Glycine, N-coco acyl derivs., sodium salts

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011-08-31 to 2011-09-21

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

study performed under GLP conditions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Glycine, N-coco acyl derivs., sodium salts

EC Number:

291-350-5

EC Name:

Glycine, N-coco acyl derivs., sodium salts

Cas Number:

90387-74-9

Molecular formula:

Molecular formula for this UVCB is not available.

IUPAC Name:

Glycine, N-coco acyl derivs., sodium salts

Test material form:

solid - liquid: aqueous solution

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 1.02 - 2.56 - 6.40 - 16.0 - 40.0 mg/L
- Sampling method: The C12- and C14-fraction 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 once every 7days.
For the old media additionally prepared replicates with test media but without daphnids and algae were prepared and incubated under test conditions until sampling.
- Sample storage conditions before analysis: All samples were stored at room temperature until start of analysis, if necessary.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution (40 mg/L test item was weighed out) was freshly prepared with dilution water.
- Eluate: Dilution water
- Differential loading: 1.02 - 2.56 - 6.40 - 16.0 - 40.0 mg/L
- Controls: 10 replicates of dilution water without test item

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna STRAUS
- Strain/clone: Clone 5
- Source: Own breeding (Origin: Institut fuer Wasser- Boden- und Lufthygiene)
- Age of parental stock (mean and range, SD): > 14 days
- Feeding during test
- Food type: Mix of Pseudokirchneriella subcapitata and Desmodesmus subspicatus
- Amount: 0.2 mg C/daphnia per day
- Frequency: daily


ACCLIMATION
- Acclimation period: 2 h in dilution water
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: During acclimation the daphnids were not fed
- Feeding frequency: Not applicable
- Health during acclimation (any mortality observed): Healthy

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES:
The parent animals were removed from the culture medium and the juveniles collected over a sieve and flushed into fresh medium

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Test conditions

Hardness:

142-171 mg/L as CaCO3

Test temperature:

20.2-21.7°C

pH:

7.58-8.53

Dissolved oxygen:

7.53-9.54

Nominal and measured concentrations:

0 (control), 1.02, 2.56, 6.40, 16.0, and 40.0 mg/L nominal

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beaker
- Type (delete if not applicable): closed with parafilm
- Material, size, headspace, fill volume: 100 mL glass beaker, filled with 50 mL test solution
- Aeration: No
- Renewal rate of test solution (frequency): 3 x per week
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control and pH-control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4, according to ELENDT (1990), modified to a total hardness of > 140 mg CaCO3/L.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h light : 8 h dark per day
- Light intensity: Max. 20 µE/m2 x s

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Adult mortality: daily
- Number of juveniles: daily
- Stillborn juveniles and aborted eggs: daily
- Appearance of first brood
- Intrinsic rate of natural increase: test end
- Growth (total length and dry weight): test end

RANGE-FINDING STUDY
- Test concentrations: range-finding study not performed

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

4 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Remarks:

and mortality

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Relevant effect levels: EC50 (24 h) = 1.70 mg/L (C.I.: 1.59 - 1.86 mg/L)

Reported statistics and error estimates:

The NOEC and the LOEC for the reproductive output and for the adult mortality were determined directly from the observation data. Significant deviations were determined in comparison to the control using statistical standard procedures as Normality Test, Equal Variance Test and Analysis of Variance.
For the determination of significant deviations of the reproductive output, the intrinsic rates of natural increase, the day of appearance of the first brood and the number of broods KRUSKAL-WALLIS, One Way Analysis of Variance on Ranks (p = 0.05) was used. The coefficients of variation around the mean number of living offspring produced per parent in the control and the test groups were evaluated.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The Lowest Observed Effect Concentration (LOEC) and the No Observed Effect Concentration (NOEC) for the reproductive output were determined directly from reduction of the reproductive output. EC10- and EC50- values for the reproductive output and the adult mortality could not be estimated because effects > 10 % were not observed.
As the concentrations in the old media (48 h) were below LOQ, the endpoints were assessed based on the nominal test item concentrations corrected with a factor of 2. Endpoint assessment based on mean measured concentrations is not possible, because the toxicity cannot be attributed to any single compound of the test item Hostapon SG containing different C-chains of the N-Cocoyl glycine sodium salt. Therefore, it was concluded that an evaluation on half of the nominal concentrations sufficiently describes a worst case scenario. Additionally the endpoints are given based on half of nominal concentrations of the active ingredient (a.i.) taking its content of 19.8 % into account.

NOECReproduction : 20 mg/L, corresponding to 3.96 mg a.i./L
LOECReproduction : > 20 mg/L, corresponding to > 3.96 mg a.i./L

NOECAdult Mortality : 20 mg/L, corresponding to 3.96 mg a.i./L
LOECAdult Mortality : > 20 mg/L, corresponding to > 3.96 mg a.i./L

Executive summary:

The long-term toxicity of the test item towards aquatic invertebrates (Daphnia magna) was investigated in a study conducted according to OECD Guideline 211 (Daphnia magna Reproduction Test). Ten daphnids each were exposed to nominal test item concentrations ranging from 1.02 to 40 mg/L for 21 d under semi-static conditions. In the frame of the test the following effect parameters were measured: adult mortality, number of juveniles, stillborn juveniles and aborted eggs, appearance of first brood, intrinsic rate of natural increase, and growth of the parental daphnids. The nominal test concentrations of the liquid test item Hostapon SG (19.8 % w/w active ingredient) were selected based on the results of an acute immobilization test.The test item concentrations of the C12- and C14 fraction were measured via LC/MS/MS.

The following results were obtained

- The average number of juveniles per parent in the control group was 131 after 21 days. The reproductive output was not statistically significant reduced in comparison to the control at any of the tested concentration levels. An EC10- or EC50-value for the reduction of the reproductive output could not be estimated, because no reduction of the reproductive output > 10 % was observed within the tested concentration range of 1.02 to 40.0 mg/L.

- The coefficient of variation of the number of living offspring produced per parent was 3 % in the control.

- The intrinsic rates of natural increase (IR) of the surviving parent animals accounting for generation time and number of offspring were used for calculation of population growth and maintenance. There was no statistically significant difference at any of the tested concentration levels of 1.02 to 40.0 mg/L when compared to the control.

- Related to the total number of produced juveniles (dead + alive) the percentage of dead juveniles and aborted eggs was ≤ 3 % in the tested concentration levels of 1.02 to 40.0 mg/L. In the control no dead juveniles or aborted eggs were observed.

- Five broods were observed during the test period at all surviving parental daphnids of the control and the concentration levels 1.02, 6.40, 16.0 and 40.0 mg/L and the control. At the concentration level of 2.56 mg/L 1 of 10 parental daphnids released four broods and 9 of 10 daphnids had five broods. There was no statistical significant difference at any of the tested concentration levels in comparison to the control. The first brood was released till day 10 and by all surviving parent animals of the control and the nominal test item concentrations of 1.02 to 40.0 mg/L.

- The test item did not induced biologically substantial adult mortality of > 20 % at any of the tested concentration levels of 1.02 to 40.0 mg/L after 21 days. All parental daphnids survived until the end of the study in the concentration levels of 1.02 to 16.0 mg/L and in the control. At the concentration level of 40.0 mg/L 1 of 10 parental daphnids died on day 20 and 9 of 10 parental daphnids survived until the end of the study. An EC10- or EC50for the adult mortality after 21 days could not be estimated because adult mortality > 10 % was not observed within the tested concentration range.

- The mean body lengths of the surviving parental daphnids at the nominal test item concentrations 1.02 to 40.0 mg/L were determined to be in the range of 5.19 to 5.60 mm per daphnid and 5.25 mm per daphnid at the control group, respectively. The mean dry body weight was determined to be in the range of 0.47 to 0.72 mg per daphnid in the test groups and 0.66 mg per daphnid in the control, respectively.

The concentrations of the C12- and C14-fraction of the test item Hostapon SG were analytically verified by LC-MS/MS of samples on days 0, 7, 14 (fresh media, 0 hours) and on days 2, 9, 16 (old media, 48 hours) at all concentration levels and the control. The recoveries in the fresh media (0 h) were in the range of 91 to 103 % of the nominal values for the C12-fraction and 87 to 106 % for the C14-fraction. Recoveries in the old media (48 h) were below the limit of quantification (LOQ) for both analytes.

As the concentrations in the old media (48 h) were below LOQ, the endpoints were assessed based on the nominal test item concentrations corrected with a factor of 2. Endpoint assessment based on mean measured concentrations is not possible, because the toxicity cannot be attributed to any single compound of the test item Hostapon SG containing different C-chains of the N-Cocoyl glycine sodium salt. Therefore, it was concluded that an evaluation on half of the nominal concentrations sufficiently describes a worst case scenario. Additionally the endpoints are given based on half of nominal concentrations of the active ingredient (a.i.) taking its content of 19.8 % into account.

NOECReproduction : 20 mg/L, corresponding to 3.96 mg a.i./L

LOECReproduction : > 20 mg/L, corresponding to > 3.96 mg a.i./L

NOECAdult Mortality : 20 mg/L, corresponding to 3.96 mg a.i./L

LOECAdult Mortality : > 20 mg/L, corresponding to > 3.96 mg a.i./L.