oligomeric reaction product of 2-hydroxybenzaldehyde with 1-chloro-2,3-epoxypropane and phenol

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:

December 4, 2015- December 11, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances):

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Medaka
- Source:Kitamura-yourijou (388 Gunchiku 12-bancho, Yatsushiro-city, Kumamoto, Japan
- Age at study initiation (mean and range, SD): Less than a half year after hatching.
- Length at study initiation (length definition, mean, range and SD):2.40 cm (2.09- 2.80 cm)
- Weight at study initiation (mean and range, SD):0.110 g (0.075 - 0.170 g)

ACCLIMATION
- Acclimation period:November 30, 2015 - December 7, 2015
- Acclimation conditions: Same as test.
- Type and amount of food during acclimation: TetraMin, TetraWerke diet. Feeding amount: 2% of the body weight/day (approx.) (until 24 hours before exposure)
- Health during acclimation (any mortality observed):Less than 5% mortality observed.

FEEDING DURING TEST: No feeding during test.

Test type:

semi-static

Water media type:

other: Dechlorinated water

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

Test fish were observed at 3, 24, 48, 72 and 96 hours after the start of exposure

Hardness:

The total hardness was 54 mg CaC03/L

Test temperature:

24±1'°C

pH:

77

Dissolved oxygen:

60% or more of the saturation (not aerated)

Nominal and measured concentrations:

100mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: 5-L glass vessel (cover: clear polyvinyl chloride (PVC) plate)
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 5L
- Aeration: not aerated
- Renewal rate of test solution (frequency/flow rate): batchwise renewal of the test solution after every 24-hour exposure.
- No. of organisms per vessel: 10 fish/test group
- No. of vessels per concentration (replicates): 1 vessel/test group
- No. of vessels per control (replicates):1 vessel/test group
- No. of vessels per vehicle control (replicates): 1 vessel/test group
- Biomass loading rate: 10 fish/test group

TEST MEDIUM / WATER PARAMETERS

A water-soluble :fraction* (WSF) was used to expose the test species to the test substance because it was considered that the test substance, which was poorly water-soluble and multi­ component, was partially soluble in water.

Dechlorinated water was used as the dilution water. Dechlorinated water was processed by treating Yokohama city tap water with an activated charcoal filter and sodium thiosulfate. Water quality parameters of the dechlorinated water have been measured periodically (about every 6 months) by MC Evolve Technologies Corporation, and confirmed to be appropriate to use for dilution water (total hardness: 10-250 mg CaC03/L, pH: 6.7-8.5). The total hardness was 54 mg CaC03/L, and pH was 7.7, both we e within the appropriate ranges.

OTHER TEST CONDITIONS
- Adjustment of pH:Not adjusted
- Photoperiod: Fluorescent light, 16-hour light (1000 lux or less)/ 8-hour dark
- Light intensity: 1000 lux or less


TEST CONCENTRATIONS
A range-finding test was conducted at loading rates" below the maximum concentration (100 mg/L) given in the Test Guideline.
The test substance was dissolved in acetone to give stock solution. From this stock solution, the required volume according to the loading rate was collected to each vessel, then acetone was dried up under nitrogen flow, and the dilution water was added. After 48-hour stirring, each solution filtrated with a filter (Millex HA 0.45 µm, Merck Millipore) was used as a test solution for the range-finding test. The stirring time was decided based on the preliminary experiment.

Since there were no significant effects when the test fish were exposed to the test solution (100 mg/Las loading rate) at the range-finding test, the definitive test was determined as a limit test. The concentration for the definitive test was determined as 100mg/L.

Reference substance (positive control):

yes

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.911 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat. (dissolved fraction)

Key result

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat. (dissolved fraction)

Details on results:

Quality and Appearance of Test Solutions
Temperatures ranged from 23.4 to 24.l °C, the D.0. ranged from 6.9 to 8.5 mg/L, and pH ranged from 7.0 to 7.8. In both the control and Cone. 1, the test solutions appeared colorless, and suspended substances, floating substances and precipitated substances were not observed in the test solutions during exposure.

Concentrations of Test Substance in Test Solutions
The mean measured concentration of Cone. 1 was 0.911 mg/L. The measured concentrations prior to renewal (and at the end of exposure) were lower than those at the preparation, namely concentration decrease was observed. Although the details were unknown, the causes for the concentration decrease was considered that the test substance was difficult to be dissolved stably in the test solutions because the test substance was multi­ component substance and the test solutions were the WSF.

Observation of Test Fish.
In both the control and Cone. 1, no dead fish and no abnormal symptoms in living fish were observed.

Sublethal observations / clinical signs:

Validity of theTest

The mortality in the control was 0%. The D.O. concentrations were ranged from 83 to 102% of the saturation, resulting 60% or more of the saturation during the exposure period. Therefore, this test is valid because these results satisfied the requirements of the test.

Validity criteria fulfilled:

yes

Conclusions:

In both the control and Conc. 1, no dead fish and no abnormal symptoms in living fish were observed.

Median Lethal Concentration (LC5O) and Median Lethal Loading Rate (LL5O)
The LC50 and LL50 values are given below, and that of 96-hour exposure are given below.

96-hour LC50: >0.911 mg/L (95% confidence limits: not available)
96-hour LL5O: >100 rng/L (95% confidence limits: not available)

Executive summary:

Method:

The study was conducted in accordance with the OECD Guideline for Testing of Chemicals No.203 "Fish, Acute Toxicity Test" (1992).

A water-soluble fraction*1(WSF) was used to expose the test species to the test substance because it was considered that the test substance, which was poorly water-soluble and multi­ component, was partially soluble in water.

*1: An aqueous fraction containing the dissolved test substance.

1) Test species: Medaka (Oryzias latipes)

2)  Dilution water: Dechlorinated water

3)   Duration: 96 Hours

4)  Exposure procedure: Semi-static (batchwise renewal of the test solution after every 24-hour exposure)

 5)  Biomass loading: 10 fish/test group

6)  Temperature: 24±1°C

7)   Light: Fluorescent light, 16-hour light (1000 lux or less)/8-hour dark

8)  Test concentrations:

Test group	Loading rate*2 (mg/L)
Control	0
Concentration 1*3	100

 

*2: Ratio of test substance to water (in mg/L) used in the preparation of the WSF

*3: Limit test at the loading rate which was the maximum concentration given in the Test Guideline

9)  Analytical method: Liquid chromatography/mass spectrometry (LC/MS)

Results

The results given below were calculated from the time-weighted mean of measured concentrations and the loading rate.

 

96-hour median lethal concentration (LC5O):

>0.911 mg/L (95% confidence limits: not available)

96-hour median lethal loading rate (LL5O):

>100 mg/L (95% confidence limits: not available)

Description of key information

96-hour LC50:       >0.911 mg/L (95% confidence limits:  not available)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.911 mg/L