Glycine max. (L.) Merr. (Fabacee) aqueous alcoholic ext., concentrate by sequential extractions and filtration

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:

June 10 - 12, 2013

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: UNI EN ISO 6341:2004 “Determination of the inhibition of the mobility of Daphnia magna Straus (Cladocera, Crustacea)”.

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: OECD Series on Testing and Assessment No 23 - Guidance Document On Aquatic Toxicity Testing Of Difficult Substances And Mixtures ENV/JM/MONO(2000)6.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Glycine max. (L.) Merr. (Fabacee) aqueous alcoholic ext., concentrate by sequential extractions and filtration

EC Number:

938-980-5

IUPAC Name:

Glycine max. (L.) Merr. (Fabacee) aqueous alcoholic ext., concentrate by sequential extractions and filtration

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Test item identification : Soybean (Glycine max) dry purified extract
Batch number : 30432/M1
Internal number : 2912125-001

Sampling and analysis

Analytical monitoring:

no

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

The study was performed on the freshwater crustacean Daphnia magna Straus. A strain of this species was supplied by MicroBioTests Inc., Belgium.
the clone has been bred in the laboratories of the Test Facility under conditions identical to those of the test, with regard to temperature, light and water quality (see below), so that no acclimation was needed prior to testing.
It was daily fed with a fixed amount of the laboratory cultured green algae Pseudokirchneriella subcapitata (formerly Selenastrum capricornutum) and with a suspension of the yeast Saccharomyces cerevisiae.
At the test start the organisms used in the test were less than 24 hours old and were not first brood progeny.

For the test concentration and for the negative control, 20 daphnids were used divided into four replicates of five daphnids each.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

The daphnids were observed for immobility after 24 and 48 hours of exposure.

Test conditions

Hardness:

140 - 250 mg/L as CaCO3

Test temperature:

the incubation temperature was continuously monitored during the course of the study by a PT100 probe installed in the environmental test chamber. It was in the range 19.9 – 20.1 °C with a mean value of 20.0 °C and a standard deviation of 0.02 °C. The temperature range provided by OECD guideline is 20 ± 2 C. The temperature values during the test were within the provided range.

pH:

The pH of the test media after preparation was 6.86 for test item solution and 7.55 for negative control. The pH did not vary by more than 1.12 unit during the test period.

Dissolved oxygen:

The dissolved oxygen concentrations in the test media and the negative control were always higher than6.0 mg/L according to OECD Guideline N 202 that provides a minimum oxygen concentration of 3.0 mg/L.

Nominal and measured concentrations:

70.0 mg/L was founded to be the maximum soluble concentration in reconstituted water.

Details on test conditions:

The test medium was prepared as follows:
prior to the test start, 0.5 litre of a 70.0 mg test item /L stock solution was prepared by direct weighting into reconstituted water (0.0350 g of test item into 500 mL of reconstituted water). The obtained solution appeared cloudy and showed insoluble particles; therefore, it was sonicated for about 20 minutes and after was mixed by the mean of a magnetic stirrer for about 48 hours in dark conditions. After stirring the solution appeared lightly brown but completely solubilised.

Light:
a 16 hours light and 8 hours darkness daily photoperiod. The measured values were in the range 893 – 920 Lux, according to provided range (700-1000 Lux).

Reference substance (positive control):

yes

Remarks:

Potassium dichromate solution

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

IC100

Effect conc.:

> 70 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

70 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 3

Duration:

48 h

Dose descriptor:

IC50

Effect conc.:

> 70 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Linear Interpolation method

Details on results:

- Results with reference substance valid? YES
- No. of immobilized daphnia: 0/20
- EC50/LC50: > 70 mg/l

Results with reference substance (positive control):

- Results with reference substance valid? YES
- No. of immobilized daphnia: 0/20
- EC50/LC50: > 70 mg/l
- Other:

Any other information on results incl. tables

At the end of test period, in the negative control 0 % of immobilization was observed and no daphnid showed signs of disease or stress (e.g. to be trapped on the test water surface). These values comply with the validity criteria of the test, that provide a maximum immobilization or a maximum number of daphnids showing desease or stress signs of 10 % in the negative control medium at the end of the test.

The obtained experimental results allowed to calculate the IC0, IC100 and the IC50 values and the NOEC at 24 and 48 hours. The results, based on the nominal concentrations of test item, were as follows:

Time (h)	IC 0 (mg/L)	IC 100 (mg/L)	IC 50 (mg/L)	NOEC (mg/L)
24	70.0	> 70.0	> 70.0	70.0
48	70.0	> 70.0	> 70.0	70.0

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The IC50 of the substance based o the nominal concentration is > 70mg/L/48h.