2-[bis(2-hydroxyethyl)amino]-2-(hydroxymethyl)propane-1,3-diol

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:

2019-10-02 to 2017-10-06

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:

July 12, 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Commission Regulation (EC) No 440/2008 of 30th May 2008 Annex Part C, C.1 (published in the Official Journal of the European Union L 142 of 31 May 2008)

Version / remarks:

May 31, 2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Version / remarks:

2016

Deviations:

yes

Remarks:

minor, without any consequences

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 100 mg/L
- Sampling method: samples of the test concentration and control were taken from the test solutions at the start and at the end of the study. Five parallel samples were analysed from the test concentration and one from the control at the start and at the end of the test.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
Test medium: ISO medium was prepared by adding 25 mL from each of four stock solutions to 900 mL deionised water proportionally.

Test item concentrations: Based on the results of the non-GLP preliminary range-finding test, a single concentration of 100 mg/L (limit concentration) was tested in a limit test. A concurrent control group ran parallel.

Preparation of test solutions: The test solution used in the test was prepared by mechanical dispersion. An amount of 0.5 g test item was dissolved in 5000 mL dilution water (ISO medium), resulting a nominal concentration of 100 mg/L. The test solution was freshly prepared in the testing laboratory just before introduction of fish.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
Species: Zebrafish (Brachydanio rerio)
Source: Akvárium magazin Kft. (Pasaréti Gyula) 1222 Budapest, Dévény u. 36.
Number of animals: 10 animals at the treated and control group respectively (no parallels were used).
Age: Juveniles.
Size of animals: size within a range of 2.0 ± 1 cm
Sex: Both female and male (were not separated).
Animal health: Apparently healthy animals were used in the test.
Acclimatization: Fish were held for at least 12 days before test initiation under the same conditions as used during the exposure period.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

none

Hardness:

approximate theoretical total hardness of 249 mg/L (as CaCO3)

Test temperature:

21.4 – 22.1 °C

pH:

7.48 – 7.83 (without adjustment)

Dissolved oxygen:

72.8 – 86.4 %

Nominal and measured concentrations:

nominal = 100 mg/L
measured = 98 - 100 % recovery

Details on test conditions:

TEST SYSTEM
- Test vessel: aquarium
- Fill volume: 5 L
- Aeration: no
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: <= 1.0 g fish / L

TEST MEDIUM PARAMETERS
- Source of dilution water: reconstituted water (ISO medium, prepared according to Annex 2 of the OECD guideline 203).

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light per day.

EFFECT PARAMETERS MEASURED: The fish were observed at approximately 4, 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality. Measurements of pH, dissolved oxygen concentration and temperature were carried out daily. The body length of each fish was measured at the end of the test in order to check their compliance with the size range recommended for the species by the test guideline (OECD No. 203). The measured values were within the range of 1.90-2.80 cm.

RANGE FINDING STUDY
- Test concentrations: 100 mg/L
- Results used to determine the conditions for the definitive study: Three fish were exposed for 96 hours in each concentration (no replicates were used). A control group was carried out concurrently. The fished exposed to the concentration at 100 mg/L was affected after 96 – hour test period. The concentration levels used and results (96 h) of the preliminary range-finding test are summarised in the section "Any other inofrmation on materials incl. tables".

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Mortality in control: None
- Any observations that might cause a difference between measured and nominal values: The dissolved oxygen concentration in the test solutions did not fall below 60 % of air saturation value during the study. No significant change in the pH (more than one unit) as well as for the temperature value was observed during the test.

Reported statistics and error estimates:

A limit test was carried out and toxic effects were not observed, therefore statistical analysis was not necessary. The endpoints of the test item were determined directly from the raw data.

Sublethal observations / clinical signs:

No mortality of sub-lethal effects were observed in the 10 fish in the control group or in the 10 fish exposed to a test concentration of 100 mg/L. The 96-h No Observed Effect Concentration (NOEC) and the LC₀ values were determined to be 100 mg/L. The LC₅₀, the LOEC, and LC₁₀₀ values were determined to be > 100 mg/L.

Table 2: Summary of the Biological Endpoints

Endpoint	Concentration [mg/L]
96-h LC50	> 100
96-h NOEC	100
96-h LC0	100
96-h LOEC	> 100
96-h LC100	> 100

Table 3: Cumulative mortality data in the definitive test

Test Group	Cumulative mortality (initial population = 10 fish / test group)
	4 h	24 h	48 h	72 h	96 h
Control	0	0	0	0	0
100 mg/L	0	0	0	0	0

 

Validity criteria fulfilled:

yes

Conclusions:

In the 96-hour acute toxicity test on Zebrafish (Brachydanio rerio) according to OECD guideline 203, the test item had no toxic effect on fish at a limit concentration of 100 mg/L. Accordingly, the 96-h LC50 value was determined to be > 100 mg/L. The 96-h NOEC was determined to be 100 mg/L.

Executive summary:

The acute toxicity of the test item on Zebrafish (Brachydanio rerio) was tested according to OECD guideline 203. Young fish were exposed in a static test to the test item in aqueous test media for 96 hours under defined conditions. Two groups of ten fish each were exposed to the test concentration or served as controls (no replicates were used). Fish were observed at approximately 4, 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality. Measurements of pH, dissolved oxygen and temperature were carried out daily. Based on the results of the non-GLP preliminary test, the test item had no toxic effect on the Zebrafish up to a concentration of 100 mg/L (limit concentration). Therefore, the main test was performed at a concentration level of 100 mg/L, including a concurrent control. The concentration of the test item was analytically determined in the test item solution at the start and at the end of the experiment. The measured test item concentration remained within ± 20 % of the nominal during the test period of 96 hours. There were no mortality and any sub-lethal effect observed in 10 fish in the control group or in 10 fish exposed to a test concentration of 100 mg/L. The 96-h No Observed Effect Concentration (NOEC) and the LC0 values were determined to be 100 mg/L. The LC50, the LOEC, and LC100 values were determined to be > 100 mg/L.

Description of key information

In a 96-hour static acute toxicity test (acc. to OECD TG 203) on Zebrafish (Brachydanio rerio) the test substance had no toxic effect on fish at a concentration of 100 mg/L (limit concentration). Accordingly, the 96-h LC50 value was determined to be > 100 mg/L. The 96-h NOEC was determined to be 100 mg/L (reference 6.1.1-1).

Additional information

The acute toxicity of the test item on Zebrafish (Brachydanio rerio) was tested according to OECD guideline 203. Young fish were exposed in a static test to the test item in aqueous test media for 96 hours under defined conditions. Two groups of ten fish each were exposed to the test concentration or served as controls (no replicates were used). Fish were observed at approximately 4, 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality. Measurements of pH, dissolved oxygen and temperature were carried out daily. Based on the results of the non-GLP preliminary test, the test item had no toxic effect on the Zebrafish up to a concentration of 100 mg/L (limit concentration). Therefore, the main test was performed at a concentration level of 100 mg/L, including a concurrent control. The concentration of the test item was analytically determined in the test item solution at the start and at the end of the experiment. The measured test item concentration remained within ± 20 % of the nominal during the test period of 96 hours. There were no mortality and any sub-lethal effect observed in 10 fish in the control group or in 10 fish exposed to a test concentration of 100 mg/L. The 96-h No Observed Effect Concentration (NOEC) and the LC0 values were determined to be 100 mg/L. The LC50, the LOEC, and LC100 values were determined to be > 100 mg/L.