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Ecotoxicological information

Long-term toxicity to fish

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Administrative data

Endpoint:
fish early-life stage toxicity
Type of information:
other: read-across to hydrolysis product (methanol)
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Meets generally accepted scientific standards, well documented and acceptable for assessment

Data source

Reference
Reference Type:
publication
Title:
An artificial fertilization method with the Japanese medaka: Implications in early life stage bioassays and solvent toxicity
Author:
Gonzáles-Doncel, M. et al.
Year:
2008
Bibliographic source:
Ecotoxicology and Environmental Safety 69: 95-103

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
Early lifestage bioassay. An in vitro fertilization method was used to study the effects on Medaka embryos reared either from 0.5 h or 6.5 h post-fertilization for 200 h.
GLP compliance:
not specified

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Methanol
- Analytical purity: >99%
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
no data

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: nominal values of 0.30, 0.50, and 2.00% v/v were measured through time (from 0 - 168 h) in 20 mL vials containing 2 mL of the respective nominal concentrations

Test solutions

Vehicle:
no
Details on test solutions:
no details

Test organisms

Test organisms (species):
Oryzias latipes
Details on test organisms:
TEST ORGANISM
- Common name: Medaka
- Strain: orange-red strain
- Source: laboratory by first author
- Age at study initiation (mean and range, SD): 0.5 (embryo stage 2) and 6.5 h (embryo stage 10) post-fertilization

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
200 h
Post exposure observation period:
72 h

Test conditions

Hardness:
no data
Test temperature:
25 ± 1°C
pH:
no data
Dissolved oxygen:
no data
Salinity:
no data
Nominal and measured concentrations:
474, 1027, 1975, 3950, 7900, 11850, 15800 mg/L (nominal) (re-calculated from % v/v data provided)
Details on test conditions:
TEST SYSTEM
- Test vessel: vials containing 2 mL test solution
- Renewal rate of test solution (frequency/flow rate): without renewal
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 5
- No. of vessels per control (replicates): 5
- No. of vessels per vehicle control (replicates): 5
- all experiments were run twice

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Rearing medium (RM)

OTHER TEST CONDITIONS
- Photoperiod: 16 h light, 8 h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
daily: mortality (absence of heartbeat and blood stagnation), hatching failure (unable to hatch by day 21 after fertilization), any deviation from normal development
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
200 h
Dose descriptor:
EC50
Effect conc.:
14 536 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
number hatched
Remarks:
for stage 2
Remarks on result:
other: 95% CL: 12798-16590
Duration:
200 h
Dose descriptor:
EC50
Effect conc.:
9 164 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: % Larva swim bladder inflation for stage 2
Remarks on result:
other: 95% CL: 7900-10586
Duration:
200 h
Dose descriptor:
EC50
Effect conc.:
10 270 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: % Larva swim bladder inflation for stage 10
Remarks on result:
other: 95% CL: 9322-11376
Duration:
200 h
Dose descriptor:
LOEC
Effect conc.:
11 850 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: % Embryo-cardiovascular for stage 2
Duration:
200 h
Dose descriptor:
LOEC
Effect conc.:
39 505 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
number hatched
Remarks:
for stage 2
Duration:
200 h
Dose descriptor:
LOEC
Effect conc.:
7 900 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: % Larva swim bladder inflation for stage 2
Duration:
200 h
Dose descriptor:
LOEC
Effect conc.:
11 850 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: % Embryo-cardiovascular for stage 10
Duration:
200 h
Dose descriptor:
LOEC
Effect conc.:
11 850 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
number hatched
Remarks:
for stage 10
Duration:
200 h
Dose descriptor:
LOEC
Effect conc.:
7 900 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: % Larva swim bladder inflation for stage 10
Duration:
200 h
Dose descriptor:
NOEC
Effect conc.:
15 800 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: % Embryo-cardiovascular for stage 2
Duration:
200 h
Dose descriptor:
NOEC
Effect conc.:
7 900 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
number hatched
Remarks:
for stage 2
Duration:
200 h
Dose descriptor:
NOEC
Effect conc.:
11 850 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: % Larva swim bladder inflation for stage 2
Duration:
200 h
Dose descriptor:
NOEC
Effect conc.:
15 800 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: % Embryo-cardiovascular for stage 10
Duration:
200 h
Dose descriptor:
NOEC
Effect conc.:
158 000 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
number hatched
Remarks:
for stage 10
Duration:
200 h
Dose descriptor:
NOEC
Effect conc.:
11 850 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: % Larva swim bladder inflation for stage 10
Duration:
200 h
Dose descriptor:
other: MATC
Effect conc.:
13 667 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: % Embryo-cardiovascular for stage 2
Duration:
200 h
Dose descriptor:
other: MATC
Effect conc.:
9 638 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: % Larva swim bladder inflation for stage 2 and 10 each
Duration:
200 h
Dose descriptor:
other: MATC
Effect conc.:
5 609 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
number hatched
Remarks:
for stage 2
Duration:
200 h
Dose descriptor:
other: MATC
Effect conc.:
13 667 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
other: % Embryo-cardiovascular for stage 10
Duration:
200 h
Dose descriptor:
other: MATC
Effect conc.:
13 667 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
number hatched
Remarks:
for stage 10
Details on results:
Based on anatomical abnormalities no differences between both time windows (stage 2 and 10) were observed at concentrations of methanol ≤ 1975 mg/L. Only hatching success (EC50 14536 mg/L) points to the earlier window of exposure as significantly more sensitive.
Reported statistics and error estimates:
In an effort to normalize the distributions and to stabilize the variances, proportional data were transformed using the arcsine square root and continuous data were log-transformed. Differences from the controls and between treatments were identified with Dunnet’s procedure and Tukey–Kramer’s analysis (P < 0.05), respectively. When data were not normally distributed, the Wilcoxon’s sign-rank test was applied (P < 0.05). The designated known control vial used in each assay as temporal reference for normal development was excluded from statistical analysis. For the analytical studies, differences between sample concentrations were identified with one-way ANOVA and the Wilcoxon’s sign-rank test (P < 0.05). These tests were done using JMP statistical software (SAS, Cary, NC).

Any other information on results incl. tables

All effect concentrations given under results and discussions were re-calculated from % v/v data provided in publication.

Applicant's summary and conclusion