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

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

December 4, 2015- July 26, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Vehicle:

not specified

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: National Institute for Environmental Studies, Japan
- Age of parental stock (mean and range, SD): Juveniles less than 24"hour-old (female)
- Feeding during test: None
- Food type: Chlorella vulgaris (unicellular green algae)
- Amount:6 mg C (organic carbon)/21/day

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES:

Test type:

semi-static

Water media type:

other: Dechlorinated water

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

Daphinds were observed imediatly after the 48h exposure period for any abnormalities.

Test temperature:

20 ± 1°C

pH:

6.0-9.0 (variation range <1.5), not adjusted

Dissolved oxygen:

3 mg/L or more, not aerated

Nominal and measured concentrations:

100mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: 100-mL glass beaker (Cover: Teflon sheet)
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 100ml
- Aeration: No aeration
- Renewal rate of test solution (frequency/flow rate): every 24h
- No. of organisms per vessel: 5 daphnids.
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates):1
- No. of vessels per vehicle control (replicates):
- Biomass loading rate: 20 daplmids/test group (5 daphnids/vessel)

TEST MEDIUM / WATER PARAMETERS
Dechlorinated water was used as the dilution water. Dechlorinated water was produced 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.
Before the exposure, the total hardness and pH were measured, resulting in 34 mg CaC03/L and 7.5 respectively. These values were within the appropriate range (total hardness: (250 mg CaC03L, pH: 6.0-9.0).


OTHER TEST CONDITIONS
- Adjustment of pH: Not adjusted
- Photoperiod: Fluorescent light, 16-hour light (800 lux or less)/ 8-hour dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Individuals that were unable to swim within 15 seconds after gentle agitation of the test vessel were considered immobile.
Daplmids showing abnormal behavior or appearance (such as discoloration) and floating daphnids on the surface of the water were recorded, if any.


RANGE-FINDING STUDY
- 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 loading daphnids were exposed to the test solution (100 mg/L as 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.

Test Procedure;
The loading daphnids were introduced into each test vessel using a glass pipette at the stat1 of exposure. The water surface was covered with a Teflon sheet to prevent evaporation of the test solutions and volatilization of the test substance. After 24-hour exposure, daphnids were transferred to the newly prepared test solutions using a glass pipette (renewal). The test ended at 48-hour exposure. During the exposure, observations of the loading daphnids, measurements of the test conditions and observations of test solutions were performed according to the following procedure:

1) Observation of the loading daphnids
Frequency: 24-hour (before renewal) and 48-hour exposure.
1: Individuals that were unable to swim within 15 seconds after gentle agitation of the test vessel were considered immobile.
2: Daplmids showing abnormal behavior or appearance (such as discoloration) and floating daphnids on the surface of the water were recorded, if any.

2) Measurement of test conditions and observation of test solutions Frequency: Two sets
Samples:
(i) At the time of preparation: The test solution in the vessel for measuring the test condition of each test group.
(ii) At the time immediately prior to renewal and at the end of exposure: The test solution in one vessel of each test group.
Items: Temperature, D.0., pH, and appearance of the test solution.

Reference substance (positive control):

yes

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.505 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat. (dissolved fraction)

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat. (dissolved fraction)

Details on results:

Measurements of Test Conditions and Observations of Test Solutions
In both the control and Cone. I, the temperature of the test solutions was 20.0-20.1°C (in the range of 20 ± 1°C), the D.O. values were 8.4-8.7 mg/L (3 mg/L or more), and the pH values were 7.4-7.7 (6.0-9.0 where variation was within the range of 1.5), fulfilling the test conditions.
In the appearance of test solutions of both the test groups, no suspended substances, no floating substances and no precipitations were observed, and the color was colorless during exposure.

Concentrations of Test Substance in Test Solutions
The mean measured concentration of Cone.I was 0.505 mg/L. The measured concentrations prior to renewal and at the end of exposure were ahnost eguivalent to those at the preparation.

Observation of Loading Daphnids
In the control, the percentage of immobilisation was 0% at the end of exposure, and no daphnids were observed floating on the surface of the water.
In Cone.I, the percentage of immobilisation was 5%, and no abnormalities were observed
on the daphnids. Since this test was the limit test at the saturated concentration, it was considered that there was no acute toxicity of the test substance on daphnids.

Results with reference substance (positive control):

- Results with reference substance valid? Acute immobilisation tests of the reference substance (potassium dichromate, guaranteed reagent) have been conducted periodically (approximately every six months). The 48-hour median effect concentration (ECSO) values are described below.
Latest result:
0.67 mg/1 (95% confidence limits: 0.59-0.76 mg/L) Exposure period: January 14, 2016 to January 16, 2016

- Limit test: The defiitive test was considered as the limit test.

Validity criteria fulfilled:

yes

Conclusions:

In the control, the percentage of immobilisation was 0% at the end of exposure, and no daphnids were observed floating on the surface of the water.
In Conc.I, the percentage of immobilisation was 5%, and no abnormalities were observed on the daphnids. Since this test was the limit test at the saturated concentration, it was considered that there was no acute toxicity of the test substance on daphnids.

The results given below were calculated from the time-weighted mean of measured concentration and the loading rate.
 
48-hour median effect concentration (EC50):
>0.505 mg/L (95% confidence limits: not available)
48-hour median effect loading rate (EL5O):
>100 mg/L (95% confidence limits: not available)

Executive summary:

Methods

The study was conducted in accordance with the OECD Guideline for Testing of Chemicals No.202"Daphnia sp., Acute Immobilisation Test" (2004). 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.

1) Test Species: Daphnia Magna

2)  Dilution Water: Dechlorinated water

3) Duration: 48 hours

4) Exposure procedure: Semi static (batchwise renewal of the test solution every 24 hours)

5) Number of loading daphnids:   20 daphnids/testgroup (5daphnids/vessel)

6) Temperature: 20 +/-1°C

7) Light: Fluorescent light, 16-hour light (800 lux or less)/8-hour dark

8) Test concentrations:

Test group	Loading rate mg/L (Ratio of test substance to water (in mg/L) used in the preparation of the WSF)
Control	0
Concentraton (Limit test at the loading rate which was the maximum concentration given inthe Test Guideline)	100

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

Results

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

 

48-hour median effect concentration (EC50):

>0.505 mg/L (95% confidence limits: not available)

48-hour median effect loading rate (EL5O):

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

Description of key information

48-hour median effect concentration (EC50):

>0.505 mg/L (95% confidence limits: not available)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.505 mg/L