Decanedioic acid, compound with ethanolamine / Decanedioic acid, compound with 2-aminoethanol

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:

01 August 2017 to 10 August 2017

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)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Samples (approximately 150 mL) of the range-finding and definitive test solutions and controls were collected into Teflon® capped glass bottles (250 mL, no preservative).
- Samples were collected immediately prior to test commencement (0 hour) and at test termination (48 hour, from pooled replicates) for analysis.
- All samples were stored at room temperature until transported to ISI and kept refrigerated by ISI until analysis was conducted.
- All samples from the range-finding and definitive tests were analysed within 24 hours of sample collection.

Vehicle:

no

Details on test solutions:

- Individual test solutions of the test material for both range-finding and definitive tests were prepared from stock solutions (1000 mg/L nominal concentration) prepared in dilution water without the use of any solubilising agent.
- Stock solutions were prepared in glass volumetric flasks and stirred until the test material was visibly dissolved.
- All stock and test solutions were corrected for purity of the test material (i.e., 88% test material, 12% water).
- All range-finding and definitive tests included a negative control consisting of dilution water with no test material added.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Strain/clone: Daphnia magna STRAUS
- Source: The organisms were originally obtained from the Ministry of the Environment and Climate Change (Toronto, Ontario, Canada), the organisms were cultured continuously from a population that has been actively reproducing in the testing laboratory since 1988.
- Age: First instar, neonate (< 24 hours old)
- Feeding during test: no

CULTURE CONDITIONS
- Test organism acclimation was not required, since test organisms were cultured under conditions identical to those of the test.
- Type and amount of food: Once/day: yeast, Cerophyll broth, Trout Chow and Pseudokirchneriella subcapitata algae

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

200 - 210 mg/L as CaCO3

Test temperature:

20 ± 2°C

pH:

8.3 - 8.5

Nominal and measured concentrations:

Nominal: 100 mg/L
Measured: 115 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beaker, 100 mL
- Material, size, headspace, fill volume: 75 mL (min)
- Aeration: none
- Renewal rate of test solution (frequency/flow rate): none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Loading rate: 15 mL/daphnid

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The source of water used for D. magna culturing and testing was moderately hard groundwater (~350 mg/L as CaCO3) obtained from an aquifer in Aberfoyle, Ontario, Canada. For culture and testing purposes with D. magna, the groundwater was filtered through a 20-micron cellulose-acetate filter, diluted with Reverse Osmosis (RO) water to achieve a hardness of ~200 mg/L (as CaCO3). A continuous supply of oil-free compressed air was provided to bring the pH and concentrations of dissolved oxygen and other gases into equilibrium with air. The concentration of dissolved oxygen in the water was maintained at > 80% of the air saturation value.
- Intervals of water quality measurement: Water quality parameters (including dissolved oxygen, pH, and conductivity) were measured in the control and all test solutions at the beginning and end of the test. Temperature was monitored and recorded daily. Water hardness was measured at the beginning of the test.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16-hour light, 8-hour dark (with 30 minute transition period)
- Light intensity: 400 to 800 lux (Ambient laboratory illumination)

EFFECT PARAMETERS MEASURED:
- For all test and controls, daphnid immobility was determined after 24 and 48 hours.
- A test organism was considered to be immobile if it was unable to swim within 15 seconds following gentle agitation of the test solution and/or unable to free itself after gentle tapping of the test vessel.

RANGE-FINDING STUDY
- Test concentrations: 0.1, 1.0, 10 and 100 mg/L
- Concentrations were prepared by adding 0.05, 0.5, 5, and 50 mL of a nominal 1000 mg/L stock solution, respectively into a 500 mL volumetric flask and making this up to volume with dilution water.
- A total of 10 daphnids (i.e., 2 replicates with 5 daphnids each) were exposed to each test level under static test conditions (i.e., no renewal of the test solution). All test organisms were transferred to fresh test and the negative control solutions using a pipette and bulb.
- Results used to determine the conditions for the definitive study: Results of the range-finding test indicated that the 48-hour EC50 for daphnid immobilisation was greater than 100 mg/L nominal concentration (i.e. the highest concentration tested). As such, the definitive test was conducted as a Limit Test at a nominal concentration of 100 mg/L. The 100 mg/L test concentration was prepared by adding 100 mL of a nominal 1000 mg/L stock solution, respectively into a 1000 mL graduated cylinder and making this up to volume in dilution water.

Reference substance (positive control):

yes

Remarks:

Sodium chloride

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

RANGE FINDING TEST
- Confirmation of Exposure Concentrations: Nominal, measured and Time-Weighted Mean (TWM) concentrations of the test material determined on samples collected at the beginning and end of the test are provided in Table 1. Measured concentrations were similar to nominal concentrations and the beginning and end of the test showing the test material was stable during the 48 hour test period
- Immobility: The test met all of the test validity criteria (i.e., immobility ≤ 10% in the control; dissolved oxygen concentration at test termination ≥ 3 mg/L in all control and test vessels). The percentage immobility of daphnids exposed to the test material during the rangefinding test along with the nominal and TWM concentrations are given in Table 2. Results of the range-finding test showed that no immobility (0%) of D. magna was observed at concentrations of the test material up to and including 100 mg/L. The TWM concentration corresponding to the nominal 100 mg/L concentration of the test material was 104 mg/L.

DEFINITIVE TEST RESULTS
- Confirmation of Exposure Concentrations: Nominal, measured and TWM concentrations of the test material determined on samples collected at the beginning and end of the test are provided in Table 3. The TWM concentration corresponding to the nominal 100 mg/L concentration of the test material was 115.0 mg/L. Measured concentrations were at the beginning and end of the test showing the test material was stable during the 48 hour test period. However, average (n=2) measured concentrations from samples collected at the beginning and end of the test were approximately 10 to 20% higher than nominal concentrations (i.e., 119 and 111 mg/L, respectively). There was no apparent explanation for the greater than nominal measured concentrations after further investigation of the procedures and the data. The results were considered likely due to sampling/analysis error. Therefore, the definitive test results were based on nominal concentrations.
- Immobility: The definitive test met all of the test validity criteria. The definitive test was conducted as a Limit Test at a nominal concentration of 100 mg/L. A summary of the test results expressed in terms of the percent daphnid immobility observed in the test after 24 and 48 hours is summarised in Table 4. Results of the definitive test confirmed the results of the range-finding test showing 0% immobility of daphnids at a concentration of 100 mg/L.
- The 24 and 48-hour EC50s for immobility of daphnids were reported to be > 100 mg/L. The 48-hour NOEC and LOEC for immobility of daphnids were reported to be 100 mg/L and > 100 mg/L, respectively.

Results with reference substance (positive control):

- Sodium chloride was the reference toxicant used in this study. The LC50 was compared to historical data and was deemed acceptable based on results falling within the 95 % confidence interval (warning limit) calculated for the last twenty reference tests.
- In conjunction with the definitive test, the 48-hour LC50 for the Daphnia magna reference toxicant test was 5.7 g/L.

Reported statistics and error estimates:

- Results of the range-finding test were based on Time-Weighted Mean (TWM) concentrations of the test material. However, results for the definitive test were based on nominal as opposed to TWM concentrations, since measured concentrations exceeded nominal by approximately 10 to 20 %.
- There was no apparent explanation for the greater than nominal measured concentrations and therefore, the definitive test results were based on nominal concentrations.
- Estimates of the 24 and 48-hour EC50 from the range-finding and definitive test results were based on visual inspection of the data, since no immobility of daphnids was observed at any concentration up to and including 100 mg/L during the study.

Table 1: Summary of the Nominal, Measured and Time-Weighted Mean Concentrations of the Test Material for the Range-Finding Test

Nominal Concentration (mg/L)	Measured Concentration (mg/L)		Time-Weighted Mean Concentration (mg/L)
	0 Hours	72 Hours
100	101.66d	105.6e	104
10	11.03	9.69	10.4
1	1.01	N/A	1.01g
0.1	0.12	N/A	0.12g
0 (Negative Control)	<MDL	<MDL	<MDL

d Average of duplicate analyses. The Relative Percent Difference (RPD) = 2.2%

e Average of duplicate analyses. RPD = 3.4%

MDL (Method Detection Level) = 0.09 mg/L

g Based on a single analysis (not a TWM)

N/A = not analysed

 

Table 2: Immobility of D. magna After 24 and 48 Hours Exposure to the Test Material During the Range-Finding Test

Nominal Concentration (mg/L)	Time-Weighted Mean (mg/L)	Immobility (%)
		T= 24 hours	T= 48 hours
100	104.0	0	10
10	10.4	0	0
1	1.01*	0	0
0.1	0.12*	0	0
0 (Negative Control)	<MDL	0	0

* Based on a single measurement at t = 0 hours (not a TWM)

 

Table 3: Summary of the Nominal, Measured and Time-Weighted Mean Concentrations of the Test Material for the Definitive Test

Nominal Concentration (mg/L)	Measured Concentration (mg/L)		Time-Weighted Mean Concentration (mg/L)
	0 hrs	48 hrs
100	118 and 120d	112 and 110	115
0 (Negative control)	<MDL	<MDL	<MDL

d Average of duplicate analyses, Relative Percent Difference (RPD) = 1.2%

MDL = 30 mg/L

 

Table 4: Immobility of D. magna After 24 and 48 Hours Exposure to the Test Material During the Definitive Test

Nominal Concentration (mg/L)	Immobility out of 20 organisms (%)
	24 h	48 h
100	0	0
0 (Negative control)	0	0

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this study, the 24 and 48-hour EC50s based on the results of the definitive test were reported to be > 100 mg/L. The 48-hour NOEC and LOEC for immobility of daphnids were estimated to be 100 and > 100 mg/L, respectively.

Executive summary:

The acute toxicity of the test material to aquatic invertebrates was investigated in accordance with the standardised guideline OECD 202, under GLP conditions in the daphnia immobilisation test.

Both the range-finding and definitive tests met all the test validity criteria. Test results were based on measured concentrations, or more specifically, Time-Weighted Mean (TWM) concentrations. All test solutions and results were corrected for purity of the test material (i.e., 88% test material; 12% water).

Results of the range-finding test indicated that the 48-h EC50 for immobility was > 104 mg/L (based on the TWM concentration). As such, the definitive test was conducted as a Limit Test at a single nominal concentration of the test material of 100 mg/L. Average measured concentrations for the range-finding test were marginally higher (i.e., 1 – 5 %) than nominal. However, for the definitive test, average (n = 2) measured concentrations from samples collected at the beginning and end of the test were approximately 10 to 20 % higher than nominal concentrations (i.e., 119 and 111 mg/L, respectively). There was no apparent explanation for the greater than nominal measured concentrations after further investigation of the procedures and the data. The results were considered likely due to sampling/analysis error. Therefore, the definitive test results were based on nominal concentrations. Results of the definitive test showed that there was no immobility of daphnids in the study.

Under the conditions of this study, the 24 and 48-hour EC50s based on the results of the definitive test were reported to be > 100 mg/L. The 48-hour NOEC and LOEC for immobility of daphnids were estimated to be 100 and > 100 mg/L, respectively.

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

 

Description of key information

Under the conditions of this study, the 24 and 48-hour EC50s based on the results of the definitive test were reported to be >100 mg/L. The 48-hour NOEC and LOEC for immobility of daphnids were estimated to be 100 and >100 mg/L, respectively.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

The acute toxicity of the test material to aquatic invertebrates was investigated in accordance with the standardised guideline OECD 202, under GLP conditions in the daphnia immobilisation test. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

Both the range-finding and definitive tests met all the test validity criteria. Test results were based on measured concentrations, or more specifically, Time-Weighted Mean (TWM) concentrations. All test solutions and results were corrected for purity of the test material (i.e., 88% test material; 12% water).

Results of the range-finding test indicated that the 48-h EC50 for immobility was > 104 mg/L (based on the TWM concentration). As such, the definitive test was conducted as a Limit Test at a single nominal concentration of the test material of 100 mg/L. Average measured concentrations for the range-finding test were marginally higher (i.e., 1 – 5%) than nominal. However, for the definitive test, average (n = 2) measured concentrations from samples collected at the beginning and end of the test were approximately 10 to 20% higher than nominal concentrations (i.e., 119 and 111 mg/L, respectively). There was no apparent explanation for the greater than nominal measured concentrations after further investigation of the procedures and the data. The results were considered likely due to sampling/analysis error. Therefore, the definitive test results were based on nominal concentrations. Results of the definitive test showed that there was no immobility of daphnids in the study.

Under the conditions of this study, the 24 and 48-hour EC50s based on the results of the definitive test were reported to be > 100 mg/L. The 48-hour NOEC and LOEC for immobility of daphnids were estimated to be 100 and > 100 mg/L, respectively.