reaction mass of cis 4-(3-methylbutyl)cyclohexanol and trans 4-(3-methylbutyl)cyclohexanol

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 29, 2007 - August 17, 2007

Reliability:

1 (reliable without restriction)

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

, 2004

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:

signed 2006-06-01

Test material

Reference

Name:

Unnamed

Type:

Constituent

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): Symrose
- Substance type: technical product
- Physical state: colourless, liquid
- Storage condition of test material: ambient temperature, dark and dry
No further details are given.

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
No further test material was used.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

The amount of test material exposed to the organisms was analysed via GC after 0 and 24 hours (new media) and 24 and 48 hours (old media).

Preparation of standards for validation of the analytical method:
- Sampling method: Five replicates of test medium fortified with 100 μg/L of the standard and two dilution water blank samples were analysed. Two dilution water blank samples were used to prove specificity and blank values being < 30 % of the LOQ.
- Sample storage conditions before analysis: All samples were stored at 7 ± 2 °C until start of analysis, if necessary.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The stock solution (100mg/L test item were weighed out) was prepared with dilution water one day prior tp application. The stock solution was stirred in a closed bottle with 110rpm for 24 hours at ambient temperature.
- Controls: Dilution water without test item tested under the same conditions as the test groups.
- Evidence of undissolved material: Undissolved particles were removed by centrifugation for 10 minutes at 3000rpm and membrane filtration (0.2µm).
No further details are given.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (20 animals, divided into 4 parallel samples, each with 5 animals)
- Strain: STRAUS (Clone 5)
- Source: Institute für Wasser-, Boden- und Lufthygiene (WaBoLu)
- Age at study initiation: 2-24 hours
- Method of breeding: In 2-3L glass vessels with approximately 1.8L culture medium, at 20 +/- 2°C, in an incubator, 16 hours illumination
- Feeding during test: The daphnids were not fed during the test.
- Acclimation period: at least 2 hours in dilution water

No further details are given.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

none

Test conditions

Hardness:

total hardness of dilution water: 246 - 253 mg CaCO3/L

Test temperature:

18-20°C +/- 1°C constant

pH:

7.8 +/- 0.2

Dissolved oxygen:

new media:
- 0 hours: 6.68 to 8.26 mg O2/L
- 24 hours: 5.91 to 8.21 mg O2/L

old media:
- 24 hours: 7.43 to 7.83 mg O2/L
- 48 hours: 7.32 to 7.87 mg O2/L

dilution water: 8.15 to 8.21 mg O2/L

Salinity:

According to Directive 92/69/EC L383A C.2. Annex
KCl: 5.76 mg/L
NaHCO3: 64.8 mg/L
CaCl2 x 2H2O: 294 mg/L
MgSO4 x 7H2O: 123 mg/L

Nominal and measured concentrations:

Nominal test item concentrations: 0.88, 1.94, 4.27, 9.39, 20.7, 45.5 and 100 mg/L (factor 2.2)
Geomeric mean measured concentration of the active ingredient: 0.60, 1.39, 3.04, 6.98, 14.9, 32.5 and 81.5 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beakers (4cm ID x 7cm H), 50mL

TEST DESCRIBTION:
A preliminary range finding test was conducted with three test item concentrations ranging from 1 to 100 mg/L set up in a geometric series with a dilution factor of 10 under semi-static conditions. Based on the results the definitive test was performed with 7 concentrations ranging from nominal 0.88 to 100 mg/L set up in a geometric series with a dilution factor of 2.2, corresponding to mean measured concentrations of the active ingredient of 0.604 to 81.9 mg/L, under semi-static conditions to enable the determination of immobilisation after 24 and 48 hours.
Water parameters were measured at 0 and 24 hours (new media) and 24 and 48 hours (old media).

TYPE AND FREQUENCY OF MEASURMENT AND OBSERVATIONS:
Prior to test start (0 hours) and after 24 hours (water renewal) pH values, dissolved oxygen concentration, temperature, conductivity and total hardness of the dilution water were measured.
In new media the water parameters were measured in one additional replicate per concentration and control. In the old media the water parameters were measured in all replicates per concentration and control. The room temperature was recorded throughout the test with a thermohygrograph. The percentage immobility was determined in all test and control groups after 24 and 48 hours. An animal was considered to be immobile, if it was not able to swim 15 second after gentle agitation of the test vessel.

Reference substance (positive control):

yes

Remarks:

potassium dichromat

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

3.04 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

1.39 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

14.9 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

32.51 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 5

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

7.81 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 7.33 - 8.33 mg/L confidence interval p= 95%

Effect concentrations 6

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

10.7 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 9.58 - 11.8 mg/l confidence interval p= 95%

Effect concentrations 7

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

5.07 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 8

Duration:

24 h

Dose descriptor:

EC10

Effect conc.:

6.95 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

no relevant observations

Results with reference substance (positive control):

Reference item: potassium dichromate p.a. (SIGMA)
Test concentration: 1.66 – 1.99 – 2.39 – 2.87 mg/L
- Results with reference substance valid? yes
- EC50: 2.04 (CI 1.88 - 2.21) after 24h
The EC50-value of the reference item potassium dichromate after 24 h is within the prescribed concentration range 0.6 mg/L to 2.1 mg/L, according to OECD 202 (2004).

Reported statistics and error estimates:

Statistical methods: EC100 -values after 24 and 48 hours were deducted directly from the dose-response-relationship. There was no mathematical calculation. The 25h and 48h EC10 and EC50-values were calculated by sigmoidal dose-response regression.

Any other information on results incl. tables

The validation of the analytical method was performed in accordance with SANCO/3029/99 rev. 4 (11/07/00):

Linearity: The analytical system gave linear response in the calibration range of 10 to 800 μg/L. The correlation coefficient (r) of the calibration curves was ≥ 0.99.

LOQ: The LOQ of the analytical method was fixed at nominal 100 μg/L of the test item and checked by means of accuracy

The mean recovery rate (fortification level 100 µg/L n=5) was 75% with a RSD of 2.89. The response of blank values of test medium control samples was lower than 30 % of LOQ

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

(i) In the control group no Daphnia were immobilised or trapped on the surface of the water (ii) The EC50 -value after 24 h of the reference item was in the range of 0.6 to 2.1 mg/L (iii) The dissolved O2 concentration was > 3 mg/L (iv) The pH-value did n

Conclusions:

Symrose was found to be toxic to Daphnia magna after 48 hours. The EC50 (48 hours) was 7.86 (7.37 – 8.38) mg/L. The NOEC after 48 hours was 1.40 mg/L. The LOEC after 48 hours was 3.06 mg/L.