butyl 4-hydroxycyclohexanecarboxylate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015-02-03 to 2015-05-04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

(2004)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentration: 100 mg/L (limit test)
- Sampling method:
- Sample storage conditions before analysis:

Vehicle:

no

Details on test solutions:

Reconstituted water according to ELENDT (1990):
Macro nutrients (mg/L):
CaCI2 x 2 H20 147
MgSO4 x 7 H20 123
NaHCO3 64.8
KCI 5.80
Na2SiO3 4.30
NaNO3 0.27
KH2PO4 0.14
K2HPO4 0.18
Na2EDTA x 2H20 5.00
FeSO4 x 7 H2O 1.99
H3BO3 0.29
MnCl2 x 4 H2O 0.36
LiCl 0.30
SrCl2 x 6 H2O 0.15
RbCl 0.071
NaBr 0.016
Na2MoO4 x 2 H2O 0.063
CuCl x 2 H2O 0.017
ZnCl2 0.013
CoCl2 x 6 H2O 0.010
KJ 0.00325
Na2SeO3 0.00219
NH4VO3 0.000575
Thiaminhydrochloride 0.075
Cyanocobalamine 0.0010
Biotin 0.00075
pH: 8.2 ± 0.8, after an aeration for 24 hours.

References:

ELENDT, B.-P. Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus.
Protoplasma 154, 25-33, 1990

















PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test item was dissolved in culture medium (ELENDT) at a concentration of 100 mg/L. The solution was stirred (approx. 1100 rpm) for 24 hours at room temperature.
- Controls: Culture medium without test item

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna STRAUS
- Strain/clone: clone 5
- Source: Institut für Wasser-, Boden- und Lufthygiene, D-14195 Berlin, Germany
- Feeding during test: No
- Age: not older than 24 hours

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

263 mg CaCO3/L

Test temperature:

18-22°C

pH:

7.54 - 7.75

Dissolved oxygen:

7.96 - 8.57 mg/L

Conductivity:

653 µS/cm

Nominal and measured concentrations:

Nominal concentration: 100 mg/L (limit test)
Measured concentrations:
Start of exposure (0 h): 104 mg/L
End of exposure (48 h): 98.1 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers, 50 mL capacity
- Type (delete if not applicable): open (loosely covered with watch glasses)
- Material, size, headspace, fill volume: glass, 4 (ID) x 7 (H) cm), 20 mL fill volume
- Renewal rate of test solution (frequency/flow rate): no
- 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: ISO test water, according to OECD 202, Annex 3
- Intervals of water quality measurement:
At the start of exposure (0 h) and end of exposure (48 h) : pH value, dissolved oxygen, temperature, conductivity and total hardness

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16/8 h light/dark cycle
- Light intensity: Diffuse light, light intensity of max. 20 µE x m-2 x s-1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Immobilisation was determined in all groups after 24 and 48 h.

VEHICLE CONTROL PERFORMED: not applicable

RANGE-FINDING STUDY
- Test concentrations: 1.00, 10.0 and 100 mg/L
- Results used to determine the conditions for the definitive study:
No significant effects were observed at any of the tested concentration levels.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

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

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Clinical findings
The nominal concentration of the preparation of the test substance and the immobilization data are shown in the following table:

--------------------------------------------------------------------
Nominal concentration
[mg/L] Immobilized/exposed Daphnia magna
---------------------------
24 hours 48 hours
--------------------------------------------------------------------
0 0/20 0/20
100 0/20 0/20
--------------------------------------------------------------------

A dose group with nominal 100 mg/L of the test item revealed no aquatic toxicity in this test system.

No remarkable observations were made concerning the appearance of the solution of the test material preparation. The test medium was a clear preparation and stayed unchanged throughout the study.


EC50 value

An aqueous solution of nominal 100 mg/L of the test substance revealed no aquatic toxicity in the test system. The 24 hours and 48 hours EC50 to Daphnia magna exceeded the limit dose of 100 mg/L of the test item and, thus, was not determined in this test.

Analysis
pH, oxygen concentrations, and temperature during the study were inconspicuous.

--------------------------------------------------------------------------------------------------------------------------
Nominal concentration
[mg/L] Start of the experiment End of the experiment
---------------------------
0 hour 48 hours (mean)
pH O2 [mg/L] pH O2 [%]
-----------------------------------------------------------------------------------------------------------------------------
0 7.75 8.57 7.58 8.48
100 7.56 7.96 7.54 8.39
------------------------------------------------------------------------------------------------------------------------------

Results with reference substance (positive control):

The percentage immobility for the reference item was determined after 24 h. The EC50-value with 95% confidence limits (CI) was calculated by sigmoidal dose-response regression. The EC50-value for the most recent of the monthly performed reference tests was: EC50 = 2.01 mg/L (CI: 1.71 - 2.26 mg/L). The EC50-value of the refernce item potassium dichromate after 24 h is within the prescribed concentration range of 0.6 - 2.4 mg/L of quality criteria according to AQS P 9/2 (02/2000) for daphnids clone 5 cultured in Elendt M4 medium. The EC50-value of the refence item is also within the the recommended range of 0.6 -2.1 mg/L according to OECD-Guideline 202.

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of a limit concentration of 100 mg/L of the test item were determined according to OECD 202 (2004).

The limit test was conducted under static conditions over a period of 48 h. The test item is a colourless liquid. The limit concentration was stirred with approximately 1100 rpm for 24 hours at room temperature before the start of the exposure and was clear throughout the exposure period.

Twenty daphnids each were exposed to the limit concentration and the control.

The concentrations of the test item were analytically verified via LC-MS/MS at the start of the exposure (0 h) and at the end of the exposure (48 h) in the limit concentration and the control. Details of the analytical method are presented in the report.

The measured concentration of the test item at the start of the exposure (0 h) was 104 mg/L. At the end of the exposure (48 h), the measured concentration of the test item was 98.1 mg/L. The measured test item concentration was within ± 20 % of the nominal concentration.

This indicates that the test item concentration was successfully maintained for the duration of the test. The effect levels given are based on the nominal concentration of the test item.

The water quality parameters (i.e. pH-value and dissolved oxygen concentration), measured at the start (0 h) and at the end of the exposure (48 h), were within the acceptable limits. The validity criteria of the test guideline were fulfilled.

Effect levels:

EC50 (24 h): > 100 mg/L

EC50 (48 h): > 100 mg/L

NOEC (48 h): 100 mg/L

LOEC (48 h) > 100 mg/L

Validity criteria fulfilled:

yes

Conclusions:

At the analytically confirmed nominal concentration of 100 mg/L of the test item, no effects were observed in Daphnia magna.

Executive summary:

see above

Description of key information

In an acute immobilisation test with Daphnia magna (STRAUS), the effects of a limit concentration of 100 mg/L of the test item were determined according to OECD 202 (2004).

The limit test was conducted under static conditions over a period of 48 h. The test item is a colourless liquid. The limit concentration was stirred with approximately 1100 rpm for 24 hours at room temperature before the start of the exposure and was clear throughout the exposure period.

Twenty daphnids each were exposed to the limit concentration and the control.

The concentrations of the test item were analytically verified via LC-MS/MS at the start of the exposure (0 h) and at the end of the exposure (48 h) in the limit concentration and the control. Details of the analytical method are presented in the report.

The measured concentration of the test item at the start of the exposure (0 h) was 104 mg/L. At the end of the exposure (48 h), the measured concentration of the test item was 98.1 mg/L. The measured test item concentration was within ± 20 % of the nominal concentration.

This indicates that the test item concentration was successfully maintained for the duration of the test. The effect levels given are based on the nominal concentration of the test item.

The water quality parameters (i.e. pH-value and dissolved oxygen concentration), measured at the start (0 h) and at the end of the exposure (48 h), were within the acceptable limits. The validity criteria of the test guideline were fulfilled.

Effect levels:

EC50 (24 h): > 100 mg/L

EC50 (48 h): > 100 mg/L

NOEC (48 h): 100 mg/L

LOEC (48 h) > 100 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L