Dodecanoic acid, ester with 1,2,3-propanetriol

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 October – 27 October 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Remarks:

Ministerium für Arbeit, Integration und Soziales des Landes Nordrhein-Westfalen, Düsseldorf, Germany

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 1.25, 2.5, 5.0, 10.0 and 20.0 mg/L (nominal)
- Sampling method: Samples of control and test solution were taken at test start, at each medium renewal of aged and renewed test solution, and at test end after 96 h. Samples of fresh test solutions were taken from the test solution preparations after distributing it to the replicates. For aged test solutions samples were taken from the test vessels.
- Sample storage conditions before analysis: Samples were stored until analysis under given storage conditions to maintain the test item stable to the best knowledge until analysis, but were measured at least seven days after sampling. All samples were discarded after finalization of the study.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Before test start and before each media renewal a stock solution was prepared by weighing in an adequate amount of test item and transferring it to Cu-reduced dilution water. Both highest test concentrations (20 and 10 mg/L) were prepared separately. Since turbidity was observed in the test solutions of 20 mg/L, the 10 mg/L solution serves as stock solution for all lower test concentrations, which were prepared separately (no serial dilution). Directly after addition of the test item the test solutions of 20 mg/L and 10 mg/L were sonicated for one hour in a sonication bath about 30°C. Afterwards the solutions were stirred for further two hours at room temperature. The test solutions were freshly prepared before test start and at each medium renewal.
- Controls: yes
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): turbidity at 20 mg/L test concentration

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish
- Source: Test facility bred
- Length at study initiation (length definition, mean, range and SD): 1.8 – 2.2 cm
- Weight at study initiation (mean and range, SD): 0.034 - 0.071 g (mean: 0.047 ± 0.014 g)
- Other: Before beginning of the study, a subsample of seven randomly chosen fish from the fish batch used in the test was measured for length and weight.

ACCLIMATION
- Acclimation period: The fish were held for at least 12 days prior to the test under generally equivalent water quality and illumination conditions to those proposed for use in the test.
- Acclimation conditions (same as test or not): same as test
- Type and amount of food during acclimation: Fish were fed ad libitum throughout the holding period with live brine shrimp (Artemia spp.) nauplii and ground flake food TetraMin® (Tetra Werke, Melle, Germany).
- Feeding frequency during acclimation: daily
- Health during acclimation (any mortality observed): Only healthy fish without diseases and abnormalities were used in the study. Mortality of the batch was less than 5 % in the week preceding the start of the study.

FEEDING DURING TEST : no

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

23.0 - 23.2°C

pH:

8.16 - 8.60

Dissolved oxygen:

oxygen saturation: 93 % - 109 %

Nominal and measured concentrations:

nominal: 1.25, 2.5, 5.0, 10.0 and 20.0 mg/L
measured (TWA): 0.257, 0.498, 0.973, 2.304 and 4.627 mg/L (20.6, 19.9, 19.5, 23.0 and 23.1% of nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: full glass beakers of 3 L
- Material, size, headspace, fill volume: 2 L of test solution fill volume
- Aeration: yes, the test solutions were slightly aerated via glass capillaries.
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.111 g/L test medium

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Cu-reduced dilution water
- Intervals of water quality measurement: Oxygen concentration, pH-value and temperature was measured directly before adding the fish and afterwards once per day.

OTHER TEST CONDITIONS
- Photoperiod: light/dark cycle 12/12 hours

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): The mortality and sublethal effects were determined after 24 h, 48 h, 72 h and 96 h. Mortality and abnormal behavior (e.g. loss of equilibrium, swimming behavior, respiratory function, and altered pigmention) was recorded after 24 h, 48 h, 72 h and 96 h. Dead animals (no visible movements, e.g. gill movement) were eliminated from the vessels as soon as they were discovered.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.0
- Range finding study
- Test concentrations: 0.4, 4.0 and 40 mg test item/L
- Results used to determine the conditions for the definitive study: 40 mg/L was turbid while the test concentrations of 4.0 and 0.4 mg/L were clear. At the end of the test 100% mortality was observed in the highest test concentration of 40 mg/L. There were no effects on the test organisms in the two lower test concentrations of 4.0 and 0.4 mg/L.
- Other: The range finder was performed with five individual specimens per control and per treatment. The test was performed under semi-static conditions for 96 hours. Renewal of test solutions was conducted daily.

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

4.3 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC10

Effect conc.:

3.7 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: After 3 h biological effects could not be detected. After 24 h seven fish at 20.0 mg/L showed an inactive swimming behavior and tail-heavy position. One fish swam on the water surface and six fish showed an orientation on the ground level. One fish showed a dark coloration of its skin. After 48 h five fish died at the highest test concentration. The two remaining fish showed an uncoordinated swimming behavior and an orientation on the ground level at 20.0 mg/L, however recovered after 72 h. No more mortality could be detected during the test duration.
- Other biological observations: No biological effects could be detected after 72 h and 96 h at all surviving fish.
- Mortality of control: no mortality
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The test solution of the highest test concentrations had a milky cloudy appearance during the test duration

Sublethal observations / clinical signs:

The measured concentrations of the test item in the freshly prepared test solutions were between 80.0 % and 101.9 % of nominal loading during the test duration. In the aged test solution the measured concentrations of the test item were between < LOQ and 37.6 %. The mean measured concentrations of fresh solutions were in the range of 83.8 % and 88.6 % of nominal, and of aged solution in the range of 5.6 % and 9.4 % of nominal. These values resulted in TWAs in the range of 19.5 % and 23.1 % of nominal concentrations.

The test solution of the highest test concentrations had a milky cloudy appearance during the test duration. Since turbidity was observed in the highest test concentration of 20 mg a.s./L it could not be excluded that the effects in the highest concentration are based on physical effects. 

Table2:         Mean Dodecanoic acid, ester with 1,2,3-propanetriol concentrations in [mg/L] and in [%] of nominal

	Dodecanoic acid, ester with 1,2,3-propanetriol
Nominal concentration [mg/L]	Fresh test solutions			Aged test solutions		TWA
	[mg/L]	[% of nominal]		[mg/L]	[% of nominal]	[mg/L]	[% of nominal]
Control	< LOQ		-	< LOQ	-	< LOQ	-
1.25	1.047		83.8	0.070	5.6	0.257	20.6
2.5	2.097		83.9	0.144	5.7	0.498	19.9
5.0	4.323		86.5	0.273	5.5	0.973	19.5
10.0	8.551		85.5	0.869	8.7	2.304	23.0
20.0	17.718		88.6	1.887	9.4	4.627	23.1
LOQ = 1.0 µg/L

Table3:         Cumulative mortality during the test period of 96 h

	Nominal concentration Dodecanoic acid, ester with 1,2,3-propanetriol [mg/L]
	Control	1.25	2.5	5.0	10.0	20.0
Fish introduced	7	7	7	7	7	7
h of application	Cumulative mortality [n]
3	-	-	-	-	-	-
24	-	-	-	-	-	-
48	-	-	-	-	-	5
72	-	-	-	-	-	5
96	-	-	-	-	-	5

-: no mortality detected

 

Table 4:         Overview Survival: Overview over the effects on survival in Danio rerio at 96 h

Treatm.[mg/L]	Total Introduced	Survived	Dead	% Mortality
Control	7	7	0	0.0
0.257	7	7	0	0.0
0.498	7	7	0	0.0
0.973	7	7	0	0.0
2.304	7	7	0	0.0
4.627	7	2	5	71.4

The pre-selected Abbott-correction is not needed since the control response is 0.0%.

 

Table 5: Validity criteria for OECD 203.

Criterion from the guideline	Outcome	Validity criterion fulfilled
The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test	0% moratality was recorded in the control after 96h	yes
The dissolved oxygen concentration must have been at least 60 per cent of the air saturation value throughout the test	The oxygen saturation in all test vessels was between 93 % and 109 %	yes
There must be evidence that the concentration of the substance being tested has been satisfactorily maintained, and preferably it should be at least 80 per cent of the nominal concentration throughout the test. If the deviation from the nominal concentration is greater than 20 per cent, results should be based on the measured concentration.	The measured concentrations of the test item in the freshly prepared test solutions were between 80.0 % and 101.9 % of nominal loading during the test duration. In the aged test solution the measured concentrations of the test item were between < LOQ and 37.6 %. The mean measured concentrations of fresh solutions were in the range of 83.8 % and 88.6 % of nominal, and of aged solution in the range of 5.6 % and 9.4 % of nominal. Thus, results were based on time weighted average (TWA) concentrations.	yes

 

 

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.

Description of key information

LC50 (96h) = 4.3 mg/L (TWA) for Danio rerio (OECD 203)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

4.3 mg/L

Additional information

One study investigating the short-term toxicity of Dodecanoic acid, ester with 1,2,3-propanetriol (CAS 37318-95-9) to Danio rerio is available. This semi-static test was conducted according to OECD 203 under GLP conditions with daily renewal of test solution. Danio rerio was exposed for 96 h to five test concentrations of 1.25, 2.5, 5.0, 10.0 and 20.0 mg/L (nominal). Analytical measurements were conducted for all test concentrations at test start, at each medium renewal of aged and renewed test solution, and at test end after 96 h. The mean measured concentrations of fresh solutions were in the range of 83.8 % and 88.6 % of nominal, and of aged solution in the range of 5.6 % and 9.4 % of nominal. These values resulted in TWAs in the range of 19.5 % and 23.1 % of nominal concentrations resulting in 0.257, 0.498, 0.973, 2.304 and 4.627 mg/L (meas. TWA) test concentrations. Thus, the effect concentrations were based on time weighted average (TWA) concentrations. Mortality was observed in the highest tested concentration (4.627 mg/L meas .TWA). Since turbidity was observed in the highest test concentration (20 mg/L) it could not be excluded that the effects in the highest concentration are based on physical effects.But as this is not proved the highest test concentration was included in the calculation of the effect value.Hence, an LC50 (96h) of 4.3 mg (meas. TWA) was determined.