Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13 Nov 2014 to 30 Jun 2015
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study similar to the guidelines, under Chinese GLP (non-adherence to OECD and no MAD to date)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
equivalent or similar to
Guideline:
other: HJ/T 153-2004, The guidelines for the testing of chemicals [S]. Beijing: SEPA, 2004 CRC-MEP. The Guidelines for the Testing of Chemicals, Effects on Biotoc Systems [M]. 2nd edition. Beijing: China Environment Press. 2013: 30-36.
Qualifier:
equivalent or similar to
Guideline:
other: GB/T 27861-2011, Chemicals-Fish acute toxicity test, Beijing: SAC, 2011. GB/T 29763-2013, Chemicals-Rare Minnow (Gobiocypris rarus) acute toxicity test Beijing: SAC, 2013.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Qualifier:
equivalent or similar to
Guideline:
EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)
GLP compliance:
no
Remarks:
according to “The guidelines of chemical testing good laboratory practices (HJ/T 155)” issued by State Environmental Protection Administration (SEPA) of the People’s Republic of China.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
Test substance: Tall oil polymd., oxidized
CAS: 68815-17-8

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
duplicate samples (50 mL) at 0, 24, 48, 72 and 96 h, extracted twice with 1,2 dichloromethane. The organic fractions were resuspended in dichloromethane (10 times concentrated) and filtered with 0.22 um millipore membrane. A 200 uL aliquot was added to 200 uL TBS t-butyldimethylsilyl (TBDMS) to allow derivatization during 30 min at 60 ºC.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION

Pretest: dilution of 0.0031, 0.0301, 0.3002 g test substance in 3 L dilution water, sonication for 30 min and stirring for 30 min. The solutions at 10 and 100 mg/L were cloudy

Main test
- Method: dilution of 1.5007 g test substance in 10 L dilution water, stirred for 24 h at 35ºC, separation (after 1 h standing) by filtration through 0.45 μm millipore membrane, the resulting 100% saturated solution was diluted further with dilution water to achieve dose levels at 10%, 20%, 40%, 60%, 80% and 100% saturated solutions (total volume 3L/dose)
- Controls: no treatment
- Evidence of undissolved material: no data

Test organisms

Test organisms (species):
other: Gobiocypris rarus
Details on test organisms:
TEST ORGANISM
- Common name: Rare minnow, Gobiocypris rarus
- Strain: NA
- Source: commercial fish supplier of Institute of Hydrobiology, Chinese Academy of Sciences
- Length at study initiation: 2.26-2.71 cm (2.48±0.15 cm)
- Weight at study initiation: 0.182-0.240 g wet weight (0.214± 0.17 g)
- Feeding during test: food witheld from 24 hours before exposure onwards

ACCLIMATION
- Acclimation period: 12 days generic accomodation (continuous flow of aerated water), 7 days thereafter under conditions of the test
- Type and amount of food: food containing Crude Protein > 36.0%, Crude Fat > 2.0%, Crude Fibre < 3.0%, Crude Ash < 12.0% and Moisture < 10.0%
- Feeding frequency: daily
- Health during acclimation: no mortality in the 7 day acclimisation period

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h

Test conditions

Hardness:
141-149 mg/L as CaCO2
Test temperature:
23.1-23.3 ºC
pH:
7.74-8.04
Dissolved oxygen:
68-95% of saturation
Nominal and measured concentrations:
nominal: 0, 10, 20, 40, 60, 80 and 100% of saturation (loading rates: 0, 15, 30, 60, 90, 120 and 150 mg/L)
measured TWA: stability of the concentrations over the exposure period > 80%
Details on test conditions:
Pretest: 5 fish/concentration at 0, 1.0, 10 and 100 mg/L in 3L dilution water during 96 h. Mortality was checked at 24, 48, 72 and 96 h.

TEST SYSTEM
- Test vessel: 5 L glass containing 3L dilution water
- Type: vessels with sealable inert lid
- Aeration: none
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- Biomass loading rate: 0.71 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: no data
- Total organic carbon: 1.3 mg/L
- Particulate matter: none
- Metals: total < 1 mg/L
- Pesticides: < 0.001 mg/L
- pH: 7.82

OTHER TEST CONDITIONS
- Adjustment of pH: none
- Photoperiod: 16/8
- Light intensity: 1000-1500 lux

EFFECT PARAMETERS MEASURED : at 24, 48, 72 and 96 h mortality, abnormal behaviour (temperature pH and dissolved oxygen daily)

TEST CONCENTRATIONS
- Range finding study: 0, 0.1, 10 and 100 mg/L
- Test concentrations: 0, 10%, 20%, 40%, 60%, 80% and 100% saturated solutions
Reference substance (positive control):
yes
Remarks:
potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
> 30 - < 60 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
0.77 mg/L
Nominal / measured:
meas. (TWA)
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
Range finding study: mortality 0 at 1.0 and 10 mg/L, 5 at 100 mg/L (at 24 h observation)
Results with reference substance (positive control):
LC50 potassium dichromate 278 mg/L (24 h)
Reported statistics and error estimates:
LC50 , based on the measured concentration) 0.77 mg/L
95% confidence limit (mg/L) 0.58~1.03

Any other information on results incl. tables

Nominal Concentration

(mg/L)

Measured Concentration

(mg/L)

Initial Number

of Fish

The Number of the Dead Fish

3 h

6 h

24h

48 h

72 h

96 h

Blank Control

ND

10

0

0

0

0

0

0

10% saturated solution

0.276

10

0

0

0

0

0

0

20% saturated solution

0.694

10

0

1

1

3

3

4

40% saturated solution

1.82

10

2

3

7

8

10

10

60% saturated solution

2.52

10

8

10

10

10

10

10

80% saturated solution

4.61

10

10

10

10

10

10

10

100% saturated solution

6.06

10

10

10

10

10

10

10

 

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The 96 h LC50 of the substance in fish is 0.77 mg/L
Executive summary:

Fish (10/concentration) were exposed to the test substance during 96 hours at concentrations of nominal: 0, 10, 20, 40, 60, 80 and 100% of saturation (loading rates: 0, 15, 30, 60, 90, 120 and 150 mg/L. measured TWA: <LOD , 0.276, 0.694 , 1.82 , 2.52, 4.61, and 6.06 mg/L). No mortality or abnormal behaviour was observed at 0, 10 and 20%. In all other concentrations all fish died before the 72 hours observation. The LC50 based on measured concentrations is 0.77 mg/L (loading rate between 30 and 60 mg/L).