (study code 88296220) the EC50 after 24 hours was determined to be 0.959 mg test item/L, indicating that the sensitivity of the daphnids was consistent with the level proposed by the OECD 202 guideline (EC50-24 h between 0.6 and 2.1 mg potassium dichromate/L).

Reported statistics and error estimates:

No statistical analysis was performed.
The EC50 could not be calculated due to the absence of toxicity of the test item and was determined to be higher than the highest tested nominal concentration. The NOEC and the LOEC were determined directly from the raw data.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

Control Immobilisation Rate was 0 % and furthermore no daphnid showed signs of disease or stress. Dissolved Oxygen Concentration was above or equal 8.7 mg O2/L in the control and test vessels at the end of the test.

Conclusions:

The toxic effect of the test item Emulsogen TO to Daphnia magna was assessed in a static limit test. The 48-hour NOEC was determined to be ≥ 100 mg test item/L. The 48-hour LOEC was determined to be > 100 mg test item/L and the 48-hour EC50 value was determined to be > 100 mg test item/L. All values refer to nominal concentrations, which was the loading rate of the only tested concentration.
The quantification of the test item Emulsogen TO in the test samples was performed using total organic carbon measurement (TOC).
The organic carbon analysis showed that the organic carbon concentrations did not decrease over the course of the test. The data was not used to calculate test item recoveries. The report is therefore based on nominal test concentrations.

Executive summary:

Title: Emulsogen TO: Acute Toxicity to Daphnia magna in a Static 48-hour Immobilisation Limit Test

Purpose:

The purpose of this study was to determine the influence of the test item Emulsogen TO on the mobility of Daphnia magna. For this purpose, young daphnids (< 24 hours old) were exposed in a static test for 48 hours to the highest concentration which can be dissolved in test water under defined conditions for 48 hours. The recorded effects were the mobility of the daphnids after 24 and 48 hours. The test method of application and the test system are recommended by the test guidelines and Daphnia magna is the recommended test species. The purpose of the analytical part of this study was to verify the concentration of the test item in the test medium.

Guidelines:

Commission Regulation (EC) No 440/2008, Annex, Part C, C.2.: "Daphnia sp. Acute Immobilisation Test", Official Journal of the European Union (EN), dated May 30, 2008

OECD Guideline for Testing of Chemicals No. 202: "Daphnia sp., Acute Immobilisation Test" adopted April 13, 2004

OECD Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures", December 15, 2000

SANCO/3029/99 rev.4 11/07/00: Residues: Guidance for generating and reporting methods of analysis in support of pre-registration data requirements for Annex II (part A; Section 4) and Annex III (part A; Section 5) of directive 91/414

Material and Methods

Test Item: Emulsogen TO; batch no.: DEB2 207167; purity: 99.5 % (w/w), according to certificate of analysis

Test Species: Daphnia magna, clone 5; 3.25 to 21 hours old

Source: The daphnids introduced in the test were taken from ibacon's in-house laboratory culture.

Test Design: This study encompassed two treatment groups (one test item concentration with a loading rate of nominal 100 mg test item/L, and one control) each containing 20 individuals. The mobility of the daphnids was determined in a static 48-hour test by visual observation after 24 and 48 hours.

Endpoints: Number of immobile organisms after 24 and 48 hours

Test Concentration: Due to the low water solubility limit of the test item a water accommodated fraction (WAF) of a supersaturated stock solution of nominal 100 mg test item/L was tested. Thus, no concentrations above the solubility limit of the test item in the used test water was tested. Additionally, a control was tested in parallel.

Test Conditions: Water temperature: 20 °C; pH value: 8.0 to 8.2; dissolved oxygen concentration: 8.7 to 9.0 mg/L; photoperiod: 16 h light - 8 h dark; light intensity: 420 to 590 lux; and thus were within the ranges requested by guideline OECD 202

Results

After 48 hours of exposure no immobilisation of the test animals was observed in the control and the only test item concentration with a loading rate of nominal 100 mg test item/L.

Conclusion: The toxic effect of the test item Emulsogen TO to Daphnia magna was assessed in a static limit test. The 48-hour NOEC was determined to be ≥ 100 mg test item/L. The 48-hour LOEC was determined to be > 100 mg test item/L and the 48-hour EC50 value was determined to be > 100 mg test item/L. All values refer to nominal concentrations, which was the loading rate of the only tested concentration. The quantification of the test item Emulsogen TO in the test samples was performed using total organic carbon measurement (TOC).The organic carbon analysis showed that the organic carbon concentrations did not decrease over the course of the test. The data was not used to calculate test item recoveries. The report is therefore based on nominal test concentrations.

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.