Reaction mass of rel-{(1R,2S)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl}methanol and rel-{(1S,2R)-1-methyl-2-[(2R)-5-methylhex-4-en-2-yl]cyclopropyl}methanol

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:

November, 2014 - April, 2015.

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

GR-50-1408 is the Givaudan identification code which was employed for ROSYFOLIA during the early, developmental and testing period.

Analytical monitoring:

yes

Remarks:

GC-FID

Details on sampling:

The main test was performed under semi-static renewal (24 hours) conditions.

All Fresh and Aged test solutions were analysed, i.e. 0 hour Fresh / 24 hour (Aged), 24 hour (Fresh) / 48 hour (Aged) for all of the Control and test concentration levels :

Control
1.0 % (nominal, % saturated solution at 100 mg/L)
1.8 % (nominal, % saturated solution at 100 mg/L)
3.2 % (nominal, % saturated solution at 100 mg/L)
5.6 % (nominal, % saturated solution at 100 mg/L)
10 % (nominal, % saturated solution at 100 mg/L)

The Fresh solutions (0 hour, 24 hour renewal) of the 100 % saturated solution were also analysed to confirm dosing and subsequent dilutions.

Vehicle:

no

Details on test solutions:

Preliminary Solubility/Stability Trial :
Based on a Fish Acute Toxicity study conducted for the test substance (Smithers
Viscient study number: 3200508), the test substance was prepared as a saturated
solution.
An initial trial was conducted to determine the concentration in a saturated solution
prepared in Elendt M4 media and the stability of the saturated solution over a 48-hour
period.
A saturated solution was prepared by stirring 200 mg/L in Elendt M4 media for
ca 24 hours. Following the stirring period, the preparation was allowed to settle for
ca 30 minutes prior to filtering through a 0.45 μm filter to give the 100% saturated
solution. This was prepared in triplicate. Samples were taken for chemical analysis.
The remainder of the 100% saturated solution was stored, in the dark, under
laboratory conditions for 48 hours after which further samples were taken for
analysis.
Analysis of the freshly prepared 100% saturated solutions at 0 hours showed
measured concentrations to range from 62 to 72 mg/L. Analysis of the saturated
solutions after 48 hours storage showed measured concentrations to range from 65 to
78 mg/L. These results indicated that the saturated solution was stable over a 48-hour
period under the storage conditions used.

Range-finding Test :
The range-finding test was conducted at nominal test substance concentrations of
0.10, 1.0, 10 and 100% saturated solution under semi-static test conditions. A control
group was also included. Duplicate test vessels were prepared for the control and each
test concentration. Based on nominal concentrations, the results of the range-finding
test suggested that the 48-hour EC50 value would be between 1.0 and 10% saturated
solution.

Definitive Test :
Based on the results of the range-finding test, for which the key results only have been
reported, the definitive test was conducted at nominal test concentrations of 1.0, 1.8,
3.2, 5.6 and 10% saturated solution.
The test was conducted using semi-static test conditions with renewal of test media
after 24 hours.
The test vessels (nominal volume 100 mL) were glass tall form beakers covered with
clear plastic petri dish lids. Four replicate test vessels were prepared for the control
and each test concentration containing approximately 100 mL of the appropriate
media.

At the start of the test, the 100% saturated solution test concentration was prepared by
weighing ca 200 mg of test substance and adding to 1000 mL of Elendt M4 medium.
This was stirred slowly for ca 24 hours, then allowed to settle for ca 1 hour and then
filtered through a 0.45 μm filter to give the 100% saturated solution test concentration. Dilutions were then prepared, in Elendt M4 medium, to give the 1.0,
1.8, 3.2, 5.6 and 10% saturated solution nominal test concentrations. A control
treatment was prepared by adding Elendt M4 medium only to the control vessels.
The remainder of the 100% saturated solution was stored in the dark under laboratory
conditions and used to prepare the test concentrations for the media renewal at
24 hours.

Test organisms (species):

Daphnia magna

Details on test organisms:

Juvenile Daphnia magna (Straus), less than 24 hours old, were taken from healthy parental laboratory cultures, held and maintained at the test facility. The culture was originally obtained from Smithers Viscient, Shawbury, UK.

Culturing of Daphnia magna :
The cultures were originally obtained from Smithers Viscient, Shawbury, UK. The
Daphnia magna are cultured in 1 litre glass beakers containing 800 mL of Elendt M4
medium. Each vessel and its contents are referred to as a 'culture'. New cultures are
initiated with juvenile Daphnia magna (less than 24 hours old), at a density of
approximately 15 daphnids per litre. The cultures are fed daily with a concentrated
suspension of Chlorella vulgaris prepared in accordance with standard operating
procedures.
The water in each culture was renewed or partially renewed at least twice a week.
Juveniles were removed when present in cultures using a sieve. Cultures were
maintained up to a maximum of 4 to 5 weeks. Juveniles for use in acute toxicity tests
were collected from the second brood onwards. Approximately 24 hours before a test
was set up, juveniles present in the cultures were removed and discarded. Over the
next 24 hours, juveniles for use in the test were removed from the culture using a
wide bore pipette and transferred to fresh culture medium. The juveniles were then
left for at least 1 hour before selecting actively swimming individuals for use. All
juveniles used to initiate a test were less than 24 hours old.
All cultures, prior to and during toxicity tests, are maintained under fluorescent
lighting on a 16-hour light:8-hour dark photoperiod with an approximate 30 minute
dawn/dusk period.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

Not indicated in Report.

Elendt M4 was prepared as the exposure medium.

Test temperature:

Temperature range : 19.2 °C - 19.7 °C

pH:

pH Range :
Control 7.48 - 7.93
0.49 mg/L (1.0% nominal dilution) 7.54 - 7.93
0.85 mg/L (1.8% 1.0% nominal dilution) 7.56 - 7.92
1.6 mg/L (3.2% nominal dilution) 7.59 - 7.90
2.7 mg/L (5.6% nominal dilution) 7.60 - 7.91
4.6 mg/L (10% nominal dilution) 7.62 - 7.90

Dissolved oxygen:

Dissolved Oxygen Range during exposure (mg O2/L) :
Control 9.16 - 9.80 mg O2/L
0.49 mg/L (1.0% nominal dilution) 9.24 - 9.79 mg O2/L
0.85 mg/L (1.8% 1.0% nominal dilution) 9.24 - 9.91 mg O2/L
1.6 mg/L (3.2% nominal dilution) 9.27 - 9.79 mg O2/L
2.7 mg/L (5.6% nominal dilution) 9.50 - 9.91 mg O2/L
4.6 mg/L (10% nominal dilution) 9.40 - 9.94 mg O2/L

Nominal and measured concentrations:

Given that a decline in measured concentration was observed over each 24-hour dosing period, it was considered justifiable to base the results on time-weighted mean measured test concentrations.
These were calculated to be 0.49, 0.85, 1.6, 2.7 and 4.6 mg/L for the 1.0, 1.8, 3.2, 5.6 and 10% saturated solution, respectively.

Details on test conditions:

Five juvenile Daphnia magna, less than 24 hours old, were added to each test vessel,
using a wide bore glass pipette to avoid damaging the animals during transfer. Four replicates per test concentration (20 daphnids, in total). The Daphnia magna were not fed during the test.

After 24 and 48 hours, the Daphnia magna in each test vessel were observed for
evidence of immobility.

The observations differentiated between mobile and immobile daphnids. An
individual was considered immobile if, when the contents of the test vessel were
briefly agitated, it did not swim during a 15-second period of observation. In addition,
Daphnia magna submerged in the body of the test media and those that were held at
the surface of the test media were also recorded.

Water Quality and Environmental Conditions :
The test was conducted with a 16-hour light: 8-hour dark photoperiod with an approximate 30 minute dawn/dusk period.

The pH, dissolved oxygen concentration (% air saturation value (ASV) and mg/L) and temperatures were determined in freshly prepared test media at the start of the test and at media renewal at 24 hours and in the old media at 24 and 48 hours. Continuous temperatures were measured using a digital (min/max) thermometer in an additional vessel maintained in the study area.

At the end of each exposure period the test water quality measurements were conducted using pooled replicate samples of old test media.

Reference substance (positive control):

yes

Remarks:

Potassium Dichromate, Batch No. 13D290030.

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

1.6 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: EC0 and also NOEC

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

4.6 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

3.4 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CI : 3.0 - 3.7 mg/L

Details on results:

After 48 hours, the concentration at which no significant (≤10%) dose related
immobility occurred was 1.6 mg/L. The NOEC was considered to be 1.6 mg/L. After
48 hours 100% immobility was recorded at the highest test concentration of 4.6 mg/L.

In terms of time-weighted mean measured concentrations the 48-hour EC50 value was
determined to be 3.4 mg/L.

The validity criteria for control immobility (≤10%) and dissolved oxygen (≥3 mg/L)
were both satisfied. The test is therefore considered valid.

Results with reference substance (positive control):

Treatment Rates: Control, 0.32, 0.56, 1.0, 1.8 and 3.2 mg/L
Study Duration: 48 hours
Test System: 5 Daphnia (<24 hours old) per vessel
Replication: 4 replicates per exposure concentration
Control Validity Criteria:Immobilisation/other sub lethal effects in the control group
did not exceed 10%

The 48-hour EC50 value and NOEC were determined to be 0.73 and 0.32 mg/L,
respectively.

This result is within the expected range (24-hour EC50 range of 0.6 to 2.1 mg/L as stated in OECD 202).

Toxicity values and the corresponding NOEC and LOEC values are presented in the following table. Values are presented in terms of time-weighted mean measured concentrations

	Time weighed mean measured concentration (mg/l)
	24hr	48hr
EC50	>4.6	3.4 (3.0-3.7)
LOEC	2.7	2.7
NOEC	1.6	1.6

Validity criteria fulfilled:

yes

Remarks:

The validity criteria for control immobility (≤10%) and dissolved oxygen (≥3 mg/L) were both satisfied. The test is, therefore, considered valid.

Conclusions:

After 48 hours, the concentration at which no significant (≤10%) dose related immobility occurred was 1.6 mg/L. The NOEC was therefore 1.6 mg/L.After 48 hours 100% immobility was recorded at the highest test concentration of 4.6 mg/L.
The 48-hour EC50 value was determined to be 3.4 mg/L.

Description of key information

 

The 48-hour acute toxicity of GR-50-1408 to the freshwater planktonic crustacean, Daphnia magna, was determined in accordance with the requirements of OECD Chemicals Testing Guideline No. 202 Daphnia sp.

Acute Immobilisation Test (adopted April 13 2004). The validity criteria for control immobility (≤10%) and dissolved oxygen (≥3 mg/L) were both satisfied. The test is, therefore, considered valid.

Based on time-weighted mean measured concentrations, the 48-hour EC50 value was determined to be 3.4 mg/L. The corresponding No Observed Effect Concentration (NOEC) was considered to be 1.6 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

3.4 mg/L