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

Bioaccumulation: aquatic / sediment

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Start: 15/05/2015 - End: 28/09/2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Tested in Accordance with Procedure 305B of the OECD Guidelines for Testing Chemicals.

Qualifier:

according to guideline

Guideline:

OECD Guideline 305 B (Bioaccumulation: Semi-static Fish Test)

Principles of method if other than guideline:

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Dietary vs Aqueous
Lipids

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Lot No.: 04698/MA
Purity > 98%

Details on sampling:

Sampling and Analysis of fish and water:
Water from the test chambers were sampled on days 0, 3, 7, 10, 14, 21, 28 to determine the test substance concentration before adding the fish, and during uptake phases. The water was sampled at the same time as the fish and before feeding.

Sampling and sample treatment:
Water samples were obtained by siphoning through inert tubing from a central point in the test chamber. Seven fish were removed from the chambers at each sampling interval. Sampled fish were rinsed quickly with water, blotted dry, killed instantly using the appropriate and humane method, and then weighed.

Treatment of extraction and concentration for the fish samples:
Seven fish and some anhydrous sodium sulfate were put in the mortar and ground. The samples were extracted by adding 5.0 mL acetonitrile and extracted after 20 min. The process was repeated twice and the extract solutions combined. The final solution was injected to UPLC-PDA after being filtered with 0.22 µm Millipore membrane.

Treatment of extraction and concentration for the water sample:
Water sample (250 mL) was extracted by 50 mL ethyl acetate each time. The process was repeated twice and the extract solutions combined. The extract solution was dried by rotary evaporation; 1.00 mL acetonitrile was used to dissolve and the sample was analyzed by UPLC-PDA.

Vehicle:

no

Details on preparation of test solutions, spiked fish food or sediment:

Preparation of test solutions:
The test solutions were prepared with saturated solutions. The aqueous saturated solution of test substance was prepared by dissolving test substance (~ 0.2 g) in 20 L test solution at 35°C for 24 h, when saturation was achieved the mixture is cooled at 23°C, subsequently, the mixture is filtered by 0.45 µm glass fiber filter membrane. Some initial filtrates (~ 100 mL) were discarded to minimize adsorption.
During the test, aeration was not used. The time interval of changing exposure solutions was 72h under the requisition of quality assurance and quality control. When changing the test solution, transfer of the fish to the fresh medium of the test substance was performed quickly. The fresh test solution was prepared 24h in advance to ensure the balance of test substance and water.

Test organisms (species):

other: Gobiocypris rarus

Details on test organisms:

TEST ORGANISM
- Common name:
Rare Minnow

- Source:
Institute of Hydrobiology, Chinese Academy of Sciences (Batch No.: F20150515G).
Fish were age synchronised but the age is not stated.

- Length at study initiation (length definition, mean, range and SD):
(3.0±0.5) cm

- Weight at study initiation (mean and range, SD):
(0.2±0.1)g

- Health status:
Fish used in tests were free from observable diseases and abnormalities.

- Description of housing/holding area:
The test fish were randomly assigned to the treatment levels and controls with 70 fish each 20 L tank. The loading rate was 0.7 g of fish (wet weight) per litre of water.

- Feeding during test
The fish were fed daily throughout the acclimation and test periods at a level of approximately 1% to 2% of body weight per day.

ACCLIMATION
-The stock population of fish was acclimated for 15 days in water at the test temperature and fed throughout on a sufficient diet (1 % ~ 2% weight of fish) and of the same type to be used during the test.

Following a 48-hour settling-in period, no fish died during the holding time of 7 d. Fish used in tests were free from observable diseases and abnormalities. No fish died during the holding time of 7d

-The ingredients of the fish food are given as follows:
Crude Protein > 36.0%, Crude Fat > 2.0%, Crude Fibre< 3.0%, Crude Ash< 12.0%, Moisture< 10.0%.

Route of exposure:

aqueous

Remarks:

Semi-static

Justification for method:

aqueous exposure method used for following reason:

Test type:

semi-static

Water / sediment media type:

other: Good quality tap water, dechlorinated for 24 hours.

Total exposure / uptake duration:

0 - <= 28 d

Hardness:

145.0 mg/L (CaCO3)

Test temperature:

23.2°C (+/- 0.2°C)

pH:

7.69 - 7.84

Dissolved oxygen:

77.5% - 90.4%

TOC:

1.62 mg/L

Salinity:

N/A

Conductivity:

N/A

Details on test conditions:

N/A

Nominal and measured concentrations:

Nominal: 0.160 mg/L

Measured: 0.157; 0.159; 0.160 mg/L

Details on estimation of bioconcentration:

BCF
A sample of water and fish was taken each day in parallel after 0, 3, 7, 10, 14, 21, 28 days. Therefore, 7 concentration values of water sample and fish sample were obtained and then 7 BCF values were calculated.

The uptake curve of the test substance was obtained by plotting its concentration in/on fish in the uptake phase against time on arithmetic scales. The steady state BCFss is calculated from the following equation:

BCFss = Cf (steady-state)/Cw (steady-state)

Where:
Cf: concentration of test substance in fish sample
Cw: concentration of test substance in water sample

Otherwise, the concentration factor is expressed as the range of BCF value.

However, since the test substance concentration in fish didn't reach equilibrium, the BCF, was used to represent biological concentration factor of fish.

Relative Standard Deviation (RSD)
The "STDEV" function of MS Office Excel was used to calculate the SD, RSD is the ratio of the SD
against the average of the concentration.

RSD(%) = SD/(average of C) *100

Where:
RSD: Relative Standard Deviation (%);
SD: Standard Deviation(%);
(average of C): Average of concentration.

Key result

Conc. / dose:

0.142 mg/L

Type:

other: BCF-28 (BCF 28 Days)

Value:

218 L/kg

Basis:

whole body w.w.

Calculation basis:

steady state

Remarks on result:

other: The BCF doesn't reach steady state by day 28. Therefore, the BCF-28 was introduced to represent the BCF for fish after 28 days.

Details on kinetic parameters:

N/A

Metabolites:

N/A

Results with reference substance (positive control):

N/A

Details on results:

The test is considered valid as all of the following performance criteria were met.
- the temperature was maintained (23±2) °C;
- the concentration of dissolved oxygen did not fall below 60% saturation;
- the concentration of the test substance in the chambers was maintained within ±20% of the mean of the measured values during the uptake phase;
- the mortality or other adverse effects/disease in both control and treated fish were less than 10% at the end of the test.

Reported statistics:

N/A

Recovery of Test Substance in Medium

Nominal Concentration (mg/L)	Measured Concentration (mg/L)	Recovery Rate (%)	Mean Recovery Rate (%)	RSD (%)
0.160	0.157	98.1	99.2	0.98
	0.159	99.4
	0.160	100

Recovery of Test Substance in Fish

Nominal Concentration (mg/L)	Measured Concentration (mg/L)	Recovery Rate (%)	Mean Recovery Rate (%)	RSD (%)
5.00	5.55	111	112	0.89
	5.66	113
	5.58	112
30.00	31.4	105	105	2.39
	32.5	108
	30.9	103

Stability of Test Substance in Test Solutions

Test Time	0 h	24 h	48 h	72 h
Measured Concentration (mg/L)	0.137	0.139	0.137	0.138

 

Conclusions:

BCF doesn't reach the steady state at the 28th day (Table 7 and Figure 7). Therefore, the BCF28, for fish was introduced to represent BCF, the mean value was 218. During the test (28 d), the BCF28 value of test substance is 218, when exposed to the test substance solution of 0.142 mg/L.

Executive summary:

Bioconcentration factor (BCFs) of the test substance in fish (Gobiocypris rarus) was tested according to "The guidelines for the testing of chemicals" (HJ/T 153-2004) and "The Guidelines for the Testing of Chemicals, Degradation and Accumulation" and with reference to Procedure 305B of the "Guidelines for Testing of Chemicals" of the OECD: "Bioconcentration: Semi-static Fish Test" (OECD TG 305B).

The recovery in the water sample was 99.2% and RSD was 0.98%, when the added concentration of the test substance is 0.16 mgL in water samples. The recovery in the fish sample was 112% and RSD was 0.89%, while the added concentration of the test substance is 5 mg/kg in fish samples. The recovery in the fish sample was 105% and RSD was 2.39%, while the added concentration of the test substance is 30 mg/kg in fish samples. Calculated from S/N>3 and S/N >/- 10, the limit of detection concentration for test substance is 0.20 mg L and the limit of quantity concentration for test substance is 0.50 mg/L.

The results show that the BCF, value of test substance is 218, when the concentration exposed to is 0.148 mg/L.

Reference 2

Endpoint:

bioaccumulation in aquatic species, other

Remarks:

QSAR

Type of information:

(Q)SAR

Adequacy of study:

key study

Study period:

2011

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data are calculated

Justification for type of information:

QSAR prediction

Qualifier:

according to guideline

Guideline:

other: REACh guidance on QSARs R6, May, July 2008

Principles of method if other than guideline:

The bioconcentration factor (BCF) was calculated by the BCFBAF subroutine (version 3.01) in the EPI SuiteTM (2011) computer model that estimates the BCF based upon chemical structure and log octanol-partition coefficient.

GLP compliance:

no

Specific details on test material used for the study:

N/A

Details on sampling:

N/A

Details on preparation of test solutions, spiked fish food or sediment:

N/A

Details on test organisms:

N/A

Hardness:

N/A

Test temperature:

N/A

pH:

N/A

Dissolved oxygen:

N/A

TOC:

N/A

Salinity:

N/A

Conductivity:

N/A

Details on test conditions:

N/A

Nominal and measured concentrations:

N/A

Details on estimation of bioconcentration:

The BCFBAF version 3.01 programme of the EPI Suite software (EPIWEB 4.1) was used to predict the BDF either regression-based estimate or Arnot-Gobas upper trophic estimate.

Key result

Type:

BCF

Value:

6.364 L/kg

Calculation basis:

other: calculation

Remarks on result:

other: regression-based method

Type:

BAF

Value:

0.9 L/kg

Calculation basis:

other: calculation

Remarks on result:

other: Arnot-Gobas method (upper trophic)

Details on kinetic parameters:

N/A

Metabolites:

N/A

Results with reference substance (positive control):

N/A

Details on results:

N/A

Reported statistics:

N/A

N/A

Validity criteria fulfilled:

yes

Conclusions:

The BCFBAF version 3.01 programme of the EPI Suite software (EPIWEB 4.01) was used to predict either a log BCF of 0.804 (BCF 6.364 L/kg) base on a regression estimate or a log BAF of -0.046 (BAF = 0.9) based on the Arnot-Gobas upper trophic model both using the smiles code.

Executive summary:

The BCFBAF version 3.01 programme of the EPI Suite software (EPIWEB 4.01) was used to predict either a log BCF of 0.804 (BCF 6.364 L/kg) base on a regression estimate or a log BAF of -0.046 (BAF = 0.9) based on the Arnot-Gobas upper trophic model both using the smiles code. Because the Arnot-Gobas upper trophic model takes also into account metabolism of the substance by the organism it is lower.

It is concluded that the test substance 1,2,4-Benzenetricarboylic acid, mixed dodecyl and octyl triesters will not accumulate in the body.

Description of key information

A BCF of 218 has been reported in an OECD 305 study. Therefore, the test substance is not considered bioaccumulative. 

Key value for chemical safety assessment

BCF (aquatic species):

6.364 L/kg ww

Additional information

A low bioconcentration factor (BCF) value (6.364 L/kg wet-wt) is estimated for of 1,2,4-Benzenetricarboxylic acid, mixed dodecyl and octyl triesters using the U. S. Environmental Protection Agency model, EPI Suite BCFBAF v.3.01.

In the study from Zhang Shenghu (2015) BCF doesn't reach the steady state at the 28th day. Therefore, the BCF28, for fish was introduced to represent BCF, the mean value was 218. During  the  test  (28  d),  the BCF28 value  of test  substance  is 218,  when  exposed  to the  test substance solution of 0.142 mg/L.

The substance is not considered as bioaccumulative following the QSAR prediction