1,2,4-Benzenetricarboxylic acid, mixed lauryl and octyl triesters

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:

2015-04-01 - 2015-11-06

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)

GLP compliance:

yes

Specific details on test material used for the study:

N/A

Analytical monitoring:

yes

Details on sampling:

The samples were analyzed directly without storage. For the analysis of the actual test substance concentrations, 30mL of test solution was taken from each test vessel at the beginning and at the end of the test.

Vehicle:

no

Details on test solutions:

The water solubility of the test substance is less than 1.0 mg/L (test data, at 25 °C the water solubility of EPI is 2.644 exp-10 mg/L). A nominal concentration of 100 mg/L of the test substance was prepared and stirred (via magnetic stirrer) in the dark for 72h at 500 rpm followed by filtration using a 0.45µm nitrocellulose membrane and glass-microfibre to make a saturated solution of the test substance in the test water as the test solution. The pH of stock solutions was 8.52 and need not to be adjusted.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: The winter eggs of the daphnias were originally supplied by MicroBioTests lnc.(ID: DM-MBT-141020).
- Age at study initiation: not older than 24 hours
- Method of breeding: Each generations of the Daphnia magna are breed in Bioassay and Safety Assessment Lab, Shanghai Academy of Public Measurement in the reconstituted test water. Cultures of adult Daphnia magna are fed concentrated freshwater suspensions of unicellular green algae Desmodesmus Subspicatus and Pseudokirchneriella subcapitata at a rate equivalent to 0.1 - 0.2mg of organic carbon per adult per day.

- Feeding during test: none

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

N/A

Post exposure observation period:

none

Hardness:

165 mg/L (as CaCO3) and it was between 140 to 250 mg/L (as CaCO3)

Test temperature:

21 - 22 °C

pH:

7.86 - 8.54

Dissolved oxygen:

7.26-9.46 mg/L

Salinity:

CaCl2 x 2 H2O: 120 mg/L; MgSO4 x 7 H2O: 120 mg/L; NaHCO3: 192 mg/L; KCl: 8 mg/L; Na2SeO3: 2 µg/L; Vitamin B12: 1 µg/L

Conductivity:

N/A

Nominal and measured concentrations:

Nominal: 100 mg/L
Measured: < LOD (0.214 mg/L)

Details on test conditions:

TEST SYSTEM
- 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: synthetic fresh water

OTHER TEST CONDITIONS
- Adjustment of pH: not adjusted
- Photoperiod: L:D = 16 h:8 h
- Light intensity: not mentioned

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilisation after 24 and 48 hours; O2 concentration, pH at the end of the test


TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable (limit test)
- Range finding study: A nominal concentration of 100 mg/L of the test substance was prepared and stirred (via magnetic stirrer) in the dark for 72h at 500 rpm followed by filtration using a 0.45 µm nitrocellulose membrane and glass-microfibre to make a saturated solution of the test substance in the test water as the test solution. The pH of stock solutions was 8.52 and need not to be adjusted. In the range-finding test, there were no immobilisation observed in the daphnids of the control group and the treatment groups during the test.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

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:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

No effects.

Results with reference substance (positive control):

- Results with reference substance valid? yes, historical control (not tested in parallel to the test substance)
- Concentrations: 0.32; 0.56; 1; 1.8; 3.2 mg/L potassium dichromate
- Results: The 24 h EC50 for potassium dichromate was 1.4 mg/L.

Reported statistics and error estimates:

N/A

Table #1: Effect of 1,2,4 -Benzenetricarboxylic acid, mixed dodecyl and octyl ester on immobilisation of Daphnia magna

Parameter		Blank control		Saturated Solution
Duration(h)ss		24	48	24	48
Number of Daphnids		20	20	20	20
amount of immobilisation	Parallel A	0	0	0	0
	Parallel B	0	0	0	0
	Parallel C	0	0	0	0
	Parallel D	0	0	0	0
Total		0	0	0	0
Effect (%)		0	0	0	0

Abnormal: no particular behavioural or morphological changes are observed.

Validity criteria fulfilled:

yes

Conclusions:

The test results showed that the 24h and 48h EC50 of the immobilisation effect for the test substance 1,2,4-BENZENETRICARBOXYLIC ACID, MIXED DODECYL AND OCTYL TRIESTERS to daphnids were higher than the concentration of the saturated solution (which measured concentration was below LOD, LOD= 0.214mg/L) under the tested conditions. Not toxic under test conditions in the range of water solubility.

Executive summary:

According to Ministry of environmental protection of the people's Republic of China, The Guidelines for the Testing of Chemicals - Effects on Biotic System, 2013, 202 Daphnia sp. EC50 Acute Immobilisation Test and OECD, Guidelines for the Testing of Chemicals, 2004, 202 Daphnia sp., Acute Immobilisation Test, the acute immobilisation toxicity of 1,2,4-BENZENETRICARBOXYLIC ACID, MIXED DODECYL AND OCTYL TRIESTERS(ZZYD20140123-0866) to Daphnia magna was determined in a 48h static test. In the limit test, a nominal concentration of 100 mg/L of the test substance was prepared and stirred (via magnetic stirrer) in the dark for 72h at 500 rpm followed by filtration using a 0.45 µm nitrocellulose membrane and glass-microfibre to make a saturated solution of the test substance in the test water (referred to as "saturated solution" in this report, since the measured concentration was below the limit of detection (LOD) in this limit test, LOD= 0.214 mg/L). Based on the results of the range-finding test, one treatment group and one control group with four replicates each were included in the limit test. The test duration was 48h. The percentage of daphnids that were immobilised was 0% of the control and treatment groups at the end of the test. The dissolved oxygen concentrations at the end of the test were higher than 3 mg/L in the controls and treatments. The test is valid. The concentrations of the test substance in the test solutions were analyzed by GC method. In the limit test, the measured concentrations of the test substance were below the LOD.

Under the test condition, there was no acute immobilisation of the saturated solution to Daphnia magna in the limit test. The EC50 of the test substance to Daphnia magna at 24h and 48h was higher than the concentration of the saturated solution (which measured concentration were below the LOD, LOD= 0.214 mg/L) under the tested conditions.

Description of key information

The test results showed that the 24h and 48h EC50 of the immobilisation effect for the test substance 1,2,4-Benzenetricarboxylic acid, mixed dodecyl and octyl triesters to daphnids were higher than the concentration of the saturated solution (which measured concentration was below LOD, LOD= 0.214mg/L) under the tested conditions. Not toxic under test conditions in the range of water solubility.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

> 100 mg/L

Additional information

Data are also available for the read across material 1,2,4-Benzenetricarboxylic acid, decyl octyl ester. The available information demonstrates a lack of toxic effects in the range of the water solubility.