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

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

Please find the attached justification.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Remarks on result:

other: Result of RA Source record CAS 1328-53-6 (BASFSE, 2000); no effects up to the limit of water solubility

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Remarks on result:

other: Result of RA Source record CAS 1328-53-6 (BASFSE, 2000); no effects up to the limit of water solubility

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 50 mg/L

Remarks on result:

other: Result of RA Source record Phthalocyanine (Jancula, 2008); likely no effects up to the limit of water solubility

Reference 2

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

No detail on procedure followed, only results presented.

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Version / remarks:

mod. for coloured substances

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (as cited): Phtalocyanin-Al-Komplex
- Identification number: 083940.0
- Batch No.: 021422D7
- Purity: 11.5%
- Appearance: liquid blue
- Solubility: miscible
- Storage: room temperature

Analytical monitoring:

not specified

Vehicle:

not specified

Test organisms (species):

other: Green Algae

Test type:

not specified

Water media type:

not specified

Limit test:

yes

Total exposure duration:

72 h

Nominal and measured concentrations:

Nominal concentration: 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: flask
- No. of vessels per concentration: 6
- No. of vessels per control: 18
- Initial cells density: 10120 cells/mL

EFFECT PARAMETERS MEASURED: density of algal cells, growth rate and inhibition (%)

Reference substance (positive control):

not specified

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 11.5 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

11.5 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Details on results:

See 'Any other information on results incl. tables'.

Table: Densities of algae cells determined during the test period of 72 hours

(Particle number x 1000/mL, at start 10120 cells/mL were incubated).

Conc. [mg/L]	Flask No.	Density of Algal cells [72h]			Growt rate [µ]	Inhibition [%]
Control	1	753	766	736
	2	828	829	833
	3	565	550	557
	4	1007	1010	876
	5	947	941	940
	6	701	697	698
	mean	791			1.45	-
	St	154.46
	n	6
100 Toxtest A	7	594	607	584
	8	540	535	548
	mean	568			1.34	7.6
	St	38.18
	n	2
100 Colortest B	9	468	474	473
	10	511	508	509
	mean	491			1.29	11.0
	St	26.16
	n	2
Inhibition A - B: -3.4%

Reference 3

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

17-2-2000 to 21-2-2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Version / remarks:

June, 1984

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

Version / remarks:

December, 1992

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.5400 (Algal Toxicity, Tiers I and II) (January 2012)

Version / remarks:

Public draft, April 1996

GLP compliance:

yes

Specific details on test material used for the study:

- Name of test substance (as cited in study report): Heliogen Grün 8730 Pulver
- Chemical name: Cl Pigment Green 7
- Batch number: BW137(Partien 61 6-61 9)
- Date of production: 24 Aug 1999
- Productnumber: 015258
- Molecular weight [g/mol]: Ca.1110
- Aggregate state: powder
- Density [g/ml]: not indicated by the sponsor
- Water-solubility: <1 mg/L
- Colour: green
- Purity of the test substance [%]: not indicated by the sponsor
- Impurities: not indicated by the sponsor
- Homogeneity: homogeneous
- lnstability against: not indicated by the sponsor
- Special storage conditions: not indicated by the sponsor
- Further remarks: none

Analytical monitoring:

no

Remarks:

The concentration control analysis was not performed, because the detection limit of the analytical method was beyond the water-solubility of the test substance

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
The test substance was stirred in demineralized water for about 20 h at approximately 20 ± 2 °C. Undissolved test substance was removed by centrifugation (c. 20 min at c. 17700 G). The eluate had a tinge of green. The nominal concentration of the eluate was 125 mg/L. The eluate was diluted with demineralized water to obtain the desired test concentrations.

Test organisms (species):

Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)

Details on test organisms:

TEST ORGANISM
- Strain: CHODAT SAG 86.81
- Source: SAG (Collection of algal cultures in Göttingen)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test temperature:

23 ± 2 °C (as described in methods)

pH:

8.0 - 8.4 (measured)

Nominal and measured concentrations:

- Nominal concentrations: 0 (control), 6.25, 12.5, 25, 50 and 100 mg/L
- Measured concentrations: not determined (the LOD was higher than the water solubility of the test substance)

Details on test conditions:

TEST SYSTEM
- Test vessel: erlenmeyer flasks (nominal volume 250 mL) plugged with gas permeable siliconsponge caps
- Test volume: 100 mL
- Pre-culture techniques: seed-culture: a seed culture was incubated for 7 days at 23 °C ± 2 °C, final cell density: 3,14E6 cells/mL; pre-culture: seed-culture was taken to inoculate a pre-culture (initial ceII density: 1E4 cells/mL). The pre-culture was incubated tor 3 days at 23 °C ± 2 °C. Final ceIl density: 0,24E6 cells/mL.
- lnoculation density: 1E4 cells/mL
- Number of replicates/conc.: 3
- Number of replicates/control: 3

TEST MEDIUM
The test medium was prepared according EEC Directive 92/69/EEC, Annex V, Part 0.3. Algal inhibition test, updating 29 December 1992 and OECD Guideilne for Testing of Chemicals, No. 201 Algal growth inhibition test, June 1984.

OTHER TEST CONDITIONS
- Measurement of temperature: continuously during the whole test period in the climate chamber
- Measurement of pH-values: after 0 h and 72 h in an additional uninoculated replicate and after 72 h in the inoculated replicate No. 1 of each concentration
- Illumination: artificial light, type universal white (e.g. OSRAM L 25), permanent illumination
- Light intensity . about 60 - 120 µE/(m2·s) at a wave length of 400 - 700 nm

EFFECT PARAMETERES MEASURED
- Test parameter: in vivo chlorophyll-a-fluorescence (pulsed excitation with Iight flashes having a wavelenght of 435 nm)
- Measurement of fluorescence: after 0, 24, 48 and 72 h
- Cell counting: after 72 h in a counting chamber (Neubauer improved) in replicate No. 2 of the inoculated control

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: no effects up to the limit of water solubility

Key result

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: no effects up to the limit of water solubility

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

12.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Remarks on result:

other: no effects up to the limit of water solubility

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

11.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

12.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Details on results:

After 72 hours, growth rate inhibition was determined to be 9.1, 4.7 and 4.5% for the nominal test concentrations 100, 50 and 25 mg/L, respectively. See 'Any other information on results incl. tables'.

Results with reference substance (positive control):

The 72-h EC50 of the control substance was 0.44 mg/L (date of the last control experiment: 24 Jan 2000)

Reported statistics and error estimates:

- Calculation of the integral of biomass growth over the total duration of the test for each concentration level and comparison of values of treated samples in relation to untreated samples.
- Calculation of the growth rate over the total duration of the study for each concentration level and comparison of values of treated samples in relation to untreated samples.
- The EC values are calculated (linear regression analysis) from the concentration-response relationship.
- The LOEC is determined by comparing the means of the fluorescence measurement of the various concentration levels with the control. The Duncan multiple range test is carried out at a 95 % significance level. Every higher tested concentration must have at least the same or a stronger effect then the LOEC.

Table: inhibition of the algal growth after 72 hours

Nominal concentration [mg/L]	biomass		growth rate
	relative	inhibition [% of the control]	relative	inhibition [% of the control]
0 (control)	1496	0.0	0.065	0.0
100	1071	28.4	0.060	9.1
50	1208	19.3	0.062	4.7
25	1255	16.1	0.062	4.5
12.5	1329	11.1	0.064	2.5
6.25	1475	1.4	0.067	-2.1

Validity criteria fulfilled:

yes

Reference 4

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

ISO 8692 (Water Quality - Fresh Water Algal Growth Inhibition Test with Scenedesmus subspicatus and Selenastrum capricornutum)

Version / remarks:

1989

Deviations:

yes

Remarks:

see 'Principles of method if other than guideline'

Principles of method if other than guideline:

The test was based on ISO 8692, but performed in transparent 96-well microplates. Initial cell concentrations were 1E+05 cells/mL. Algal densities were measured at 518 nm due to strong absorption of the substance at the standard 680 nm. The incubation temperature was 27 °C.

GLP compliance:

no

Specific details on test material used for the study:

The test substance was synthesized by Research Institute of Organic Synthesis (VUOS PLC), Rybitvi, Czech republic.

Analytical monitoring:

no

Vehicle:

not specified

Test organisms (species):

Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM
- Strain: CCALA 433
- Method of precultivation: erlenmeyer flasks at 27 °C in a light intensity of 100 µmol/m/s (6000 lux)
- Medium during cultivation: a mixture of 1:1 medium Z and medium BB, diluted to 50% with water

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test temperature:

27 °C

Nominal and measured concentrations:

- Nominal concentrations: 0 (control), 0.01, 0.1, 1, 10 and 50 mg/L
- Measured concentrations: concentrations were not analytically verified

Details on test conditions:

TEST SYSTEM
- Test vessel: 96-well microplates
- Volume: 250 µL
- Initial cells density: 10E5 cells/mL
- No. of vessels per concentration: 5
- No. of vessels per control: 5

TEST MEDIUM / OTHER TEST CONDITIONS
Similar as used for cultivation (see 'Details on test organisms').

EFFECT PARAMETERS MEASURED: density of algal cells
- Method of detection: microplate reader (Sunrise, Tecan, Austria) at a wavelength of 518 nm.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: likely no effects up to the limit of water solubility

Details on results:

No inhibition was observed.

Results with reference substance (positive control):

Not reported

Reported statistics and error estimates:

Linear regression analysis to determine the inhibition percentage

Validity criteria fulfilled:

not specified

Description of key information

No effects were observed up to the limit of water solubility.

Additional information

A study (report) performed with the substance itself was considered to contain insufficient documentation and is thus not suitable for classification purposes. Therefore, data from the following two close structural analogues are used as part of a weight of evidence approach: polychloro copper phthalocyanine (CAS 1328-53-6) and phthalocyanine with SO3Na groups on rest group 1 and 2 and an H-atom on rest group 3 and 4. In these studies, no analytical verification of test concentrations was performed. The stability of these molecules play an important role for their applications; since no hydrolysis, photolysis, adsorption, biodegradation or evaporation of these molecules in/from the test medium is expected, it may be assumed that test organisms were exposed to the water-soluble fraction of test substances for the entire exposure period.

The toxicity towards freshwater algae was determined with structural analogue Polychloro copper phthalocyanine (CAS 1328-53-6) in a study according to OECD TG 201 and in compliance with GLP criteria (BASF 2000). In this study, exponentially growing freshwater alga (Desmodesmus subspicatus) were exposed to nominal test substance concentrations of 0 (control), 6.25, 12.5, 25, 50 and 100 mg/L for 72 hours under static conditions. The cell concentration in each flask was determined at the start of the test and after 0, 24, 48 and 72 hours exposure. The mean value of the cell concentration for each test concentration was plotted against time to produce growth curves from which the percentage inhibition of biomass (i.e.cell growth as calculated by comparison of Areas Under the Growth curves (AUG)) and the average specific growth rate for exponentially growing cultures was calculated. The 72-h EC50 values for growth rate (ErC50) and biomass (EbC50) were determined at >100 mg/L, respectively. The 72-h EC10 values for these endpoints were >100 (growth rate) and 11.5 mg/L (biomass), respectively.

 Furthermore, phthalocyanine was tested for its toxicity towards freshwater algae in a (published) study that assessed algicidal activity of several Phtalocyanines (Jancula, 2008). The test was based on ISO 8692, but performed in transparent 96-well microplates. Exponentially growing freshwater alga (Pseudokirchneriella subcapitata) were exposed to the test substance of interest (with SO3Na groups on rest group 1 and 2 and an H-atom on rest group 3 and 4) in concentrations of 0, 0.01, 0.1, 1, 10 and 50 mg/L for 72 hours. Test concentrations were not analytically verified. The 72-h ErC50 was determined to be >50 mg/L. Additionally, a study with the substance itself was performed where the toxicity towards freshwater algae was determined in a limit test according to OECD TG 201 modified for coloured substances (BASF SE, 1998). In this study, green algae were exposed to nominal test concentrations of 0 and 100 mg/L for 72 hours. Information on any analytical verification of test concentrations is absent. The cell concentration in each flask was determined at the start of the test and after 72 hours exposure. Taking purity into account, the 72-h ErC50 and NOEC values were determined to be >11.5 mg/L (nominal concentration 100 mg/L). These studies show no adverse effects on the growth rate of algae. Overall, no effects were observed up to the limit of water solubility and thus the substance is not considered to be harmful to algae.