Reaction products of 2,2-dimethylpropane-1,3-diol with 1-choloro-2,3-epoxypropane

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

Council Regulation (EC) No 440/2008, Annex Part C, C.1., May 30, 2008
OECD Guideline for Testing of Chemicals, No. 203, adopted 18 June 2019
OCSPP 850.1075 of the United States Environmental Protection Agency (EPA), October 2016
OECD guidance document No. 23 (ENV/JM/MONO(2000)6/REV1; 8 February 2019)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Chemical name: Reaction products of 2,2-dimethylpropane-1,3-diol with 1-chloro-2,3-epoxypropane
CAS No.: 17557-23-2
EC / List no.: 701-333-0

Analytical monitoring:

yes

Details on sampling:

Three samples were taken from the ecotoxicological medium of different concentration level.
Samples were properly diluted with ultrapure water.
Diluted samples were stored at 2 - 8 oC until analysis.
Dilution of the ecotoxicological medium samples of the Test Item
Nominal concentration Dilution factor Dilution scheme
Control 40 25 µL → 1 mL
10 mg/L 50 20 µL → 1 mL

Vehicle:

no

Details on test solutions:

The test solution used in the test was prepared at each water renewal period by mechanical dispersion without using any solubilising agent, as the substance is water soluble.
An appropriate amount* of test item was dissolved in the dilution water (ISO medium) in order to obtain the nominal concentration of 10 mg/L at each water renewal period.
After the formulation procedure the test animals were immediately introduced into the test solutions at each water renewal period.
* weighted amounts:
1st renewal: 0.0102 g test item dissolved in 1020 mL ISO medium
2nd renewal: 0.0108 g test item dissolved in 1080 mL ISO medium
3rd renewal: 0.0108 g test item dissolved in 1080 mL ISO medium
4th renewal: 0.0104 g test item dissolved in 1040 mL ISO medium

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: Zebrafish (Danio rerio)
Origin: Toma István e.v.
Hungary, 2146 Mogyoród, Gödöllői u. 41/B.
Number of animals: There were 10 animals in the treated and control group respectively (no replicates were used).
Age and size of animals: Juveniles were used. The total body length of each fish used was within the range of 1-2 cm. The total body weight of the test animals per groups was recorded on the day 0 (at the start of the test). The loading of the test aquaria was calculated (g fish / liter test solution). The loading did not exceed 0.8 g/L.
Sex: Both female and male (were not separated).
Animal health: Fish were bred in a well-known fish farm, under disease- and parasite-controlled conditions. Apparently healthy animals were used in the test.

Test type:

semi-static

Water media type:

freshwater

Remarks:

ISO medium, prepared according to the recommendation of OECD guideline 203

Limit test:

yes

Total exposure duration:

96 h

Hardness:

The reconstituted water (ISO medium) has an approximate theoretical total hardness of 249 mg/L (as CaCO3), calculated based on the concentration of calcium and magnesium ions in ISO medium.

Test temperature:

Temperature: The measured temperatures were within a range of 22.6 – 23.4°C in the test aquariums (the maximum deviation did not exceed ± 1°C) during the test.

pH:

pH: The test was carried out without adjustment of pH. The measured pH values were in the range of 7.76 – 8.09 during the test.

Dissolved oxygen:

Oxygen Concentration: The dissolved oxygen concentration was in the range of
75.4 – 88.1 % of the air saturation value at the temperature used. Aeration of test solution or control was not necessary during the experiment.

Nominal and measured concentrations:

measured

Details on test conditions:

Aquariums were kept during the test in a climate chamber with controlled environmental conditions. The water temperature, pH value and dissolved oxygen concentration were determined at the start and at the end of each water renewal period in the test concentration and in the control.
Test type: Semi-static test. Based on the analytical results obtained during the acute daphnia test (Study number: 555-202-6264) the test item was not expected to be stable for the duration of the experiment in ISO Medium, therefore a semi-static test was performed using 24-h water renewal periods.
Test Duration: 96 hours
Loading: The loading of the test aquaria did not exceed 0.8 g fish/L.
Temperature: The measured temperatures were within a range of 22.6 – 23.4°C in the test aquariums (the maximum deviation did not exceed ± 1°C) during the test.
Oxygen Concentration: The dissolved oxygen concentration was in the range of
75.4 – 88.1 % of the air saturation value at the temperature used. Aeration of test solution or control was not necessary during the experiment.
pH: The test was carried out without adjustment of pH. The measured pH values were in the range of 7.76 – 8.09 during the test.
Light: Photoperiod: 16 h light : 8 h dark; Intensity: 565 lux (measured at the start of the test)
Hardness: The reconstituted water (ISO medium) has an approximate theoretical total hardness of 249 mg/L (as CaCO3), calculated based on the concentration of calcium and magnesium ions in ISO medium.
Feeding: The fish were not fed during the test.

Test Units

Type and Size: Glass aquariums with appropriate size for holding the fish up to the loading of 0.8 g fish/L.
Identification: Each test unit was uniquely identified with study number and test group.

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

> 7.6 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: no mortality or any sub-lethal effect were observed.

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

ca. 7.6 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: no mortality or any sub-lethal effect were observed.

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 7.6 mg/L

95% CI:

>

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: no mortality or any sub-lethal effect were observed.

Results with reference substance (positive control):

no reference substance

Reported statistics and error estimates:

A limit test was carried out and no toxic effects were observed, therefore statistical analysis was not necessary.

Sublethal observations / clinical signs:

No mortality or any sub-lethal effects were observed during the 96-h test period either in the exposure concentration of 7.6 mg/L or in the concurrent control.

Validity criteria fulfilled:

yes

Conclusions:

In this 96-hour acute toxicity test on Zebrafish (Danio rerio) the test item Reaction products of 2,2-dimethylpropane-1,3-diol with 1-chloro-2,3-epoxypropane had no toxic effect on fish at the exposure concentration of 7.6 mg/L, which is higher than the threshold concentration (EC50 of 7.488 mg/L, (see details in section 5.6.2 and 5.9.1)). Accordingly, the test item has no toxic effect on fish up to at least the threshold concentration of 7.488 mg/L.


Biological endpoints of the study are summarised in table below:


Summary of the biological endpoints (same as Table 1)
Endpoint Concentration
(based on measured concentrations)
96-h LC50 > 7.6 mg/L
(> 7.488 mg/L; threshold concentration)
96-h NOEC 7.6 mg/L
(> 7.488 mg/L; threshold concentration)
96-h LOEC > 7.6 mg/L
(> 7.488 mg/L; threshold concentration)

Executive summary:

Test design:
Test item:	Reaction products of 2,2-dimethylpropane-1,3-diol with 1-chloro-2,3-epoxypropane
Test species:	Zebrafish (Danio rerio)
Dilution water:	ISO medium, prepared in the laboratory of TOXI-COOP ZRT.
Concentrations:	Limit test; using a single concentration of 10 mg/L (nominal) based on data obtained from acute Algae and Daphnia studies (threshold concentration; see section 5.6.2); untreated control ran in parallel. The corresponding calculated exposure concentration (based on analytical measurements) was 7.6 mg/L.
Test design:	Fish were exposed to aqueous test media containing the test item for 96 hours in a semi-static test (24-h water renewal periods). Ten fish were exposed to the test concentration and control (no replicates were used). Fish were observed twice a day (in the morning and afternoon) during the exposure at approx. 2-3, 5-6, 24, 30, 48, 54, 72, 78 and 96 hours after start of the test for signs of intoxication and mortality. Measurements of pH, dissolved oxygen concentration and water temperature were carried out daily (at the start and end of each water renewal period).
Analytics:	For determination of the test item concentrations, samples were taken from the treatment and the control group at the start and at the end of each water renewal period. Concentrations were determined using LC-MS/MS.
Endpoints:	LC50, NOEC and LOEC for 96-h test period.
Statistics:	Statistical analysis was not necessary to determine test endpoints.

 

Results:

No mortality or any sub-lethal effects were observed during the 96-h test period either in the exposure concentration of 7.6 mg/L or in the concurrent control. Accordingly, the 96-h NOEC was determined to be 7.6 mg/L. The 96-h LC50 and LOEC were determined to be higher than 7.6 mg/L. All validity criteria were met and therefore the study was considered to be fully valid  The biological endpoints are summarised below in Table 1.

 

 

Table 1: Summary of the biological endpoints

Endpoint	Concentration (based on measured concentrations)
96-h LC50	> 7.6 mg/L (> 7.488 mg/L; threshold concentration)
96-h NOEC	7.6 mg/L (> 7.488 mg/L; threshold concentration)
96-h LOEC	> 7.6 mg/L (> 7.488 mg/L; threshold concentration)

 

 

Conclusion:

In this 96-hour acute toxicity test on Zebrafish (Danio rerio) the test item Reaction products of 2,2-dimethylpropane-1,3-diol with 1-chloro-2,3-epoxypropane had no toxic effect on fish at the exposure concentration of 7.6 mg/L, which is higher than the threshold concentration (EC50 of 7.488 mg/L, (see details in section 5.6.2 and 5.9.1)). Accordingly, the test item has no toxic effect on fish up to at least the threshold concentration of 7.488 mg/L.

Description of key information

Short-term toxicity to fish (LC50): NON-toxic (> 100 mg/L)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

> 7.6 mg/L