[No public or meaningful name is available]

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:

19 January 2022

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2022

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

No amendment was raised and one deviation occurred during the conduct of the study that WAF Trial was conducted on 31 January 2022 instead of 28 January 2022 and this deviation does not have any impact on outcome of the study.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Enzymatically produced steviol glycosides

IUPAC Name:

Enzymatically produced steviol glycosides

Test material form:

solid: particulate/powder

Test solutions

Vehicle:

yes

Remarks:

reconstituted water

Test organisms

Test organisms (species):

Daphnia magna

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

no

Test conditions

Hardness:

Dose Range Finding Study (Day 0): 205.3 mg CaCO3/L;
Limit Test (Day 0): 230.3 mg CaCO3/L

Test temperature:

Dose Range Finding Study: 20.1°C to 20.3°C;
Limit Test: 20.7°C to 20.9°C

pH:

Dose Range Finding Study: 7.36 to 7.40;
Limit Test: 7.26 to 7.28

Dissolved oxygen:

Dose Range Finding Study: 7.88 to 7.96 mg/L;
Limit Test: 7.63 to 7.84 mg/L

Reference substance (positive control):

yes

Remarks:

potassium dichromate (K2Cr2O7)

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

Dose Range Finding Study
Dose range finding study was conducted at the nominal loading rate of 0.01, 0.1, 1.0, 10.0, 50.0 and 100.0 mg/L. Daphnids were observed for the clinical signs of lethargy, erratic movement and localized on bottom of container. However, no such clinical signs and no immobility were observed up to highest loading rate of 100.0 mg/L and in control group during the 48 hours for exposure period.
During the dose range finding study, samples from all the nominal loading rates were collected at 0 hour (fresh) and at 48 hours (spent). The samples were analyzed for the determination of Total Organic Carbon (TOC) content.
Total organic carbon (TOC) content was found to be 0.4901, 0.5506, 1.390, 5.468, 24.85 and 48.14 ppm in fresh samples and 0.8361, 0.4862, 1.634, 4.813, 23.37 and 46.08 ppm in spent samples at the nominal loading rates of 0.01, 0.1, 1.0, 10.0, 50.0 and 100.0 mg/L respectively.
Limit Test
Based on results of the dose range finding study, limit test was performed as main study at the nominal loading rate of 100.0 mg/L along with control group.
Daphnids were observed for the clinical signs of lethargy, erratic movement and localized on bottom of container. However, no such clinical signs and no immobility were observed up to highest loading rate of 100 mg/L and in control group during the 48 hours for exposure period.
During the limit test, samples from nominal loading rate (100 mg/L) were collected at 0 hour (fresh) and at 48 hours (spent). The samples were analyzed for the determination of Total Organic Carbon (TOC) content.
Total organic carbon (TOC) content was found to be 44.29 and 46.39 ppm in fresh and spent sample respectively.

Results with reference substance (positive control):

EC50=1.03 mg/L (24h);
EC50=0.68 mg/L (48h)

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 Enzymatically produced steviol glycosides was >100 mg/L.