trans-1-(1-oxohexadecyl)-4-[(1-oxohexadecyl)oxy]-L-proline

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:

2013-06-09 to 2013-07-02

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
- Sampling method: Water samples (duplicate per condition) were taken from the control and all surviving test groups at 0 (frexh media), 24 (olf media), 72 (fresh media) and 96 (old media) hours for quantitative analysis.
- Sample storage conditions before analysis: -20°C

Vehicle:

no

Details on test solutions:

Amounts of test item (210, 378, 672, 1176 and 2100 mg) were each serarately added to the surface of 21 liters of dechlorinated tap water to give the 10, 18, 32, 56 and 100 mg/L loading rates respectively. After the addition of the test item, the dechlorinated tap water was stirred by magnetic stirrer using a stirring rate such that a vortex formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixtures allowed to stand for 1 hour. After standing, the WAFs were removed by filtration through a 0.2 µm Sartorius Sartropore filter (initial 1 liter discarded) to give the 10, 18, 32, 56 and 100 mg/L loading rates respectively.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Strain: Oncorhynchus mykiss
- Source: Brow Well Fisheries Limited, Hebden, near Skipton, Yorkshire, UK
- Age at study initiation (mean and range, SD):
- Length at study completion (length definition, mean, range and SD): 4.7 cm (sd=0.1)
- Weight at study completion (mean and range, SD): 1.37 cm (sd=0.11)
- Feeding during test
- Food type: commercial fish food
- Amount: on demand
- Frequency: regularly, the diet was discontinued 24 hours prior testing

ACCLIMATION
- Acclimation period: 12days
- Acclimation conditions (same as test or not): same
- Health during acclimation (any mortality observed): 0 mortality during 7 days prior to the start

QUARANTINE (wild caught)
Fish were maintained in a glass fiber tank with a "single pass" water renewal system.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

determined 3 and 6 hours after the start of exposure and then daily throughout the test until termination after 96 hours.

Hardness:

approximately 140 mg/L as CaCO3

Test temperature:

14°C to 16°C

pH:

between 7.5 and 8.3

Dissolved oxygen:

between 9.1 and 10.2 mg O2/L

Salinity:

not availbale

Nominal and measured concentrations:

nominal concentrations : 10, 18, 32, 56 and 100 mg/L
Measured concentrations : < LOQ

Details on test conditions:

no additional detail

Reference substance (positive control):

no

Duration:

3 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

6 h

Dose descriptor:

LL50

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Mortalities (7) were only observed at 56 mgL

Duration:

24 h

Dose descriptor:

LL50

Effect conc.:

>= 18 - <= 32 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LL50

Effect conc.:

>= 18 - <= 32 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LL50

Effect conc.:

>= 18 - <= 32 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

24 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: with 95% confidence limits of 18 -32 mg/L loading rate WAF.

Details on results:

no additional detail

Results with reference substance (positive control):

not applicable

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test item to the freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated and gave a 96-hour LL50 value of 24 mg/L loading rate WAF with 95% confidence limits of 18-32 mg/L loading rate WAF. The Lowest Observed Effect Loading rate was 32 mg/L loading rate WAF. The no Observed Effect Loading rate was 18 mg/L loading rate.

Executive summary:

Introduction

A study was performed to assess the acute toxicity of the test item to rainbow trout (Oncorhynchus mykiss). The method followed was designed to be compatible with the OECD Guidelines for testing of Chemicals (1992) N° 203, "Fish, Acute Toxicity test" referenced as Method C.1 of Commission Regulation (EC) N° 440/2008.

 

Method

Due to the low aquous solubility and the complex mixture of the test item for the purposes of the test, the test item was prepared as a Water Accommodation Fraction (WAF).

Following a preliminary range-finding test, fish were exposed, in groups of seven, to Water Accomodation Fractions (WAFs) of the test item over a range of nominal loading rates of 10, 18, 32, 56 and 100 mg/L for a period of 96 hours at a temperature of 14°C to 16 °C under semi-static test conditions. The number of mortalities and any sub-lethal effects of exposure in each test and control vessel were determined 3 and 6 hours after the start of exposure and then daily throughout the test until termination after 96 hours.

 

Results

The 96 -hours LL50 based on nominal loading rates was 24 mg/L loading rate WAF with 95% confidence limits of 18 -32 mg/L loading rate WAF. The No Observed Effect Loading rate was 18 mg/L loading rate WAF.

Chemical analysis of the test preparations at 0, 24, 72 and 96 hours showed measured test concentrations of less than the limit of quantification (LOQ) of the analytical mathod employed were obtained which was determined to be 0.078mg/L. This does not infer that no test item was in solution, just that any dissolved test item was at a concentration of less than the LOQ.

Given that toxicity cannot de attributed to a single component or a mixture of components but to the test item as a whole, the resultst were based on nominal loading rates only.

Description of key information

LC50 = 24mg/L 

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

24 mg/L