Chlorinated Fatty Acid Methyl Ester

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:

2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Methyl octadecanoate, chloro

Molecular formula:

C17H30Cl4COOCH3, C17H29Cl5COOCH3 , and C17H28Cl6COOCH3 (primary isomers based on 35-40% Cl by weight)

IUPAC Name:

Methyl octadecanoate, chloro

Constituent 2

Reference substance name:

Methyl hexadecanoate, chloro

Molecular formula:

C15H28Cl4COOCH3 and C15H27Cl5COOCH3 (primary isomers based on 35-40% Cl by weight)

IUPAC Name:

Methyl hexadecanoate, chloro

Constituent 3

Reference substance name:

Methyl dodecanoate, chloro

Molecular formula:

C19H31Cl5COOCH3 and C19H30Cl6COOCH3 (primary isomers based on 35-40% Cl by weight)

IUPAC Name:

Methyl dodecanoate, chloro

additive 1

Reference substance name:

Soybean oil, epoxidized

EC Number:

232-391-0

EC Name:

Soybean oil, epoxidized

Cas Number:

8013-07-8

Molecular formula:

C57H98O12

IUPAC Name:

Epoxidized Soybean Oil

Test material form:

liquid

Details on test material:

A complex combination of chlorinated fatty acid mtethyl esters that are predominantly C18 (~ 70 – 85%) and C16 (~ 15 - 30%) in length. The chlorination levels by weight are between 35-40%. The chlorination process is largely random, though chlorination will occur first by direct addition at unsaturated sites in the unsaturated methyl esters present in the starting material.

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

yes

Remarks:

acetone

Details on test solutions:

Before conducting the preliminary range finding study, Doverguard E35 was checked for solubility. Doverguard E35 formed emulsion in test media (OECD media) at the concentration of 200 mg/mL. Further, solubility was checked in acetone, dimethyl sulfoxide (DMSO) and dimethylformamide (DMF). In acetone, test item was observed soluble at the concentration level of 1000 mg/mL. This stock solution was serially diluted to prepare the concentration levels of 500, 250, 125, 62.5, and 31.25 mg/mL. To check the precipitation, these stock solutions were added at the concentration of 0.1 mL/L in test media. Precipitation was observed at 100, 50, 25, 12.5 mg/L. Visually clear solution was observed but very tiny droplets were observed under microscope at 6.25 mg/L. No precipitation was observed at 3.125 mg/L. A test solution of 6.25 mg/L for the study was selected as the upper limit.

Test organisms

Test organisms (species):

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

Details on test organisms:

Strain: ATCC 22662
Source: American Type Culture Collection, Manassas, USA

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Remarks on exposure duration:

samples were evaluated up to 96 hours

Test conditions

Test temperature:

22 to 23 °C

pH:

7.49 to 8.12

Nominal and measured concentrations:

0.0 (control), 0.0 (vehicle control), and 6.25 mg/L

Details on test conditions:

Initial Mean Number of Cells/mL: 5292

Reference substance (positive control):

no

Remarks:

solvent control

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

6.25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 2

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

> 6.25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Remarks on result:

not determinable

Remarks:

No effects observed.

Details on results:

There was no effect on algal growth by either the test material or the solvent (acetone) at the upper limit of 6.25 mg/L, which was the maximum concentration that could be tested just prior to the onset of significant test material precipitation.

Applicant's summary and conclusion

Conclusions:

C16 and C18 chlorinated fatty acid methyl esters do not inhibit the growth of alga (Pseudokirchneriella subcapitata) up to 6.25 mg/L, the maximum concentration that could be tested.The EC50, NOEC and LOEC value were greater than 6.25 mg/L

Executive summary:

This study was performed to assess effects of C16 and C18 chlorinated fatty acid methyl esters on the growth of the alga Pseudokirchneriella subcapitata,caused by exposure under static conditions.

Exponentially growing cultures of P. subcapitata were exposed to the nominal test concentrations of 0.0 (control), 0.0 (vehicle control), and 6.25 mg/L. The highest test concentration was selected based on the solubility, precipitation and result of range finding study. Algal cultures were assessed for their growth by visual cell count at 24, 48, 72, and 96 h.

There was no significant alteration observed in the algal growth (biomass), yield, and growth rate of the group exposed to the test material when compared with that of the vehicle control group. C16 and C18 chlorinated fatty acid methyl esters do not inhibit the growth of alga (Pseudokirchneriella subcapitata) up to 6.25 mg/L.The EC₅₀, NOEC and LOEC value were greater than 6.25 mg/L.