Rhatany, Krameria triandra, ext.

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:

13.08.2018 - 06.09.2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The study was performed in compliance with the Principle of Good Laboratory Practice, confirmed by Statement of GLP Compliance.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Principles of method if other than guideline:

The temperature for the study ranged from 22.5 to 23.5 °C and was therefore higher than stated in the guideline. As no immobility occurred in the blank control during the test and all validity criteria were met, this deviation was stated as uncritical.
The deviation was assessed and signed by the study director on 25. Sep. 2018.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

Rhatany, Krameria triandra, ext.

EC Number:

283-919-1

EC Name:

Rhatany, Krameria triandra, ext.

Cas Number:

84775-95-1

Molecular formula:

not available

IUPAC Name:

Extractives and their physically modified derivatives such as tinctures, concretes, absolutes, essential oils, oleoresins, terpenes, terpene-free fractions, distillates, residues, etc., obtained from Krameria triandra, Krameriaceae.

Test material form:

liquid: viscous

Specific details on test material used for the study:

Krameria triandra extract obtained from Rhatany root by hydroalcoholic extraction. Inhomogenous (warm up to 60 °C and stir) reddish brown viscous substance.Batch no. PES180014
CAS No. 84775-95-1
EINECS-No. 283-919-1
Molecular weight not applicable, UVCB
Purity not applicable, UVCB
Vapour pressure unknown
Stability H2O: 96h; EtOH: 96h; acetone: 96h; CH3CN: 96h; DMSO: 96h
Solubility H2O: unknown; EtOH: >1 g/L; acetone: >1 g/L; CH3CN: >1 g/L; DMSO: >1 g/L
Production date Jan. 2018
Expiry date Jan. 2019
Storage Fridge (2 - 8 °C)

Sampling and analysis

Analytical monitoring:

no

Details on sampling:

Storage:
The test item was stored in a closed vessel in the fridge (2 - 8 °C). Aliquots from the original vessel were made on 29. May 2018. They were also stored in the fridge at 2 - 8 °C.

Test solutions

Vehicle:

no

Details on test solutions:

The newborn daphnia, aged between 0 and 20.25 hours, were were sieved from the medium and immediately placed into a beaker containing dilution water. After the settling-in period, animals which showed no apparent damage were used for the test.

Dilution water (Daphnia medium) with the following specification was used (stated in SOP 11800201):
Parameter Concentration in mg/L
CaCl2*2H2O 293.80
MgSO4*7H2O 123.30
NaHCO3 64.80
KCl 5.80
Resulting hardness in mmol/L: 2.502
Resulting hardness in mg CaCO3/L: 250

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Species Daphnia magna
Authority STRAUS
Strain Berlin
Sex female
Age between 0 and 24 hours
Origin Umweltbundesamt Berlin
In-house breeding since 27. September 2007
Selection of the test system was made following the proposal of the guidelines.

Animal Husbandry:
Daphnia magna is bred in the LAUS GmbH throughout the year. The animals are kept for the use in toxicity tests. They multiply by parthenogenesis, thus being genetically identi-cal. The husbandry is performed similar to the method described in the OECD guideline, following SOP 115 002 01 („Zucht und Hälterung von Daphnia magna STRAUS“), version 12 from 02. Feb. 2015.
Vessels preserving glasses, nominal volume 2 L
Medium M4-Medium (recipe of ELENDT)
Food green algae (Desmodesmus subspicatus)
Medium renewal twice a week
Photo period 16/8 hours, using neon tubes
Temperature 20 +/- 2 °C

Study design

Test type:

static

Water media type:

other: M4-medium (recipe of ELENDT)

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

After 24 and 48 hours, the immobilised Daphnia were counted.

Test conditions

Hardness:

Resulting hardness in mmol/L: 2.502
Resulting hardness in mg CaCO3/L: 250

Test temperature:

22.5 – 23.5°C

pH:

7.9

Dissolved oxygen:

Nominal Concentration in mg/L O2-Concentration in mg/L
0 h 48 h
Blank control 8.9 8.6
4.6 8.8 8.5
10 8.8 8.5
22 8.7 8.5
46 8.5 8.4
100 8.6 8.5

Reference substance (positive control):

yes

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 21 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

element

Remarks:

carbon content

Basis for effect:

mobility

Details on results:

As no toxicity occurred during the test no statistical evaluation was conducted and the biological results are given in a range.
The biological results are presented in the following table:
Parameter Minimum measured C-content of test item (40.39%) Maximum measured C-content of test item (50.51%)
24h EC50 > 22.8 mg/L > 19.2 mg/L
48h EC50 > 22.8 mg/L > 19.2 mg/L
48h NOEC ≥ 22.8 mg/L ≥ 19.2 mg/L
48h LOEC > 22.8 mg/L > 19.2 mg/L

Any other information on results incl. tables

Immobility

Nominal Concentration in mg/L	Immobility 24 hours					Immobility 48 hours
	absolute				in %	absolute				in %
Blank control	0	0	0	0	0	0	0	0	0	0
4.6	0	0	0	0	0	0	0	0	0	0
10	0	0	0	0	0	0	0	0	0	0
22	0	0	0	0	0	0	0	0	0	0
46	0	0	0	0	0	0	0	0	0	0
100	0	0	0	0	0	0	0	0	0	0

As no toxicity occurred during the test no statistical evaluation was conducted and the biological results are given in a range.

Biological Results Test Item

The biological results are presented in the following table:

Parameter	Minimum measured C-content of test item (40.39%)	Maximum measured C-content of test item (50.51%)
24h EC50	> 22.8 mg/L	> 19.2 mg/L
48h EC50	> 22.8 mg/L	> 19.2 mg/L
48h NOEC	≥22.8 mg/L	≥ 19.2 mg/L
48h LOEC	> 22.8 mg/L	> 19.2 mg/L

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

Immobilisation in the controls may not exceed 10 %. Immobilisation in the controls was 0 %. The concentration of dissolved oxygen at the end must be at least 3 mg/L. The lowest concentration of dissolved oxygen at the end of the test was 8.4 mg/L.

Conclusions:

None of the tested concentrations showed immobilisation. None of the animals was immobilised in the blank control.

Executive summary:

This study was performed in order to evaluate the toxicity of the test item Krameria triandra extract obtained from Rhatany root by hydroalcoholic extractionin an acute immobilisation test under static conditions towards Daphnia magna. Daphnia magna STRAUS was chosen in the guideline as a typical species of zoo-plankton.

Young daphnia, aged less than 24 hours at the start of the test, were exposed to the test item for a period of 48 hours. Immobilisation was recorded at 24 hours and 48 hours and compared with control values. Not more than 10% of the control daphnia should show immobilisation or other signs of disease or stress, for example, discoloration or unusual behaviour such as trapping at surface of water. The results were analysed in order to calculate the EC₅₀at 24 h and at 48 h.

A positive control was tested in a separate study to assure that the test conditions are reliable.

One valid experiment was performed.

The study was performed using 5 concentrations ranging from 4.6 to 100 mg/L. For each test concentration and the blank control, 20 Daphnia were exposed to the test item for 48 hours in a static test system. After 24 and 48 hours, the immobilised Daphnia were counted.

None of the tested concentrations showed immobilisation. None of the animals was immobilised in the blank control.

Potassium dichromate K₂Cr₂O₇(CAS No. 7778-50-9) was used as positive controlin a current reference study to assure that the test conditions are reliable.

 

At the start and at the end of the test, the content of the test item in the test solutions was determined using measurement of dissolved organic carbon (DOC). The measured concentration based on DOC measurement was between 0 and 23% of the nominal concentration at the beginning and at the end of the test when considering the minimum measured carbon-content (C-content) of 42.39% of the test item. Therefore, the biological results were stated on the geometric means of the measured concentrations.

When considering the maximum measured C-content of 50.51% of the test item, the measured concentration based on DOC measurement was between 0 and 19% of the nominal concentration at the beginning and at the end of the test. Therefore, the biological results were stated on the geometric means of the measured concentrations.

No observations were made which might cause doubts concerning the validity of the study outcome. All validity criteria were met.

The result of the test is considered valid.