Veratrole

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:

From12 June 2012 to 01 August 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study, OECD 202 compliant.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

11 April 2011

Test material

Test material information

Constituent 1

Reference substance name:

Veratrole

EC Number:

202-045-3

EC Name:

Veratrole

Cas Number:

91-16-7

Molecular formula:

C8H10O2

IUPAC Name:

1,2-dimethoxybenzene

Test material form:

other: liquid

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampling method: 10 mL of a solution in medium containing 100 mg/L test item were added to 90 mL demin-eralised water with 5 g NaCl; then, the solution was extracted two times with the solvent dichloromethane (4 mL, 3 mL), the organic phase was collected after drying with Na2SO4 into a 10 mL flask. 1000 µL internal standard solution (1000 mg/L 1,3,5-trimethylbenzene in dichloromethane) was added to the flask, the flask was filled to 10 mL with dichloro-methane and the solution was measured via GC/FID.

Test solutions

Vehicle:

no

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Species: Daphnia magna
- Variety: STRAUS
- Strain: Berlin
- Sex: female
- Age: between 0 and 24 hours
- Origin: Umweltbundesamt Berlin
- Arrival of Strain: 27. Sep. 2007
- Selection of the test system was made following the proposal of the guidelines.
- Age at study initiation (mean and range, SD): 0 and 23 hours

- Method of breeding: 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 identical. The husbandry is performed similar to the method described in EN ISO 6341, following SOP 115 002 01 („Zucht und Hälterung von Daphnia magna STRAUS“), edition 10, adopted 04. Nov. 2010.
Vessels: preserving glasses, nominal volume 2 l
Medium: M4-Medium (recipe of ELENDT), composition see annex
Food: unicellular green algae (Desmodesmus subspicatus)
Medium renewal: twice a week
Photo period: 16/8 hours, using neon tubes
Temperature: 20+/- 2°C

- Feeding during test: no

ACCLIMATION
- Acclimation period: 30 mn
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: none
- Health during acclimation (any mortality observed): animals which showed no apparent damage were used for the test.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

250 mg/L

Test temperature:

22.3 - 22.6°C

pH:

7.7 - 7.9

Dissolved oxygen:

8.2 - 8.5 mg/L

Nominal and measured concentrations:

100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beakers, nominal volume 50 mL, tall shape
- Type (delete if not applicable): open
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Deionised water with an enrichment of certain minerals (as demanded in the guidelines) is used in the test.
Dilution water specification:
CaCl2*2H2O: 293.80 mg/L
MgSO4*7H2O: 123.30 mg/L
NaHCO3: 64.80 mg/L
KCl: 5.80 mg/L
Resulting hardness in mmol/L: 2.502
Resulting hardness in mg CaCO3/L: 250
After preparation, the dilution water is aerated, and the pH was adjusted to 7.8 +/- 0.2.

- Culture medium different from test medium: yes, Culture medium was M4 medium

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: none (darkness)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mobility

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: there was no mortality in a previous test, so a limit test was launched.
- Test concentrations: 100 mg/L

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

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

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The tested concentration showed no toxicity. None of the animals were immobilised in the control.
At the beginning and at the end of the test, the content of the test item in the test solutions was determined using GC-determination. The recovery after 48 hours was 83.1 % of the start concentration. The measured start concentration was 127 % of the nominal concentration. After 48 h the measured concentration was 106 % of the nominal concentration, therefore the correlation between nominal and measured concentration was stated as good. Thus, the determination of the biological results was based on the nominal concentration.

Results with reference substance (positive control):

- Results with reference substance valid: yes
- Mortality: yes
NOEC 24 h: 0.625 mg/L
24h EC50i: 1.3 mg/L (CL: 1.2 – 1.4 mg/L)
24h EC100i: 3.0 mg/L

Other: The 24h-EC50i of potassium dichromate was tested in a current reference test. The value was determined as 1.3 mg/L, lying within the demanded range of 0.6 – 1.7 mg/L.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

Immobilisation in the controls was 0 %. The pH-value in the test solutions should not vary more than 1 unit during the test. The highest variation was 0.2 units. The lowest concentration of dissolved oxygen at the end of the test was 8.2 mg/L (>= 3 mg/L)

Conclusions:

Veratrole is not Harmful to Daphnia magna.

Executive summary:

In a study (Muckle M, 2012), the toxicity of Veratrole with Daphnia magna was evaluated in a limit test at 100 mg/l. This study was GLP and followed the standard guidelines OECD 202 and EC C.2.

Twenty 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.

The tested concentration showed no toxicity. None of the animals were immobilised in the control.

The EC50 determined was higher than 100 mg/L.

The NOEC was >= 100 mg/L

In these conditions, Veratrole is not considered harmful to Daphnia magna.