Registration Dossier

Ecotoxicological information

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
21 July 2008 – 14 October 2009
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study has been conducted according to OECD Guideline 211. No claim for GLP compliance is made for this study. However, the study was carried out in the spirit of GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Deviations:
no
GLP compliance:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Name of test material as cited in study report: SLI (76) Stripped
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not applicable

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: Nominal: 0, 560, 1000, 1800, 3200, 5600 (ug/L); Measured total: <0.5, 103, 192, 289, 752, 1307 ug/L
- Sampling method: Test item concentrations were determined by LC/MS
- Sample storage conditions before analysis: no data

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A 200 mg/L stock solution was prepared daily by adding 50 mg of S2849001 in 250 mL of purified water. The resulting dispersion was heated to approx 40 - 50 oC with stirring until completely dissolved.
- Eluate: no data
- Differential loading: no data
- Controls: yes
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): not applicable (water)
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: daphnia magna
- Strain/clone: not stated
- Justification for species other than prescribed by test guideline: not applicable (prescribed by guideline)
- Source: In-house cultures
- Age of parental stock (mean and range, SD): no data
- Feeding during test: yes
- Food type: chlorella vulgaris & microfeast PZ20 supplement
- Amount: chlorella vulgaris: 3.4 x 10^5 cells/mL/day; microfeast: 1.4mL of a 1g/L solution per day
- Frequency: Daily

ACCLIMATION
- Acclimation period: no

QUARANTINE (wild caught)
- not applicable

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: not applicable

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
21 d
Post exposure observation period:
No

Test conditions

Hardness:
average of 224mg/L as CaCO3
SD 9.5
(highest concentration and controls measured at various time points; n=39)
Test temperature:
average of 19.6 degrees C
SD 0.31
(measured in 6 vessels daily)
pH:
average 8.42
SD 0.44
(measured in all concentrations on 11 different days)
Dissolved oxygen:
average 9.34 mg/L
SD 0.94
(measured in all concetrations on 11 different days)
Salinity:
not measured
Nominal and measured concentrations:
see tables 1 & 2 below
Details on test conditions:
TEST SYSTEM
- Test vessel: 120mL glass bottle
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: ~100mL
- Aeration: no
- Type of flow-through (e.g. peristaltic or proportional diluter): semi-static
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 1/vessel
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10
- No. of vessels per vehicle control (replicates): not applicable
- Biomass loading rate: no data

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M7 Media
Made up following OECD Guideline 211; no other measurements taken
- Culture medium different from test medium: no
- Intervals of water quality measurement: every 2-3 days

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light/8 hours dark
- Light intensity: no data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : time to first brood, size of first brood, number of offspring per surviving adult, mortality

VEHICLE CONTROL PERFORMED: not applicable

RANGE-FINDING STUDY
- Test concentrations: no data
- Results used to determine the conditions for the definitive study: used to determine test concentrations & analytical method
Reference substance (positive control):
no

Results and discussion

Effect concentrations
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 154 µg/L
Nominal / measured:
meas. (TWA)
Conc. based on:
dissolved
Basis for effect:
reproduction
Remarks on result:
other: no data
Details on results:
- Mortality of parent animals: no data
- No. of offspring produced per day per female: no data
- Body length and weight of parent animals: no data
- Type and number of morphological abnormalities: no data
- Type and number of behavioural abnormalities: no data
- Time to first brood release or time to hatch: 8.7 days (controls), 9.4 (560ug/L), 9.4 (1000ug/L), 9.6 (1800ug/L), 9.8 (3200ug/L), 11 (5600ug/L)
- Egg development time: no data
- Brood size: First brood only: 9.1 (control); 11.5 (560ug/L), 10.5 (1000ug/L), 10.2 (1800ug/L); 10.9 (3200ug/L); 9.2 (5600ug/L)
- Time to sexual maturity: no data
- Type and magnitude of biochemical changes: no data
- Other biological observations: no data
- Effect concentrations exceeding solubility of substance in test medium: not applicable
Results with reference substance (positive control):
not applicable
Reported statistics and error estimates:
The data were analysed using an analysis of variance followed by Dunnett’s test to obtain the 21 day NOEC for mean juvenile production, time to release of first brood and size of first brood.

Any other information on results incl. tables

Table 3: Summary of Effect Data

Nominal concentration (µg/L

Mean Measured Concentration (µg/L)

Mean Number of Neonates at Day 21#

Percentage Adult Mortality at Day 21

Mean size of first brood

Mean time to release of 1st brood (days)

Total

Dissolved

0

<5.0

<5.0

70

0

9.1

8.7

560

102

77.7

99.6*

0

11.5

9.4

1000

192

154

104.9*

0

10.5

9.4

1800

289

260

121.3*

0

10.2

9.6*

3200

752

457

112.3*

10

10.9

9.8*

5600

1307

717

102.1*

30

9.2

11*

#  including dead neonates

* Significantly different to the control (P<0.05) 

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The lowest value determined for an adverse effect was time to first brood. The 21 day NOEC for this endpoint based on dissolved concentrations was 154 μg/L (192 μg/L total concentration)
Executive summary:

The purpose of this study was to determine the effects of SLI (76) stripped on survival and reproduction of Daphnia magna.

SLI (76), provided by Unilever Mannheim was stripped of its fatty acid content via Soxhlet extraction at the Materials Science Laboratory, Trumbull, USA. The resulting material, SLI (76) stripped was used for this study.

Daphnia were exposed to a range of concentrations, nominally 0.0, 560, 1000, 1800, 3200 and 5600 μg/L. Test item analysis was performed to determine total and dissolved concentrations. The dilution medium used was Elendt M7 artificial medium. A 3.4 x 105 cells/mL aliquot of Chlorella vulgaris and 1.4 mL of a 1 g/L Microfeast PZ20 supplement was added to the test media containing the appropriate concentration of SLI(76) stripped. The stock solution and test media were renewed daily.

The data were analysed using an analysis of variance followed by Dunnett’s test to obtain the

21 day NOEC for mean juvenile production, time to release of first brood and size of first

brood.

The lowest value determined for an adverse effect was time to first brood. The 21 day NOEC for this endpoint based on dissolved concentrations was 154 μg/L (192 μg/L total concentration).

There was no statistically significant effect at any concentration on size of first brood therefore the NOEC for this endpoint based on dissolved concentrations can be quoted as >717 μg/L (>1307 μg/L total concentration)

A statistically significant stimulatory effect was observed at all concentrations based on number of neonates per surviving adult. The number of neonates rose steadily up to a dissolved concentration of 260 ug/L (289 μg/L total concentration) after which it then began

to decline. The NOEC for this endpoint cannot therefore be determined with any confidence. The 21 day NOEC for adult mortality is 260 μg/L based on mean measured dissolved concentrations (289 μg/L as total concentration).