Reaction mass of Ammonium bisethanedioato bisaquo niobate(V) (anhydrous form) and Ammonium hydrogen ethanedioate ethanedioic acid (anhydrous form)

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:

15 Jan - 12 Feb 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

October, 2012

Deviations:

no

GLP compliance:

yes

Radiolabelling:

no

Details on sampling:

- Sampling intervals/frequency for test organisms: Test fish were analysed 5 times during uptake phase; 4 fish per treatment group at each sampling time. Test fish analsis was performed at intervals more than 48 hours, and final analsis was conducted after 28 days.Test fish of control was analysed at test start and at test end.
- Sampling intervals/frequency for test medium samples:Test water was analyzed once before addition of the test fish and thereafter.
- Details on sampling and analysis of test organisms and test media samples:
45 mL water sample and 1 mL sulfuric acid were added and concentrate to 1 mL (electric griddle), ultrasonic irradiation (ultrasonic bath), filled up to 10 mL, filtrate and used for ICP AES analysis
1 g fish sample and 2 mL sulfuric and nitric acid and 2 mL, wet ashing (organic sythesizer 140 °C 2 h) cooling, filled up to 50 mL and filtrate (membrane filter pore size 0.45 µm) and used for ICP AES analysis

Vehicle:

no

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

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
For level 1: stock solution of 14.0 mg/L was prepared and appropriate amount was used to achieve the taget test item concentration of 700 µg/L
For level 2: stock solution of 1.40 mg/L was prepared and appropriate amount was used to achieve the taget test item concentration of 70 µg/L
- Controls: yes

PREPARATION OF SPIKED FISH FOOD
- Details on fish food: Feed for carp Composition : Proteins content 2:30.0%, Lipid content: 2:4.0% Manufacturer; amount corresponding to approximately 3% of total body weight was fed twice a day in halves (once aday in all at holiday); fish were fasted for 24 hours before sampling
- Details of spiking: a stock solution was prepared with ion-exchange water; those were used to achieve the required test item concentration.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Common carp
- Strain: Cyprinus carpio
- Source: CERI Kurume
- Age at study initiation: yearling fish
- Length at study initiation: 7.0 - 7.4 cm (at the beginning of exposure)
- Lipid content at test initiation: 3.10%
- Health status: Mortality during acclimatization was less than 5%
- Feeding during test
- Food type: Feed for carp(proteins content ≥30%
- Amount: approximately 3% of total body weight
- Frequency: twice a day in halves (once a day in all at holiday)

ACCLIMATION
- Acclimation period: After rearing in the underground water for 16 days, the fish were acclimatized in the dechlorinated water for 21 days.
- Acclimation conditions (same as test or not): 25±2 °C
- Health during acclimation (any mortality observed): The external disinfection was carried out in an aqueous solution containing OTC for fisheries (Oxytetracyclinehydrochloride, Kyoritsu Seiyaku) and sodium chloride (The Salt Industry Center of Japan); Mortality during acclimatization was less than 5%

Route of exposure:

aqueous

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

28 d

Hardness:

41.1 mg CaCO3/L (Level 1)
40.2 mg CaCO3/L (Control)

Test temperature:

Before uptake phase:
23.9 - 24.0 °C (Level 1)
23.9 - 24.0 °C (Level 2)
23.8 - 23.9 °C (Control)
uptake phase:
23.9 - 24.2 °C (Level 1)
23.9 - 24.2 °C (Level 2)
23.8 - 24.1 °C (Control)

pH:

Before uptake phase:
7.6 (Level 1, Level 2 and Control)
uptake phase:
7.4, 7.5 (Level 1)
7.4, 7.5 (Level 2)
7.6, 7.6 (Control)

Dissolved oxygen:

Before uptake phase:
7.9 mg/L (Level 1)
7.9 mg/L (Level 2)
7.5 mg/L (Control)
uptake phase:
7.7 - 8.0 mg/L (Level 1)
7.6 - 7.9 mg/L (Level 2)
7.6 - 8.0 mg/L (Control)

TOC:

before uptake phase:
<1.00 mgC/L, <1.00 mgC/L (Level 1, Level 2 and Control)
uptake phase:
<1.00 mgC/L (Level 1, Level 2 and Control)

Conductivity:

150 µS/µcm (dilution water)

Details on test conditions:

TEST SYSTEM
Test vessel:
- Type: open
- Material, size, headspace,fill volume: Test vessel: 70 L glass tank, Fill volume: no information
- Aeration:The test tanks were supplied with air during the exposure
- Type of flow-through: Flow-through system assembled at the laboratory was used
- Renewal rate of test solution (frequency/flow rate): 0.04 mL/min for stock solution and 800 mL/min for dilution water, 1152 L/day of test water were supplied
- No. of organisms per vessel: at the beginning of exposure: 28 (Level 1 and 2), 12 (Control)
- No. of vessels per concentration (replicates): 1 test tank per level
- No. of vessels per control / vehicle control (replicates): 1 test tank

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dechlorinated water [City water was treated by charcoal bed system (ORGANO Co., Ltd.)]
- pH: 7.7
- Particulate matter (suspended solids): < 1 mg/L
- Total organic carbon: < 0.5 mg/L
- Chemical oxygen demand: < 1 mg/L
- Metals (mg/L): Total mercury < 0.0005, Cadmium < 0.001, Chromium (VI) <0.01, Lead < 0.001, Arsenic < 0.005, Iron <0.01, Copper 0.001, Cobalt < 0.001, Manganese < 0.005, Aluminium <0.02, Zinc <0.1, Nickel < 0.001, Silver < 0.0001
- Pesticides (mg/L): 1,2-Dichloropropane <0.002, Chlorothalonil < 0.001, Propyzamide < 0.0008, Chlomitrofen < 0.0001, Simazine < 0.0003, Thiobencarb < 0.001, Diazinon < 0.0005, Isoxathion < 0.0008, Fenitrothion <0.0003, EPN <0.0006, Dichlorvos <0.001, Iprobenfos <0.0008, PCB < 0.0005, Fluorine 0.4, Anionic surfactant <0.02
- Chlorine (residual): < 0.02 mg/L
- Alkalinity: 38 mg/L
- Total hardness (Ca,Mg): 35 mg/L
- Intervals of water quality measurement: Date of Analysis January 5, 2016: lt was confirmed that the dilution water meets the criteria as stipulated in the standard operation procedure
- Intervals of test medium replacement: once a week

OTHER TEST CONDITIONS
- Photoperiod:14 h light/10h dark
- Light intensity: artificial light of white fluorescent lamp

RANGE-FINDING / PRELIMINARY STUDY: no range-finding test was performed

Nominal and measured concentrations:

Control, 70 µg/L (low exposure level, L2) and 700 µg/L (high exposure level, L1)

Reference substance (positive control):

no

Lipid content:

3.1 %

Time point:

start of exposure

Lipid content:

2.87 %

Time point:

end of exposure

Conc. / dose:

716 µg/L

Temp.:

>= 23.9 - <= 24.2 °C

pH:

7.5

Type:

BCF

Value:

> 2.3 - <= 12 dimensionless

Basis:

not specified

Time of plateau:

28 d

Remarks on result:

not determinable

Conc. / dose:

70.9 µg/L

Temp.:

>= 23.9 - <= 24.2 °C

pH:

7.5

Type:

BCF

Value:

< 23 dimensionless

Basis:

not specified

Time of plateau:

28 d

Rate constant:

growth rate constant (d-1)

Value:

0.019

Remarks on result:

other: Level 1

Rate constant:

growth rate constant (d-1)

Value:

0.02

Remarks on result:

other: Level 2

Rate constant:

growth rate constant (d-1)

Value:

0.015

Remarks on result:

other: Control

Details on results:

- Mortality of test organisms: Mortality or other adverse effects/disease in both control and test group was not observed
- Behavioural abnormalities: no abnormal behaviour was noted

Validity criteria:

The temperature variation is less than ±2 °C of set water temperature 25 °C.

The concentration of dissolved oxygen does not fall below 60% of saturated concentration 8.1 mg/L at 25 °C.

The concentration of the test item in the test tank maintained within ±20% of the mean of the measured values during the test period.

Mortality or other adverse effects/disease in both control and test group is not observed.

Analytical Results:

Concentrations of the test item were maintained 2: 94% of nominal concentrations and the variations were within ±20% of the average measured concentrations. Concentrations of the test item in test water before the uptake phase were 736 µg/L (Level 1) and 63.0 µg/L (Level 2). The measured concentration of the test item in test water of control before and after the uptake phase were less than the LOQ of the test item.

Table 1: Measured concentrations of test item in test water (Unit : µg/L)

Level	After 5days	After 13days	After 18days	After 21days	After 28days	Average (Standard deviation)
1	733	795	688	691	674	716 (49.3)
2	66.1	80.1	67.4	72.8	68.1	70.9 (5.73)

 

Table 2: BCFs     Values in the parenthesis shows the average.

Level	After 5days	After 13 days	After 18 days	After 21 days	After 28 days
1	<2.3	4.9	12	8.0	5.8
	2.5	4.4	8.4	8.7	7.0
		(4.6)	(10)	(8.4)	(6.4)
2	<23	<23	<23	<23	<23
	<23	<23	<23	<23	<23

 

Validity criteria fulfilled:

yes

Description of key information

The substance can be regarded as not bioaccumulative (Cyprinus carpio, OECD 305, nominal)

Additional information

One experimental study (CBMM Europe BV, Key, 2016, Bioaccumulation) is available investigating uptake, depuration and metabolism of the test item in fish and water according OECD 305 and GLP. 10 fish were exposed to two levels of test item concentration 700 µg/L (Level 1) and 70 µg/L (Level 2) in a flow-through system for 28 days. Test water and tested fish were analysed. Test water was analyzed once before addition of the test fish and thereafter, analyzed at the same time as analysis of test fish during the uptake phase. Test water of the control was analyzed before addition of the test fish and after the experimental completion. Tested fish were analyzed five times during the uptake phase.Test fish analysis was carried out at intervals of more than 48 hours, and the final analysis was conducted after 28 days. The lipid content in fish was 3.10% before experimental start and 2.87% after the experimental completion. Concentrations of the test item were maintained 2: 94% of nominal concentrations and the variations were within ±20% of the average measured concentrations. Concentrations of the test item in test water before the uptake phase were 736 µg/L (Level 1) and 63.0 µg/L (Level 2). The BCFs were between <2.3 and 12 for Level 1 and < 23 for Level 2. The kg calculated from the all individual test fish weight data were 0.0195 (Level 1), 0.0204 (Level 2) and 0.0155 (Control). The values are substantially lower than the threshold values for bioaccumulating (or very bioaccumulating) substances, according to Japan's Chemical Substances Control Law and Regulation (EC) No 1907/2006 (REACh). Therefore, the substance can be regarded as not bioaccumulative.