Fatty acids, C16-18, reaction products with diethanolamine

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-03-12 to 2013-03-15

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: The water accommodated fraction (WAF) was prepared with a nominal loading of the test item of 100 mg/L as specified below.
- Sampling method: The WAF and the control were analytically verified by LC-MS/MS at the start (0 h) and at the end of the exposure (48 h) as specified below.
At the start of the exposure (0 h), samples were taken immediately after preparation of the limit concentration and analysed.
At the end of the exposure (48 h), samples were taken directly from the test vessels, which contained the daphnids.
- Sample storage conditions before analysis: All original samples were stored at room temperature until sample preparation. Prepared samples were stored at room temperature until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The water accommodated fraction (WAF) was prepared with a nominal loading of the test item of 100 mg/L as specified below. 100 mg/L of the test item were weighed out and transferred with an appropriate amount of dilution water into a brown glass flask. This dispersion was shaken for 24 h with 20 rpm at room temperature. After a separation phase of 1.5 h, the aqueous phase or WAF was removed by siphoning from the approximate centre of the glass flask to give the 100 mg/L loading rate.
- Eluate: Dilution water
- Differential loading: None, limit test

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (STRAUS)
- Strain: Clone 5
- Source: Institut für Wasser-, Boden- Lufthygiene (WaBoLu), further own breeding
- Age at study initiation (mean and range, SD): 2 to 24 h
- Method of breeding: In 2-3 L glass vessels with appr. 1.8 L culture medium, at 20 +/- 2 °C, in an incubator, 16 h illumination, illumination strength max. 20 µExm-2 x s-1
- Feeding during test: No feeding

ACCLIMATION
- Acclimation period: 2 h
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: No feeding
- Health during acclimation (any mortality observed): Healthy, no mortality observed

DILUTION WATER:
Dilution water ISO test water, according to OECD 202, Annex 3:
Component Concentration [mg/L]
KCl 5.76
NaHCO3 64.8
CaCl2  2 H2O 294
MgSO4  7 H2O 123
pH-value: 7.8  0.2

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

No

Hardness:

Dilution water at test start: 242 (mg CaCO3/L)

Test temperature:

20.2 °C (Water)

pH:

Water Quality Parameters at the Start of the Exposure (0 h)
(measured in one additional replicate of the WAF and the control)
Nominal loading level of the
water accommodated fraction
(WAF)
[mg/L] pH-Value
100 7.74
Control 7.80

Water Quality Parameters at the End of the Exposure (48 h)
(measured in all replicates)
Nominal loading level of the
water accommodated fraction
(WAF)
[mg/L] pH-Values
Replicates
1 2 3 4
100 7.72 7.70 7.69 7.69
Control 7.79 7.78 7.77 7.74

Dissolved oxygen:

Water Quality Parameters at the Start of the Exposure (0 h)
(measured in one additional replicate of the WAF and the control)
Nominal loading level of the
water accommodated fraction
(WAF)
[mg/L] Dissolved
O2-Concentration
[mg/L]
100 8.28
Control 8.73

Water Quality Parameters at the End of the Exposure (48 h)
(measured in all replicates)
Nominal loading level of the
water accommodated fraction
(WAF)
[mg/L] Dissolved O2 –Concentration [mg/L]
Replicates
1 2 3 4
100 7.75 7.75 7.84 7.75
Control 8.46 8.39 8.25 8.22

Salinity:

Not measured, freshwater

Nominal and measured concentrations:

please refer to "Any other information on materials and methods"

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers, 50 mL
- Type (delete if not applicable): closed with parafilm
- Material, size, headspace, fill volume: Glass beakers (4 cm ID x 7 cm H), 50 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): static test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): None


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: see above
- Conductivity: Dilution water at test start:632 µS/cm
- Culture medium different from test medium: Elendt M4 acc. to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
- Intervals of water quality measurement: At test start and test end


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16:8 light:dark cycle
- Light intensity: Diffuse light, illumination strength max. 20 µE x m-2 x s-1


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Immobility was determined in all groups after 24 h and 48 h.

TEST CONCENTRATIONS
- Range finding study:
Immobilisation Rates in the non GLP preliminary Test
(n = 20, divided into 2 replicates with 10 daphnids each)
Nominal loading level of the
water accommodated fraction
(WAF)
[mg/L] IMMOBILISATION [%]
24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
100 0 0 0 0 0 0
Control 0 0 0 0 0 0

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Mortality of control: 0 %
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- EC50/LC50: 1.60 (CI 1.54 - 1.65) mg/L

Reported statistics and error estimates:

The effects of the WAF were deduced directly from the observed immobilisation rates.

Immobilisation Rates of the definitive Test after 24 and 48 hours of Exposure

              (n = 20, divided into 4 replicates with 5 daphnids each)

Nominal loading level of the water accommodated fraction (WAF) [mg/L]	IMMOBILISATION [%]
	24 h					48 h
	Replicates					Replicates
	1.	2.	3.	4.	MV	1.	2.	3.	4.	MV
100	0	0	0	0	0	0	0	0	0	0
Control	0	0	0	0	0	0	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

At the water accommodated fraction (WAF) with a nominal loading of 100 mg/L of the test item Fatty acids, C16-18-, reaction products with diethanolamine, no effects were observed on Daphnia magna.

Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the water accommodated fraction (WAF)* with a nominal loading of 100 mg/L of the test item Fatty acids, C16-18-, reaction products with diethanolamine were determined according to OECD 202 (2004). The limit test was conducted under static conditionsover a period of 48 hours from 2013-03-12 to 2013-03-15, with the definitive exposure phase from 2013-03-13 to 2013-03-15, at Dr. U. Noack-Laboratorien, Käthe-Paulus-Str. 1,31157 , .

The test item is solid and has a water solubility of ≤ 4 mg/L. In view of the difficulties associated with the evaluation of aquatic toxicity of poorly water soluble test items, a modification of the standard method for the preparation of aqueous media was performed. An approach endorsed by several important regulatory authorities in the EU and elsewhere (ECETOC 1996 and OECD 2000), is to expose the organisms to a Water Accommodated Fraction (WAF) of the test item in cases where the test item is a complex mixture and is poorly soluble in water and in the permitted solvents.
Using this approach, aqueous media were prepared by shaking the test item with dilution water at20 rpm for 24 hours at room temperature to ensure equilibration between the test item and the water phase. 100 mg/L of the test item were weighed out and transferred with an appropriate amount of dilution water into a brown glass flask. After the completion of shaking and following of a 1.5-h settlement period, the test item phase was separated by siphon and the test organisms exposed to the clear aqueous phase, the WAF. Twenty daphnids were exposed to the WAFand the control.

The test item Fatty acids, C16-18-, reaction products with diethanolaminewas analytically verified by LC-MS/MS at the start of the exposure (0 h) and at the end of the exposure (48 h) of the WAF with a nominal loading of 100 mg/L and the control.

The measured concentration of the test item in the WAF at the start of the exposure (0 h) was 0.208 mg/L. The measured concentration in the WAF at the end of the exposure (48 h) was 0.119 mg/L.

Exposures are expressed in terms of the original concentration of the test item Fatty acids, C16-18-, reaction products with diethanolamine in water at the start of the mixing period (loading rate), irrespective of the actual concentration of the test item in the WAF.

Water quality parameters (i.e. pH-value and dissolved oxygen concentration), measured at the start (0 h) and at the end of the exposure (48 h), were within the acceptable limits. The validity criteria of the test guidelines were fulfilled.

   EL-Values, NOEL and LOEL

(based on the nominal loading level of the water accommodated fraction)

Fatty acids, C16-18-, reaction products with diethanolamine
	Test duration [h]	Nominal loading level of the water accommodated fraction (WAF) [mg/L]	Confidence interval p = 95% [mg/L]
EL10 / EL50 / EL100	24	> 100	Not applicable
	48	> 100	Not applicable
NOEL	48	100	---
LOEL	48	> 100	---

Description of key information

At the water accommodated fraction (WAF) with a nominal loading of 100 mg/L of the test item Fatty acids, C16-18-, reaction products with diethanolamine, no effects were observed on Daphnia magna.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

In the acute immobilisation test withDaphnia magna(STRAUS), the effects of thewater accommodated fraction(WAF)* with a nominal loading of 100 mg/L of the test itemFattyacids, C16-18-, reactionproducts with diethanolamine were determined according to OECD 202 (2004).The limit test was conductedunder static conditionsover a period of 48 hours from2013-03-12 to 2013-03-15, with the definitive exposure phase from2013-03-13 to 2013-03-15, at Dr. U. Noack-Laboratorien, Käthe-Paulus-Str. 1,31157,.

The test item is solid and has a water solubility of≤4 mg/L.In view of the difficulties associated with the evaluation of aquatic toxicity of poorly water soluble test items, a modification of the standard method for the preparation of aqueous media was performed. An approach endorsed by several important regulatory authorities in the EU and elsewhere (ECETOC 1996 and OECD 2000), is to expose the organisms to a Water Accommodated Fraction (WAF) of the test item in cases where the test item is a complex mixture and is poorly soluble in water and in the permitted solvents.
Using this approach, aqueous media were prepared by shaking the test item with dilution water at20 rpmfor 24 hoursat room temperatureto ensure equilibration between the test item and the water phase.100 mg/L of the test item were weighed out and transferred with an appropriate amount of dilution water into a brown glass flask.After the completion of shaking and following of a 1.5-h settlement period, the test item phase was separated by siphonand the test organisms exposed to the clear aqueous phase, the WAF.Twenty daphnids were exposed to the WAFand the control.

The test itemFattyacids, C16-18-, reactionproducts with diethanolaminewas analytically verified by LC-MS/MSat the start of the exposure(0 h) and at the end of the exposure (48 h) of the WAF with a nominal loading of 100 mg/L and the control. Details for the analytical method are presented in part 13.

The measured concentration of the test item in the WAF at the start of the exposure (0 h) was 0.208 mg/L. The measured concentration in the WAF at the end of the exposure (48 h) was 0.119 mg/L. The analytical results are presented inTable4.

Exposures are expressed in terms of the original concentration of the test itemFatty acids, C16-18-, reactionproducts with diethanolaminein water at the start of the mixing period (loading rate), irrespective of the actual concentration of the test item in the WAF. The effect levels are presented inTable1.

Water quality parameters (i.e. pH-value and dissolved oxygen concentration), measured at the start (0 h) and at the end of the exposure (48 h), were within the acceptable limits. The validity criteria of the test guidelines were fulfilled.

 

 

 

At thewater accommodated fraction (WAF)* with a nominal loading of 100 mg/L
of the test itemFattyacids, C16-18-, reactionproducts with diethanolamine,

no effects were observed on Daphnia magna.

 

 

 

*) Water accommodated fraction (WAF): The aqueous medium containing only the fraction of a multi-component

  test item that is dissolved and / or present as a stable dispersion/emulsion under test conditions in the test

  medium, acc. to OECD Series, No. 23.

Table1:     EL-Values, NOEL and LOEL

(based on the nominal loading level of the water accommodated fraction)

Fattyacids, C16-18-, reaction products with diethanolamine
	Test duration   [h]	Nominal loading level of the water accommodated fraction (WAF) [mg/L]	Confidence interval p = 95%   [mg/L]
EL10/ EL50/ EL100	24	> 100	Not applicable
	48	> 100	Not applicable
NOEL	48	100	---
LOEL	48	> 100	---