REAKTIV-GELB F-68 072 FW

Bioaccumulation: aquatic / sediment

Link to relevant study record(s)

Reference

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

03 May 2007 - 10 August 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 305 (Bioconcentration: Flow-through Fish Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not applicable.

Radiolabelling:

no

Details on sampling:

Analytical Procedure – Analysis of Samples
Fish samples from the biological test [2 control fish and 4 fish of the test solution (concentration 1 mg/L) at the end (after 168h)] were prepared
according to the above described procedure (analytical procedure without test item).

During the uptake phase the water characteristics as temperature, dissolved oxygen, TOC and pH were measured. Analytical monitoring in both water and fish samples was carried out using a HPLC-method. On day two, five and seven, 4 fish were taken from the control and the test item concentration 1 mg/L and stored deep frozen at -70°C. After 168 hours of exposure 2 control fish and 4 fish of the test item concentration 1 mg/L were analysed. As all values were below the limit of quantification (17.9 μg/g fish fresh-weight) none of the earlier fish samples was analysed.

Vehicle:

no

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

See above "Details on Analytical Methods".

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Test species
Name : Zebra fish (Danio rerio)
Source : Aqua KlöGer (Germany)
Date of birth : August-October 2006
Date supplied : 2007-02-13
Mortalities during
acclimatisation period : < 5 %
Medication : none
Mean standard length
(n = 20) : 3.35 cm (S.D. = 0.16 cm)
Mean loading (n = 20) : 0.75 g fish/L

Keeping conditions
Max. 500 animals were held in 100 L chambers in synthetic fresh water with about 5 volume replacements per day at 22 ± 2 °C
Fish were fed throughout on a sufficient diet (Tetra Min® commercial fish food ground) at least once a day, occasionally supplemented by live Daphnia and frozen bloodworms (Chironomus larvae). Temperature was measured daily; dissolved oxygen concentration and pH was measured once a week.
Mortalities were recorded daily, dead and/or diseased fish were discarded. If necessary, chambers were cleaned from faeces and food residues. Once a week, glass of chambers was cleaned thoroughly

Holding and dilution water
Synthetic fresh water was prepared in accordance with ISO 7346-1 (1996):
Water quality – Determination of the acute lethal toxicity of substances to a fresh-water fish [Brachydanio rerio Hamilton-Buchanan (Teleostei,
Cyprinidae)]-Part 1: Static method

Stock solutions of the following substances were prepared:

The corresponding amounts of the solutions were added to deionised water of a constant quality to achieve a concentration of
294.0 mg/L CaCl2 • 2 H2O
123.3 mg/L MgSO4 • 7 H2O
63.0 mg/L NaHCO3
5.5 mg/L KCL
Since the keeping water was of constant quality, the composition of the ions, acidity and hardness was determined every 3 months. Once a year
determinations of heavy metals, pesticides and chloride aromatics was carried out. This freshly prepared standard keeping water was used for both, the maintenance of the test animals under flow-through conditions and the preparation of stock and test solutions of the test item.

Route of exposure:

aqueous

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

168 h

Hardness:

The hardness of the dilution water used was 15.9 °dH (= 283.8 mg/L CaCO3) in the control and 15.6 °dH (= 278.5 mg/L CaCO3) in the system with test item.

Test temperature:

The temperature was in the range of 20 – 25 °C at beginning of the test and did not vary by more than ± 2 °C.

pH:

7.8 - 8.0

Dissolved oxygen:

The dissolved oxygen concentration was > 60% of the air-saturation value throughout the study.

TOC:

< 2 mg/l

Salinity:

Not applicable.

Details on test conditions:

Exposure conditions
Test vessels : glass aquaria holding 40 L of test media
Experimental design : 1 test concentration plus 1 control 40 animals per test concentration/control flow-through system (frequency of water renewal: about 6 times per day).
Method of initiation : animals were placed in prepared media
Loading : 0.1 – 1 g fish (wet weight) per litre test medium
Photoperiod : 16 h light: 8 h dark
Temperature : mean < +/- 2 °C (in the temperature
range 20 – 25 °C)
Aeration : gentle aeration via narrow glass tubes
Test item concentration/s : 1 mg/L
Medium renewal : flow-through system
Duration of Uptake phase: 168 hours
Food and feeding rate : Tetra Min® fish food, ground (8% lipid, 48% raw protein, 2% fibre, raw ash 11%, water 6%)
2 % of wet weight per day, supplied in 2 portions daily (10.00 a.m., 2.00 p.m.)
Parameters measured during the test :
- temperature and dissolved oxygen values were measured at the beginning of the test and every 24 hours thereafter
- pH was measured once a week
- mortality was recorded every 24 hours.
- analysis of the test item in each test system and in fish
- analysis of the lipid content of fish at test start and end
- at the end of the test the fish was weighed and the length was measured

Dosing system
The test was run under constant flow-through conditions. The stock solution of the test item and the synthetic fresh water were dosed into a mixing chamber with a volume of 0.8 L by means of membrane pumps. From this mixing chamber, the test solution was directly introduced into the test vessel. The control vessel received synthetic fresh water only, the flow rate being the same as in the test vessel.
The flow rates of the pumps were checked daily during the acclimation phase (except on weekends) and daily during the uptake phase.

Nominal and measured concentrations:

1 mg/l

Reference substance (positive control):

no

Details on estimation of bioconcentration:

The bioconcentration factor was calculated as the ratio of concentration in the whole fish (Cf) and in the water (Cw) at steady-state (BCFss) according to equation (1):

BCFss = Cf / Cw (1)

The bioconcentration factor referring to the lipid content at steady-state (BCFss) was calculated according to equation (2):

BCFss wet weight / lipid content * 100 = BCFss lipid content (2)

BCFss wet weight : BCF referring to wet weight
lipid content : mean value of lipid content [%] at each test concentration
BCFss lipid content: BCF referring to the lipid content

Lipid content:

15.3 %

Time point:

end of exposure

Remarks on result:

other: BCFss referring to lipid content of wet weight < 117

Type:

BCF

Value:

< 17.9 dimensionless

Basis:

total lipid content

Time of plateau:

168 h

Calculation basis:

steady state

Remarks on result:

other: The BCF determinations referring to the lipid content confirmed that REAKTIV GELB F68072 FW does not have a potential for bioconcentration.

Remarks:

Conc.in environment / dose:1 mg/l

Details on kinetic parameters:

Not measured.

Metabolites:

Not measured.

Results with reference substance (positive control):

Not applicable.

Details on results:

Test concentration [mg/L]: 1
Mean value of lipid content [%] 15.3
BCFSS referring to: lipid content of wet weight <117

Compared with the nominal concentration of 1 mg/L it can be concluded that the BCF was < 17.9 and that Reaktiv Gelb F68072 FW does not have a
significant bioaccumulation potential.

See tabulated data below.

Reported statistics:

None.

ABIOTIC PARAMETERS Acclimation phase

Test item concentration [mg/L]:   Control

Day	Date 2007	Tempera­ture [°C]	Oxygen [mg/L]	Oxygen [% saturation]	pH-value	TOC [mg/L]	Comments
-6	05-03	22.8	8.4	98	7.9	—	none
-5	05-04	22.9	8.5	99	—	—	none
-4	05-05	—	—	—	—	—	—
-3	05-06	—	—	—	—	—	—
-2	05-07	23.0	8.3	96	8.0	< 2	none
-1	05-08	22.8	8.5	100	—	—	none

Comments: The acclimation phase was run without fish.

ABIOTIC PARAMETERS AND MORTALITY : Uptake phase

Test item concentration [mg/L]:   Control

 

Day	Date	Tempera­ture	Oxygen		PH value	TOC		Mortality
	2007	[°C]	[mg/L]	[% saturation]		[mg/L]	abso­lute	cumu­lative	cumu­lative [%]
0	05-09	22.7	8.5	99	8.0	—	0	0	0
1	05-10	22.7	8.5	98	—	< 2	0	0	0
2	05-11	22.8	8.2	97	—	—	0	0	0
3	05-12	22.9	8.3	98	—	—	0	0	0
4	05-13	22.9	8.2	98	—	—	0	0	0
5	05-14	22.9	8.5	100	—	—	0	0	0
6	05-15	22.8	8.3	97	—	—	0	0	0
7	05-16	22.8	8.4	98	7.9	—	0	0	0

Comments:Loading rate at test start was 40 animals (total wet weight: 14.1 g)

Acclimation phase

Test item concentration [mg/L]:   1

Day	Date 2007	Tempera­ture [°C]	Oxygen [mg/L]	Oxygen [% saturation]	pH-value	TOC [mg/L]	Comments
-6	05-03	23.1	8.4	99	7.9	—	none
-5	05-04	23.1	8.4	99	—	—	none
-4	05-05	—	—	—	—	—	—
-3	05-06	—	—	—	—	—	—
-2	05-07	23.5	8.3	97	8.0	< 2	none
-1	05-08	23.2	8.6	101	—	—	none

Comments: The acclimation phase was run without fish.

 

Uptake phase

Test item concentration [mg/L]:   Control

 

Day	Date	Tempera­ture	Oxygen		pH value	TOC		Mortality
	2007	[°C]	[mg/L]	[% saturation]		[mg/L]	abso­lute	cumu­lative	cumu­lative [%]
0	05-09	23.2	8.4	99	8.0	—	0	0	0
1	05-10	23.1	8.4	99	—	< 2	0	0	0
2	05-11	23.2	8.2	98	—	—	0	0	0
3	05-12	23.4	8.2	98	—	—	0	0	0
4	05-13	23.4	8.3	98	—	—	0	0	0
5	05-14	23.4	8.5	101	—	—	0	0	0
6	05-15	23.2	8.3	98	—	—	0	0	0
7	05-16	23.2	8.3	98	7.8	—	0	0	0

 

Comments:Loading rate at test start was 40 animals (total wet weight: 14.7 g)

 

TEST RESULTS - CHEMICAL ANALYSIS IN WATER (C_w) Acclimation phase

Test item concentration [mg/L]:   Control

 

Hour	Day	Date	Control mediu HPLC values	m(Cw)[mg/L] mean
-44	2	2007-05-07	< 0.060/<0.060	< 0.060
-41	2	2007-05-07	< 0.060/<0.060	< 0.060
-24	1	2007-05-08	< 0.060/<0.060	< 0.060

Comments:none

 

 

TEST RESULTS - CHEMICAL ANALYSIS IN WATER (C_w) Acclimation Phase:

Test item concentration [mg/L]:   Control

 

Hour	Day	Date	Test medium (Cw) [mg/L]
			HPLC values	mean
-44	2	2007-05-07	1.532/ 1.540	1.536
-41	2	2007-05-07	1.461 / 1.460	1.461
-24	1	2007-05-08	1.240/ 1.257	1.249

Comments:none

 

 

Test item concentration [mg/L]:   1

 

Hour	Day	Date	Test medium (Cw) [mg/L]
			HPLC values	mean
-44	2	2007-05-07	1.532/ 1.540	1.536
-41	2	2007-05-07	1.461 / 1.460	1.461
-24	1	2007-05-08	1.240/ 1.257	1.249

Comments:none

 

TEST RESULTS - CHEMICAL ANALYSIS IN WATER (C_w) AND FISH (C_F) Uptake phase

Test item concentration [mg/L]:   Control

 

Hour	Day	Date	Test item concentrations in:
			control medium (Cw) [mg/L]		control fish (Cf) [μg/g]
			HPLC Values	Mean	HPLC Values *	Mean
4	0	2007-05-09 (4 h after test start)	<   0.060/ <   0.060	< 0.060	—	—
24	1	2007-05-10 (24 h after test start)	<   0.060/ <   0.060	< 0.060	—	—
48	2	2007-05-11 (48 h after test start)	<   0.060/ <   0.060	< 0.060	—	—
52	2	2007-05-11 (52 h after test start)	<   0.060/ <   0.060	< 0.060	—	—
120	5	2007-05-14 (120 h after test start)	<   0.060/ <   0.060	< 0.060	—	—
124	5	2007-05-14 (124 h after test start)	<   0.060/ <   0.060	< 0.060	—	—
168	7	2007-05-16 (168 h after test start)	<   0.060/ <   0.060	< 0.060	<   17.9/ <   17.9	< 17.9

 

Comments:*) The above analytical values were obtained from two individual specimens.

 

TEST RESULTS – CHEMICAL ANALYSIS IN WATER (CW) AND FISH (CF)

Uptake phase

Test item concentration [mg/L]: 1

 

Hour	Day	Date	Test item concentrations in:
			control medium (Cw) [mg/L]		control fish (Cf) [μg/g]
			HPLC Values	Mean	HPLC Values *	Mean
4	0	2007-05-09 (4 h after test start)	1.375/ 1.381	1.378	—	—
24	1	2007-05-10 (24 h after test start)	1.442 / 1.438	1.440	—	—
48	2	2007-05-11 (48 h after test start)	1.385/ 1.399	1.392	—	—
52	2	2007-05-11 (52 h after test start)	1.401 / 1.390	1.396	—	—
120	5	2007-05-14 (120 h after test start)	1.400/ 1.407	1.404	—	—
124	5	2007-05-14 (124 h after test start)	1.355/ 1.384	1.370	—	—
168	7	2007-05-16 (168 h after test start)	1.341 / 1.347	1.344	<   17.9/ <   17.9/ <   17.9/ <   17.9	< 17.9

Comments:*) The above analytical values were obtained from four individual specimens.

 

TOTAL WET WEIGHT AND BODY LENGTH OF TEST FISH AT THE END OF THE STUDY

 

Test fish	Body length [cm] at various test concentrations [mg/L]
		Control	1
1	3.5	3.5
2	3.5	3.0
3	3.3	3.4
4	3.4	3.4
5	3.1	3.5
6	3.3	3.4
7	3.4	3.3
8	3.0	3.2
9	3.5	3.4
10	3.5	3.4
Mean value of body length	3.35	3.35
Standard deviation	0.18	0.15
Wet weight [g] of 10 fish	3.51	3.94
Loading [g fish/L]	0.70	0.79

 

Comments:At the end of the test, ten fish were taken at random from both, the test and the control vessels in order to check their compliance with the size range recommended for the zebra fish by the OECD test guideline.

ANALYSIS OF THE LIPID CONTENT OF THE FISH

[% of wet weiqht:

 

Date	Control	Test item concentration 1 mg/L
2007-05-09 Start of the uptake phase	13.2	13.2
2007-05-16 End of the depuration phase	10.7	17.3
Mean value of lipid content*	12.0	15.3

Validity criteria fulfilled:

yes

Conclusions:

On the basis of the fact that the BCF was < 17.9, Reaktiv Gelb F68072 FW does not have a significant bioaccumulation potential.

Executive summary:

A study was performed to assess the bioconcentration potential Reaktiv Gelb F68072 FW in fish.

 

The study was conducted in agreement with the German Federal Environment Agency (UBA) and, with adaptations, according to the general provisions of OECD TG 305 – Bioconcentration: Flow-through Fish Test (June 1996). This guideline

describes a procedure for characterising the bioconcentration potential of items in fish under flow-through conditions.

 

During the uptake phase of the test, a group (n= 40) of Danio rerio was exposed to Reaktiv Gelb F68072 FW at a nominal concentration of 1 mg/L. The test concentration was based on the outcome of an acute fish test using the same test

species with a LC50of > 100 mg/L. Recoveries for the test item concentration in water during the 7 d uptake phase according to analytical HPLC determinations were between 134.4 and 144.0 %. Fat content of fish was 13.2 % at the beginning of the

test and 17.3 % at the end of the test.

 

After an exposure period of 168 h (with a LOQ of 17.9 μg/g fish fresh-weight) no test item could be detected in the fish. Thus the test was ended without depuration phase.

 

Compared with the nominal concentration of 1 mg/L it can be concluded that the BCF was < 17.9 and that Reaktiv Gelb F68072 FW does not have a significant bioaccumulation potential.

Description of key information

On the basis of the fact that the BCF was < 17.9, Reaktiv Gelb F68072 FW does not have a significant bioaccumulation potential.

Key value for chemical safety assessment

BCF (aquatic species):

8 dimensionless

Additional information

A study was performed to assess the bioconcentration potential Reaktiv Gelb F68072 FW in fish.

 

The study was conducted in agreement with the German Federal Environment Agency (UBA) and, with adaptations, according to the general provisions of OECD TG 305 – Bioconcentration: Flow-through Fish Test (June 1996). This guideline describes a procedure for characterising the bioconcentration potential of items in fish under flow-through conditions.

 

During the uptake phase of the test, a group (n= 40) of Danio rerio was exposed to Reaktiv Gelb F68072 FW at a nominal concentration of 1 mg/L. The test concentration was based on the outcome of an acute fish test using the same test species with a LC50of > 100 mg/L. Recoveries for the test item concentration in water during the 7 d uptake phase according to analytical HPLC determinations were between 134.4 and 144.0 %. Fat content of fish was 13.2 % at the beginning of the test and 17.3 % at the end of the test.

 

After an exposure period of 168 h (with a LOQ of 17.9 μg/g fish fresh-weight) no test item could be detected in the fish. Thus the test was ended without depuration phase.