2-{4-[4-[4-fluoro-6-(2-(2-vinylsulfonylethoxy)ethylamino)-1,3,5-triazin-2-ylamino]phenylazo]phenylazo}naphthalene-4,6,8-trisulfonate, trisodium salt

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 March 2001 to 10 April 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 A (Ready Biodegradability: DOC Die Away Test)

Deviations:

yes

Remarks:

DOC Determination, only one sample from each flask for DOC-determination instead of the measurement in duplicate.

Qualifier:

according to guideline

Guideline:

EU Method C.4-A (Determination of the "Ready" Biodegradability - Dissolved Organic Carbon (DOC) Die-Away Test)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40'800/A
Batch number: WP 2/01
Active ingredient: ca. 70%
Aggregate state at room temperature: Solid / powder
Color: Dark orange
Storage conditions: At room temperature at about 20 °C, away form direct sunlight
Expiration Date: February 14, 2007

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

Mixture of polyvalent bacteria (activated sludge) collected on March 12, 2001 from the aeration tank of a domestic sewage treatment plant, PRO RHENO Basel communal STP. The activated sludge was washed 4 times with tap water and the last time with test medium. A sample of the activated sludge was taken to determine the dry weight of the Suspended Solids (SS). An amount corresponding to 2 - 3 g per litre of suspended solids was mixed with test medium. The pre-treated sludge was aerated until use. The pH value of the activated sludge was 7.0. A bacteria concentration of 28 mg/L suspended solids was obtained by adding 10 mL of the pre-treated sludge (2.8 g/L SS) per 1000 mL test medium.

Duration of test (contact time):

28 d

Initial conc.:

38.1 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

For each test series the following number of test vessels was set up:
- Test item 2 vessels with inoculum + test medium + test item
- Inoculum control 2 vessels with inoculum + test medium
- Procedure control 2 vessels with inoculum + test medium + a readily biodegradable reference compound
- Abiotic sterile control 1 vessel with test medium + test item without inoculum, sterilised by ultra-filtration
- Inhibition control 1 vessel with inoculum + test medium + test item + reference compound

A measured volume of inoculated mineral medium, containing a known concentration of the test item as the nominal source of organic carbon, was aerated in the dark or diffuse light at 22±2 °C nominal. The degradation was followed by DOC analysis at frequent intervals over a period of 28 days. The degree of biodegradation was calculated by expressing the concentration of DOC removed (corrected by the blank inoculum control) as a percentage of the concentration initially added. The degree of primary biodegradation could also be calculated from supplemental chemical analysis made at the beginning and at the end of the test.

Reference substance:

other: D(+)-Glucose anhydrous for biochemistry [Merck No. 1.08337]

Parameter:

% degradation (DOC removal)

Value:

0

Sampling time:

28 d

Results with reference substance:

Procedure control Biodegradation after 14 days 96 %

Inhibition control Biodegradation after 14 days 48 %

Abiotic control Biodegradation after 28 days 2 %

Biodegradation in the inhibition control was >35 % after 14 days, hence the test item FAT 40800/A was not inhibitory on substrate biodegradation.

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

FAT 40800/A was not biodegradable after 28 days of incubation.

Executive summary:

A GLP-compliant ready biodegradability test was performed following the OECD 301A and EU C.4 guideline. The test item was exposed to a mixture of polyvalent bacteria (activated sludge obtained from a communal wastewater treatment plant) for 28 days. The design was a static test at 20.0 - 22.0 °C, test volume 200 mL, continuously shaken, in darkness. With test concentrations: Test item 38.1 mg/L DOC, Reference item 37.9 mg/L DOC. No degradation (0 %) of the test substance was observed, so, FAT 40800/A was not biodegradable after 28 days of incubation. Biodegradation of reference material after 14 days was 96 %.

Reference 2

Endpoint:

biodegradation in water: screening test, other

Remarks:

BOD5

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28 March 2001 to 03 April 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

ISO 5815 (Water quality - Determination of Biochemical Oxygen Demand after 5 Days (BOD5) - Dilution and Seeding Method)

Deviations:

yes

Remarks:

Prior the test the seeded dilution water was aerated during the night.

Principles of method if other than guideline:

Internal method: SOP N-A.211.101.01

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40'800/A
Batch number: WP 2/01
Active ingredient: ca. 70%
Aggregate state at room temperature: Solid / powder
Color: Dark orange
Storage conditions: At room temperature at about 20 °C, away form direct sunlight
Expiration Date: February 14, 2007

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

Seeding water taken from the aeration tank of a domestic sewage treatment plant, ARA PRO RHENO Basel communal collected on March 28, 2001. A part of seeding water was filtrated over glasfibre. The filtrat was aerated until use. 100 mL of the filtrat were added per 10 litres dilution water.

Duration of test (contact time):

5 d

Initial conc.:

6.3 mg/L

Based on:

test mat.

Initial conc.:

12.5 mg/L

Based on:

test mat.

Initial conc.:

25 mg/L

Based on:

test mat.

Initial conc.:

50 mg/L

Based on:

test mat.

Initial conc.:

100 mg/L

Based on:

test mat.

Initial conc.:

200 mg/L

Based on:

test mat.

Initial conc.:

400 mg/L

Based on:

test mat.

Initial conc.:

800.3 mg/L

Based on:

test mat.

Details on study design:

A mixture of a total volume of 1 litre containing the neutralised test item and seeded dilution water was mixed well. Then 2 incubation flasks were filled up and the oxygen concentration was measured. The remaining 0.5 litre was refilled up to 1 litre with seeded dilution water and the next 2 flasks were filled up. This procedure was repeated eight times. After 5 days of incubation at 20 °C ± 1°C in the dark, the remaining oxygen concentration in the flasks was measured.

Reference substance:

other: D(+)-Glucose anhydrous for biochemistry [Merck No. 1.08337] / L-Glutamic acid for biochemistry [Merck No. 1.00291]-solution.

Remarks on result:

other: The results show a BOD5 of 0 mg/g.

Parameter:

BOD5

Value:

0 g O2/g test mat.

Results with reference substance:

BOD5 of the reference item 187 mg O2/L.

Validity criteria fulfilled:

yes

Interpretation of results:

other: BOD5 of FAT 40800 is 0 mg/g

Conclusions:

The BOD5 of FAT 40800 is 0 mg/g.

Executive summary:

A study was performed determining the Biochemical Oxygen Demand of [BOD5] following the ISO 5815 guideline under GLP-regulations in Switzerland, ISO 9001 Quality management. The reference substance was D(+)-Glucose and L-Glutamic acid.

As innoculum Filtrated seeding water obtained from a communal wastewater treatment plant. The test substance was tested in concentration from 6.3 -800.3 mg/L in a static test. The results show a BOD5 of 0 mg/g.

Reference 3

Endpoint:

biodegradation in water: screening test, other

Remarks:

COD

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28 March 2001 to 29 March 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.6 (Degradation: Chemical Oxygen Demand)

Deviations:

yes

Remarks:

mean of 2 determinations instead of 3

Qualifier:

according to guideline

Guideline:

other: DIN 38409 - H 41-1 (1980)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40800/A
Batch number: WP 2/01
Active ingredient: ca. 70 %
Aggregate state at room temperature: Solid / powder
Color: Dark orange
Storage conditions: At room temperature at about 20 °C, away form direct sunlight
Expiration Date: February 14, 2007

Oxygen conditions:

not specified

Inoculum or test system:

not specified

Duration of test (contact time):

2 h

Initial conc.:

1 g/L

Based on:

test mat.

Details on study design:

A predetermined amount of the test item FAT 40800/A in water was oxidised by potassium dichromate in a strong sulphuric acid medium with silver sulphate as a catalyst under reflux for 2 hours at 148 ± 3 °C. Afterwards the test flasks were cooled down. The residual dichromate was determined by titration with standardised ferrous ammonium sulphate solution.

Reference substance:

other: potassium hydrogenphthalate

Remarks on result:

other: COD only

Parameter:

COD

Value:

945 mg O2/g test mat.

Results with reference substance:

COD of Potassium hydrogen phthalate 197 mg O2/L.

Validity criteria fulfilled:

yes

Interpretation of results:

other: COD has been determined

Conclusions:

COD of FAT 40800/A 945 mg O2/g.

Executive summary:

A study was performed to investigate the Chemical Oxygen Demand of the test substance following the DIN 38409 - H 41-1 and 92/69/EEC, C.6. GLP regulations and ISO 9001 Quality Management L01-001185. The test was performed with a reaction mixture of Potassium dichromate in a strong sulphuric acid medium with silver sulphate as a catalyst. This was boiled with the test item under reflux at 148 ± 3 °C for 2 hours. The Sample weight was 5.0 mg of the test item and 20 mL of a 0.17 g/L solution of the reference item potassium hydrogenphthalate. The COD for the test substance was determined to be 945 mg O2/g.

Reference 4

Endpoint:

biodegradation in water: inherent biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 March 2001 to 06 June 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 302 B (Inherent biodegradability: Zahn-Wellens/EMPA Test)

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

other: Commission Directive 87/302/EEC, Annex L 133, Part. C, pp 99-105 (edition 1988).

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40800/A
Batch number: WP 2/01
Active ingredient: ca. 70 %
Aggregate state at room temperature: Solid / powder
Color: Dark orange
Storage conditions: At room temperature at about 20 °C, away form direct sunlight
Expiration Date: February 14, 2007

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, adapted

Details on inoculum:

Mixture of polyvalent bacteria (activated sludge) collected on May 08, 2001 from the aeration tank of a domestic sewage treatment plant, ARA Pro Rheno Basel communal. The activated sludge was washed twice with tap water one day before the test was started. A sample of the activated sludge was taken to determine the dry weight of the Suspended Solids (SS). The pre-treated sludge was aerated until use. 0.47 g/L suspended solids. A bacteria concentration of 0.47 g/L suspended solids was obtained by adding 88 mL of the pre-treated sludge (11.4 g/L SS) per 2 litres test medium.

Duration of test (contact time):

28 d

Initial conc.:

150 mg/L

Based on:

DOC

Initial conc.:

155.7 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

A mixture of a total test volume of 2 litres each containing test item, mineral medium and activated sludge was agitated and aerated at 20 - 25 °C in two glass vessels for 28 days. A reference control running in duplicate containing activated sludge, mineral medium and the reference item, and a blank control containing activated sludge and mineral medium were run under the same test conditions. The biodegradation process was monitored by determination of the dissolved organic carbon concentrations (DOC). The ratio of eliminated DOC, corrected for the blank after each time interval, to the initial DOC value was expressed as the percentage biodegradation at each sampling time.

Reference substance:

diethylene glycol

Remarks:

[Fluka No 32160]

Parameter:

% degradation (DOC removal)

Value:

8

Sampling time:

28 d

Details on results:

- Adsorption of FAT 40800/A after 3 hours: 0 %
- Biodegradation of FAT 40800/A after 28 days: 8 %
- Total elimination of FAT 40800/A after 28 days (regarding adsorption and biodegradation): 7 %

Results with reference substance:

Biodegradation of the reference item after 19 days: 99 %.

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

The biodegradation of FAT 40800/A after 28 days was 8 % while adsorption after 3 hours was 0 %.

Executive summary:

A inherently biodegradability test was performed following the OECD 302 B and 87/302/EEC, Part C guidelines under GLP. The test substance was exposed to a mixture of polyvalent bacteria (activated sludge obtained from a communal wastewater treatment plant) for 28 days at a test concentration of 150.0 and 155.7 mg/L DOC, The reference substance, diethylene glycol, at 151.2 and 153.3 mg/L DOC. Based on the test results, adsorption of FAT 40800/A after 3 hours was 0 % while biodegradation after 28 days was 8 % and total elimination (regarding adsorption and biodégradation) after 28 days was 7 %. The substance can be regarded as non-biodegradable.

Description of key information

The test substance is neither readily nor inherently biodegradable.

Key value for chemical safety assessment

Biodegradation in water:

not biodegradable

Type of water:

freshwater

Additional information

The ready biodegradability of the test substance was determined in a GLP-compliant study according to OECD Guideline 301A. The initial test substance concentration was 38.1 mg/L based on DOC. Biodegradation of the test substance after 28 days was found to be 0 %. The inherent biodegradability of the test substance was determined in a GLP-compliant test according to OECD Guideline 3026. The initial test substance concentration was 150 and 155.7 mg/L based on DOC. In this test the total elimination of test substance after 28 days (regarding adsorption and biodegradation) was 7 %. Based on these findings the substance can be regarded as non-biodegradable. The biological oxygen demand (BOD5) was examined in a GLP-compliant study according to ISO 5815 (corresponding to EC method C.5). The BOD5 of the test substance was found to be 0 mg O2/L. The chemical oxygen demand (COD) was examined in a GLP-compliant study according to DIN 38409- H 41-1 (corresponding to EC method C.6. The COD of the test substance is 945 mg O2/L. The BOD5/COD ratio supports the conclusion that the test substance is not readily biodegradable.