fatty acid chloride, C12, reaction product with sodium glutamate and sodium hydroxide

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

Justification is provided in the separate statement.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

6.5 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

act. ingr.

Executive summary:

No study is available on the test item. Two studies from supporting substance (structural analogue or surrogate) are available.

 

Based on the nominal concentrations of the test item Sodium N-cocoyl glycinate the 48-h EC50 for Daphnia magna was estimated to be 33.0 mg/L (95 % confidence limits: 30.2 - 37.2 mg/L) or 6.5 mg/L based on active ingredient.

The NOEC after 48 h was determined to be 10.2 mg/L as the highest tested concentration level with no biologically significant effect. The LOEC after 48 h was determined to be 25.6 mg/L as the lowest tested concentration level with a biologically significant effect.

 

In the acute immobilization test with Daphnia magna the effects of the test item Sodium N-octoyl glutamate were determined according to the Guidelien Din 38 412 Part 1 and Part 11. Based on the nominal concentration of the test item the m48 h EC50 for Daphnia magna was estimated to be 18 mg/L based on test material.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

05.1994

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: DIN 38412, Part 1 and Part 11

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Daphnia magna

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Details on test conditions:

1 - 5 - 10 - 20 - 50 - 100 mg/L

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Validity criteria fulfilled:

not specified

Conclusions:

EC50 (48h) = 18 mg/L (nominal) for Daphnia magna (DIN 38 412)

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2011-06-15 to 2011-06-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed under GLP according to OECD guideline

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 C12- and C14 fractions of the test item Hostapon SG were analytically verified by LC-MS/MS in the fresh media (0 h) and old media (48 h) of all concentration levels and the control.
- Sampling method: For the old media, separate replicates without test organisms were prepared and incubated under test conditions.
- Sample storage conditions before analysis: All samples were stored at room temperature until start of analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
Hostapon SG is a long chain carboxylic acid forming Calcium salt with very low solubility removing most of the test item from the aqueous solution. The recovery of the test item at the end of the test (48 h) is therefore very low and the guideline quality criteria are no longer fulfilled. To overcome this testing issue Ca2+ ions are complexed by Na2EDTA. Na2EDTA has a very low acute toxicity to Daphnia magna which is confirmed by a Na2EDTA control in the preliminary range finding test. As no effects were expected in the ISO test water containing 800 mg/L of Na2EDTA, no additional control without Na2EDTA was necessary.
- Method: The stock solution (160 mg/L test item was weighed out) was freshly prepared with dilution water which contained Na2EDTA. Disppersion treatment: Agitation. For preparation of the further test concentrations of 1.64 to 64.0 mg/L aliquots of the stock solution were taken while stirring with approx. 600 rpm.
- Eluate: ISO Test water (following OECD 202, Annex 3) as described below. By addition of 800 mg/L of Ethylenediaminetetraacetic acid disodium salt dihydrate (Na2EDTA * 2 H2O) the Ca2+ ions were completely complexed and therefore most of the test item was still in the aqueous solution.
The pH of the dilution water was adjusted to pH 7.0 ± 0.2 by addition of 1 M NaOH.
- Differential loading: 6 test item concentrations were tested, set up in a geometrical series with a dilution factor of 2.5:
1.64 - 4.10 - 10.2 - 25.6 - 64.0 - 160 mg/L, prepared with dilution water. 20 g of the test solution each was weighed out into the test vessels. The daphnids were inserted with a small amount of dilution water by using a pipette.
- Controls: Na2EDTA control: Dilution water containing 800 mg/L Na2EDTA without test item was tested under the same conditions as the test groups. As no Na2EDTA related effects were observed in the preliminary test no separate control without Na2EDTA was included.

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 glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20+/- 2 °C, in an incubator, 16 h illumination, illumination strength max. 20 µE*m-2*s-1. Culture medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L. Culture Feeding: At least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata, Desmodesmus subspicatus and Chlorella vulgaris, with an algae cell density of > 106 cells/mL.
- 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

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

No

Hardness:

Dilution water: 21.4 (mg CaCO3/L), This corresponds to 14 mg Ca2+/L. Ca2+ ions were complexed by Na2EDTA * 2 H2O.

Test temperature:

19 - 20 °C.

pH:

Water Quality Parameters at Test Start (0 h, Fresh Media)
(measured in one replicate per loading rate, pH-control and control)
Hostapon SG
Nominal Test Item Concentration
[mg/L] pH-Value
160 6.95
64.0 6.86
25.6 6.83
10.2 6.85
4.10 6.91
1.64 6.91
Na2EDTA Control 7.08

Water Quality Parameters at Test End (48 h, Old Media) (measured in all replicates)
Hostapon SG
Nominal Test Item Concentration
[mg/L] pH-Values
Replicates
1 2 3 4
160 6.92 6.91 6.89 6.86
64.0 6.93 6.89 6.92 6.88
25.6 6.81 6.80 6.81 6.87
10.2 6.80 6.83 6.85 6.85
4.10 6.81 6.81 6.81 6.84
1.64 6.96 6.95 6.95 6.93
Na2EDTA Control 7.08 7.04 6.99 6.98

Dissolved oxygen:

Water Quality Parameters at Test Start (0 h, Fresh Media) (measured in one replicate per loading rate, pH-control and control)
Hostapon SG
Nominal Test Itemn Concentration [mg/L]
Dissolved O2-Concentration[mg/L]
160 8.91
64.0 8.92
25.6 8.92
10.2 8.91
4.10 8.95
1.64 8.96
Na2EDTA Control 9.10
Water Quality Parameters at Test End (48 h, Old Media)
(measured in all replicates)
Hostapon SG
Nominal
Test Item Concentration [mg/L] Dissolved O2 -Concentration [mg/L]
Replicates
1 2 3 4
160 8.83 8.80 8.88 8.86
64.0 8.74 8.81 8.84
25.6 8.82 8.84 8.81 8.86
10.2 8.87 8.84 8.86 8.88
4.10 8.88 8.86 8.87 8.91
1.64 8.87 8.90 8.93 8.90
Na2EDTA Control 8.88 8.71 8.85 8.86

Salinity:

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
- 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): One application at test start, 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: ISO Test water (following OECD 202, Annex 3) as described below. By addition of 800 mg/L of Ethylenediaminetetraacetic acid disodium salt dihydrate (Na2EDTA * 2 H2O) the Ca2+ ions were completely complexed and therefore most of the test item was still in the aqueous solution. The pH of the dilution water was adjusted to pH 7.0 ± 0.2 by addition of 1 M NaOH.
- Conductivity: Dilution water at test start: 886 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) :
IImmobility was determined in all groups after 24 and 48 h. An animal was considered to be immobile, if it was not able to swim within 15 sec after gentle agitation of the test vessel.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.64 - 4.10 - 10.2 - 25.6 - 64.0 - 160 mg/L (dilution factor 2.5)
- Range finding study
Immobilization Rates of the Preliminary Test after 24 and 48 h of Exposure
(n = 20, divided into 2 replicates with 10 daphnids each)
Hostapon SG
IMMOBILIZATION [%]
Nominal Test Item Concentration
[mg/L] 24 h 48 h
Replicates Replicates
1. 2. MV 1. 2. MV
1001) 2) 20 50 35 80 90 85
10.01) 2) 10 0 5 30 0 15
1.001) 2) 0 0 0 20 0 10
Na2ETA Control 1) 2) 0 0 0 0 0 0
Control 3) 0 0 0 0 0 0
1) = Prepared with ISO test water (acc. to OECD 202), containing 800 mg/L of Ethylenediaminetetraacetic acid
disodium salt dihydrate (Na2EDTA * 2 H2O)
2) = The pH was adjusted with 1 M NaOH from pH = 3.9 to pH = 7
3) = Pure ISO test water acc. to OECD 202

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

6.5 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: 6.0-7.4 (95% CI)

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

3.4 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: 2.8-4.0 (95% CI)

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

5.1 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: n.a.

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

2 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: n.a.

Details on results:

- Behavioural abnormalities: None
- Mortality of control: 0%
- Other adverse effects control: None
- Abnormal responses: None
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- EC50: 1.44 mg/L (CI 1.31 - 1.55 mg/L)

Reported statistics and error estimates:

EC100-values after 24 and 48 h and the NOEC and LOEC after 48 h were estimated directly from the observation data, without mathematical calculation. EC10- and EC50-values were calculated by sigmoidal dose-response regression. The confidence intervals were calculated from the best-fit values, the standard error and the t-distribution with the software GraphPad prism. The concentration effect relationships are shown graphically.

Immobilization Rates of the Definitive Test after 24 and 48 h of Exposure

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

Hostapon SG	IMMOBILIZATION [%]
Nominal Test Item Concentration [mg/L]	24 h					48 h
	Replicates					Replicates
	1.	2.	3.	4.	MV	1.	2.	3.	4.	MV
160	80	80	80	100	85	100	100	100	100	100
64.0	20	0	40	60	30	80	80	100	100	90
25.6	0	0	0	0	0	20	20	40	40	30
10.2	0	0	0	0	0	0	0	0	0	0
4.10	0	0	0	0	0	0	0	0	0	0
1.64	0	0	0	0	0	0	0	0	0	0
Na2EDTA Control	0	0	0	0	0	0	0	0	0	0

MV    = Mean value

Validity criteria fulfilled:

yes

Conclusions:

Based on the nominal concentrations of the test item Hostapon SG the 48-h EC50 for Daphnia magna was estimated to be 33.0 mg/L (95 % confidence limits: 30.2 - 37.2 mg/L) or 6.5 mg/L based on active ingredient.
The NOEC after 48 h was determined to be 10.2 mg/L as the highest tested concentration level with no biologically significant effect. The LOEC after 48 h was determined to be 25.6 mg/L as the lowest tested concentration level with a biologically significant effect.

Executive summary:

In the acute immobilization test with Daphnia magna (STRAUS) the effects of the test item Hostapon SG (batch number: ESD 0006364) were determined according to OECD 202 (2004) from 2011-06-15 to 2011-06-17 at Dr.U.Noack-Laboratorien, Käthe-Paulus-Str.1, 31157 Sarstedt, Germany.

Hostapon SG is a long chain carboxylic acid forming Calcium salt with very low solubility removing most of the test item from the aqueous solution. The recovery of the test item at the end of the test (48 h) is therefore very low and the guideline quality criteria are no longer fulfilled. To overcome this testing issue Ca²⁺ ions were complexed by Na₂EDTA. Na₂EDTA has a very low acute toxicity to Daphnia magna which is confirmed by a Na₂EDTA control in the preliminary range finding test. As no effects were expected in the ISO test water containing 800 mg/L of Na₂EDTA, no additional control without Na₂EDTA was necessary.

The study was conducted under static conditions over a period of 48 h with 6concentrations of the test item Hostapon SG ranging from 1.64 to 160 mg/Lset up in a geometric series with a dilution factor of 2.5. Twenty daphnids were exposed to each concentration level and the control.

The concentrations of the C12- and C14-fraction of the test item Hostapon SG were analytically verified by LC-MS/MS in the fresh (0 h) and old media (48 h) of all concentration levels and the control.
The recoveries of the C12-fraction were in the range of 94 to 99 % of the nominal values in the fresh media (0 h) and 83 to 109 % in the old media (0 h). Recoveries of the C14-fraction were in the range of 89 to 98 % of the nominal values in the fresh media (0 h) and 66 to 94 % in the old media (0 h).

All effect values are based on the nominal concentrations of the test item Hostapon SG as it contains different C-chains of the N-Cocoyl glycine sodium salt, and toxicity cannot be attributed to any single compound.  

The water quality parameters (pH-value and dissolved oxygen concentration), measured at 0 and 48 h, were determined to be within the acceptable limits. The validity criteria of the test guideline were fulfilled.

EC₁₀-, EC₅₀- (with Confidence Interval), EC₁₀₀ - Values, NOEC and LOEC

              (based on the nominal concentrations of the test item Hostapon SG)

	Test Duration [h]	Nominal Concentration of the Test Item Hostapon SG [mg/L]	Confidence Interval p = 95% [mg/L]
EC10	24	40.7	31.4 - 53.5
	48	17.3	14.3 - 20.4
EC50	24	86.7	74.6 - 103
	48	33.0	30.2 - 37.2
EC100	24	> 160	---
	48	160	---
NOEC	48	10.2	---
LOEC	48	25.6	---

Results on acitive ingredient can be seen in the result table above.

Description of key information

EC50 (48h) = 6.5 mg/L (measured) for Daphnia magna (OECD 202, read across)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

6.5 mg/L

Additional information

No study is available on the test item. Two studies from supporting substance (structural analogue or surrogate) are available.

 

Based on the nominal concentrations of the test item Sodium N-cocoyl glycinate the 48-h EC50 for Daphnia magna was estimated to be 33.0 mg/L (95 % confidence limits: 30.2 - 37.2 mg/L) or 6.5 mg/L based on active ingredient.

The NOEC after 48 h was determined to be 10.2 mg/L as the highest tested concentration level with no biologically significant effect. The LOEC after 48 h was determined to be 25.6 mg/L as the lowest tested concentration level with a biologically significant effect.

 

In the acute immobilization test with Daphnia magna the effects of the test item Sodium N-octoyl glutamate were determined according to the Guidelien Din 38 412 Part 1 and Part 11. Based on the nominal concentration of the test item the m48 h EC50 for Daphnia magna was estimated to be 18 mg/L based on test material.