Reaction mass of oxybispropanediol and tetraglycerol and triglycerol, esterification products with lactic acid and lauric acid

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:

effects on growth of green algae

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experiment start date: December 23, 2020
Experiment completion date: February 19, 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 (Alga, Growth Inhibition Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of oxybispropanediol and tetraglycerol and triglycerol, esterification products with lactic acid and lauric acid

Molecular formula:

not applicable - UVCB substance

IUPAC Name:

Reaction mass of oxybispropanediol and tetraglycerol and triglycerol, esterification products with lactic acid and lauric acid

Specific details on test material used for the study:

Test item name: Finestar 2009
CAS Number: 2225876-48-0
Water solubility: soluble in water
Molecular formula: C12 H22 O8
Molecular weight: 294.30
Batch/Lot number: 3253082031
analyzed purity: 100% active
Saponification value: 45.87
Date of manufacture: August 27, 2020
Date of expiry: August 26, 2021
Appearance: Pale yellow viscous liquid
Storage temperature: Room temperature (15 to 30°C)
Storage condition: Keep away from light, moisture, and oxidizing - reducing chemicals
Storage container: Keep in original container

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

For the main study, six replicates for treatment and control were prepared. 10 mL of test samples from each replicate were drawn and mixed together for each group. The collected samples were centrifuged at 2000 rpm for 10 minutes in order to remove the algal cell. The representative samples were divided into two portions. One portion was sent for test concentration analysis at 0 h and 72 h. the second portion was stored at -20 +/- 5°C.

Test solutions

Vehicle:

no

Details on test solutions:

Solubility test
Test item was found to be soluble in algal medium (OECD medium) at the concentration of 200 mg/mL (Stock A).

Preliminary range-finding study
A preliminary study was conducted with the test concentrations of 0.0 (control), 0.01, 0.1, 1.0, 10.0 and 100 mg Finester 2009/L. two replicates were taken for each concentration of the test item and three replicates for the control group.

The percent inhibition of biomass was 0.67, 0.97, 1.38, 1.98 and 3.40 for the test concentrations of 0.01, 0.1, 1.0, 10.0 and 100 mg/L respectively.

The percent inhibition of growth rate was 0.00, 0.00, 0.00, 0.00 and 0.14 for the test concentrations of 0.01, 0.1, 1.0, 10.0 and 100 mg/L, respectively.

Test organisms

Test organisms (species):

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

Details on test organisms:

Strain number: ATCC 22662
Origin and source of culture: American Type Culture collection, Manassas, USA
Date of receipt: March 21, 2018
Date of the last sub-culture: February 08, 2021
Method of cultivation: static condition

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

72 h

Post exposure observation period:

All test and control cultures were inspected microscopically at 72 hours

Test conditions

Test temperature:

22 - 23°C

pH:

8.00 - 8.08

Salinity:

Not applicable as freshwater study

Nominal and measured concentrations:

Based on the results of the preliminary range-finding study, the main study was restricted to a limit study. the limit study was conducted with 0.0 (control) and 100 mg/L of test item.

Details on test conditions:

Pre-culture
The pre-culture was prepared 2 days prior to the commencement of the study by transferring 0.6 mL from the latest subculture to a final volume of 100 mL into a new culture vessel. The pre-culture was incubated under the same conditions as those required for the test and were used when growing exponentially. The temperature for pre-culture ranged between 22 and 23°C.

Initial cell concentration
Mean cell concentration was derived from 3 samples taken from the pre-culture. The cell concentration of the algal stock was determined as 750,000 cells/mL by manual counting using a haemocytometer and a microscope. Each replicate was inoculated with 0.9 mL of algal culture to obtain the required cell concentration of 5663 cells/mL.

Culturing apparatus
250 mL conical flask

Test culture maintenance
All pre-culture flasks were maintained between 22 and 23°C and within 7033 and 7490 lux (+/- 15% of the mean value) with a universal UV-free white, fluroescent lamp. Illumination and temperature were recorded daily using a light meter and a minimum-maximum thermometer, respectively. The cells in the cultured flasks were maintained in suspension by agitating the test flasks continuously at 100 repm, using an orbital shaker during the test period.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 2

Key result

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 3

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

growth rate

Details on results:

Algal cell count
The cell concentration in the control culture increased by a factor of 97.3 times within the three-day test period.

The coefficient of variation of average specific growth rate during the whole test period (0-3) in replicate control was 0.65%. The mean coefficient of variation for days 0-1, 1-2 and 2-3 in control was 10.27%.

Active ingredient concentration and stability analysis
The stability of the test material in test media was performed during the method validation, and was stable up to 72 hours (>80% of the nominal concentration). The test solution was aalysied for Finester and stability at 0 and 72 h during the limit study and were found to be within the acceptable limit (>80% of the nominal concentration).

Growth inhibition, EC 60 Values, NOEC and LOEC
There was no significant alteration observed in the algal growth (biomass), yield, and the growth rate of the test item exposed in the treatment group as compared to the control group. Finester 2009 did not inhibit the growth of algae up to 100 mg/mL.

Any other information on results incl. tables

Mean Values of Algal Biomass, Yield, specific Growth Rate

Group	Test concentration (mg/L)	Mean number of cells Count/mL at					Specific growth rate (0-3_
		0 h	24 h	48 h	72 h	Yield
G1	0	5663	25000	111250	550833	545170	1.53
G2	100		23333	110833	549583	543920	1.53

Validation criteria of study

The coefficient of variation of the average specific growth rates during the test period in the replicate control was 0.65%.

The cell concentration in the control cultures within the test period was increased exponentially by a factor of 97.3%.

The mean coefficient of variation for days 0-1, 1-2 and 2-3 in control cultures was 10.27%.

Thus, the validity criteria were met.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Results of the study indicate that Finestar 2009 exposure caused no effect in the algae growth. Based on concentration-effect values for the biomass inihibition, growth rate inhibition and yield inhibition for cells, it can be concluded that finestar 2009 exposure caused no inhibitory effect on the growth. EC50, NOEC and LOEC values were greater than 100.0 mg/L.

Executive summary:

This study was performed to assess effects on the growth of the alga Pseudokitchneriella subcapitata, caused by exposure under static conditions, of Finestar 2009.

Exponentially growing cultures of alga were exposed to the nominal test concentrations of 0.0 (Control) and 100 mg/L. Algal cultures were assessed for their growth by visual cell count at 24, 48 and 72 hours.

The stability of finester 2009 in test media was performed during the method validation and Finester 2009 in the test media was stable up to 72 hours (>80% of nominal concentration). The test solution was analysed for chlorantraniliprole and novaluron concentration and stability at 0 and 72 hrs during the limit study and were found to be within the acceptable limnit (>80% of nominal concentration).

there was no significant alteration observed in the algal growth (biomass), yield, and growth rate of the group exposed to finester 2009 when compared to the control group. finester 2009 does not inhibit the growth of alga (P. subcapitata) up to 100 mg/L. The EC50, NOEC and LOEC values were greater than 100 mg/L.