Extract from the seeds of Trigonella foenum-graecum (Fabaceae) obtained by extraction with polar solvents

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 September 2019 - 23 December 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identification/Synonym FENUGREEK EXTRACT
Substance type UVCB substance
EC No. 950-727-0
Recommended storage Ambient (18-20°C) and protected from temperatures below 4°C; dry (< 70% rh);
conditions to be stored under dark conditions

Analytical monitoring:

yes

Details on sampling:

Samples for analysis were taken from the control and all test solutions (from a replicate of each test concentration without algae dedicated exclusively to chemical analyses) at the start and the end of the test. Concentration of dissolved organic material was checked by analysis of Total Organic Carbon (TOC) in the control medium and the WAFs. TOC analysis was not performed in compliance with the OECD GLP principles but was adapted to fit the specific parameters of the test item, in accordance with ISO 17025.

Vehicle:

no

Details on test solutions:

Nominal loading rates 1.0, 3.2, 10.0, 32.0 and 100.0 mg.L-1

Test organisms (species):

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

Details on test organisms:

Pseudokirchneriella subcapitata, CCAP 278/4

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test temperature:

23°C +- 2°C

pH:

8.1 +- 0.2

Nominal and measured concentrations:

1.0, 3.2, 10.0, 32.0 and 100.0 mg.L-1

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL, all-glass closed flasks with ground glass stopper, completely filled with test solution with minimum headspace.
- Type (delete if not applicable): closed
- Initial cells density: 5.10^3 cells.mL-1

GROWTH MEDIUM
- Standard medium used: yes

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: yes
- Photoperiod: Continuous illumination
- Light intensity and quality:4,440-8,880 lux

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3.16

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

72 h

Dose descriptor:

EL50

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

growth rate

Key result

Duration:

72 h

Dose descriptor:

EL50

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

biomass

Key result

Duration:

72 h

Dose descriptor:

NOELR

Effect conc.:

ca. 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

growth rate

Key result

Duration:

72 h

Dose descriptor:

NOELR

Effect conc.:

ca. 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

biomass

Details on results:

In any case, based on biological results, the 72-hour ErL50 and EyL50 values were estimated to be ≥ 100.0 mg.L-1. The NOErLR and NOEyLR value determined by ToxRat was 1.0 mg.L-1.

Results with reference substance (positive control):

On August 26, 2019 (KA19-002; most recent test), the 72h-EC50 was 0.719 mg.L-1 for the parameter growth rate. Hence, the sensitivity of this batch of Pseudokirchneriella subcapitata was consistent with the level proposed by the ISO 8692 (6) (expected 72h-ErC50: 0.65 mg.L-1 to 1.73 mg.L-1).

Reported statistics and error estimates:

Shapiro Wilk`s test on normal distribution
Levene´s Test on Variance Homogeneity
Williams Multiple Sequential t-test Procedure

Validity criteria fulfilled:

yes

Conclusions:

The toxic effect of test item to the unicellular algal species Pseudokirchneriella subcapitata was investigated in a closed static test using Water Accommodated Fractions. Under the experimental conditions and based on nominal loading rates, the 72-hour EL50 for the parameters growth rate and yield were therefore considered to be higher than 100.0 mg.L-1. The 72-hour NOELR value for both growth rate and yield was 1.0 mg.L-1.

Description of key information

ErLx and EyLx ≥ 100.0 mg.L-1.

NOErLR and NOEyLR = 1.0 mg.L-1.

Key value for chemical safety assessment

EC50 for freshwater algae:

100 mg/L

EC10 or NOEC for freshwater algae:

1 mg/L