Octadecanoic acid, reaction products with diethylenetriamine, di-Me sulfate-quaternized

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:

03 December 2018 - 02 May 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

13 April 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Octadecanoic acid, reaction products with diethylenetriamine, di-Me sulfate-quaternized

EC Number:

291-707-5

EC Name:

Octadecanoic acid, reaction products with diethylenetriamine, di-Me sulfate-quaternized

Cas Number:

90459-62-4

Molecular formula:

C24H55N3O6S

IUPAC Name:

bis(2-aminoethyl)amine octadecanoic acid dimethyl sulfate

Test material form:

solid

Remarks:

paste

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all concentrations

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
The solubility of the test item in the test medium was found to be low in reconstituted water according to an internal method.
Therefore, individual WAFs were prepared by adding the appropriate amount of test item to dilution water (reconstituted water) at loading rates of 0.01, 0.1, 1.0, 10.0, 50.0 and 100.0 mg/L during the dose range finding study and at loading rates of 0.53, 0.95, 1.71, 3.09, 5.56 and 10.0 mg/L during the main study. Each mixing vessel was filled to achieve minimal headspace given the constraints of the vessels and closed with foil-covered stoppers. The mixture was stirred using a magnetic stirrer at 100 RPM for 6 hours (the duration of mixing was selected based on the WAF equilibration trial).
During the range finding study and the main study, samples were collected on day 0 (0 hour, fresh) and day 2 (48 hours, aged) and the TOC content was analyszed TOC content, using the TOC analyzer (TOC Analyzer/TNML; Make: Shimadzu; Model: LCPH E200 ROHS).
The mixtures were allowed to settle and equilibrate to test temperature (before removing the test solution). The aqueous portions of the WAFs were collected using a steri- pipette and transferred to the test vessels. The same method was employed for the control group using reconstituted water.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: Straus
- Age at study initiation (mean and range, SD): Young daphnids less than 24 hours old (Derived from healthy stock showing no signs of stress, such as high mortality rate, discolored daphnids, presence of male and ephippia, delay in the production of the first brood) were selected for the treatment.
- Source: In house maintained Daphnia magna procured from Environmental and Molecular Toxicology Division, Department of Zoology, Karnataka University, Karnataka, India.
- Feeding during test : No feeding during the test.


ACCLIMATION
- Acclimation period:
The 25 gravid female daphnids were acclimatiszed in reconstituted water for 48 hours prior to test (similar for range finding and main study). Daphnids were fed with live algal cells at the beginning and on Day 2 of acclimatiszation. Off springs produced on the first day are not used. Progenies produced on day two (less than 24 hours old) are transferred into the test containers and these daphnids were used for range finding as well as main study respectively.
During acclimatiszation of the dose range-finding test, the dissolved oxygen was between 7.43 andto 7.66 mg/L, the temperature betweenof 20.8 and 21.6°C, the pH betweenof 7.09 andto 7.21 and the hardness wasof 220 mg CaCO3/L.
During acclimatiszation of the main test, the dissolved oxygen was between 7.45 andto 7.86 mg/L, the temperature betweenof 20.3 and 20.9°C, the pH betweenof 7.32 andto 7.54 and the hardness wasof 220 mg CaCO3/L.
Prior to acclimatiszation, the glass jars were labeled for identification (study no., study code, no. of gravid females acclimatiszed, acclimatiszation start and acclimatiszation end dates).
- Type and amount of food: Daphnids were fed with live algal cells (Pseudokirchneriella subcapitata) of 2 mL per liter during the acclimatiszation.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

220 mg/L (as CaCO3)

Test temperature:

20.1 to 20.9°C

pH:

7.20 to 7.61

Dissolved oxygen:

7.45 to 7.86 mg/L

Nominal and measured concentrations:

nominal loading rates: control + 0.53, 0.95, 1.71, 3.09, 5.56 and 10.00 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beakers with a capacity of 100 mL
- Volume of solution: 100 mL
- Aeration: The dilution water was aerated prior to use for the test so that the dissolved oxygen concentration has reached saturation.
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4


OTHER TEST CONDITIONS
- Photoperiod: 16 h light/8 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Each test vessel was observed for immobilised daphnids at 24 and 48 hours after the beginning of the test. Those animals not able to swim within 15 seconds after gentle agitation of the test beaker were considered to be immobilised. In addition to immobility, any abnormal behavior or appearance was recorded.

RANGE-FINDING STUDY
- Test concentrations: 1.0, 10.0, 50.0 and 100 mg/L
- Results used to determine the conditions for the definitive study:
No clinical signs of toxicity were observed in the control group and at the nominal loading rates of 0.01 and 0.1 mg/L during the 48 hours observation period.
Clinical signs of daphnids localised on the bottom of the container and lethargy wereas observed during the 48 hours observation period at the nominal loading rate of 1.0 mg/L.
Clinical signs of daphnids localised on the bottom of the container and lethargy wereas observed during the 24 hours observation period at the nominal loading rates of 10.0, 50.0 and 100.0 mg/L.
The immobiliszation valuespercentages of 20, 100, 100 and 100 % were observed at the nominal loading rates of 1.0, 10.0, 50.0 and 100 mg/L, respectively, during the 48 hours observation period. No immobiliszation was observed in the control group and at the nominal loading rates of 0.01 and 0.1 mg/L during the 48 hours observation period.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

3.29 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% c.i. 2.87 - 3.78 mg/L

Details on results:

- Immobilisation of control: No clinical signs of toxicity and immobility were observed in the control group and at the nominal loading rate of 0.53 mg/L during the 48 hours observation period.

Clinical signs of daphnids localised on the bottom of the container and lethargy were observed during the 24 hours and 48 hours observation periods at nominal loading rates of 0.95, 1.71, 3.09 and 5.56 mg/L.
Clinical signs of daphnids localised on the bottom of the container and lethargy were observed during the 24 hours observation period at nominal loading rate of 10.0 mg/L.
Immobiliszation values of 10, 25, 40, 75 and 100 % were observed at the tested nominal loading rates of 0.95, 1.71, 3.09, 5.56 and 10.0 mg/L, respectively, during the 48 hours observation period.

Results with reference substance (positive control):

- Results with reference substance valid?
- Relevant effect levels: EC50 = 0.58 mg/L; within the acceptance range of the guideline (0.50 to 2.12 mg/L)

Reported statistics and error estimates:

Data are analysed by Finney’s probit analysis to calculate the slope of the curve and the EL50 with confidence limits. The 48 hours LL50 calculated was found to be 3.29 mg/L. The 95% confidence interval with lower limit of 2.87 mg/L and upper limit of 3.78 mg/L. The 48 hours NOEL and LOEL was found to be 1.71 and 3.09 mg/L loading rate, respectively.

Any other information on results incl. tables

Validity criteria

Immobilisation in the control group was 0 % at the termination of the test during main study (validity criterion: should not exceed 10 %).

Dissolved oxygen concentration at the end of the test was 6.95 mg/L (Validity criterion: should be ≥ 3 mg/L).

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Based on the above results of the experiment and under experimental conditions employed, it is concluded that the 48 hours acute median Effective Loading Rate (EL50) of Octadecanoic acid, reaction products with diethylenetriamine, di-Me sulfate-quaternized on Daphnia magna is 3.29 mg/L. The 95% confidence interval has a lower limit of 2.87 mg/L and an upper limit of 3.78 mg/L. The 48 hours NOEL and LOEL were found to be 1.71 mg/L and 3.09 mg/L loading rate, respectively.

Executive summary:

The test item Octadecanoic acid, reaction products with diethylenetriamine, di-Me sulfate-quaternized was evaluated for acute toxicity on Daphnia magna as per the OECD Guideline No. 202, “Daphnia sp., Acute Immobilisation Test”.

Daphnids were exposed to the test item by aqueous exposure (treatment) under static condition. Daphnids were observed at 24 and 48 hours for signs of toxicity and immobilisation.

 

Range Finding Study

No clinical signs of toxicity or immobilisation were observed in the control group and at the nominal loading rates of 0.01 and 0.1 mg/L during the 48 hours observation period.

Clinical signs of daphnids localised on the bottom of the container and lethargy were observed during the 48 hours observation period at the nominal loading rate of 1.0 mg/L.

Clinical signs of daphnids localised on the bottom of the container and lethargy were observed during the 24 hours observation period at the nominal loading rates of 10.0, 50.0 and 100.0 mg/L.

Immobilisation values of 20, 100, 100 and 100 % were observed at the nominal loading rates of 1.0, 10.0, 50.0 and 100 mg/L, respectively, during the 48 hours observation period. No immobilisation was observed in the control group and at the nominal loading rates of 0.01 and 0.1 mg/L during the 48 hours observation period.

 

Main Study

Based on the results of the range finding study, the main study was conducted at the nominal loading rates of 0.53, 0.95, 1.71, 3.09, 5.56 and 10.00 mg/L of the test item along with the control.

No clinical signs of toxicity and immobility were observed in the control group and at the nominal loading rate of 0.53 mg/L during the 48 hours observation period.

Clinical signs of daphnids localised on the bottom of the container and lethargy were observed during the 24 hours and 48 hours observation periods at the nominal loading rates of 0.95, 1.71, 3.09 and 5.56 mg/L.

Clinical signs of daphnids localised on the bottom of the container and lethargy were observed during the 24 hours observation period at the nominal loading rate of 10.0 mg/L.

Immobilisation values of 10, 25, 40, 75 and 100 % were observed at the tested nominal loading rates of 0.95, 1.71, 3.09, 5.56 and 10.0 mg/L, respectively, during the 48 hours observation period.

TOC-Analysis of the fresh test preparations at 0 hours showed measured TOC values to range from 0.8 to 4.2 mg TOC/L. TOC-Analysis of the aged test preparations at
48 hours showed measured TOC values to range from 0.8 to 4.3 mg TOC/L. The change in TOC content from 0 hours to 48 hours was smaller than 10% for each of the 6 different WAF loading rates.

As the concentration of the test substance has been satisfactorily maintained within 20% of the measured initial concentration throughout the test, the results are based on nominal values.

 

Conclusion

Based on the above results of the experiment and under experimental conditions employed, it is concluded that the 48 hours acute median Effective Loading Rate (EL50) of Octadecanoic acid, reaction products with diethylenetriamine, di-Me sulfate-quaternized on Daphnia magna is 3.29 mg/L. The 95% confidence interval has a lower limit of 2.87 mg/L and an upper limit of 3.78 mg/L. The 48 hours NOEL and LOEL were found to be 1.71 mg/L and 3.09 mg/L loading rate, respectively.