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

Toxicity to aquatic plants other than algae

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to aquatic plants other than algae

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:

7 d

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Basis for effect:

growth rate

Remarks on result:

other: Result of RA Source CAS 28901-96-4 (BASFSE, 2015); no effects up to the limit of water solubility

Duration:

7 d

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Basis for effect:

growth rate

Remarks on result:

other: Result of RA Source CAS 28901-96-4 (BASFSE, 2015); no effects up to the limit of water solubility

Duration:

7 d

Dose descriptor:

EC50

Effect conc.:

> 50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

frond number

Remarks on result:

other: Result of RA Source record Phthalocyanine (Jancula, 2012); likely no effects up to the limit of water solubility

Reference 2

Endpoint:

toxicity to aquatic plants other than algae

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 221 (Lemna sp. Growth Inhibition Test)

Version / remarks:

March, 2006

Qualifier:

according to guideline

Guideline:

other: Commission Regulation (EC) No 761/2009, Annex, Part C, C.26.: "Lemna sp. Growth Inhibition Test", Official Journal of the European Union (EN)

Version / remarks:

August, 2009

Qualifier:

according to guideline

Guideline:

other: OECD Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures"

Version / remarks:

December, 2015

Qualifier:

according to guideline

Guideline:

other: SANCO/3029/99 rev.4 11/07/00: Residues: Guidance for generating and reporting methods of analysis in support of pre-registration data requirements for Annex II (part A; Section 4) and Annex III (part A; Section 5) of directive 91/414

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Name of test material (as cited in study report): Pigment Additive PAG/Cu-phthalocyanine monosulfonic acid
- Chemical name: Cuprate(1-), [29H,31H-phthalocyanine-C-sulfonato(3-)- N29,N30,N31,N32]-, hydrogen
- Lot No.: 130001/RA gemahlen (Laborjournalnr. V12424/ RA)
- Aggregate State at Room Temperature: Solid
- Colour: Blue
- Expiry Date: February 17, 2034
- Storage: At room temperature (20 ± 5 °C), in the dark
- Purity: as the test item is only soluble in 1-Methyl-2-pyrrolidone at a very low concentration level, no purity of the test can be stated by NMR or HPLC
- Water Solubility:
1. 5 mg/L ± 1 mg/L at 50 mg/L (20.0 °C ± 0.5 °C)); 8 mg/L ± 2 mg/L at 100 mg/L (20.0 °C ± 0.5 °C) with TOC determination;
2. 4 mg/L ± 1 mg/L at 50 mg/L (20.0 °C ± 0.5 °C); 6 mg/L ± 1 mg/L at 100 mg/L (20.0 °C ± 0.5 °C) with copper determination

Analytical monitoring:

yes

Details on sampling:

At 0 hour and 7 days

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
A defined amount of the test item was added directly to the test water for each test concentration and was carefully stirred for 24 hours at room temperature in the dark to dissolve as much of the test item as possible. The highest test item concentration of 100 mg test item/L was prepared by mixing 100.8 mg test item into 1008 mL test water, for the test item concentration of 33.3 mg test item/L, 33.0 mg test item were mixed into 991 mL test water, for the concentration of 11.1 mg test item/L, 10.8 mg were mixed into 973 mL test water. The concentration of 3.7 mg test item/L was prepared by mixing 3.6 mg into 973 mL test water and for 1.2 mg test item/L, 3.6 mg were mixed into 3000 mL test water. Then, the undissolved test item was separated by filtration (0.45 μm cellulose acetate filter) and the filtrates were used as test concentrations. The test concentrations were prepared just before introduction of the Lemna (= start of the test).

Test organisms (species):

Lemna gibba

Details on test organisms:

TEST ORGANISM
- Species: Lemna gibba G 3
- Origin: the fronds introduced in the test were taken from ibacon's in-house laboratory culture.
- Breeding Conditions: the plants are cultured in the laboratories of ibacon under standardised conditions according to the test guidelines.
- Pre-Culture: for at least 7 days under test conditions with weekly media exchange.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

7 d

Hardness:

3.55 mmol/L (as Carbon hardness)

Test temperature:

22 - 25 2°C

pH:

- Test start: 7.6 - 7.9
- Test end: 8.8 - 8.9

Salinity:

0.472 mmol/L

Conductivity:

1461 μS/cm

Nominal and measured concentrations:

- Water soluble fractions of nominal loadings of 0, 1.2, 3.7, 11.1, 33.3 and 100 mg test item/L

Details on test conditions:

TEST SYSTEM
- Type and size: glass vessels of 250 mL volume with approximately 200 mL test medium, covered with glass dishes.
- Identification: each test unit was uniquely identified with the serial number, treatment and replicate number.
- Replicates: 3 replicates per treatment.
- Exposure: the test vessels were placed in a random order and were repositioned on days 3 and 5 to minimize differences in light intensity.
- Introduction of Plants: colonies consisting of 4 fronds were transferred from the inoculum culture. Each test vessel contained a total of 12 fronds.

TEST MEDIUM / WATER PARAMETERS
- Test medium: Reconstituted Water (20x AAP-Growth Medium). The test water was prepared 1 day before test start to allow pH to stabilise.
- Source/preparation of dilution water: Analytical grade salts were added at the following nominal concentrations in deionised water (conductivity < 5 μScm-1)
- NaHCO3: 300 mg/L
- K2HPO4 × 3 H2O: 30 mg/L
- MgSO4 × 7 H2O: 290 mg/L
- NaNO3 510 mg/L MgCl2 × 6 H2O: 240 mg/L
- CaCl2 × 2 H2O: 90 mg/L
- H3BO3: 3.7 mg/L
- MnCl2 × 4 H2O: 8.3 mg/L
- ZnCl2: 0.066 mg/L
- CoCl2 × 6 H2O: 0.029 mg/L
- CuCl2 × 2 H2O: 0.00024 mg/L
- Na2MoO4 × 2 H2O: 0.145 mg/L
- FeCl3 × 6 H2O: 3.2 mg/L
- Na2EDTA × 2 H2O: 6.0 mg/L

OTHER TEST CONDITIONS
- Adjustment of pH: yes, the pH was adjusted with 2 M HCl to 7.4
- Light Regime: continuous illumination
- Light Intensity: 8115 Lux (mean value) with a range of 7540 to 8840 lux. The illumination was achieved by fluorescent tubes, installed above the test vessels. Measurements were performed once during the test at 6 places distributed over the experimental area at the surface of the test media. Differences in light intensity over the test area did not exceed ± 15 % of the mean.


EFFECT PARAMETERS MEASURED
- Number of Fronds: at test start frond and colony numbers were recorded. At days 3, 5 and 7 frond numbers and appearance of colonies were observed.
- Dry Weight of Fronds: at test start the dry weight of a sample of fronds identical to those used to inoculate the test vessels was determined. At the end of the test the dry weight of all plants from each vessel was determined. Accordingly, the plants were collected at the end of the test from the test vessels and dried at 60 °C to a constant weight.
- Recording: test conditions were recorded with suitable instruments and documented in the raw data.
- Other: water temperature: the temperature was measured daily in a test vessel filled with water and incubated under the same conditions as the test vessels; pH-values: the pH-value of each treatment was measured at test start and at test end.

RANGE-FINDING STUDY
- Test concentrations: 100 and 10 mg/L
- Results used to determine the conditions for the definitive study: no effects observed in the higher WAF

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol (tested twice a year)

Key result

Duration:

7 d

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:

7 d

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:

7 d

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

Duration:

7 d

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:

7 d

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:

7 d

Dose descriptor:

EC10

Effect conc.:

48.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Duration:

7 d

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:

7 d

Dose descriptor:

EC10

Effect conc.:

88.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Details on results:

- Shape of fronds: the shape of fronds and colonies after the test period of 7 days was not different from those in the control up to and including the nominal test concentration of 100 mg test item/L.
- Other: no effects on growth rate were observed. The only effects observed were based on frond number [yield] and dry weight [yield], from which the 7-d EyC10 of 48.2 and 88.3 mg/L values were determined, respectively..

Results with reference substance (positive control):

The 7-d ErC 50 was determined to be 7.68 mg/L (growth rate, based on frond number) and the 7-d EyC50 to be 7.68 mg/L (yield, based on frond number) are xx mg/L. The 7-d ErC10 and EyC10 values are 4.27 mg/L and 2.18 mg/L, respectively. These effect values are based on nominal concentrations.

Reported statistics and error estimates:

- The ErC50 and the EyC50 and the corresponding EC10 values and where possible their 95 %-confidence limits were calculated by Probit analysis.
- For the determination of the 7-day LOEyC and NOEyC values significant differences at the test concentrations compared to the control values were tested by the Williams t-test.
- For the determination of the 7-day LOErC and NOErC values significant differences at the test concentrations compared to the control values were tested by the Williams t-test.
- The software used to perform the statistical analysis was ToxRat Professional, Version 2.10.05, ToxRat Solutions GmbH.

Validity criteria fulfilled:

yes

Reference 3

Endpoint:

toxicity to aquatic plants other than algae

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

other: ISO 20079, 2005. Water Quality – Determination of the Toxic Effect of Water Constituents and Waste Water on Duckweed (Lemna minor) – Duckweed Growth Inhibition Test. International Organization for Standardization, Geneve, Switzerland

Principles of method if other than guideline:

Effect values (frond number) not based on growth rate, but the (quantitative) amount.

GLP compliance:

no

Remarks:

the experiment described in this publication did not comply with GLP-criteria.

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:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION: A stock solution of 50 mg/L was repeatedly diluted by factor 2.

Test organisms (species):

Lemna minor

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

7 d

Test temperature:

24 °C

Nominal and measured concentrations:

- Nominal concentrations: 0 (control), 6.25, 12.5, 25 and 50 mg/L
- Measured concentrations: concentrations were not analytically verified

Details on test conditions:

TEST SYSTEM
- Test vessel: 250-mL beakers containing 150 mL of growth medium
- Replicates per concentration and control: 3

GROWTH MEDIUM
- Type of medium: Steinberg medium

OTHER TEST CONDITIONS
- Light intensity: 100 µmol (m2/s)
- Photoperiod: 24h/day
- Lamp: Phillips, TLD 36 W/33

EFFECT PARAMETERS MEASURED: frond number, root length and dry weight

Reference substance (positive control):

no

Duration:

7 d

Dose descriptor:

EC50

Effect conc.:

> 50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

frond number

Remarks on result:

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

Duration:

7 d

Dose descriptor:

EC50

Effect conc.:

>= 37.27 - <= 50.11 mg/L

Conc. based on:

test mat.

Basis for effect:

other: root length

Remarks on result:

other: 43.69 ± 6.42 mg/L (value and standard deviation)

Duration:

7 d

Dose descriptor:

EC50

Effect conc.:

>= 37.44 - <= 44.1 mg/L

Conc. based on:

test mat.

Basis for effect:

biomass

Remarks on result:

other: 40.77 ± 3.33 mg/L (value and standard deviation)

Details on results:

No effects on frond number were observed. The 7-d EC50 values for root length and dry weight were determined to be 43.69 and 40.77 mg/L, respectively.

Reported statistics and error estimates:

Effect concentrations were determined using linear regression analysis.

Description of key information

EC50/EC10(7d) > 100 mg/L for frond number of Lemna minor (OECD 221, read across)

Additional information

No data is available for the substance itself. Therefore, data from the following two close structural analogues are used as part of a weight of evidence approach: Pigment Additive PAG (CAS 28901-96-4) and phthalocyanine with SO3Na groups on rest group 1 and 2 and an H-atom on rest group 3 and 4.

The toxicity towards freshwater aquatic plants was determined for the substance Pigment Additive PAG (CAS 280901-96-4) in a study according to OECD 221 and in compliance with GLP criteria (BASF 2015). In this study, duckweed (Lemna gibba) was exposed to water soluble fractions of nominal loadings of 0 (control), 1.2, 3.7, 11.1, 33.3 and 100 mg/L for 7 days under static conditions. Test concentrations were analytically verified at the start and the end of exposure, based on carbon content. These measured values were uniform over time and thus effect values are reported as nominal concentrations. At days 3, 5 and 7 frond numbers and appearance of colonies were observed. Additionally, at the test start the dry weight of a sample of fronds identical to those used to inoculate the test vessels was determined and at the end of the test the dry weight of all plants from each vessel was determined. The 7-d EC50 values for growth rate (ErC50) and yield (biomass, EyC50) based on frond number were determined at >100 mg/L (based on either frond number or dry weight). The 7-d ErC10 was determined to be >100 mg/L (based on either frond number or dry weight) and the 7-d EyC10 values were 48.2 and 88.3 mg/L considering biomass frond number and dry weight, respectively.

Additionally, the toxicity towards freshwater aquatic plants was determined for phthalocyanine (R1,2 =SO3Na, R3,4 = H) in a bioassay that was designed according to ISO 20079 (Jancula, 2012). In this study, duckweed (Lemna minor) was exposed to nominal concentrations of 0 (control), 6.25, 12.5, 25 and 50 mg/L for 7 days under static conditions. Test concentrations were not analytically verified. At day 7 the frond numbers, root length and dry weight were determined. The 7-d EC50 values for frond number, root length and dry weight were >50 mg/L, 43.69 ± 6.42 mg/L and 40.77 ± 3.33 mg/L, respectively.

No effect on (the growth rate of) frond numbers was observed. Based on the close resemblance between the structural analogues and the target substance, its E(r)C50 is also assumed to be >50 mg/L. Overall, no effects on growth were observed up to the limit of water solubility and thus the substance is not considered to be harmful to aquatic plants other than algae.