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Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012-12-10 to 2013-02-08
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study, no deficiencies
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 0.156 - 0.313 - 0.625 - 1.25 - 2.5 mg/L
- Sampling method:
Analytical evaluation of Sodium methyl oleyl taurate was carried via LC-MS/MS on a reversed phase column in gradient mode from freshly prepared media after 0 h and 72 h and from corresponding 24 h aged test media after 24 h and 96 h. An electrospray tandem mass spectrometer operating in positive ion mode was used as detector.
- Sample storage conditions before analysis:
All original samples were stored at room temperature until sample preparation. Prepared samples were stored at room temperature until analysis.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Solutions were directly weighed into dilution water. The test solutions were stirred with an ultraturrax at 17000 rpm for 1 min directly prior to test start and application of the test fish.
- Eluate: Dilution water
- Differential loading: 0.156 - 0.313 - 0.625 - 1.25 - 2.5 mg/L
- Controls: 7 fish in dilution water without test item were tested under the same test conditions as the test replicates.
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: 2.59 cm
- Weight at study initiation (mean and range, SD): Average body weight: 0.14 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.


Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
Total Hardness at day 0: 48 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): daily renewal (after 0, 24, 48, 72 hours)
- 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
5.00 100 --- --- ---
0.50 0 0 0 0
Control 0 0 0 0








Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
1.32 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
is UVCB means 100% act.ingr.
Basis for effect:
mortality
Remarks on result:
other: 95% CI 1.31-1.32 mg/L,
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 hours 0 % mortality occurred. Therefore no regression analysis was carried out..
After 48 hours a mortality of 71 % occurred in the highest tested concentration. The LC10,20,50-values were calculated by non linear regression (sigmoidal dose response, variable slope). After 72 hours only 0 and 100 % mortality occurred. Hence the LC50 after 72 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 could only be given as a range of those concentrations.
The LC10,20,50-values after 96 hours of exposure were calculated by non linear regression (0-100, sigmoidal dose response, variable slope).
The chosen methods for the 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 raw data.

Observations in the Test Vessels

Nominal test item
concentration
[mg/L]

Effect *



Number of fish effected at observation time [hours]

24

48

72

96

2.50

(E)

-

5/7

2/2

-

(4.1)

7/7

2/7

-

-

(2.7)

5/7

-

-

-

(2.5)

5/7

-

-

-

(2.4)

2/7

-

-

-

(2.1)

-

2/7

-

-

1.25

(E)

-

-

-

1/7

(4.1)

-

7/7

7/7

6/7

(2.6)

2/7

-

-

-

(2.5)

-

6/7

3/7

3/7

(2.4)

2/7

-

-

-

(2.1)

-

1/7

4/7

3/7

(1)

5/7

-

-

-

0.625

(1)

7/7

7/7

7/7

7/7

0.313

(1)

7/7

7/7

7/7

7/7

0.156

(1)

7/7

7/7

7/7

7/7

Control

(1)

7/7

7/7

7/7

7/7

Measured Concentrations of Sodium methyl oleyl taurate (C18:1-part) in Aqueous Phase and at Glass Walls of Test Solutions at 0 and 24 hours

Nominal concentration

Aqueous Phase

(Volume: 10 L)

Adsorbed

to Glass

Test item

[mg/L]

a.i.

[mg/L]

Fresh Media

(0 hours)

Old Media

(24 hours)

after Exposure

of 24 hours

Meas. conc. a.i.

[mg/L]

%

Meas. conc. a.i.

[mg/L]

%

Meas. conc. a.i.

[mg/L]

%

1.25

0.763

0.872

114

0.628

  82

0.0283

4

0.625

0.381

0.453

119

0.387

102

0.0298

8

0.313

0.191

0.209

109

0.165

  86

0.00494

3

Measured Concentrations of Sodium methyl oleyl taurate (C18:1-part) in Aqueous Phase and at Glass Walls of Test Solutions at 72 and 96 hours

Nominal concentration

Aqueous Phase

(Volume: 10 L)

Adsorbed

to Glass

Test item

[mg/L]

a.i.

[mg/L]

Fresh Media

(72 hours)

Old Media

(96 hours)

after Exposure

of 24 hours

Meas. conc.

[mg/L]

%

Meas. conc.

[mg/L]

%

Meas. conc.

[mg/L]

%

1.25

0.763

0.775

102

0.702

  92

0.00848

1

0.625

0.381

0.359

  94

0.397

104

0.00939

2

0.313

0.191

0.178

  93

0.198

104

0.00616

3

Validity criteria fulfilled:
yes
Conclusions:
All effect levels are given based on the nominal concentrations of the test item Sodium methyl oleyl taurate and its active ingredient. The test item was found to be toxic to zebrafish with a LC50 after 96 hours of 1.32 (1.31 – 1.32) mg test item/L, corresponding to 0.810 (0.800 – 0.810) mg a.i./L. At the nominal test item concentration of 0.625 mg/L (0.381 mg a.i./L) no mortality or non lethal effects were observed.
Executive summary:

The acute toxicity of the test item Sodium methyl oleyl taurate(batch no.: DEB4003485)to fish (zebrafish) was determined according to OECD-Guideline for Testing of Chemicals No. 203 (1992) from 2012-12-10 to 2013-02-08 with the definitive exposure phase from 2013-02-04 to 2013-02-08 at Dr.U.Noack-Laboratorien, 31157 Sarstedt, Germany.

A semi-static test with daily renewal of the test media was performed with nominal test item concentrations of 0.156 - 0.313 - 0.625 - 1.25 - 2.5 mg/L. Duration of the test was 96 hours. 7 test organisms were exposed to each concentration and the control. Water quality parameters pH-value, temperature and oxygen-saturation, measured after 0, 24, 48, 72 and 96 h, were determined to be within the acceptable limits.

The concentrations of the active ingredient of Sodium methyl oleyl taurate(C18:1-part) were analytically verified by LC-MS/MS at the start of the exposure (0 h and 72 h) and at the end of the exposure (24 h and 96 h) of all concentration levels and the control. The measured concentrations of active ingredient at the start of the exposure were in the range of 91 to 119 % of the nominal values. The measured concentrations of the active ingredient at the end of the exposure were in the range of 61 to 119 % of the nominal values, indicating that the concentrations of the active ingredient were successfully maintained for the duration of the test. The endpoints were based on the nominal concentrations of the test item Sodium methyl oleyl taurate and additionally given on the nominal active ingredient (Table 1 and Table 2).

Adsorption of the test item to the test vessels were analytically measured (refer to 4.2.4). The measured amount of adsorbed test item was below 10 % in all cases.

IMPORTANT NOTE TO AVOID CONFUSION:

As the registration substance is a UVCB this means that the test item is 100% active ingredient.

All effect levels are given based on the nominal concentrations of the test item

LC-Values (0 – 96 hours)

                   Based on nominal test item concentrations [mg/L]

Test duration
[hours]

LC10

LC20

LC50

24

> 2.50

> 2.50

> 2.50

48

2.32 (1.25* - 2.50**)

2.37 (1.25* - 2.50**)

2.45 (1.25* - 2.50**)

72

1.25* LC10 2.50***

1.25* LC20 2.50***

1.77 (1.25* - 2.50***)

96

1.24 (1.24 – 1.24)

1.27 (1.27 – 1.27)

1.32 (1.31 – 1.32)

LC0 =

Highest test item concentration with
0 % mortality after
96 hours

0.625

LC100 =

Lowest test item concentration with
100 % mortality after
96 hours

2.50

*Concentration with 0 % mortality

**Concentration with 71 % mortality

***Concentration with 100 % mortality

Description of key information

All effect levels are given based on the nominal concentrations of the test item Sodium methyl oleyl taurate and its active ingredient. The test item was found to be toxic to zebrafish with a LC50 after 96 hours of 1.32 (1.31 – 1.32) mg test item/L, corresponding to 0.810 (0.800 – 0.810) mg a.i./L. At the nominal test item concentration of 0.625 mg/L (0.381 mg a.i./L) no mortality or non lethal effects were observed.

Key value for chemical safety assessment

LC50 for freshwater fish:
1.32 mg/L

Additional information