Amines, C12-14-branched alkyl, dodecylbenzenesulfonates (1:1)

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:

11 December 2017 to 09 Febraury 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: All test concentrations, including control
- Sampling method: Analytical samples were taken from all test item loading rates and control at test start and after 48 hours from aged solutions. For each sampling event, a retained sample was also taken.
- Sample storage conditions before analysis: All samples were stored deep frozen until they were transferred to the analytical laboratory.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Warwe Accommodated Fraction (WAF) - Stock solutions (S1-S4) were prepared by directly weighing 10.0, 5.00, 2.50 and 1.25 mg in 1000 mL test medium, respectively. These stock solutions were stirred in the dark for 48 h, at room temperature and let settle down for 1 h (based on OECD Series on Testing and Assessment No. 23). The solution was clear and transparent but not the entire test item went into solution, as flakes of substance were observed at the water surface in solutions S1 and S2. Additionally in stock solution S1, white crumbs were observed at the bottom of the flask. After settling no tyndall effect was observed and the necessary volumes for the test were withdrawn via a Teflon tube from the medium level of the stock solutions. The stock solutions were centrifuged at 5 000 rpm for 1 hour. After centrifugation no tyndall effect was observed. The test item loading rate V1 was made by diluting stock solution S4 to give the required test loading rate. ~ 50 mL of the prepared solutions were transferred to each test vessel
- Eluate: Elendt M4 medium
- Differential loading: 10.0, 5.00, 2.50, 1.25, 0.625 mg/L
- Controls: Yes - test medium only
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Not applicable
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): Not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): Not in utilised test WAF

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia
- Strain/clone: Clone V
- Source: Originally purchased in a healthy condition from the Federal Environment Agency in Berlin/Germany
- Age of parental stock (mean and range, SD): Not reported
- Feeding during test: No

ACCLIMATION
- Acclimation period: The animals are continuously bred in the laboratory
- Acclimation conditions (same as test or not): Yes
- Type and amount of food: The animals were fed with single cell green algae (Desmodesmus subspicatus, formerly Scenedesmus subspicatus).
- Feeding frequency: At least three times a week
- Health during acclimation (any mortality observed): No adverse health reported

QUARANTINE (wild caught)
- Duration: Not applicable
- Health/mortality: Not applicable

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: Not applicable

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Remarks on exposure duration:

Standard guideline exposure duration

Hardness:

13°dH (232 mg/L as CaCO3)

Test temperature:

19.0 – 21.4 °C

pH:

7.57 – 7.92

Dissolved oxygen:

≥ 8.5 mg/L

Salinity:

Not applicable

Conductivity:

Not reported

Nominal and measured concentrations:

Nominal Concentrations (WAF): 10.0, 5.00, 2.50, 1.25, 0.625 mg/L and control

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL glass beakers
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: Glass, 100 mL, 50 mL
- Aeration: No
- Type of flow-through (e.g. peristaltic or proportional diluter): Not applicable
- Renewal rate of test solution (frequency/flow rate): Not applicable
- 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): Not applicable
- Biomass loading rate: Not applicable

TEST MEDIUM / WATER PARAMETERS
Elendt M4 medium was used as test medium. At test start the pH-value of the control (untreated test medium) was 7.78, the dissolved oxygen concentration was 9.1 mg/L and the total hardness was 13°dH (232 mg/L as CaCO3).
- Culture medium different from test medium: Yes
- Intervals of water quality measurement: 24 h

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h light: 8 h dark
- Light intensity: Not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation

VEHICLE CONTROL PERFORMED: Not applicable

RANGE-FINDING STUDY
- Results used to determine the conditions for the definitive study: Yes

Reference substance (positive control):

yes

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

6.83 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95 % CL: 6.38 - 7.31)

Details on results:

After 24 hours of exposure no immobilisation was observed in the control and at test item loading rates of 0.625 mg/L and 2.50 mg/L. No immobilisation higher than the allowed control immobilisation was observed at 1.25 mg/L and 5.00 mg/L. 60 % immobilisation was observed at the highest test item loading rate of 10.0 mg/L.

After 48 hours of exposure no immobilisation was observed in the control. No immobilisation higher than the allowed control immobilisation was observed at 0.625, 1.25, 2.50 and 5.00 mg/L. At the highest test item loading rate of 10.0 mg/L all daphnids were immobile.

After 48 hours the mobile daphnids at the test item loading rate 5.00 mg/L appeared to be sluggish. No other symptoms of toxicity were observed during the course of the study.

Results with reference substance (positive control):

The results indicate an EC50 (24 h) of the reference item potassium dichromate between 1.00 and 2.00 mg/L. Since the results are in accordance with the requirements of the OECD guideline 202 and fall within the historical data generated with the reference item at the testing facility, the daphnids were suitable for the determination of the toxicological effects of the test item.

Reported statistics and error estimates:

The values for EL50 were determined by Weibull analysis using linear max. likelihood regression. The evaluation of data was performed by ToxRat Professional 3.2.1.
The NOELR was established based on the highest loading rate at which the immobilisation is not higher than the allowed control immobilisation (≤ 10 % immobilisation).

Table 1. Immobilisation after 24 h

Nominal Test Item Loading Rate (mg/L)	Control	0.625	1.25	2.50	5.00	10.0
Replicate 1	0	0	1	0	0	3
Replicate 2	0	0	0	0	0	2
Replicate 3	0	0	0	0	1	4
Replicate 4	0	0	0	0	0	3
Sum	0	0	1	0	1	12
%	0	0	5	0	5	60

Table 2. Immobilisation after 48 h  

Nominal Test Item Loading Rate (mg/L)	Control	0.625	1.25	2.50	5.00	10.0
Replicate 1	0	1	2	0	0	5
Replicate 2	0	0	0	1	0	5
Replicate 3	0	0	0	0	1	5
Replicate 4	0	0	0	0	0	5
Sum	0	1	2	1	1	20
%	0	5	10	5	5	100

Table 3. Determined concentration of C12-14 branched alkyl, dodecylbenzenesulfonates (1:1) via an indicator component

Test Item Nominal Concentration (mg/L)	Sampling Time (h)	Concentration (mg/L)	% of nominal concentration
Control	0 h fresh	not detected	-
Control	48 h aged	not detected	-
0.625	0 h fresh	0.644	103
0.625	48 h aged	0.698	112
1.25	0 h fresh	1.36	109
1.25	48 h aged	1.37	110
2.50	0 h fresh	2.28	91
2.50	48 h aged	2.50	100
5.00	0 h fresh	4.80	96
5.00	48 h aged	4.84	97
10.0	0 h fresh	9.43	94
10.0	48 h aged	11.3	113

Validity criteria fulfilled:

yes

Conclusions:

According to the results of the test, the EL50 (48 h) for immobilisation was determined to be 6.83 mg/L (nominal). The corresponding NOELR (48 h) was 5.00 mg/L (nominal).

Executive summary:

The study was conducted to determine the immobilisation effect of the test item to Daphnia magna, in accordance with OECD 202. Following a static GLP range-finding test, a static main test with nominal loading rates of 10.0, 5.00, 2.50, 1.25, 0.625 mg/L and control was performed. The test item is a complex substance (UVCB) composed of constituents with varying water solubility, therefore the test item is applied as water accommodated fraction (WAF). Temperature, pH-value and oxygen concentration of the test solutions measured after 0, 24 hours aged and 48 hours are reported. Hardness of the test medium (untreated control) was measured on the day of application. Analytical samples taken at 0 hours (initial value) and 48 hours from aged test solutions were analysed from control and all test item loading rates. The values for EL50 were determined by Weibull analysis using linear max. likelihood regression. The NOELR was established based on the highest loading rate at which the immobilisation is not higher than the allowed control immobilisation (≤ 10 % immobilisation). The total hardness (as CaCO3) of the test medium (untreated control) was determined to be 13°dH (232 mg/L CaCO3); the mean pH-value of the untreated control was determined to be 7.71 ± 0.12 (Std. Dev.), the mean temperature of the control and all test item loading rates was measured to be 19.7 ± 0.6 °C (Std. Dev.) and the mean oxygen concentration was determined to be 8.8 ± 0.2 mg/L (Std. Dev.). The initial measured content of active ingredient was between 91 and 109 % of nominal and the aged measured content was between 97 and 113 % of nominal. Since the content of active ingredient in the samples was between 80 and 120 % of nominal the toxicological endpoints were evaluated using nominal loading rates. According to the results of the test, the EL50 (48 h) was determined to be 6.83 mg/L (nominal). The corresponding NOELR (48 h) was 5.00 mg/L (nominal).

Description of key information

48h EL50 = 6.83 mg/L; OECD 202 (Daphnia magna): Kümmich, 2018

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

6.83 mg/L

Additional information

OECD 202 (2018): A study was conducted to determine the immobilisation effect of the test item to Daphnia magna, in accordance with OECD 202. Following a static GLP range-finding test, a static main test with nominal loading rates of 10.0, 5.00, 2.50, 1.25, 0.625 mg/L and control was performed. The test item is a complex substance (UVCB) composed of constituents with varying water solubility, therefore the test item is applied as water accommodated fraction (WAF). Temperature, pH-value and oxygen concentration of the test solutions measured after 0, 24 hours aged and 48 hours are reported. Hardness of the test medium (untreated control) was measured on the day of application. Analytical samples taken at 0 hours (initial value) and 48 hours from aged test solutions were analysed from control and all test item loading rates. The values for EL50 were determined by Weibull analysis using linear max. likelihood regression. The NOELR was established based on the highest loading rate at which the immobilisation is not higher than the allowed control immobilisation (≤ 10 % immobilisation). The total hardness (as CaCO3) of the test medium (untreated control) was determined to be 13°dH (232 mg/L CaCO3); the mean pH-value of the untreated control was determined to be 7.71 ± 0.12 (Std. Dev.), the mean temperature of the control and all test item loading rates was measured to be 19.7 ± 0.6 °C (Std. Dev.) and the mean oxygen concentration was determined to be 8.8 ± 0.2 mg/L (Std. Dev.). The initial measured content of active ingredient was between 91 and 109 % of nominal and the aged measured content was between 97 and 113 % of nominal. Since the content of active ingredient in the samples was between 80 and 120 % of nominal the toxicological endpoints were evaluated using nominal loading rates. According to the results of the test, the EL50 (48 h) was determined to be 6.83 mg/L (nominal). The corresponding NOELR (48 h) was 5.00 mg/L (nominal).