Dihydro-3-(tripropenyl)furan-2,5-dione

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-02-11 to 2013-02-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study, no deficiencies

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Tripropenyl succinic acid

Cas Number:

147384-84-7

Molecular formula:

R-C4H6O4 , whereas R=C8-C10-alkyl-(branched, unsaturated)

IUPAC Name:

Tripropenyl succinic acid

Details on test material:

- Name of test material (as cited in study report): Tripropenyl succinic acid
- Physical state: Viscous liquid, pale brown
- Analytical purity: 99.3 %
- Purity test date: 2013-03-04
- Lot/batch No.: 12-SK002
- Expiration date of the lot/batch: January 2014
- Stability under test conditions: Not applicable
- Storage condition of test material: Room temperature, protected from light, in original container

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: A limit concentration of 100 mg/L was tested.
- Sampling method: Analytical evaluation of Tripropenyl succinic acid was carried out via LC-MS/MS at test start and test end.
- Sample storage conditions before analysis: All test samples were stored at room temperature before preparation and until start of analysis.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Directly weighing of the stock solution. The test item dispersion was stirred in a brown glass flask for
24 hours with a magnetic stirrer at room temperature.
- Eluate: Reconstituted soft water
- Differential loading: None, limit test concentration of 100 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: 2.4 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.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test conditions

Hardness:

Total Hardness at day 0: 4 mg CaCO3/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: Reconstituted fresh water (soft) acc. to ASTM E 729, modified to a total hardness of approx. 4 mg CaCO3/L*, by reducing the content of Ca2+ and Mg2+ about 90 %
Composition of Dilution Water
Component Concentration [mg/L]
NaHCO3 48.0
CaSO4  2 H2O 3.00
MgSO4  7 H2O 6.14
KCl 2.00
pH-value 7 - 8

- 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
100 0 0 0 0
Control 0 0 0 0

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

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:

The concentrations leading to 0 and 100 % mortality (LC0 and LC100) after 96 h were determined directly from the test results if applicable. LC10,20,50-values have not to be calculated in a limit test.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Observations in the Test Vessels

Nominal test item concentration [mg/L]	Effect*	Test Duration [h]
		24	48	72	96
100	(1)	7/7	7/7	7/7	7/7
Control	(1)	7/7	7/7	7/7	7/7

                              *)  The numbers in brackets correspond to the following observation:

                   (1)  = behaviour

Table:    Cumulative Mortality [%] in the Test Vessels

Nominal test item concentration [mg/L]	Test Duration [h]
	24	48	72	96
100	0	0	0	0
Control	0	0	0	0

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In this study Tripropenyl succinic acid was found to cause no toxic effects to Zebrafish after 96 hours when tested with the nominal limit test concentration of 100 mg/L.
The LC50 after 96 hours is > 100 mg Tripropenyl succinic acid/L. All effect levels are given based on the nominal test item concentration of the test item.

Executive summary:

The acute toxicity of the test item Tripropenyl succinic acid (batch no.: 12-SK002) to fish (zebrafish) was determined according to OECD-Guideline for Testing of Chemicals No. 203 (1992) from 2013-02-11 to 2013-02-15, at Dr.U.Noack-Laboratorien, 31157 Sarstedt, Germany. A static test with a limit test concentration of 100 mg/L was performed.

The study was done in reconstituted soft water (acc. to ASTM E 729), modified to a hardness of 4 mgCaCO₃/L, in order to avoid precipitations of the test item in the presence of higher concentrations of calcium and magnesium ions.

The duration of the test was 96 h. Seven test organisms were exposed to the limit 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.

Analytical evaluation of the limit concentration of Tripropenyl succinic acid and the control was carried out via LC-MS/MS from freshly prepared test media (0 h) and old media (96 h). The measured concentration of Tripropenyl succinic acid in the freshly prepared medium was 120 mg/L. In the old test medium the measured concentration was 110 mg/L. For details of the analytical results please refer to part 8. All effect levels are given based on the nominal concentration of the test item Tripropenyl succinic acid

LC-Values with 95 % Confidence Intervals

                         based on nominal test item concentration [mg/L]

Test duration [h]	LC 50	p = 95 %
24	> 100	n.a.
48	> 100	n.a.
72	> 100	n.a.
96	> 100	n.a.
LC100 = Lowest test item concentration with 100 % mortality after 96 h	> 100
LC0 = Highest test item concentration with 0 % mortality after 96 h	100