Brominated chlorinated copper, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, wherein the number of bromines is equal to or more than 10 and less than 16, and the number of chlorines is equal to or more than 0 and less than 7

Biodegradation in water and sediment: simulation tests

Some information in this page has been claimed confidential.

Administrative data

Endpoint:

biodegradation in water: sediment simulation testing

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

read-across source

Reference

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

From 2003-02-10 to 2003-04-09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Data from reliable source. Though no detailed test report is available, all tests from the Japanese Authorities are performed according to an OECD guideline and GLP incl. analytics.

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Name of test material (as cited in study report): K-1680
- Lot/batch No.: DLM9599
- Stability under test conditions: stable as confirmed by infrared spectrum
- Storage condition of test material: stored in dark and cool place

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, non-adapted

Details on inoculum:

- Source of inoculum/activated sludge : Sludges were sampled from 10 sites including sewage plant, river, lake and sea in Japan. Filtrates of sludge samples collected at above sites were mixed together. This mixture of filtrates (5L) was mixed with filtrate of the old sludge (5L) to make a new sludge suspension (10L). This suspension was adjusted to pH of 7.0±1.0 and aerated.
- Method of cultivation: After stopping the aeration of the incubation tank to let the sludge settle for approximately 30 minutes, about 1/3 of the supernatant was replaced by equal amount of dechlorinated water before resuming aeration. After more than 30 minutes of aeration, synthetic sewage water was added to make the concentration in renewed supernatant to be 0.1%. This procedure was repeated daily to prepare the activated sludge culture. The incubation temperature was 25±2 °C.
- Storage conditions: Appearance of the supernatant and formation of the activated sludge was observed, and precipitability, pH, temperature and dissolved oxygen concentration of the activated sludge were recorded. Activated sludge used for the test was observed under an optical microscope as appropriate to confirm that no abnormalities were found in biota.
- Concentration of sludge: Concentraion of suspended sludge was 4900 mg/L.

Duration of test (contact time):

28 d

Initial conc.:

100 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Remarks:

BOD

Parameter followed for biodegradation estimation:

test mat. analysis

Details on study design:

TEST CONDITIONS
- Composition of medium: A, B, C and D solutions were composed according to Japanese standard testing method for industrial waste water, biochemical oxygen demand (JIS K 0102-1998-21). Basal medium was prepared by mixing 3mL portions of these solutions and puridied water (Takasugi Pharmaceutical Co., Ltd. The japanese Pharmacopeia) to make a total of 1L. The pH was adujusted to 7.0.
- Test temperature: 25±1°C
- Suspended solids concentration: 30mg/L


TEST SYSTEM
- Culturing apparatus: 300mL glass bottle
- Number of culture flasks/concentration: 3 bottles (activated sludge + test substance)
- Method used to create aerobic conditions: stirring by magnetic stirrer
- Measuring equipment: coulometer
- Details of trap for CO2 and volatile organics if used: soda lime


CONTROL AND BLANK SYSTEM
- Inoculum blank: activated sludge + basal medium
- Abiotic sterile control: test substance + purified water
- Other ( activity control): aniline + activated sludge + basal medium

Reference substance:

aniline

Test performance:

The degree of degrafability based on BOD measurement of aniline were 57% (day 7) and 72% (day 14).

Parameter:

% degradation (O2 consumption)

Value:

5

Sampling time:

28 d

Remarks on result:

other: BOD

Parameter:

% degradation (test mat. analysis)

Value:

0

Sampling time:

28 d

Remarks on result:

other: gravimetric analysis

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Cross-reference 2

Reason / purpose for cross-reference:

read-across source

Reference

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)

Principles of method if other than guideline:

Test procedures according to EU Directive 79/831/EEC Annex V, part C: Methods for the Determination of the ecotoxicity degradation - biodegradation, Manometric respirometry test. This method is comparable to the OECD guideline 301 F.

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): Kupferphthalocyanin
- Analytical purity: 99.3 %

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic (adaptation not specified)

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): Municipal activated sludge from laboratory wastewater treatment plant fed with municipal sewage and synthetic wastewater
- final dry solids concentration: 30 mg/l
- Laboratory culture: yes

Duration of test (contact time):

28 d

Initial conc.:

107 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

TEST CONDITIONS
- pH: at the end 7 - 7.5
- pH adjusted: no
- CEC (meq/100 g):


SAMPLING
- Sampling frequency: daily


CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control: yes
- Toxicity control: yes

Reference substance:

aniline

Key result

Parameter:

% degradation (O2 consumption)

Value:

< 1

Sampling time:

28 d

Results with reference substance:

> 80% biodegradation, based on THSB

	BW1	BW2	KS	IH	PS1	PS2	PS3	PS4	PS5	PS6	PS7	PS8
Water(mL)	240	240	230	230	240	240	240	240	240	240	240	240
Testsubstance(mg)	0	0	0	26	29	29	28	29	25	25	25	25
Refernecesubstance(mL)	0	0	10	10	0	0	0	0	0	0	0	0
Inoculum(mL)	10	10	10	10	10	10	10	10	10	10	10	10
Volume (mL)	250	250	250	250	250	250	250	250	250	250	250	250
O2 Consumption (%)	-	-	2.7	-1	-4	-4	-4	-4	-1	0	-3	-2

 

BW: Biotic control

KS: Reference substance control

IH: Reference substance + Test substacne control

PS: Test substance replicate

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Cross-reference 3

Reason / purpose for cross-reference:

data waiving: supporting information

Reference

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Data from reliable source. Though no detailed test report is available, all tests from the Japanese Authorities are performed according to an OECD guideline and GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))

GLP compliance:

no

Specific details on test material used for the study:

- Name of test material (as cited in study report): C.I. Pigment Blue 15
Purity: 99.3 %

Oxygen conditions:

aerobic

Inoculum or test system:

mixture of sewage, soil and natural water

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): In March, June, September, and December, sludge was sampled at the following 10 places in Japan: 1. Fukogawa city sewage plant, 2. Fukashiba industry sewage plant, 3. Nakahama city sewage plant, 4. Ochiai city sewage plant, 5. Kitakami river, 6. Shinano river, 7. Yoshino river, 8. Lake Biwa, 9. Hiroshima bay, 10. Dookai bay; sampling: 1. City sewage: Returned sludge from sewage plants was taken. 2. Rivers, lake and sea: Surface water and surface soil which were in contact with atmosphere were collected.
- Method of cultivation: About 30 minutes after ceasing aeration to the sludge mixture, supernatant corresponding to about 1/3 of the whole volume was removed. Then the equal volume of dechlorinated water was added to the remaining portion and aerated again, followed by addition of synthetic sewage at a concentration of 0.1% (w/v). This procedure was repeated once every day. The culturing was carried out at 25 ± 2 °C. 5 L of the filtrate of the supernatant of old activated sludge was mixed with 500 mL of the filtrate of the supernatant of new sludge and cultured at pH 7.0 ± 1.0 under sufficient aeration using prefiltered open air. During the cultivation, appearance of the supernatant, precipitability, formation of flock, pH, dissolved oxygen concentration in the solution and temperature were checked and necessary adjustments were made, Microflora in the activated sludge was microscopically observed and sludge with no abnormal symptom was used for the test.
- Concentration of sludge: 30 mg/L

Duration of test (contact time):

14 d

Initial conc.:

100 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

TEST CONDITIONS
- Composition of medium: 3 mL each of four stock solutions, as described in JIS K 0102-1986-21, are diluted to 1000 mL with purified water
- pH: 7.0
- pH adjusted: yes
- Suspended solids concentration: determined according to Method Japanese Industrial Standards (JIS) K 0102-1986-14.1

TEST SYSTEM
- Culturing apparatus: Closed system oxygen consumption measuring apparatus (Coulometer: Ohkura Electric Co., Ltd.); 300 mL vessel, absorbent for evolving carbon dioxide Soda lime No .l (extra pure reagent, Wako Pure Chemical Industries, Ltd.).
- Number of culture flasks/concentration: 1
- Measuring equipment: Coulometer, Okhura Electric Co., Ltd.
- Test performed in open system: no
- Details of trap for CO2 and volatile organics if used: soda lime, extra pure, Wako Pure Chemical Industries, Ltd.)

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control: yes
- Toxicity control: no

Reference substance:

aniline

Parameter:

% degradation (O2 consumption)

Value:

0

Sampling time:

14 d

Results with reference substance:

71% biodegradation, based on O2 consumption at the day 7

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Cross-reference 4

Reason / purpose for cross-reference:

data waiving: supporting information

Reference

Endpoint:

dispersion stability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Guideline:

OECD Guideline 318 (Dispersion Stability of Nanomaterials in Simulated Environmental Media)

GLP compliance:

no

Remarks:

The study does not estimate the toxicological effect of a substance and thus, the conduction of this test under GLP is not required.

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: Heubach Colour Private Limited
- Expiration date of the lot/batch: 14J0836
- Purity: ca. 100%
- State of aggregation: Green-yellowish to green powder

Analytical monitoring:

yes

Details on sampling:

after 6, 15 and 24 hours

Vehicle:

no

Details on test sample/test medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
Sample preparation for testing:
- Pretreatment of dry nanomaterial: sonicating with the sonicator brand/type Branson 550, Model 102C
- Nanomaterial pre-wetted: yes
- Preparation of stock dispersion: The stock suspension was prepared at 0.16 g pigment and a volume of 40 g ultrapure water, sonicated as mentioned above, stored at 5°C overnight. NOM was prepared at 0.032 g per 200 mL ultrapure water. The resulting NM concentration is 4 g/L.

Stock media
Testing in the presence of
- natural organic matter (NOM): 5 g
- electrolyte at different pH: Considering all dilution steps, the final DOC concentration was 10 mg/L. The influence of the concentration of Ca was tested at 0 mM, 1 mM and 10 mM Ca.
Storage of stock dispersion: Due to the quasi-continuous measurement (two triplicates = 6 samples, each measured in 1-minute intervals), no dilution was applied, and no storage occurred

Test media
- Loss of nanomaterial to test vessel wall and deviation of test vessel material: no
- Agglomeration/settling behaviour of nanomaterial investigated: yes

Total observation duration:

24 h

Details on test conditions:

TEST CONDITIONS ACCORDING TO MATRIX TESTING
- Pre-defined number concentration of test material in the test media
- Volume of stock dispersion added to test media
- pH : 4, 7 and 9
- Concentration of electrolyte in test vessels: Ca(NO3)2: 0, 1 and 10 mM
- Chemical composition of NOM : 2R101N Suwannee River NOM (SRNOM)
- NOM concentration (as DOC) in the test vessels : 10 mg/L
- Replicates:
- Buffer used:
- Equilibration time: 1 h
- Centrifugation and conditions after 6h: yes
- Sampling: after 6, 15 and 24 hours

Key result

Observation Time:

24 h

Test material concentration:

mg/L

Concentration:

nominal

Dispersion Stability:

3.8 %

St. deviation:

0.3 %

pH:

7

Medium:

10 mM NOM, 0 mM Ca(NO3)2

At any of the time points mentioned in the TG-318, the influence of Ca is critical. Regardless of pH, the pigment is categorized at the 24h-sampling time as “unstable” in 10 mM Ca, representing high water hardness.
After 6h, the stability for the samples in 0 mM Ca and at pH 4, 7 and 9 was high.

For the samples in 1 mM Ca at pH 4 the stability increased to an intermediate level after 24 hours, whereas at pH 7 and 9 the pigment was stable.

The Full results of the dispersion stability in the presence of NOM is presented in following table

Table 1: Full results of the dispersion stability in the presence of NOM

	Ca(NO3)2	Stability after 6h	Standard deviation	Stability after 15h	Standard deviation	Stability after 24h	Standard deviation
	[mM]	[%]	[%]	[%]	[%]	[%]	[%]
pH 4	0	92.5	2.4	56.7	7.3	42.6	9.0
pH 4	1	61.3	3.5	14.1	1.3	6.3	0.7
pH 4	10	13.7	0.6	4.4	0.4	4.3	0.8
.
pH 7	0	95.3	0.2	80.1	2.5	18.6	1.8
pH 7	1	36.9	1.2	7.3	0.4	4.1	0.4
pH 7	10	13.8	1.3	5.0	0.2	3.8	0.3
.
pH 9	0	94.7	0.3	84.0	1.9	83.5	1.7
pH 9	1	24.7	2.6	6.2	0.7	4.4	0.6
pH 9	10	13.7	2.0	4.5	0.5	3.5	0.5

To rationalize the observed dispersion stability, we finally checked the particle size distribution directly in the environmental medium (exact same sample preparation as for the UV/VIS measurements). We applied the NanoDefine method of Analytical Ultracentrifugation (SOP AUC-RI, published by 3). The centrifugation parameters are given in the methods section.
The observed size distributions confirm the moderate agglomeration at 1 mM Ca, pH7, with NOM (Figure 4). If the particles would have been significantly dissolved, no size distribution would be observable at all by this method, which relies on the detection of the movement of particles during centrifugal separation.
Additionally, the centrifugation methods include a determination of the remaining absorption after centrifugation, fully consistent with the conventional determination of the dissolved fraction after centrifugation as recommended by the TG-318. The remaining absorption was measured at c.a 0.014. This is a fraction of 1.7% of the initial absorption, but actually is close to the LOD of the built-in UV/Vis detector. Considering the LOD, between 0% and 1.7% of the sample may have been dissolved.
All evidence combined, the results after centrifugation confirm that at least 98.3% of the observed dispersion stability has to be attributed to the particles, not to dissolution.

 

 

 

 

 

Results and discussion

Transformation products:

no

Remarks:

not expected according to substance properties