Reaction product of Chloro[29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]aluminium, sulphuric acid and caustic soda

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:

1-10-1981 to 31-12-1981

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

A standard bioaccumulation test was performed without refering to aspecific guideline. This test was carried out in flowing water and the decree to which the test compound is accumulated in fish bodies was examined.

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): Tinolux 4204

Details on sampling:

SAMPLING
- Sampling intervals/frequency for test organisms: after 3, 7, 14, 21, 28, 42 and 56 days

EXTRACTION PROCEDURE
Section 1:
- Body weight determination and addition of 160 mL acetone-chloroform (1:1)
- Homogenization for 10 minutes
- Centrifugation (6000 rpm, 20 min)
The steps in section 1 were repeated 3 times.

Section 2:
- Washing with tetrahydrofuran, water and acetone x 2 and addition of 30 mL dimethylsulfoxide
- Shaking for 30 mintues
- Centrifugation (3000 rpm, 10 minutes)
The steps in section 2 were repeated 4 times

Section 3:
- Purification on alumina column (15 mm x 30 cm)

After all three sections, samples were analytically verified by HPLC.

Vehicle:

no

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

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test substance was directly added to the test vessels.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: carp
- Length at study initiation (length definition, mean, range and SD): about 10 cm
- Weight at study initiation (mean and range, SD): 20 - 40 g
- Health status: < 5% died
- Method of breeding: reared in a suitable fish pond
- Feeding during test
- Food type: pelleted feed
- Amount: ad libitum
- Frequency: 2-3- times daily

ACCLIMATION
- Acclimation period: 5 - 10 days
- Acclimation conditions (same as test or not): same

Route of exposure:

aqueous

Test type:

flow-through

Total exposure / uptake duration:

8 wk

Test temperature:

25 +- 2 °C

Dissolved oxygen:

7 ppm

Details on test conditions:

Actual details are not given, but a full description of the method is given here:

METHOD OF TEST ON THE DEGREE OF ACCUMULATION
This test is carried out in flowing water and the decree to which the test compound is accumulated in fish bodies is examined.

(1) Apparatus and equipment.
The apparatus is outlined as follows:
1. Active carbon tank for dechlorination,of city water
2. Diluting-water tank in which temperature is controlled and water aerated
3. Tank for stock of solution of the test compound
4. Flow meter
5. Mixing tank
6. Aquarium of volume of 3 to 10 liters per fish, provided with temperature regulator

The aquarium should be glass made, clean and of sufficient volume for rearing the test fish.

The apparatus used for water supply or dilution of the test compound should be glass made and clean as possible. Use of pasties should be limited to the parts where it is inevitable, such as joints.

(2) Test fish
Carp weighing 20 to 40 g and measuring about 10 cm in body length are used.

(3) Rearing and acclimating
The carp (Cyprinus carpio) are reared in a suitable fish pond. While rearing the diseased, the weakened and those showing other abnormal signs should be eliminated. Then external or internal parasitic pathogenies are examinated by sterilizer bathing or medication to keep the fish in good body condition and after that they are transferred to an acclimating tank. If there is a difference between the test temperature and the water temperature of the fish pond, the fish are acclimated in the acclimating tank by the method provided above and in the meantime the fish with damages in the gills or the skin or diseased are eliminated. It is desirable that the water in the fish pond and acclimating tank is always flowing.
- If the test temperature is higher than the water temperature of the fish pond, fish are acclimated for not less than a day in a temperature higher by 5 °C or less than the water temperature of the fish pond. Then the temperature is gradually raised at the rate not more than 3 °C/day and. finally the fish are reared in the test temperature for 5 to 7 days.
- If the test temperature is lower than the water temperatureof the fish pond, fish are acclimated for not less than a day in a temperature lower by 3 °C or less than the water temperature of the fish pond, then the temperature is gradually lowered at the rate not more than 2 °C/day and finally fish are reared in the test temperature for 7 to 10 days.

(4) Feeding
Pelleted feed is used. Fish are fed 2 to 3 times daily. In order to ayoid any leftover, the feed should be given slowly so that the portion of the feed floating on the water surface is entirely eaten by the test fish before it reaches the bottom of the aquarium.

(5) Assessment of adequacy of test fish
The carp in the acclimating tank in which not less than 5 per cent of the whole fish have weakened or died within one week before the start of the test should not be used.

(6) Test procedure
- Test water and the concentration of the test compound: two levels of concentration as low as possible within the analytical limit (one of the two levels of concentration is ten times higher than the other) should be prepared. The test is carried out in each of two concentration levels mentioned above, and at the same time blank test is made in the water in which the test compound is not present. During the test period, test water is controlled so that the concentration of dissolved oxygen is kept at about 7 ppm.
- Test temperature: the test water is kept at 25 ± 20 °C.
- Test period: the test should be continued for 8 weeks.
- Procedure: the test fish are reared under the conditions provided earlier using the apparatus and equipment provided earlier. Excretions of the carp and smudges on the wall of the aquarium should be removed about once a day throughout the test period.

(7) Analysis of test fish
- Every one or two weeks two to three test fish are taken out not less than ten hours after feeding and subjected to analysis. The fish from the water concentration level in which the test fish have showed distinct abnormal signs during the test should not be served for analysis.
- The surface of fish body is blotted dry with gauze and each fish is weighed accurately to the unit of 0.1 g. In the next place, the whole body of each fish, in principle, is homogenated and a suitable pretreatment such as extraction and concentration, depending upon the chemical properties of the test compound, is made to prepare a sample for quantitative analysis. The analysis is made as directed in the General Rules of Analysis provided in the JIS (gas and liquid chromatography, mass spectrometry, atomic absorption spectrophotometry , etc.).

Nominal and measured concentrations:

Measured concentrations: 0.53 ± 0.02 ppm and 0.055 ± 0.003 ppm

Reference substance (positive control):

no

Details on estimation of bioconcentration:

CALCULATION METHOD
Formula: CFn = (Fn - Fb) / W

- CFn: concentration factor
- Fn: Concentration of the test compound in the fish body after at the end of the test period
- Fb: Arithmetical mean concentration of the test compound in the fish body at the start of and at the termination of the blank test
- W: mean concentration of the test compound in the aquarium

Key result

Conc. / dose:

0.53 other: ppm

Temp.:

25 °C

Type:

BCF

Value:

0.6 dimensionless

Basis:

whole body w.w.

Time of plateau:

56 d

Remarks on result:

other: highest determined BCF in 56 days for this concentration

Key result

Conc. / dose:

0.055 other: ppm

Temp.:

25 °C

Type:

BCF

Value:

1.7 dimensionless

Time of plateau:

56 d

Remarks on result:

other: highest determined BCF in 56 days for this concentration

Validity criteria fulfilled:

not applicable

Conclusions:

The study indicate that the substance does not bioaccumulate in fish.

Description of key information

The test substance does not significantly accumulate in organisms.

Key value for chemical safety assessment

BCF (aquatic species):

1.7 L/kg ww

Additional information

Bioaccumulation was determined in a study exposing freshwater fish (Cyprinus carpio) to concentrations of 0.53 +- 0.02 ppm and 0.055 +- 0.003 ppm of test substance for 8 weeks. After 3, 7, 14, 21, 42 and 58 days the bioconcentration factor was determined. The BCF of the 0.53 ppm treatment did not exceed the the factor of 0.6 during the test period. At 0.055 ppm the BCF did not exceed a factor of 1.7.