Trioctyl benzene-1,2,4-tricarboxylate

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant study conducted according to internationally recognised test methods.

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

- Lot/batch No.: 001101TMO

Analytical monitoring:

yes

Details on sampling:

- Nominal concentrations: 0, 5.29, 9.53, 17.1, 30.9, 55.6 and 100.0 mg/L
- Sampling method: 0.5 - 5.0 L sampled on 4 occasions - immediately after preparing test solution (fresh medium on Day 0), and 2 days later before renewal (exhausted medium)
- Sample storage conditions before analysis: no data

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test substance solution at 3000 or 5000 mg/L concentration (in medium with HCO-50 solubiliser) was diluted to the required nominal concentration using medium (with solubilizer HCO-50 at 100 mg/L in final test/control media).
- Eluate: Dilution media
- Controls: yes, control & dispersant control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): solubilizer HCO-50
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 1.0 wt .% dispersant, 100 mg/l
- Evidence of undissolved material (e.g. precipitate, surface film, etc): No - test solutions were clear, colourless and with no observed precipitate

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Justification for species other than prescribed by test guideline: N/A
- Source: NIES Japan
- Age of parental stock (mean and range, SD): within 24h of age
- Feeding during test, yes
- Food type: Chlorella vulgaris
- Amount: 0.1-0.2 mgC/day/individual
- Frequency: daily

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

None

Hardness:

244-255 mg/L as CaCO3

Test temperature:

19.1-20.2 °C

pH:

7.1-7.8.

Dissolved oxygen:

7.1-8.8 mg/L

Salinity:

N/A

Nominal and measured concentrations:

Nominal concentrations: 0, 5.29, 9.53, 17.1, 30.9, 55.6 and 100.0 mg/L
Measured concentrations: See Results table

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beaker
- Material, size, headspace, fill volume: 80 mL
- Aeration: No data
- Renewal rate of test solution (frequency/flow rate): 3 times/week
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10
- No. of vessels per vehicle control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4(OECD guideline No.211 Annex 2)
- Intervals of water quality measurement: 3 times, fresh and exhausted medium

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light:8 hours dark
- Light intensity: =<1,200 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : See results tables below

VEHICLE CONTROL PERFORMED: Yes

RANGE-FINDING STUDY: yes
- Test concentrations: for parent Daphnia (Acute immobilization test) was >100mg/L. Dispersant control was also tested
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

LC50

Effect conc.:

99.3 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 58.9 - 134.0

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

10.4 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other: 9.1 - 11.6

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

7.18 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

4.04 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

- Mortality of parent animals: See Results table below
- No. of offspring produced per day per female: See Results table below
- Number of males and females (parental) : 10

Results with reference substance (positive control):

N/A

Reported statistics and error estimates:

Eco-Statics (Version 1.1)
Method of calculating mean measured concentrations: Time-weighted mean.

Analysis by HPLC: Within 58.0-182.3% of the nominal concentration. Time-weighted mean of the measured concentrations of test chemical during the 21-day exposure were 4.04, 7.18, 14.9, 28.1, 62.3 and 134.0 mg/L.

   

Nominal	Measured concentration (mg/L)
Concentration	Day 0	Day 2	Day 2	Day 4	Day 6	Day 8	Day 13	Day 15
(mg/L)	New	Old	New	Old	New	Old	New	Old
Control	<0.006	<0.006	<0.006	<0.006	<0.006	<0.006	<0.006	<0.006
Dispersant control	<0.006	<0.006	<0.006	<0.006	<0.006	<0.006	<0.006	<0.006
5.29	4.80	3.07	4.46	4.11	4.12	4.03	4.21	3.66
9.53	7.17	6.32	7.59	7.90	7.18	7.29	7.19	6.86
17.1	15.5	13.7	16.5	15.6	14.1	14.4	15.0	14.3
30.9	31.6	27.1	28.7	28.4	27.1	26.9	28.4	26.5
55.6	88.6	80.0	59.3	49.9	49.7	56.0	65.3	50.8
100.0	214.3	182.3	145.9	127.0	94.1	109.2	98.4	102.9
new: freshly prepared test solutions. old: test solution after 2-3 days exposure

 

 

   

Measured	Cumulative Number of Dead ParentalDaphniaon Day:
Concentration	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
(mg/L)
Control	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Dispersant control	0	0	0	0	0	0	0	0	0	0	0	0	0	0	1	1	1	1	1	1	1
4.04	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	1	2	2	3	3
7.18	0	0	1	1	1	1	1	1	1	1	1	1	1	2	2	2	3	4	5	5	5
14.9	0	2	2	2	2	2	2	2	2	2	3	3	3	3	3	3	5	5	5	5	5
28.1	0	0	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	1	2
62.3	0	0	2	2	2	2	2	2	2	2	2	2	2	2	2	2	2	2	2	2	2
134.0	0	2	2	2	3	5	5	5	6	6	7	7	7	7	7	7	7	7	7	7	7

 

 

 

Measured	Mean Cumulative Number of Juveniles produced/adult on Day:
Concentration	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
(mg/L)
Control	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	18.2	19.8	19.9	68.3	71.6	71.6	106.5	110.3	110.3	153.7	161.8	161.8	197.7
Dispersant control	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	1.2	2.6	14.7	18.9	36.8	64.7	78.2	88.9	107.9	120.1	123.0	150.0	162.8
4.04	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	2.0	2.9	5.1	16.4	36.0	43.6	59.9	84.9	90.7	105.9	115.6	122.9	135.4
7.18	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.2	0.2	31.4	34.6	34.6	51.0	65.2	65.2	70.6	93.0	93.0
14.9	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	1.2	1.2	1.2	12.4	12.4	12.4	30.8	36.0	36.0	38.6	51.4	51.4
28.1	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	1.4	3.0	4.6	9.4	10.3	10.3	25.5	28.6	28.6	39.0	44.1
62.3	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.3	1.8	1.8	3.9	5.1	5.1	9.6	12.9
134.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0	0.0

 

 

Time (days) to first brood production
Vessel Number	Measured concentration (mg/L)
	Control	Dispersion control	4.04	7.18	14.9	28.1	62.3	134.0
1	10	13	9	-	-	-	20	>21
2	9	13	-	13	13	13	14	-
3	9	10	10	-	-	11	20	>21
4	9	11	-	-	-	-	-	-
5	9	11	13	-	17	12	-	>21
6	9	9	12	13	-	12	20	-
7	9	-	11	11	10	12	15	-
8	9	11	-	13	-	11	15	-
9	9	11	10	13	10	11	17	-
10	9	9	10	-	13	11	15	-
Mean	9.1	10.9	10.7	12.6	12.6	11.6	17.0	>21

 

   

 

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions that this test was conducted the following values were calculated:
NOEC (21-d, reproduction) : 4.04 mg/L,
LOEC (21-d, reproduction) : 7.18 mg/L,
EC50 (21-d, reproduction) : 10.4 mg/L
LC50 for parental Daphnia (21-d) : 99.3 mg/L

Executive summary:

Chronic toxicity to Daphnia has been investigated in a 21 -day reproduction test according to OECD test methods. Dissolution/dispersion agents were used to increase concentration of the substance in the test medium. The following values were calculated:

NOEC (21-day, reproduction): 4.04 mg/L;

LOEC (21 -day, reproduction): 7.18 mg/L;

EC50 (21 -day, reproduction): 10.4 mg/L;

LC50 parental (21 -day): 99.3 mg/L

Reference 2

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Study period:

1997-06-18 to 1997-07-09

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions. For read-across justification see Section 13.

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 202 Part 2

Deviations:

yes

Remarks:

In the development of new EC-methods the test conditions deviate from the OECD-Guideline. The evaluation is adoated to the EC-guideline. No influence on the results is expected

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

yes

Remarks:

In the development of new EC-methods the test conditions deviate from the OECD-Guideline. The evaluation is adapted to the EC-guideline. No influence on the results is expected

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

For determination of stability of the test substance solution, a solution in deionized water was made and analyzed after 0 and 72 hours.
The stock solution for the test was deep-frozen at -18°C and reanalyzed every week during the test.

Vehicle:

no

Details on test solutions:

As the test substance is poorly soluble in water, a 1 g/L mixture in M4 medium without NaHCO3 and FeSO4 containing Na2EDTA was prepared. This mixture was stirred for 18 hours and filtrated afterwards. The filtrate showed an initial content of 4.1 mg/L DOC. The filtrate was used as Stock solution.
For changes of test solutions every second or third day during the test the stock solution was deep-frozen at -18°C. One day before application the required amount of the frozen stock solution was thawed and after sampling for analytical purpose mixed with NaHCO3 and FeSO4 containing Na2EDTA.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain: Daphnia magna Straus, clone 5
- Source: laboratory breed
- Age at study initiation: not older than 24 hours
- Method of breeding: The females were maintained in synthetic fresh water (M4 medium, Elendt, 1990). A reddish dark-brown coloration of the animals indicated that they were healthy.
- Feeding during test: daily with Scenedesmus subspicatus (days 0 to 6: 0.5x10E7 cells/daphnid; days 7 - 21: 1x10E7 cells/daphnid)


ACCLIMATION
- Acclimation period: Females were separated from the offspring 24 hours before the start of the test. The offspring produced overnight were transferred into test water and were used for the test.
- Type and amount of food: not mentioned
- Feeding frequency: not mentioned

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Post exposure observation period:

none

Hardness:

not mentioned

Test temperature:

20 ± 1 °C

pH:

7,6 - 8,2 over all test vessels. In one case a pH deviation of > 0.3 Units was found between the freshly prepared solution and the 2-3 day old solutions. As the reproduction rate was not impaired by this deviation, it was considered as not affecting the validity of this test.

Dissolved oxygen:

92% to 107% of saturation

Salinity:

according to guideline (M4 medium, Elendt)

Nominal and measured concentrations:

Nominal: WSF
Measured: 4.5 mg/L (mean of measured concentrations,concentration of test solutions remained stable during 72 hours: deviations < 20%)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: Glass beakers, 100 mL filled with 60 mL test solution
- Aeration: not mentioned
- Renewal rate of test solution (frequency/flow rate): 3 times per week
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10
- Biomass loading rate: 1 organism/60 mL


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Synthetic fresh water according to Elendt M4-Medium
- Culture medium different from test medium: no
- Intervals of water quality measurement: Day 0 (fresh samples), 2, 9, 12, 14, 16


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16h light, 8h dark
- Light intensity: light: 600 lux, dark: 100 lux


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
mortality and offspring on day 2, 5, 7, 9, 12, 14, 16, 19, 21

VEHICLE CONTROL PERFORMED: no

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 4.5 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

dissolved

Basis for effect:

reproduction

Reported statistics and error estimates:

As the reproduction rate with test substance was higher than in the control samples, no statistic has been performed.

Table #1: Number of living parent animals

Conc. [mg/L]	Parallel	2d	5d	7d	9d	12d	14d	16d	19d	21d	Day 21
control	1	1	1	1	1	1	1	1	1	1	Total parent animals             9
	2	1	1	1	1	1	1	1	1	1
	3	1	1	1	1	1	1	1	1	1
	4	1	1	1	1	1	1	1	1	1
	5	0	0	0	0	0	0	0	0	0
	6	1	1	1	1	1	1	1	1	1
	7	1	1	1	1	1	1	1	1	1
	8	1	1	1	1	1	1	1	1	1
	9	1	1	1	1	1	1	1	1	1
	10	1	1	1	1	1	1	1	1	1
4.5	1	1	1	1	1	1	1	1	1	1	Total parent animals                 9
	2	1	1	1	1	1	1	1	1	1
	3	1	1	1	1	1	1	1	1	1
	4	1	1	1	1	1	1	1	1	1
	5	1	1	1	1	1	1	1	1	1
	6	1	1	1	1	1	1	1	1	1
	7	1	1	1	1	1	1	1	1	1
	8	1	1	1	1	1	1	1	1	1
	9	1	1	1	1	1	1	1	1	1
	10	1	1	1	1	1	1	1	0	0

Table #2: Number of living offspring

conc. [mg/L]	parallel	21 day total	21 day mean
control	1	89	mean value offspring per parent animal 74
	2	61
	3	63
	4	88
	5	-
	6	84
	7	69
	8	76
	9	70
	10	66
4.5	1	80	mean value offspring per parent animal 78
	2	69
	3	85
	4	79
	5	75
	6	79
	7	78
	8	79
	9	80
	10	-

Validity criteria fulfilled:

yes

Conclusions:

The highest tested concentration at which no negative effects were observed on the reproduction rate or survival of the parent animals is:
NOEC >= 4.5 mg/L (highest concentration under test conditions)
The lowest tested concentration at which the first significant effects were observed on the reproduction rate or survival of the parent animals is:
LOEC > 4.5 mg/L (highest concentration under test conditions)
The concentrations were based on the product.

Executive summary:

The effect of 1,2,4-Benzenetricarboxylic acid, mixed decyl and octyl triesters on the immobilisation of Daphnia magna had been studied in accordance with OECD Guideline for Testing of Chemicals No. 202 part II.

Daphnia magna was exposed to 1,2,4-Benzenetricarboxylic acid, mixed decyl and octyl triesters in a semistatic reproduction test over 21 days. As the test substance is poorly soluble in water, a 1 g/L mixture with synthetic freshwater was prepared. This mixture was stirred for 18 hours and filtrated afterwards. For changes of test solutions during the test the stock solution was deep-frozen at -18°C. The analytical determination of the defrosted stock solutions used for the test showed that they were stable within the 3 days of exposure (deviation > 20%), therefore the mean of the measured values (4.5 mg/L) were used for evaluation.

The highest tested concentration at which no negative effects were observed on the reproduction rate or survival of parental animals is: NOEC >= 4.5 mg/L (highest test concentration under test conditions).

The lowest tested concentration at which the first significant effects were observed on the reproduction rate or survival of the parent animals is: LOEC > 4.5 mg/L (highest concentration under test conditions).

The concentrations were based on the product.

Description of key information

Long-term toxicity to Daphnia: NOEC (21-d reproduction) - 4.04 mg/L; EC50 (21-d reproduction) - 10.4 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

NOEC

Effect concentration:

4.04 mg/L

Additional information

Chronic toxicity to Daphnia has been investigated in a 21-day reproduction test according to OECD test methods and using dissolution/dispersion agents to increase concentration of the substance in the test medium. The NOEC (21-day, reproduction) was 4.04 mg/L and the EC50 (21 -day, reproduction) was 10.4 mg/L.

Tests with a structural analogue of the substance, a trimellitate ester with mixed C8-C10 side chains, showed no toxicity when tested at a concentration of 4.5 mg/L, this finding being in agreements with the observed NOEC/LOEC from testing with the substance itself. This lower value compared with the first study reflect the achieved concentrations of the substance in the test media, this influenced by the agents used to achieve dissolution/dispersion.