3-tert-butyladipic acid

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:

2017-05-23 to 2017-05-25

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:

April 2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

May 2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Version / remarks:

1996

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Changzhou Sunlight Pharmaceutical Co., Ltd., China; 20160505 / 116727
- Expiration date of the lot/batch: May 2017
- Purity test date: 30.01.2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature
- Stability under test conditions: stable

Analytical monitoring:

yes

Details on sampling:

- Concentrations: For the analysis of the test item concentrations, samples of each test concentration (100 mg/L) and control were taken from the test solutions (4 x 5 mL at the start and 4 x 5 mL at the end of the study).
- Sampling method: Four parallel samples were taken from the 100 mg/L concentration level and from the control sample. The samples were prepared immediately after sampling.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test solution used in the test was prepared by mechanical dispersion. An appropriate amount of test item was dissolved in the dilution water (ISO medium) in order to obtain the concentration of 100 mg/L and this solution was placed into ultrasonic bath for 15 minutes. The resulted solution was the test concentration. Untreated control was included parallel in the test. The test solution was prepared immediately before introduction of daphnids.
- Controls: Yes. The dilution water (ISO-medium) was used without addition of the test item.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea (Daphnia magna)
- Strain/clone: Daphnia magna (Straus)
- Source: Laboratory of Hydrobiology (Central Agricultural Office, Directorate of Plant-, and Soil Protection) 2100 Gödöllő, Kotlán S. u. 3. Hungary
- Sex: Female
- Age of parental stock: Less than 24 hours old
- Feeding during test: None

ACCLIMATION
- Acclimation: Test animals were bred under similar conditions as that used during the exposure period, therefore additional acclimatisation before the test was not necessary
- Food: The animals were fed with centrifuged green alga suspension

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

249 mg/L (as CaCO3)

Test temperature:

19.8 – 20.1 °C (measured in the test vessels)
19.2 – 20.1 °C (within the climate chamber)

pH:

6.00– 8.14

Dissolved oxygen:

7.21 – 8.21 mg/L

Nominal and measured concentrations:

Nominal: 100 mg/L
Measured: The analytically determined concentration of the tested substance was maintained within ± 20 % of the nominal concentrations throughout the test. Therefore, the analysis of the results was based on the nominal concentration value.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers of approximately 40 mL test medium
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO medium (according to OECD 202) was used as dilution water in the experiment.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours light (artificial illumination) and 8 hours darkness

EFFECT PARAMETERS MEASURED:
The immobility or mortality of the Daphnia was determined by visual observation after 24 and 48 hours of exposure. Those animals not able to swim within 15 seconds after gentle agitation of the test beaker were considered to be immobile.

RANGE-FINDING STUDY
- Test concentrations: A non-GLP preliminary test was performed at the nominal concentation of 100 mg/L in order to check the toxicity of the test item on the daphnids up to at least this concentration.
- Results used to determine the conditions for the definitive study: In the preliminary test no immobility or any sub-lethal effect was detected in the treated concentration and in the control group during the 48-h exposure period. Based on the results of the preliminary test, a single test concentration at 100 mg/L nominal concentration and one control group were tested in a limit test.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

other: EC20

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

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

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:

No immobilisation or any abnormal behaviour of test animals was observed in the control group and 10% immobilization was observed at the test concentration of 100 mg/L after 48 hours of exposure. No significant immobilisation was observed in the test concentration of the test item 3-Tert Butyladipic Acid during the 48-hour exposure of the test.

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: The 24h EC50 is 1.41 mg/L (95% conf. limits: 1.19 – 1.68 mg/L)

Table 1: Immobilisation of the test animals (summary table)

Nominal concentration [mg/L]	No. of Daphnia tested	No. of immobilized Daphnia after		% of immobilized Daphnia after
		24 h	48 h	24 h	48 h
Control	20	0	0	0	0
100	20	2	2	10	10

Validity of the Study
Control: The immobilisation rate of the introduced Daphnia did not exceed 10 %. Immobility was not observed in the untreated control and noDaphnia were trapped at the water surface or showed any sign of stress.
Dissolved Oxygen Concentration: The dissolved oxygen concentration at the end of the test was more than 3 mg/L in control and test vessels.
All validity criteria were within acceptable limits and therefore the study was considered as valid.

Analytical Results:
The measured concentration of the test item in the test solution was 102 % of the nominal value at the start and 102 % of the nominal value at the end of the test. There is evidence that the concentration of the tested substance was maintained within ± 20 % of the nominal concentration throughout the test. Therefore the analysis of the results was based on the nominal concentration values.

Table 2: Summarised measured concentration of the test substance in formulation samples

Sample date	Nominal concentration (mg test item/L	Mean of the measured concentrations (mg test item/L)	Measured concentrations in percentage of the nominal	Relative Standard Deviation % (n=4)
23 May 2017	Control	Not detected	-	-
	100	101.7	102	0
25 May 2017	Control	Not determined	-	-
	100	101.8	102	1

Validity criteria fulfilled:

yes

Conclusions:

In this 48-hour acute toxicity test on Daphnia magna, the obtained results showed that the test item had no significant toxic effects on the mobility of Daphnia up to the highest tested concentration of 100 mg/L. Thus, the 48h-EC50 was determined to be >100 mg/L and the NOEC was determined to be 100 mg/L. The results are based on the nominal test item concentration.

Description of key information

In an acute toxicity study with Dahnia magan an EC50 of >100 mg/L and a NOEC of 100 mg/L were determined after 48 h of exposure to the test item.

Additional information

An acute toxicity study accordingto OECD 202 was performed to evaluate the influence of the test item, 3 -Tert Butyladipic, on the mobility and survival of Daphnia magna. In a limit-test, conducted under static conditions, young Daphnia were exposed to aqueous test medium containing the test item at 100 mg/L nominal concentration for an exposure time of 48 hours. The obtained results showed that the test item had no significant toxic effect on the mobility of Daphnia at the limit concentration of 100 mg/L. Thus, the 48h-EC50 was determined to be >100 mg/L and the NOEC was determined to be 100 mg/L. The analytically determined concentration of the tested substance was maintained within ± 20 % of the nominal concentrations throughout the test. Therefore, the analysis of the results was based on the nominal concentration value.