Fatty acids, C14-18 and C16-18 unsatd., compds. with triethanolamine

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:

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)

Specific details on test material used for the study:

Batch no.: 2016100027

Analytical monitoring:

no

Vehicle:

yes

Remarks:

acetone was used as solvent

Details on test solutions:

Acetone was used as solvent. A solvent control was run along with an untreated control. The maximum acetone concentration on test solutions was identical in all treatment (except untreated control) and did not exceed 0.01 % (v/v).
Stock solution A to E were prepared prior test start. Stock solution B to E were made diluting stock solution A.
1:10000 dilution of these stock solutions were used to result in the anticipated test item concentrations in test solution.
All stock solutions and their dilutions were prepared using acetone and in graduated flasks as follows:
- weighting 530 mg test item into a graduated flask, filling up to 10 mL (stock solution A = 53000 mg/L test item)
- stock solution B = 29404 mg/L test item: 2.774 mL stock solution A were filled to 5 mL
- stock solution C = 16358 mg/L test item: 1.543 mL stock solution A were filled to 5 mL
- stock solution D = 9084.2 mg/L test item: 0.857 mL stock solution A were filled to 5 mL
- stock solution E = 5045.6 mg/L test item: 0.476 mL stock solution A were filled to 5 mL
Test concentration: 0.50, 0.91, 1.64, 2.94, 5.30 mg/L test item nominal.
Test solutions were prepared directly in test vessels. After adding of 1 mL stock solution, test solutions were stirred thoroughly. Each test vessel contained a total volume of 10 L of test solution.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

rainbow trout - Oncorhynchus mykiss
mean length: 5.00 +/- 0.19 cm
mean weight: 1.09 +/- 0.17 g
mean loading: 0.76 g fish/L
Time for acclimatisation: 55 days
Daily light-dark-cycle: 16 hours light:8 hours dark
Oxygen concentration: at least 80 % of air saturation value
Temperature: 15.0 °C
Feeding: daily (fish food), until 24 hours before test start no feeding during the test
During the acclimatisation period, fish were not exposed to the test item. Mortality of fish during the acclimatisation period was 0.00 %. 7 days before test start, fish were transferred into fresh test medium. Equal environmental conditions as used during the test were given.

Test type:

static

Water media type:

other: reconstituted water according to Guideline 203

Limit test:

no

Total exposure duration:

96 h

Hardness:

Water hardness: 10 - 250 mg CaCO3/L
water is aerated until oxygen saturation is achieved and then stored for about two days without further aeration before use

Test temperature:

14.3 – 15.5°C, recorded continuously

pH:

pH of the test medium before start: 7.58
pH values of test solutions: 7.44 - 7.88

Dissolved oxygen:

Oxygen concentration: 8.45 to 8.95 mg/L O2, corresponding to 86.4 - 91.5 % saturation of the maximum saturation.
Test medium was aerated until oxygen saturation was achieved, then stored for quarantine days without aeration before use. Permanent aeration was employed during the test.

Nominal and measured concentrations:

Test concentration: 0.50, 0.91, 1.64, 2.94, 5.30 mg/L test item nominal

Details on test conditions:

Test vessel (pool): stainless steel container (approx. 13 L volume)
Test volume : 10 L
Number of fish/vessel (= replicate): expsoure of 7 fish/replicate
Number of replicates/treatment: 1
Loading: maximum loading of 1.0 g fish/L

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 5.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: water solubility = 5.3 mg/L

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

> 5.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: water solubility = 5.3 mg/L

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 5.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: water solubility = 5.3 mg/L

Details on results:

Under given conditions, Triethanolamine trioleate did cause mortality exclusively at the highest employed test concentration of 5.30 mg/L test item nominal. An effect of 28.57 % mortality at test end was observed. Significant effects were determined as LOEC > 5.30 mg/L test item nominal. The corresponding NOEC was ≥ 5.30 mg/L test item nominal.

Reported statistics and error estimates:

The determination of lowest observed effects concentration (LOEC) was carried out by hypothesis testing using Fisher`s Exact Binomial Test (p ≤ 0.05, one-sided greater) for binomial distributed data. Lethal concentrations LCx were determined by concentration-response modelling (Probit analysis, maximum likelihood). Statistical evaluation was carried out using ToxRat Professional 3.2.1 (RATTE, 2015).
The statistical test determined a LOEC higher than the highest employed test item concentration. Different settings at the statistical software could not improve this situation. For effect concentration estimates LCx, confidence intervals could only be partially be determined. The LC50 was determined outside the range of applied concentrations (LC50 = 5.71 mg/L test item nominal).

Sublethal observations / clinical signs:

Observations

	Test item (mg/L)	Test item (mg/L)	Test item (mg/L)	Test item (mg/L)	Test item (mg/L)	Test item (mg/L)
	control 2	0.50	0.91	1.64	2.94	5.30
Time after application	mortality (%)	mortality (%)	mortality (%)	mortality (%)	mortality (%)	mortality (%)
3 h	0.00	0.00	0.00	0.00	0.00	0.00
6 h	0.00	0.00	0.00	0.00	0.00	0.00
24 h	0.00	0.00	0.00	0.00	0.00	0.00
48 h	0.00	0.00	0.00	0.00	0.00	14.29
72 h	0.00	0.00	0.00	0.00	0.00	28.57
96 h	0.00	0.00	0.00	0.00	0.00	28.57

Validity criteria fulfilled:

yes

Conclusions:

In the OECD 203 test, the test item caused mortality exclusively at the highest employed test concentration of 5.30 mg/L test item nominal (maximun water solubility). An effect of 28.57 % mortality at test end was observed. Significant effects were determined as LOEC > 5.30 mg/L test item nominal. The corresponding NOEC was ≥ 5.30 mg/L test item nominal.

Description of key information

The test item caused mortality exclusively at the highest employed test concentration of 5.30 mg/L test item nominal (maximun water solubility). An effect of 28.57 % mortality at test end was observed.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

5.3 mg/L

Additional information

LC50 > 5.3 mg/L/96h. Solubility of the substance: 5.3 mg/L