Reaction mass of Benzenepropanal, 4-ethyl-α,α-dimethyl- and 3-(2-ethylphenyl)-2,2-dimethylpropanal

Bioaccumulation: aquatic / sediment

Link to relevant study record(s)

Reference

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 May 2014 to 9 June 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 305 (Bioaccumulation in Fish: Aqueous and Dietary Exposure) -I: Aqueous Exposure Bioconcentration Fish Test

Qualifier:

according to guideline

Guideline:

other: Japanese national guideline: "Method for Testing the Degree of Accumulation of Chemical Substances in Fish Body" stipulated in the "Testing Methods for New Chemical Substances".

GLP compliance:

yes (incl. QA statement)

Radiolabelling:

no

Details on sampling:

SAMPLING AND FREQUENCY OF ANALYSIS
- Test water: The test water of Level 1 and Level 2 were analyzed once before the first analysis of test fish and at the same time as analysis of test fish thereafter. One sample was analyzed at each sampling time. Test water samples were pretreated before GC-MS analysis.
- Test fish: Analysis of test fish was performed five times during the exposure period. Four fish per treatment level were taken out at each sampling time and divided into two groups (two fish per group) because one fish was not enough for the analytical sensitivity of the test item. Test fish collection at final three successive analyses was carried out at intervals of more than 48 hours, and the last analysis was performed 28th days. Analysis of control fish was performed before the experimental start and after the experimental completion. Four fish were taken out at the sampling time and divided into two groups, and analyzed individually.

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

PREPARATION OF STOCK SOLUTIONS
- Dispersant: N,N-Dimethylformamide was used to prepare stock solutions.
- Level 1: Test sample (1000 mg) was dissolved in N,N-Dimethylformamide to prepare 1000 mg/L stock solution (1 L).
- Level 2: Test sample (100 mg) was dissolved in N,N-Dimethylfonnamide to prepare 100 mg//L stock solution (1 L).
- Control: N,N-Dimethylformamide was supplied.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

- Common name: common carp
- Supplier: CERl Kunume
- Lot No.: TFC-140318
- Length: 7.0 - 9.3 cm (at the beginning of exposure: 7.0 - 7.5 cm)
- Age: Yearling fish
- Feeding: Feed: Feed for fry of carp; Composition: Proteins content ≥43.0%, Lipid content ≥3.0%; Manufacturer: Nippon Formula Feed Mfg.; Feeding amount and interval: Amount corresponding to about 3% of total body weight was fed twice a day in halves (once a day in all at holiday). The fish were starved for 24 hours before sampling.
- Conditions for acclimatization: The external disinfection was carried out in an aqueous solution containing OTC for fisheries (oxytetracycline hydrochloride, Kyoritsu Seiyaku) and sodium chloride (The Salt Industry Center of Japan). Thereafter fish were acclimatized in the following conditions: Period: 23 days; Temperature: 25 ± 2°C; Mortality during acclimatization was less than 5%.
- Reason for selection: The previous data conducted with this species can be compared and the size of this species is adequate for handling.

Route of exposure:

aqueous

Justification for method:

aqueous exposure method used for following reason: log Kow <5

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

28 d

Hardness:

- Dilution water (underground water): 12 mg/L (as Ca, Mg)

Test temperature:

- Level 1: 24.8 - 25.2 °C
- Level 2: 24.5 - 25.1 °C
- Control: 24.5 - 25.1 °C

pH:

- Level 1: 7.9
- Level 2: 7.9
- Control: 7.9 - 8.0

Dissolved oxygen:

- Level 1: 7.7 - 8.2 mg/L
- Level 2: 7.8 - 8.1 mg/L
- Control: 7.8 - 8.2 mg/L

TOC:

- Dilution water (underground water): 0.7 mg/L

Conductivity:

- Dilution water (underground water): 280 µS/cm

Details on test conditions:

TEST MEDIUM / WATER PARAMETERS
- Origin of dilution water for test: Groundwater from the premises of CERI Kurume;
- Test water quality assessment: Except for the alkalinity and electric conductivity, it was confirmed that the dilution water met the requirements of at least one of the following standards: 1. Ministerial ordinance of the Ministry of Health, Labour and Welfare No.101 (Revised May 30, 2003); 2. OECD Guidelines for Testing of Chemicals, No.210, July 17, 1992, "Fish, Early-life Stage Toxicity Test"; 3. Water quality criteria for fisheries (Japan Fisheries Resource Conservation Association, March 1983); 4. Environmental Quality Standards for Water Pollutants No.14 (Revised February 22, 1999, Environment Agency); 5. OECD Guidelines for Testing of Chemicals, No 305, June 14, 1996, "Bioconcentration : Flow-through Fish Test".

TEST CONDITIONS / CIRCUMSTANCES
- Test tank: 70-L glass tank;
- Flow rate of test water: 0.04 mL/min for stock solution and 2000 mL/min for dilution water, 2880 L/day of test water, were supplied. Flow rate was measured once per day during the xposure period.
- Stock solution bottle: 1-L brown glass bottle (frequency of renewal once per two weeks);
- Aeration: The test tanks were supplied with air during the exposure;
- Number of fish (at the beginning of exposure): Level 1and 2 exposure: 24; Control: 16
- Reason for duration of exposure / uptake: It was evaluated that a steady-state was reached after 28 days.
- Cleaning of tanks: Excreta of fish and other dirt on test tank were removed about once a day druing the test period.

OTHER TEST CONDITIONS
- Photperiod: 14 hours light / 10 hours dark (artificial light of white fluorescent lamp).

Nominal and measured concentrations:

- Nominal concentrations: 20 µg/L (Level 1) and 2 µg/L (Level 2).
- Measured concentrations: Concentrations of the test item were maintained ≥81% of nominal concentrations and the variations were within ± 20% of the average measured concentrations. See 'Any other information on materials and test methods incl. tables' for detail.

Reference substance (positive control):

no

Lipid content:

3.02 %

Time point:

start of exposure

Lipid content:

3.48 %

Time point:

end of exposure

Key result

Conc. / dose:

20 µg/L

Temp.:

25 °C

pH:

7.9

Type:

BCF

Value:

< 132 L/kg

Basis:

whole body w.w.

Calculation basis:

steady state

Remarks on result:

other: < the concentration in the fish was < 160 ng/g fish (detection limit) and using this limit the BCF was calculated as a < value but it is corrected for 5% lipid content.

Remarks:

Based on 5% l

Conc. / dose:

20 µg/L

Temp.:

ca. 25 °C

pH:

7.9

Type:

BCF

Value:

< 9.4 L/kg

Basis:

whole body w.w.

Calculation basis:

steady state

Remarks on result:

other: Peak 1

Conc. / dose:

20 µg/L

Temp.:

ca. 25 °C

pH:

7.9

Type:

BCF

Value:

< 8.3 L/kg

Basis:

whole body w.w.

Calculation basis:

steady state

Remarks on result:

other: Peak 2

Conc. / dose:

20 µg/L

Temp.:

ca. 25 °C

pH:

7.9

Type:

BCF

Value:

< 8.9 L/kg

Basis:

whole body w.w.

Calculation basis:

steady state

Remarks on result:

other: Peak 3

Conc. / dose:

2 µg/L

Temp.:

ca. 25 °C

pH:

7.9

Type:

BCF

Value:

< 92 L/kg

Basis:

whole body w.w.

Calculation basis:

steady state

Remarks on result:

other: Peak 1

Conc. / dose:

2 µg/L

Temp.:

ca. 25 °C

pH:

7.9

Type:

BCF

Value:

< 81 L/kg

Basis:

whole body w.w.

Calculation basis:

steady state

Remarks on result:

other: Peak 2

Conc. / dose:

2 µg/L

Temp.:

ca. 25 °C

pH:

7.9

Type:

BCF

Value:

< 87 L/kg

Basis:

whole body w.w.

Calculation basis:

steady state

Remarks on result:

other: Peak 3

Details on results:

- BCF at steady-state (BCFss): Because the test item in test fish at the last three successive analyses were not more than the LOQ, BCFss was not calculated. However, because all BCFs were less than 100, it was evaluated that a steady-state was reached after 28 days.
- Lipid content: The change of lipid contents during the test was + 15% i.e. within ± 25%.
- Fish observations: No abnormality in behavior or appearance was noted.
- In the report in the summary tables the concentrations versus the BCFs were switched around. The low concentration had the higher BCFs. In the final tables and graphs presented in the report the concentrations versus BCFs were correctly reported. In the key results the correct values are presented.

Validity criteria fulfilled:

yes

Remarks:

See 'Any other information on materials and methods incl. tables'.

Conclusions:

In fish (C. carpio), at test concentrations of 20 µg/L and 2 µg/L, the BCFs were determined to be <9.42 L/kg and <92 L/kg, respectively as no Floralozone could be determined at the starting day of depuration. The numbers presented are the limit of detection in fish 160 ng/g. (ug/kg fish).

Executive summary:

The bioaccumulation of the substance was determined in a study according to Japanese national guidelines equivalent to OECD TG 305 and in compliance with GLP criteria (IFF, 2014). In this study groups of 24 fish (carp: Cyprinus carpio) were exposed for 28 days to nominal test substance concentrations of 20 and 2 µg/L in a flow-through system. Test water nominal concentrations were analytically verified and recoveries within ± 20% of nominal. After the exposure period, fish were measured for weight and body length, chopped into pieces and homogenised with tetrahydrofuran for analysis (whole body ww only). Fish lipid content was measured before and after the 28 -day exposure period and determined to increase from 3.02% to ca. 3.48% throughout the test which was attributed to growth of the fish. Depuration could not be examined because the test item in test fish at the last three successive analyses were below the LOQ. After the 28 -day exposure period, the BCF values at test concentrations of 20 and 2 µg/L were determined to be <92 and < 9.4 L/kg ww, respectively as no Floralozone could be determined at the starting day of depuration. The numbers presented are the limit of detection in fish 160 ng/g. (ug/kg fish).

Taking into account the detection limit the worst case BCF of 92 L/kg ww for whole fish can be converted to a standard fish lipid content of 5%, which would result in a BCF <= <132 L/kg ww (92/3.48*5).

Description of key information

BCF in an OECD TG 305 is < 132 L/kg based on 5% lipid content, the value is based on absence of substance in fish at the start of depuration and the detection limit in fish of 160 ng/g (ug/kg) fish.

Key value for chemical safety assessment

BCF (aquatic species):

132 L/kg ww

Additional information

The bioaccumulation of the substance was determined in a study according to Japanese national guidelines equivalent to OECD TG 305 and in compliance with GLP criteria (IFF, 2014). In this study groups of 24 fish (carp: Cyprinus carpio) were exposed for 28 days to nominal test substance concentrations of 20 and 2 µg/L in a flow-through system. Test water nominal concentrations were analytically verified and recoveries within ± 20% of nominal. After the exposure period, fish were measured for weight and body length, chopped into pieces and homogenised with tetrahydrofuran for analysis (whole body ww only). Fish lipid content was measured before and after the 28 -day exposure period and determined to increase from 3.02% to ca. 3.48% throughout the test which was attributed to growth of the fish. Depuration could not be examined because the test item in test fish at the last three successive analyses were below the LOQ. After the 28 -day exposure period, the BCF values at test concentrations of 20 and 2 µg/L were determined to be <92 and < 9.4 L/kg ww, respectivelyas no Floralozone could be determined at the starting day of depuration. The numbers presented are the limit of detection in fish 160 ng/g. (ug/kg fish).

Taking into account the detection limit the worst case BCF of 92 L/kg ww for whole fish can be converted to a standard fish lipid content of 5%, which would result in a BCF <= <132 L/kg ww (92/3.48*5).