Reaction mass of 1,3-dioxane-5,5-dimethanol and 1,3-Propanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]-

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

02 - 06 February 2009

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

Analytical monitoring:

yes

Details on sampling:

Portions of the test solution were sampled before test and after 24 hours for analysis in the Sponsor's laboratory

Vehicle:

no

Details on test solutions:

Test media were prepared daily, 100 g of POLYOL PX was dissolved per one litre of aerated dilution water as a stock solution

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
Noted in report (page 8), but no explicit details of test organisms provided

ACCLIMATION
- Acclimation period: Fish were placed in an internal quarantine for 14 days prior to the commencement of the test.

The fish ranged between 0.15 and 0.51 g and the length ranged between 26 and 33 mm.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not applicable

Hardness:

250 ± 25 mg calcium carbonate/L

Test temperature:

22 ± 1°C

pH:

Measured values - 6.7 - 7.9

Dissolved oxygen:

Measured values - 77 - 101%

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal: Control, 400, 800, 1600, 3200 & 6400 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open / closed Not reported
- Material, size, headspace, fill volume: Not reported
- Aeration: Not reported
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate): every 24 hours
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dilution water
The composition of the dilution water:
1. Calcium chloride
Dissolve 11.76 g CaCl2x 2H2Oin water and dilute to 1 litre.
2. Magnesium sulphate
Dissolve 4.93 g MgS04 x 7H2Oin water and dilute to 1 litre.
3. Sodium bicarbonate
Dissolve 2.59 g NaHC03 in water and dilute to 1 litre.
4. Potassium chloride
Dissolve 0.23 g KCI in water and dilute to 1 litre.
Mix 1-4in water and dilute to 40 litres.
- Total organic carbon: Not reported
- Particulate matter: Not reported
- Metals: Not reported
- Pesticides: Not reported
- Chlorine: Not reported
- Alkalinity: Not reported
- Ca/mg ratio: Not reported
- Conductivity: Not reported
- Culture medium different from test medium: No
- Intervals of water quality measurement: Not reported but portions of the test solution were sampled before test and after 24 hours for analysis in the Sponsor's laboratory.


OTHER TEST CONDITIONS
- Adjustment of pH: with HCL or NaOH
- Photoperiod: 16h light 8h dark
- Light intensity: Not reported but test performed in 'normal laboratory lighting'


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality at 0, 3, 24, 48, 72 and 96 hours
Bodyweights and body length at termination (test groups vs control group)
Visible abnormalities: e.g. darker pigmentation, loss of equilibrium, disturbance in swimming behaviour or increased respiration rate

Reference substance (positive control):

yes

Remarks:

Disodium heptachromate

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

800 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1 800 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: loss of equilibrium, disturbance in swimming behaviour in all surviving fish at 1600 mg/L
- Other biological observations: dark pigmentation, increased respiration rate in all surviving fish at 1600 mg/L
- Mortality of control: None
- Other adverse effects control: None

Results with reference substance (positive control):

The report states the following: The fish batch was qualified for testing purposes with disodium heptachromate. The 24 hour LC50 value fulfilled the aceptable criterion of 200-400 mg/L (SS 028162).

Reported statistics and error estimates:

Not detailed

Sublethal observations / clinical signs:

Mortality in the test and control group during the 96 hours of exposure.

Concentration (mg/L)	0	3	24	48	72	96	Total
Control	0	0	0	0	0	0	0
400	0	0	0	0	0	0	0
800	0	0	0	0	0	0	0
1600	0	0	0	0	3	3	3
3200	0	0	7	7	7	7	7
6400	0	0	7	7	7	7	7

 

Validity criteria fulfilled:

yes

Conclusions:

A 96 hour acute toxicity study in zebra fish (Danio rerio) was conducted to assess the toxicity of Polyol PX. The 96 hour LC50 was calculated to be 1800 mg/L (based nominal exposure concentrations)

Executive summary:

In a 96-h acute toxicity study, zebra fish (Danio rerio) were exposed to Polyol PX at nominal concentrations of 0 (control), 400, 800, 1600, 3200 & 6400 mg/L under semi-static conditions. The 96-h LC50 was 1800 mg a.i./L. The NOEC value, based on mortality, was 800 mg /L. Sublethal effects were not observed. Based on the results of this study, Polyol PX would be classified as practically nontoxic to zebra fish in accordance with the classification system of the U.S. EPA.

This toxicity study is classified as supplementary and satisfies the guideline requirement for fish, short-term toxicity study.

Results Synopsis

Test organism size/age (mean wet weight or length):
Test Type (Flowthrough, Static, Static Renewal):

LC50: 1800 mg/L
NOEC: 800 mg/L
Endpoint Effected: mortality

 

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 15 NOV 2021 to 19 NOV 2021

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:

yes

Remarks:

On day 4 of the test, the vessels were inspected only once per day in the morning. Because the test was started in the morning and was finished after 96 hours, and no mortality was observed, this can be stated as uncritical for the outcome of the study.

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

Commission Regulation (EC) No. 440/2008, Method C.1: “Acute Toxicity for Fish”
adopted 30. May 2008

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 50-1000 mg/L
- Sampling method: For the calibration, a dilution series in water was prepared, using a derivatized stock solution. 10 mL of the test item in matrix or blank matrix were evaporated via nitrogen evaporation system. The residue was silylated by dissolving in 0.8 mL pyridine and 0.2 mL BSTFA via ultrasonic bath and heating at 101.0 – 106.9 °C for 15 minutes.
- Sample storage conditions before analysis: The test item was stored in a closed vessel at room temperature (20 ± 5 °C), in a dry area protected from direct sunlight and the elements.

Vehicle:

no

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Strain: not reported
- Source: The original animals were obtained in 2019 from the University Heidelberg and used for breeding and production of eggs. The offspring of the original animals was used in the test. During the last 7 days, mortality was below 5 %.
- Age at study initiation: sexually immature young fish of the same age (specific age not reported)
- Length at study initiation: mean (1.8 cm); range (1.4 - 2.0 cm); SD (0.23 cm)
- Weight at study initiation: mean (108.68 mg); range (44.4 - 190.2 mg); SD (52.33 mg)
- Method of breeding: not reported
- Maintenance of the brood fish: not reported

ACCLIMATION
The test fish were kept following SOP 115 005 04 edition 9, valid from 26. Apr. 2021 under following conditions:
Medium: reconstituted freshwater
Food: two times each working day; warm-water fish food and / or artemia
Photo period: 12/12 hours, using LED’s
Temperature: 25 ± 2 °C
Before the fish were set into the aquaria, temperature adaptation has taken place. Temperature, pH and O2-concentrations were recorded every 24 ± 1 hours in the new and the old medium. Hardness, Nitrate and TOC were measured at the beginning of the test in the Blank control.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Remarks on exposure duration:

Observations were made after 1 h, 4 h and 24 ± 1 h after the start of the exposure. On days 2-3 of the test, all vessels were inspected twice per day. On day 4 of the test, all vessels were inspected only once per day in the morning.

Post exposure observation period:

None

Hardness:

53.4 mg CaCO3/L (measured only on Day 0 in Blank control)

Test temperature:

22.6 – 23.8 °C

within one test, temperature did not vary more than 2 °C
between test vessels and between successive days

pH:

7.5 - 7.7

Dissolved oxygen:

8.1 – 8.6 mg/L

Nominal and measured concentrations:

Nominal: 0 (control), 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquaria - total volume 12 L
- Aeration: Accomplished with glass tubes
- Renewal rate of test solution: Every 24 ± 1 hours (semi-static)
- No. of organisms per vessel: 7
- No. of vessels per concentration: 1
- No. of vessels per control: 1
- Biomass loading rate: 1 fish/L

TEST MEDIUM / WATER PARAMETERS
- Intervals of water quality measurement: The content of the test item in the test solutions was measured at the beginning, at each medium renewal (in the old and new test solutions) and at the end of the test. In the control solution the content of the test item was measured at the beginning and at the end of the test.

OTHER TEST CONDITIONS
- Adjustment of pH: None
- Photoperiod: 12/12 hours using LED’s
- Light intensity: 426 lux (stated range in the study plan: 400 – 800 lux) Light intensity is lower than stated in the guideline (540 – 1000 lux). The range 400 – 800 lux is the light intensity in the husbandry protocol. Because husbandry is successful since many years and the fish are in very good health, this deviation can be stated as uncritical.

TEST CONCENTRATIONS
- Range finding study: The concentration to be tested was based on the result of a non-GLP pre-test.
- Test concentrations: 0 (control), 100 mg/L (nominal concentration)

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

NOEC

Remarks:

Limit test

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Remarks:

Limit test

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Reported statistics and error estimates:

The tested concentration showed no toxicity. Therefore, no statistical evaluation was performed and the biological result was reported as a range.

Validity criteria fulfilled:

yes

Remarks:

No mortality occurred in the blank control. The concentration of dissolved oxygen was at least 8.1 mg/L or 93.9 % throughout the test. At the start and at the end of the test, and at each medium renewal the content of the test item was determined.

Conclusions:

One valid experiment was performed.
The toxicity towards Danio rerio was tested using a semi-static test design.
The study was performed as a limit test at the concentration 100 mg/L (nominal). The tested concentration showed no toxicity. No fish died or showed any signs of abnormal behavior in the control and the treatment.

Executive summary:

In a 96-h acute toxicity study, zebrafish (Danio rerio) were exposed to Polyol PX at nominal concentrations of 0 (control) and 100 mg/L under semi-static conditions. The 96-h LC50 was > 100 mg/L. Sublethal effects were not observed. Based on the results of this study, Polyol PX would be classified as practically nontoxic to fish in accordance with the classification system of the U.S. EPA.

This toxicity study is classified as acceptable and satisfies the guideline requirement for “short-term toxicity to fish” study.

Description of key information

The LC50 for Danio rerio based on mortality was determined to be >100mg/l based on nominal test concentrations. Nominal test concentrations were confirmed analytically.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

> 100 mg/L

Additional information

Two studies are available investigating the short-term toxicity of Polyol PX to fish.

 

The key study, a klimisch 1 study assessing toxicity to Danio rerio determined an LC50 for mortality of >100mg/l based on nominal concentrations (analytically confirmed).

 

Another klimisch 1 study is used as supporting infomation and also investigated the toxicity towards Danio rerio. This study determined a 96h LC50-value of 1800 mg/L for mortality. The concentration was not analytically confirmed.