Octadecanoic acid, reaction products with triethylenetetramine, chloromethane-quaternized

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From July 01 to July 19, 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.5400 (Algal Toxicity, Tiers I and II) (January 2012)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

Octadecanoic acid, reaction products with triethylenetetramine, chloromethane-quaternized

EC Number:

291-716-4

EC Name:

Octadecanoic acid, reaction products with triethylenetetramine, chloromethane-quaternized

Cas Number:

90459-71-5

Molecular formula:

Not applicable - UVCB substance

IUPAC Name:

Octadecanoic acid, reaction products with triethylenetetramine, chloromethane-quaternized

Test material form:

solid

Specific details on test material used for the study:

Test item name: Octadecanoic acid, reaction products with triethylenetetramine, chloromethane-quaternized
Batch number: I1-0000196125
Manufacturing date: July 28, 2018
Expiry date : July 28, 2019
C.A.S. number: 90459-71-5
EEC number : 291-716-4
Molecular weight: UVCB substance

Sampling and analysis

Analytical monitoring:

no

Test solutions

Details on test solutions:

0.01, 0.03, 0.10, 0.30 and 1.00 mg test item/L.

Test organisms

Test organisms (species):

Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)

Details on test organisms:

The study was performed with the unicellular, fresh-water green algae Pseudokirchneriella subcapitata strain (formerly known as Selenastrum capricornutum), cultured in the laboratories of the Test Facility and originally purchased from the Institute of Plant Physiology of the University of Göttingen, Germany.
The algae were cultured in a climatic chamber at 24 °C ± 2 °C under continuous uniform illumination in the range 4440 - 8880 Lux in the spectral range 400-700 nm. The culture medium was the same of the test medium; the stock cultures were weekly transferred to fresh medium and maintained in continuous shaking to ensure the necessary amount of CO2 and to keep algae in suspension. Cultures containing deformed or abnormal cells were discarded. Only exponentially growing algal cultures have been used to start the test.

Study design

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test conditions

Test temperature:

23.9 – 24.1 °C.

pH:

7.93 – 7.96 at the test start
7.08 – 7.65 at the test end

Nominal and measured concentrations:

The biological results were referred to nominal concentrations of test item.
The main final test included five nominal test item concentration (constant factor 3.2) corresponding to 0.01, 0.03, 0.10, 0.30 and 1.00 mg test item/L.

The test medium was prepared as follows:
prior to the test start, a 10.0 mg test item/L stock solution was prepared by direct weighting of 0.0050 g of test item into 500 mL of algal medium. The obtained solution appeared clear with particles if test item. This solution was sonicated for about five minutes and after was stirring with magnetic stirrer for about 2 hours. After stirring the stock solution appeared clear and transparent. Start by this stock solution the other concentrations were prepared by dilution as reported in the following table.

Details on test conditions:

All the described procedures were conducted under a laminar flow bench to ensure the sterility of the algal cultures.
For the test concentrations and for the negative control, 100 mL capacity conical glass flasks capped with air permeable stoppers were used.
Three replicates for test concentrations and six for negative control were prepared. At test start, 50 mL of test solutions were poured into the test flasks and inoculated with algae to obtain a starting cell density of about 104 cells/mL.
The algal inoculum was taken from a pre-culture, started 3 days before the beginning of the test in order to ensure that the assay was performed with exponentially growing algal population. The cell density of the inoculum was around 106 cells/mL.

The test flasks were incubated in a climatic chamber, under continuous illumination and constant shaking. The flasks were randomly distributed under the light and were replaced every day during the test.
Light conditions:
continuous illumination; light intensity was ranged between 6340 and 6850 Lux, within the OECD recommended range (4440 - 8880 Lux).

Reference substance (positive control):

yes

Remarks:

3,5-Dichlorophenol pestanal

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

0.44 mg/L

Nominal / measured:

nominal

Basis for effect:

growth rate

Effect concentrations 2

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

0.09 mg/L

Nominal / measured:

nominal

Basis for effect:

cell number

Effect concentrations 3

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

0.03 mg/L

Nominal / measured:

nominal

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The toxicity of test item Octadecanoic acid, reaction products with triethylenetetramine, chloromethane-quaternized was tested on Pseudokirchneriella subcapitata.

The 72-hour EC10, EC20 and EC50, the NOEC and LOEC, calculated in terms of the nominal concentrations of test item, for the two end-points, are reported in the following table:


Endpoint 0 - 72 h EC10 (mg/L) 0 - 72 h EC20 (mg/L) 0 - 72 h EC50 (mg/L) 0 - 72 h NOEC (mg/L) 0 - 72 h LOEC (mg/L)
Growth rate 0.08 (0.05 – 0.16)* 0.14 (0.07 – 0.23)* 0.44 (0.21 – 0.58)* 0.03 0.10
Yield** 0.03 (0.00 – 0.05)* 0.04 (0.02 – 0.07)* 0.09 (0.06 – 0.20)* 0.03 0.10

(*) 95% confidence limits.