Cyclopropanemethanol, 1-methyl-2-[(1,2,2-trimethylbicyclo[3.1.0]hex-3-yl)methyl]-

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15 February, 1999 - 14 October, 1999

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Version / remarks:

21 September, 1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1300 (Daphnid Chronic Toxicity Test)

Version / remarks:

Publoc Draft of April, 1996.

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

JAVANOL :
Batch No. : 8825
Purity : 88.4% (sum of 2 isomers)
Date of Expiry : 27 November, 2000

Analytical monitoring:

yes

Remarks:

GC-FID

Details on sampling:

Singular samples were taken from each vessel at each concentration for Days 0, 7, 14 and 21 of the exposure phase and on Day 2 from the lowest, middle and highest exposure concentrations.

A volume of 1 L was taken for all exposure levels, except for the 0.20 mg/L (nominal) exposure level where 500 mL were sampled.

Samples were analysed on the day of sampling.

Vehicle:

yes

Remarks:

Acetone (maximum concentration at 0.1 mL/L)

Details on test solutions:

A stock solution was prepared of JAVANOL at a concentration of 100 mg/mL in acetone. A volume of 0.1 mL of this stock solution was added per Litre of test medium and this solution stirred overnight. Two days prior to use in the test, this solution was filtered through a 0.2 micron filter to obtain the fraction actually dissolved. This solution was prepared weekly and a total of 5 L was required for 7 days of dosing. A sample for analysis was taken at each weekly renewal of the stock, i.e. from both the "old" and the "fresh" solution. This solution was stored in the dark at 7 - 10°C under con-tinuous stirring. On a daily basis, the required amounts to be dosed were taken from this solution.

Dosing System :
The solutions with test substance were dosed via a computer controlled dosing system consisting of 6 dispensers. Via this system the dosed volume entered a mixing flask separately from the dilution water supply. The dilution water was also supplied via a dispenser connected with a six-way valve. The flow-rate was 300 mL per hour. The dosed volume was mixed with the dilution water in a mixing flask under continuous stirring. The whole system was verified daily.

The system comprised of two stainless-steel vessels of 1.5 L per concentration, each with 2 mesh containers of stainless-steel.

The test solutions in the test vessels were fortified on day 0 prior to introduction of the test organisms.

The target Nominal concentrations to be studied were :
0.20 mg/L
0.10 mg/L
0.05 mg/L
0.02 mg/L
0.01 mg/L
Solvent Control (with a maximum concentrtion of acetone of 0.1 mL/L).

Test organisms (species):

Daphnia magna

Details on test organisms:

Breeding of Daphnia magna :
Start of each Batch : With newborn animals (i.e. less than 3 days old), by placing them individually in 50 mL M7 medium
Max Age of the Cultures : 4 Weeks
Renewal of the Medium : 3 times per week
Temperature of Medium : 18 - 22°C, within +/- 1°C
Feeding : Daily, suspension of freshwater algae (Chlorella pyrenoidosa)
Medium : M7

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Hardness:

180 mg CaCO3/L

Test temperature:

20.8 +/- 1°C

pH:

pH range 7.9 - 8.0

Dissolved oxygen:

Dissolved O2 range : 7.3 - 9.5 mg O2/L

Nominal and measured concentrations:

The Nominal and Geometric mean measured exposure concentrations were, as follows :

Nominal (mg/L) Mean Measured (mg/L)
Solvent Control Not Detected
0.01 0.009
0.02 0.018
0.05 0.031
0.1 0.069
0.2 0.131

Details on test conditions:

Test Type : Flow-Through
Test Duration : 21 days
Test Chambers : The test system comprised of two stainless-steel vessels of 1.5 L per concentration , each with 2 mesh containers of stainless-steel.
Test Medium : M7
Experimental Design : Each mesh contained 10 neonate (< 24 h old) daphnids ate the start of the study. Test solutions were stirred continuously.
Light Regime : 16 hours light / 8 hours dark (600 Lux)
Feeding : 3 times daily (Chlorella pyrenoidosa suspension). Ration was 0.2 - 0.3 x 10^8 cells/mL/hour to maintain the average ration at approximately 0.2 mg/C/daphnid/day

Measurements and Recordings :
For Parental Daphnids -
Immobility and Mortality : Every workday the number of living, immobile and dead parental daphnids were recorded. dead daphnids were removed when observed;
Presence of Eggs in Brood Pouch : Every workday;
Body Length : At the end of the test period.

For F1 Generation -
Appearance first Brood : When observed;
Newborn Daphnids : Every workday the number of newborn young was counted and the condition of the young recorded. Thereafter, the young were removed;
Presence of Unhatched Eggs : When observed ;
Incidence of Immobility : When observed.

PhysicoChemical Measurements :
pH : 3 times per week in the Control and the highest exposure concentration;
Oxygen : 3 times per week in all test concentrations;
Temperature : Daily, in all test solutions.

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.031 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks:

The most sensitive end-points leading to an NOEC of 0.031 mg/L were based on reproduction and body length.

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

> 0.131 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

Endpoint based on parental survival - only 5% Parent mortality was observed at the 0.131 mg/L exposure concentration, the hoighest test concentration tested.

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.131 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

Endpoint based on parental survival - only 5% Parent mortality was observed at the 0.131 mg/L exposure concentration, the hoighest test concentration tested.

Duration:

21 d

Dose descriptor:

EC10

Effect conc.:

0.037 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

Parental mortality ranged between 2.5 and 10% in the JAVANOL treated test solutions. The reproduction capacity of the two higher treatment levels appeared to be affected. However, the average reproduction at 0.131 mg/L was higher than the average at 0.069 mg/L. The variation of the total reproduction was rather high between the replicates of the Control group (61 to 126 young per parent). During the total expsoure period of 21-days, no significant (> 10%) immobilisation (including mortality) of new-born young was observed at any of the test concentrations. The first broods were observed on Day 7 at all treatments. No biologically significant numbers of unhatched eggs were observed during the reproduction phase in any of the test groups.

The mortality of the parental daphnids in the controls did not exceed 20%. The average cumulative number of young per female in the controls after 21-days was > 60 (actual = 92).

Validity criteria fulfilled:

yes

Conclusions:

The most sensitive end-points observed in the study were for Reproduction and Body Length.

The 21-day overall NOEC is 0.031 mg/L.

The 21-day EC50 and NOEC for Parental Survival are, respectively, > 0.131 mg/L and 0.131 mg/L.

The 21-day EC10 for Reproduction is 0.037 mg/L.

Description of key information

The most sensitive end-points observed in the study were for Reproduction and Body Length.

The 21-day overall NOEC is 0.031 mg/L.

The 21-day EC50 and NOEC for Parental Survival are, respectively, > 0.131 mg/L and 0.131 mg/L.

The 21-day EC10 for Reproduction is 0.037 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.031 mg/L

Additional information

The most sensitive end-points observed in the study were for Reproduction and Body Length.

The 21-day overall NOEC is 0.031 mg/L.

The 21-day EC50 and NOEC for Parental Survival are, respectively, > 0.131 mg/L and 0.131 mg/L.

The 21-day EC10 for Reproduction is 0.037 mg/L.