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

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:

February 17th 2021 to March 24th 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation 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:

Analysis of Test Concentration
10 mL of test samples from each replicates were drawn and mixed together for each group at 0 and 48h. The samples were divided into two portions. One portion (20 mL) was sent for test concentration analysis and the second portion (20 mL) was stored at -20 ± 5 ºC temperature till the study completion.

Test solutions

Vehicle:

yes

Details on test solutions:

Diluent Water
The diluent water was aerated, prior to use, for the test so that dissolved oxygen concentration has reached saturation. Reconstituted water was prepared by adding different chemicals to the distilled water.

Solubility Test
Solubility of test item was performed by dissolving 200 mg of test item in 1.0 mL of reconstituted
water and the test item was completely soluble at the test concentration of 200 mg/mL. Hence,
reconstituted water was used as vehicle for the study.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Source
Healthy Daphnia magna (showing no signs of stress such as presence of males and ephippia, delay in the production of the first brood, discolouration, etc. and having normal behavior) less than 24 h old [second progeny] cultured in the Laboratory of Ecotoxicology, Jai Research Foundation, were used for the study. The master culture of Daphnia magna was originally procured from the MicroBioTests Inc, Kleimoer 15, 9030 Mariakerke (Gent), Belgium. The sensitivity of the test system and reliability of the experimental technique has been previously validated with potassium dichromate as a positive control.

Gravid females were acclimatised to the test conditions for a minimum period of 48 h. During this
period, Daphnia were fed ad libitum with 1 mL of concentrated stock of live algal cells
(Pseudokirchneriella subcapitata). Freshly hatched daphnids belonging to the same stock (less than 24 h-old) were collected with the help of a micropipette and was used for the study. Feed was not provided during the exposure period.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

Daphnia culture water with a total hardness of 173.6 mg/L as CaCO3.

Test temperature:

The whole study was conducted in a specially designed low temperature water bath present to a water temperature of 20.0 °C (constant within ± 2 °C).

pH:

Between 7.21 and 7.83

Dissolved oxygen:

Between 8.17 and 8.95 mg/L

Nominal and measured concentrations:

The concentrations selected for the preliminary screening were 0.0 [control], 0.01,
0.1, 1.0, 10.0, and 100.0 mg finester 2009/L.

Based on the results of the preliminary range finding study, the main study was restricted to limit
study. The limit study was conducted with control (without finester 2009) and 100.0 mg finester
2009/L.

Details on test conditions:

Test Conditions
Temperature, pH, and dissolved oxygen content (using Multi portable meter) of reconstituted water were recorded at 0 and 48 h. Total hardness of reconstituted water was measured once during acclimatisation and test period using the titrimetric method (IS 3025, 1983). The pH did not vary by more than 1.5 units in any of the tests. Temperature, dissolved oxygen, and total hardness were within the guideline limits.

Each replicate comprising of five daphnids in 100 mL of test media.
Glass beaker of 600 mL capacity.
20 daphnids per group. Each group consisted of four replicates of 5 daphnids per replicate.

The whole study was conducted in a specially designed low temperature water bath present to a water temperature of 20.0 °C (constant within ± 2 °C). Uniform temperature in the water chamber of the water bath was obtained by maintaining a slow flow of water by a motor. The beakers containing the test solutions with daphnids were kept in the water bath. The beakers were held in position by special clamps provided within the chamber of the water bath.

A photoperiod of 16 h light and 8 h darkness cycle was maintained with an automatic timer attached to the water bath. Lighting was provided via fluorescent tubes attached on the lid of the low temperature water bath.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Immobility and Behavioural Observation
No immobilisation was observed at 100.0 mg finester 2009/L as well as in the control (without finester 2009) 24 and 48 h. Daphnids exposed to 100.0 mg finester 2009/L and control (without finester 2009 ) did not exhibit any abnormal behavioural symptom at 24 and 48 h.

Analysis of Test Concentrations
The stability of Finester 2009 in test media was performed, during the method validation (JRF Study N° 228-2-13-26945) and Finester 2009 in test media was stable up to 48 h (>80% of the nominal concentration). Test media was analysed for Finester 2009 active ingredient concentration and the stability at 0 and 48 h, during the main study. The active ingredient concentration of Finester 2009 in test media was within the acceptable limit (>80% of the nominal concentration).

Results with reference substance (positive control):

The sensitivity of the test system and reliability of the experimental technique was assessed using a positive control potassium dichromate.

The per cent immobilisation observed at 24 h was 0., 0, 20, 30, and ti0 at te.~1 concentration levels of 0.08, 0.15, 0.28, 0.53, and 1.00 mg potassium dichromate/L, respectively. No immobility was observed in any replicant of the control group.

At 24 h, lethargy was observed in G3 (0.15 mg/L), G4 (0.28 mg/L), GS (0.53 mg/L), and G6 (1.00 mg1L). No immobility or any behavioural symptom was observed in G 1 (control), and li2 (0.08 mg/[.) groups.

At 48 h, lethargy was observed in G3 (0.15 mg/L), G4 (0.28 mg,'L) and GS (0.53 mg,L). Complete
immobilisation was observed in all replicates of (,6 ( 1.00 mg,1). No immobility or behavioural symptom was observed in (G1 (control) and G2 (0.08 mg/L) groups.

Results are within the range of historical control data of JRF and the published value (EC50) of positive control.
This result proves the efficiency of test system and reliabilities of conditions.

Any other information on results incl. tables

Immobility Data at 0, 24 and 48 h Exposure Period.

Group	Test concentration (mg/L)	N° of Replicates	N° of Daphnia/group	Immobility N° and % at 0 hours	Immobility N° and % at  24 hours	Immobility N° and % at   48 hours
G1	0.0 (Control)	4	20	0	0	0
G2	100.0	4	20	0	0	0

Statistical Analysis
As the study was conducted as a limit test, probit analysis was not carried out.

Validity Criteria of the Study
Immobilisation in the control was 0% (not exceed 10%) at the termination, thus the validity criterion was met. Dissolved oxygen concentration was ≥ 8.17 mg/L in the control and test vessels at the end of the test. Thus the validity criterion was met. Daphnia in the control groups did not show immobilisation or other signs or disease or stress, e.g., discolouration or unusual behaviour such as trapping at the surface of water.

 

Euthanasia of Daphnia
At the end of the study, daphnia were euthanised by 2-step euthanasia procedure which was chemical induction of anesthesia (5% ethanol for 5 minutes), followed by heat induction in boiling water and disposed as per in-house SOP.

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Results of this study indicate that finester 2009 exposure caused no effect in the daphnia magna. The 48 h EC50, No Observed Effect Concentration (NOEC) and Lowest Observed Effect Concentration (LOEC) of finester 2009 were greater than 100.0 mg/L.

Executive summary:

The present study was conducted according to:
OECD, 2004 : The Organisation for Economic Co-operation and Development (OECD) Guidelines for the Testing of Chemicals, OECD 202, Daphnia sp., Acute Immobilisation Test, adopted by the Council on April 13, 2004.

Immobility and Behavioural Observation
No immobilisation was observed at 100.0 mg finester 2009/L as well as in the control (without finester 2009) 24 and 48 h. Daphnids exposed to 100.0 mg finester 2009/L and control (without finester 2009 ) did not exhibit any abnormal behavioural symptom at 24 and 48 h.
Test Conditions
Reconstituted water was used as test media. Water quality parameters, viz., temperature (ranged
between 20.0 and 20.9 °C), pH (ranged between 7.21 and 7.83), and dissolved oxygen (8.17 and 8.95 mg/L) were measured at 0 and 48 h. Hardness (173.6 mg/L as CaCO3) was measured at the beginning of the test. Light intensity (ranged between 1040 and 1360 Lux) was measured daily. All water quality parameters were found to be within the specified limits, as per the OECD guidelines

Results-

Analysis of Test Concentrations
The stability of finester 2009 in test media was performed, during the method validation (JRF Study N° 228-2-13-26945) and finester 2009 in test media was stable up to 48 h (>80% of the nominal concentration). Test media was analysed for finester 2009 active ingredient concentration and the stability at 0 and 48 h, during the main study. The active ingredient concentration of finester 2009 in test media was within the acceptable limit (>80% of the nominal concentration).

Statistical Analysis
As the study was conducted as a limit test, probit analysis was not carried out.

Positive Control
The sensitivity of the test system and reliability of the experimental technique was assessed using a positive control potassium dichromate.

Validity Criteria of the Study
Immobilisation in the control was 0% (not exceed 10%) at the termination, thus the validity criterion was met. Dissolved oxygen concentration was ≥ 8.17 mg/L in the control and test vessels at the end of the test. Thus the validity criterion was met. Daphnia in the control groups did not show immobilisation or other signs or disease or stress, e.g., discolouration or unusual behaviour such as trapping at the surface of water.