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Ecotoxicological information

Short-term toxicity to fish

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Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
2012-06-04 to 2012-06-09
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study under GLP, no deficiencies
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Polyglycol BB 300
- Physical state: Liquid, colourless
- Storage condition of test material: Room temperature, protected from light, in original container.

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: 20.0 – 40.0 – 80.0 – 160 – 320 mg/L
- Sampling method: Analytical evaluation of various concentrations of Polyglycol BB 300 and the control were carried out via GC-MS at start and end of the exposure or whenever 100 % mortality occurred.
- Sample storage conditions before analysis: All original samples were stored at 6 ± 2 °C until sample preparation, if necessary. Prepared samples were stored at room temperature until analysis.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Directly weighing and mixing with an ultraturrax (1 min, 17000 rpm)
- Eluate: Dilution water
- Differential loading: 20.0 – 40.0 – 80.0 – 160 – 320 mg/L
- Controls: 7 fish in dilution water (without test item) were tested under the same test conditions as the test replicate.

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebrafish
- Source: All fish used in the test originate from a single brood stock at DR.NOACK-LABORATORIEN from a single brood stock. (supplier: Niedersächsischer Landesbetrieb für Wasserwirtschaft, Küsten- und Naturschutz, An der Scharlake 39, D-31135 Hildesheim)
- Length at study initiation (length definition, mean, range and SD): Average body length: 2.24 cm
- Weight at study initiation (mean and range, SD): Average body weight: 0.246 g
- Method of breeding: Holding was performed at the test facility at 23+/-2 °C and diffuse light (0.1 - 10 µmol photons • m-2 • s-1, natural photoperiod). The water was changed at least once per week. The dissolved oxygen concentration was more than 80 % of the air saturation value. No disease treatments were administered throughout holding and testing. Food was provided 3 times per week. The amount of food was 4 % of the fish body weight per feeding day. The test fish were not fed 24 h before the test started.
- Feeding during test: No feeding during test



ACCLIMATION
- Acclimation period: Zebrafish with at least 12 days of acclimatisation and mortality < 5 % within these days before the study starts were used in the test.
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: 4 % of the fish body weight per feeding day. The test fish were not fed 24 h before the test started. Food: Sera Vipan; SERA GMBH, D-52518 Heinsberg
- Feeding frequency: 3 times per week.
- Health during acclimation (any mortality observed): No mortality observed, no disease treatments were administered throughout holding and testing.


Study design

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

Test conditions

Hardness:
Total Hardness at day 0: 39 mg/L
Test temperature:
Please refer to "Any other information on materials and methods"
pH:
Please refer to "Any other information on materials and methods"
Dissolved oxygen:
Please refer to "Any other information on materials and methods"
Salinity:
Not measured, freshwater
Nominal and measured concentrations:
Please refer to "Any other information on materials and methods"
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass aquaria
- Type (delete if not applicable): open, loosely covered by glass tops
- Material, size, headspace, fill volume: glass, 10 L test volume per vessel
- Aeration: Gentle aeration
- Renewal rate of test solution (frequency/flow rate): application at test start
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: less than 1 g fish per L test solution


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Same as for holding
- Culture medium different from test medium: Tap water of local origin was used for holding. The water was filtered on activated charcoal and aerated for at least 24 h to remove chlorine.
Nominal water parameters:
Total hardness: 10 - 250 mg CaCO3/L
pH-value: 6.0 - 8.5
- Intervals of water quality measurement: pH-value, temperature and oxygen saturation were measured in all vessels at the beginning of the test and every 24 h (from old and new media). Total hardness of the water was determined at the beginning out of the test control. During the test the water temperature was recorded continuously with a data logger (once per hour).


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: natural
- Light intensity: 0.1 - 10 µmol photons x m-2 x s-1


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Fish were considered dead if there was no visible movement (e.g. gill covers movement) and if touching of the caudal peduncle produced no reaction. Records were kept of visible abnormalities (e.g. loss of equilibrium, swimming behaviour, respiratory function, pigmentation, etc.). Observations were made after 24, 48, 72 and 96 h.


TEST CONCENTRATIONS
- Range finding study:
Cumulative Mortality [%] in the Preliminary Test
(n = 5)
Nominal test item concentration
[mg/L] Test Duration [hours]
24 48 72 96
10000 100#) --- --- ---
400 100#) --- --- ---
80.0 0 0 0 0
10.0 0 0 0 0
Control 0 0 0 0








Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
210 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
139 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Behavioural abnormalities: Not observed
- Mortality of control: 0%
- Other adverse effects control: Not observed
- Abnormal responses: Not observed
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The test item was clearly dissolved in the tested loading levels during exposure.
Results with reference substance (positive control):
No reference substance tested
Reported statistics and error estimates:
After 24 – 96 hours only 0 and 100 % mortality occurred. Hence the LC50 after 24 – 96 hours is the geometric mean value of the nominal concentrations with 0 and 100 % mortality. The concentrations causing 0 and 100 % were chosen as lower and upper confidence limit, respectively. Therefore LC10,20-values were not applicable and were given as a range of those concentrations.
The chosen methods for determination of LCx-values were in accordance with the OECD series on testing and assessment No. 54 (2006). The concentrations leading to 0 and 100 % mortality (LC0 and LC100) after 96 h were determined directly from the test results.

Any other information on results incl. tables

Observations in the Test Vessels

Mean measured test item concentration

Effect *

Test duration [hours]

[mg/L]

24

48

72

96

317

(E)

7/7

---

---

---

139

(E)

-

-

-

-

(4.1)

-

-

7/7

7/7

(3.3)

-

-

-

7/7

(2.5)

4/7

-

-

-

(2.1)

3/7

7/7

7/7

7/7

  85.8

(2.5)

-

-

3/7

3/7

(2.4)

2/7

3/7

2/7

2/7

(1)

5/7

4/7

2/7

2/7

  42.0

(1)

7/7

7/7

7/7

7/7

  19.9

(1)

7/7

7/7

7/7

7/7

Control

(1)

7/7

7/7

7/7

7/7

Cumulative Mortality [%] in the Test Vessels

Mean measured
test item concentration

Test duration [hours]

[mg/L]

24

48

72

96

317

100

100

100

100

139

0

0

0

0

  85.8

0

0

0

0

  42.0

0

0

0

0

  19.9

0

0

0

0

Control

0

0

0

0

LC-Values with 95 % Confidence Intervals (0 – 96 hours)

                                Based on mean measured test item concentrations [mg/L]

Test duration [hours]

LC10  (p = 95 %)

LC20  (p = 95 %)

LC50  (p = 95 %)

24

139* LC10 317**

139* LC20 317**

210 (139* – 317**)

48

139* LC10 317**

139* LC20 317**

210 (139* – 317**)

72

139* LC10 317**

139* LC20 317**

210 (139* – 317**)

96

139* LC10 317**

139* LC20 317**

210 (139* – 317**)

LC0 =

139

Highest test item concentration with
0 % mortality after
96 hours

LC100 =

317

Lowest test item concentration with
100 % mortality after
96 hours

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
In this study Polyglycol BB 300 was found to cause lethal effects to Zebrafish after 96 hours at a mean measured concentration of 317 mg/L (highest test concentration). The mean measured concentration LC50 (96 hours) was 210 (139 – 317) mg/L.
All effect levels are given based on the mean measured concentrations level of the test item Polyglycol BB 300.
Executive summary:

The acute toxicity of the test item Polyglycol BB 300 (batch no.: DEG4155254)to fish (zebrafish), was determined according to OECD-Guideline for Testing of Chemicals No. 203 (1992) from 2012-06-04 to 2012-06-09, with the definitive exposure phase from 2012-06-04 to 2012-06-08, at Dr.U.Noack-Laboratorien, 31157 Sarstedt, Germany.

A static test with one application at start of the exposure phase conducted with the nominal test item concentrations of 20.0 – 40.0 – 80.0 – 160 – 320 mg/L (factor 2), corresponding to mean measured concentrations of 19.9 – 42.0 – 85.8 – 139 – 317 mg/L. Duration of the test was 96 hours. 7 test organisms were exposed to each test concentration and control. Water quality parameters temperature, pH-value and O2-saturation measured at 0, 24, 48, 72 and 96 hours were determined to be within the acceptable limits.

The determination of various concentrations of the test item Polyglycol BB 300 and the control was carried out via GC-MS from freshly prepared media after 0 hours and from old test media after 96 hours.

The recovery rates of Polyglycol BB 300 media were in the range of 94 – 115 % in freshly prepared and in the range of 79 – 103 % in the old test media of the nominal values, respectively. All effect levels are given based on the mean measured concentrations of the test item Polyglycol BB 300

LC-Values with 95 % Confidence Intervals (0 – 96 hours)

                   Based on mean measured test item concentrations [mg/L]

Test duration [hours]

LC10  (p = 95 %)

LC20  (p = 95 %)

LC50  (p = 95 %)

24

139* LC10 317**

139* LC20 317**

210 (139* – 317**)

48

139* LC10 317**

139* LC20 317**

210 (139* – 317**)

72

139* LC10 317**

139* LC20 317**

210 (139* – 317**)

96

139* LC10 317**

139* LC20 317**

210 (139* – 317**)

LC0 =

139

Highest test item concentration with
0 % mortality after
96 hours

LC100 =

317

Lowest test item concentration with
100 % mortality after
96 hours