trisodium 3-amino-4-[4-[4-(2-(2-ethenylsulfonylethoxy)ethylamino)-6-fluoro-1,3,5-triazine-2-ylamino]-2-sulfophenylazo]-5-hydroxynaphthalene-2,7-disulfonate

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:

Study initiation date - 15 February 1999; Experiment start date - 09 March 1999; Experiment completion date - 06 April 1999; Study completion date - 22 April 1999.

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:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40'574/B
Batch: WP 23/99
Purity: Approx. 75 %
Appearance: Solid, dark-red powder
Storage: At room temperature at about 20 °C
Expiration Date: 08 February 2006

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic (adaptation not specified)

Details on inoculum:

Strain / Inoculation: Mixture of polyvalent bacteria collected on March 07, 1999 from the aeration tank of a domestic sewage treatment plant, PRO RHENO Basel communal.

Sludge pretreatment: 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. pH value of the activated sludge: 7.21;

Bacteria concentration: 24 mg/l Suspended Solids (SS). The bacteria concentration was obtained by adding 10 ml of the pre-treated sludge (2.4 g/1 SS) per 1000 ml test medium.

Duration of test (contact time):

28 d

Initial conc.:

33.7 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

Experimental conditions
Temperature: room temperature (20 - 21.5 °C)
Test duration: 28 days
Lighting: Indirect daylight

Stock solutions and test concentrations
Stock solutions: The test concentrations were prepared from stock solutions of 5.76 g/l for the test substance and 8.60 g/l for the reference substance.

Test concentrations: 33.7 mg/l DOC for the test substance; 34.0 mg/l DOC for the reference substance.

Reference substance:

other: D(+)-Glucose

Parameter:

% degradation (DOC removal)

Value:

ca. 0

Sampling time:

28 d

Details on results:

Points of degradation plot (test substance):
2 % degradation after 1 d;
3 % degradation after 3 d;
0 % degradation after 7 d;
0 % degradation after 10 d;
0 % degradation after 14 d;
0 % degradation after 21 d;
0 % degradation after 28 d;

Results with reference substance:

Points of degradation plot (reference substance):
34 % degradation after 1 d
96 % degradation after 3 d
95 % degradation after 7 d
97 % degradation after 10 d
95 % degradation after 14 d
94 % degradation after 21 d
99 % degradation after 28 d

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

The test substance FAT 40574/B was not readily biodegradable after 28 days of incubation.

Executive summary:

This report describes the experimental techniques and the results obtained in a study for ready biodegradability performed with FAT 40574/B. The study employs method specified according to OECD test guideline 301A in a GLP-certified laboratory. A mixture of polyvalent bacteria (activated sludge obtained from a communal wastewater treatment plant) were used as inoculum. D-Glucose was used as reference standard. Test duration was 28 days. Based on the findings of the study, the biodegradation after 28 days was 0 % and the reference substance degraded 95 % in 14 days.

Reference 2

Endpoint:

biodegradation in water: inherent biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Study initiation date - 25 February 1999; Experiment start date - 10 March 1999; Experiment completion date - 07 April 1999; Study completion date - 27 April 1999.

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:

no

Qualifier:

according to guideline

Guideline:

EU Method C.9 (Biodegradation: Zahn-Wellens Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40'574/B
Batch: WP 23/99
Purity: Approx. 75 %
Appearance: Solid, dark-red powder
Storage: At room temperature at about 20 °C
Expiration Date: 08 February 2006

Oxygen conditions:

aerobic

Inoculum or test system:

sewage, domestic (adaptation not specified)

Details on inoculum:

Activated sludge of a communal sewage treatment plant (ARA Pro Rheno Basel) collected on March 09, 1999 was washed twice with tap water one day before the test was started.

Duration of test (contact time):

28 d

Initial conc.:

147.5 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

Temperature: 20 - 25 °C
pH: 7.0 - 8.0
O2: 6.8 - 8.7 mg/l
Sludge dry weight: 0.51 g/l
DOC of distilled water: 0 mg/l
DOC content of test substance: 30.4 %
Amount of test substance (cone.): 493 mg/l
Test volume: 2 litres
Test duration: 28 days
Aeration: yes
Agitation: yes
Lighting: diffuse day light

Stock solution and test concentrations
Stock solution: The day prior to the start of the test the DOC content of the stock solution was determined. From this value the actual start concentration was calculated. 296 and 294 mg/l DOC of the test substance 304 and 304 mg/l DOC of the reference substance
Test concentration: The stock solution was diluted 1 : 2 with test water including activated sludge to get the final test concentration of:
148 and 147 mg/l DOC of the test substance
152 and 152 mg/l DOC of the reference substance

Test medium
For each stock solution the mineral salts were dissolved in water (distilled water, previously
checked for purity by DOC analysis) and made up to 1 litre.
Stock solution A
0.5 g - NH4Cl
33.4 g - Na2HPO4 . 2 H2O
8.5 g - KH2PO4
21.75 g - K2HPO4
Stock solution B 27.5 g - CaCl2
Stock solution C 22.5g - MgSO4. 7 H2O
Stock solution D 0.25g - FeCl3. 6 H2O

Preparation of the mineral medium
10 ml of solution A was mixed with 800 ml of distilled water, 1 ml of solutions B, C, D was added and made up to 1 litre.

Reference substance:

diethylene glycol

Parameter:

% degradation (DOC removal)

Value:

ca. 2

Sampling time:

28 d

Details on results:

Experimental values (test substance):
0 % degradation after 1 d;
0 % degradation after 5 d;
0.5 % degradation after 8 d;
1 % degradation after 14 d;
1.5 % degradation after 21 d;
3.5 % degradation after 27 d;
1.5 % degradation after 28 d;

Results with reference substance:

Experimental values (reference substance):
0 % degradation after 1 d
13 % degradation after 5 d
92 % degradation after 8 d
99 % degradation after 9 d

Validity criteria fulfilled:

yes

Interpretation of results:

not inherently biodegradable

Conclusions:

FAT 40574/B was found to be not inherently biodegradable

Executive summary:

In a GLP study, the inherent biodegradation of FAT 40574/B was evaluated according to OECD test guideline 302B. Activated sludge obtained from a communal wastewater treatment plant was used as inoculum. DOC removal was measured in 28 days. The test concentration was 148 and 147 mg/L DOC. Diethylene glycol was used as positive control. Based on the findings of this study, the biodegradation after 28 days was 2 %. The reference substance degraded 99 % after 9 days.

Reference 3

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Experiment start date - 05 March 1999; Experiment end date - 10 March 1999; Study completion date - 24 March 1999.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.5 (Degradation: Biochemical Oxygen Demand)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

Identity: FAT 40'574/B
Batch: WP 23/99
Purity: Approx. 75 %
Appearance: Solid, dark-red powder
Storage: At room temperature at about 20 °C
Expiration Date: 08 February 2006

Oxygen conditions:

other: Titration study

Inoculum or test system:

sewage, domestic (adaptation not specified)

Details on inoculum:

Seeding water taken from the aeration tank of a communal sewage treatment plant (ARA Pro Rheno Basel, Switzerland) collected on
March 04, 1999.

Duration of test (contact time):

5 d

Initial conc.:

2.3 mg/L

Based on:

test mat.

Initial conc.:

4.6 mg/L

Based on:

test mat.

Initial conc.:

9.3 mg/L

Based on:

test mat.

Initial conc.:

18.5 mg/L

Based on:

test mat.

Initial conc.:

37 mg/L

Based on:

test mat.

Initial conc.:

74 mg/L

Based on:

test mat.

Initial conc.:

148 mg/L

Based on:

test mat.

Initial conc.:

296 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

- Calculation of BOD5: The BOD5 of a test substance is calculated from the O2 consumption, determined by the difference of the O2- concentration at the begin and end of the test (corrected by the blank control) and the corresponding concentration of the test substance in the test flask.
(Otcs-Ob) - Otce . 1000 / C

Where:
Otcs = O2-concentration at the test start (test flask with substance concentration C)
Ob = O2-consumption of the blank in 5 days
Otce = O2-concentration after 5 days (test flask with substance concentration C)
C= concentration of the test substance

Reference substance:

other: Glucose

Test performance:

Reference substance: 188 mgO2/L

Details on results:

The BOD5 of the test substance, corrected by the blank control is 5 mg/g O2.

Parameter:

BOD5

Value:

5 mg O2/g test mat.

Results with reference substance:

The BOD5 of Glucose/Glutamic acid corrected by the blank control is 188 mg/g O2.

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

The BOD5 of the test substance is 5 mg/g O2.

Executive summary:

The biochemical oxygen demand (BOD5) of the test substance was determined in a titration study according to EEC Directive 79/831, Annex V, C.5. The nominal test item concentrations were 2.3, 4.6, 9.3, 18.5, 37, 74, 148 and 296 mg/L and the test was performed at the temperature of 20 ± 1 °C for the duration of 5 days. The oxygen consumption was measured with an oxygen-sensitive electrode at the start and at the end of the test. Additionally, also a blank and a positive control (Glucose) were used to assess the validity of the determination. The BOD5 of the test substance was found to be 5 mg/g O2. In another study, the COD was found to be 765 mg/g O2. The resultant BOD:COD ratio will be 0.006, indicating that the substance is not readily biodegradable. The BOD5 of Glucose was found to be 188 mgO2/L.

Description of key information

FAT 40574 is not biodegradable.

Key value for chemical safety assessment

Biodegradation in water:

not biodegradable

Type of water:

freshwater

Additional information

Ready biodegradation

This report describes the experimental techniques and the results obtained in a study for ready biodegradability performed with FAT 40574/B. The study employs method specified according to OECD test guideline 301A in a GLP-certified laboratory. A mixture of polyvalent bacteria (activated sludge obtained from a communal wastewater treatment plant) were used as inoculum. D-Glucose was used as reference standard. Test duration was 28 days. Based on the findings of the study, the biodegradation after 28 days was 0 % and the reference substance degraded 95 % in 14 days. 

Inherent biodegradation

In a GLP study, the inherent biodegradation of FAT 40574/B was evaluated according to OECD test guideline 302B. Activated sludge obtained from a communal wastewater treatment plant was used as inoculum. DOC removal was measured in 28 days. The test concentration was 148 and 147 mg/L DOC. Diethylene glycol was used as positive control. Based on the findings of this study, the biodegradation after 28 days was 2 %. The reference substance degraded 99 % after 9 days.

BOD:

The biochemical oxygen demand (BOD5) of the test substance was determined in a titration study according to EEC Directive 79/831, Annex V, C.5. The nominal test item concentrations were 2.3, 4.6, 9.3, 18.5, 37, 74, 148 and 296 mg/L and the test was performed at the temperature of 20 ± 1 °C for the duration of 5 days. The oxygen consumption was measured with an oxygen-sensitive electrode at the start and at the end of the test. Additionally, also a blank and a positive control (Glucose) were used to assess the validity of the determination. The BOD5 of the test substance was found to be 5 mg/g O2. In another study, the COD was found to be 765 mg/g O2. The resultant BOD:COD ratio will be 0.006, indicating that the substance is not readily biodegradable. The BOD5 of Glucose was found to be 188 mg O2/L.

COD

The chemical oxygen demand (COD) of the test substance was determined in a titration study according to EEC Directive 79/831, Annex V, C.6. The nominal test item concentration was 5.0 mg and the test was performed at the temperature of 148± 3 °C for the duration of 2 h. The titration was performed with potassium dichromate. Additionally, also a blank and a positive control (potassium hydrogen phthalate) were used to assess the validity of the determination. The COD of the test substance is 765 mg/g O2, while the COD of KHP is 199 mg/g O2 (criteria: 200 ± 8 mg/g O2).The BOD of test substance, determined in another experiment was found to be 5 mg/g O2, the resultant BOD COD ratio was found to be 0.006 indicating the test substance to be not readily biodegradable.