Pentasodium 7-(4-(4-(5-amino-4-sulfonato-2-(4-((2-(sulfonato-ethoxy)sulfonyl)phenylazo)phenylamino)-6-chloro-1,3,5-triazin-2-yl)amino-2-ureidophenylazo)naphtalene-1,3,6-trisulfonate

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:

22 March 1996 to 13 June 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.4-D (Determination of the "Ready" Biodegradability - Manometric Respirometry Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Directive 92/69 EEC of July 31, 1992 C.6: Degradation - Chemical Oxygen Demand (EEC Publication No. L 383 A, December 1992).

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: DIN 38409, H-41, December 1980: Bestimmung des chemischen Sauerstoffbedarfs (CSB) im Bereich über 15 mg/l.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40549/A
Batch No: TV1
Expiration date: 01 Jan 2001
Stability in water: for atleast 48 hours
Solubility in water: approximately 90 mg/L at 20 degree celcius.
Aggregate state under storage conditions: Solid (powder)
Colour: Yellow brown.
Storage conditions: At room temperature
Safety precautions: Routine hygenic procudures are sufficient to assure personnel health and safety.

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

Type: Bacterial sludge
Source: Domestic waste water treatment plant (ARA Ergolz II, Füllinsdorf/Switzerland)
Conditioning: The sludge used for this study was washed three times with tap water and amounts corresponding to 4 g dry material per litre (±10 %) were mixed with Sörensen buffer solution (pH 7) and then aerated until use. The pH of the sludge was adjusted to 7 ± 1 with NaOH or HCl, and filtered through cotton wool prior to use.
The final concentration of the inoculum corresponded to 30 mg suspended solids per litre.

Duration of test (contact time):

28 d

Initial conc.:

100 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

Incubation:
Apparatus: The test flasks were incubated on a SAPROMAT D12 (Voith GmbH) equipped with an evaluation program.
Principle: Electro-chemical analysis process: The reaction processes consume the dissolved oxygen in liquid and generate CO2. The CO2 is adsorbed by soda lime and the total pressure decreases. The drop in pressure is detected and converted into an electrical signal by means of an electrode type manometer. The consumed oxygen is replaced by electrolytically generated oxygen from copper sulfate solution.
Duration: 28 days
Light Regimen: Darkness
Temperature: 22 °C, maintained with a built-in thermostat and checked once per week.
pH: was measured in all incubation flasks at the end of the incubation period.

Reference substance:

aniline

Remarks:

purity: 99.5%

Parameter:

% degradation (O2 consumption)

Value:

2.7

Sampling time:

28 d

Details on results:

The BOD of the non-inoculated flask (flask 3) containing FAT 40549/A and sterile test medium was 0 mg Og/I by the end of the test. Therefore, no abiotic degradation occurred over the 28-day exposure period.

The BOD of the toxicity control (flask 4) containing both FAT 40549/A and the reference compound Aniline showed a similar course of biodégradation over the 28-day exposure period as the two procedure controls (flasks 9 and 10) containing the reference compound Aniline, only. At the end of the test (day 28), the BOD of flasks 4, 9 and 10 was 194, 194 and 222 mg/L, respectively.

Parameter:

BOD5

Value:

1 other: mg O2/l;

Parameter:

COD

Value:

759 mg O2/g test mat.

Results with reference substance:

The percentage degradation of Aniline in the procedure controls (flasks 9 and 10) was 73.4 % and 87.1 % on exposure day 14.

The COD of FAT 40'549/A was determined to be 75.0, 76.3 and 76.3 mg O2/100 mg, resulting in a mean of 75.9 mg O2/100 mg. The COD of the positive controls was determined to be 200.1 mg O2/l on average and less than 10 % of the oxidizing agent was used up in the blank controls.

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

FAT 40549/A was found to be not readily-biodegradable (2.7 %) in the "Manometric Respirometry Test" over the 28-day exposure period.

Executive summary:

FAT 40'549/A was investigated for its biodegradability in the "Manometric Respirometry Test" over a period of 28 days. This test was conducted in accordance with OECD TG 301F in a GLP certified laboratory.

The reference compound Aniline was readily biodegraded by an average of 80.3 % and 81.8 % on exposure day 14 and at the end of the 10-day window between exposure day 15 and 16, respectively. Aniline reached an average biodegradation rate of 86.3 % by the end of the test (day 28). In the toxicity control, containing both FAT 40'549/A and the reference compound Aniline, no inhibitory effect on the microorganisms was observed.

Only a very slight biochemical oxygen demand (2 mg O2/L was measured for the test article in the two test flasks over the 28-day exposure period. Therefore, FAT 40549/A was found to be practically non-biodegradable (2.7 %) under the test conditions over 28 days.

Reference 2

Endpoint:

biodegradation in water: inherent biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09 April 1996 to 12 June 1996

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:

other: Directive 87/302/EEC of November 18,1987, p. 99, Zahn-Wellens Test.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40'549/A
Batch No: TV1
Expiration date: 01 Jan 2001
Stability in water: for atleast 48 hours
Solubility in water: approximately 90 mg/L at 20 degree celcius.
Aggregate state under storage conditions: Solid (powder)
Colour: Yellow brown.
Storage conditions: at room temperature
Safety precautions: Routine hygenic procudures are sufficient to assure personnel health and safety.

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

Test Organisms: Microorganisms from a domestic waste water treatment plant (ARA Ergolz II, Füllinsdorf / Switzerland)

Conditioning: The sludge was centrifuged, the supernatant liquid phase decanted, the solid material resuspended in tap water and again centrifuged.
This procedure was repeated two times. An aliquot of the final sludge suspension was weighed, thereafter dried and the ratio (R) of wet sludge to dry sludge calculated. Finally, the amount of wet sludge corresponding to 4 g ±10 % dry weight basis was added per one litre of Soerensen buffer (pH 7) and kept aerated until use.

Inoculum concentration: An amount of sludge from a domestic waste water treatment plant, corresponding to 0.2 g dry material, was added per litre final test medium. The ratio of inoculum and test article (as DOC) was within 2.5:1 and 4 : 1

Duration of test (contact time):

28 d

Initial conc.:

200 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

TEST CONDITIONS
- Composition of medium: The stock solutions according to the guidelines were:
a) 8.5 g KH2P04, 21.75 g K2HP04, 33.4 g NagHPO« x 2H20, 0.5g NH4CI dissolved in bidistilled water and made up to 1000 ml with bidistilled water. The pH was 7.4.
b) 22.5 g MgS04 x 7H20 dissolved in bidistilled water and made up to 1000 ml with bidistilled water.
c) 36.4 g CaCI2 x 2H20 dissolved in bidistilled water and made up to 1000 ml with bidistilled water.
d) 0.25 g FeCI3 x 6H20 dissolved in bidistilled water and made up to 1000 ml with bidistilled water. Note: In order to avoid having to prepare this solution immediately before use, 1 (one) drop of concentrated HCl per litre was added.
10 ml of stock solution a) and 1 ml each of stock solutions b)-d) were combined and made up to 1000 ml with bidistilled water. The pH was adjusted to 7.4 with diluted hydrochloric acid.
- Test temperature: 20.5 - 22.0 °C
- pH: 7.2 to 8.0
- Aeration of dilution water: yes The oxygen concentration was checked at the start of the test and at the respective sampling intervals, resulting in oxygen concentrations in the range from 7.7 -10.4 mg 02 per litre.

TEST SYSTEM
- Culturing apparatus: Aerated 2.5 L cylindrical reactor flasks
- Method used to create aerobic conditions: Aeration

Reference substance:

aniline

Preliminary study:

The DOC of a solution containing 100 mg FAT 40'549/A per 1000 ml test medium was 31.1 mg C/l.

Parameter:

% degradation (DOC removal)

Value:

-3

Sampling time:

28 d

Details on results:

Expressed as percentage removal, average biodegradation ranged from -3 % to -11 % over the exposure period of 28 days to microorganisms from a domestic waste water treatment plant.

Results with reference substance:

The initial DOC concentration (measured after 3 hours of exposure) of the test solution containing the reference compound Aniline rapidly decreased by 92 % within the first seven days of exposure. Consequently, the validity criterion of 70 % DOC-removal within 14 days of exposure was fulfilled. At the end of the test (day 28), Aniline was completely biodegraded (100%).

Throughout the entire exposure period of 28 days, the mean DOC concentration varied from 57.0 mg/l to 61.9 mg/l and was higher in comparison to the initial mean DOC concentration of 56.2 mg/l measured on day 0 after 3 hours of exposure. Practically no DOC-removal was observed during the first three hours of exposure.

Validity criteria fulfilled:

yes

Interpretation of results:

not inherently biodegradable

Conclusions:

FAT 40549/A was non-biodegradable in the "Zahn-Wellens / EMPA Test" over the 28-day exposure period to microorganisms from a domestic waste water treatment plant.

Executive summary:

FAT 40549/A was investigated for its biodegradability in the "28-Day Zahn-Wellens / EMPA Test" according to method OECD guideline 302B.

The mean concentration of DOC (Dissolved Organic Carbon) in the test flasks containing the test article varied from 57.0 mg/L to 61.9 mg/L over the exposure period of 28 days and was higher in comparison to the initial mean DOC concentration of 56.2 mg/L measured on day 0 after 3 hours of exposure.

Practically no DOC-removal was observed during the first three hours of exposure.

Consequently, FAT 40549/A was found to be non-biodegradable over the 28-day test period. The reference compound Aniline ultimately biodegraded by 92 % within seven days of exposure. At the end of the test (day 28), positive control aniline was completely biodegraded (100 %), indicating the test system being valid.

Description of key information

FAT 40549/A was non-biodegradable.

Key value for chemical safety assessment

Biodegradation in water:

under test conditions no biodegradation observed

Type of water:

freshwater

Additional information

The ready biodegradability of FAT 40'549/A was investigated in the "Manometric Respirometry Test" over a period of 28 days. The reference compound Aniline was readily biodegraded by an average of 80.3 % and 81.8 % on exposure day 14 and at the end of the 10-day window between exposure day 15 and 16, respectively. Aniline reached an average biodegradation rate of 86.3 % by the end of the test (day 28). In the toxicity control, containing both FAT 40'549/A and the reference compound Aniline, no inhibitory effect on the microorganisms was observed. Only a very slight biochemical oxygen demand (2 mg O2/l) was measured for the test article in the two test flasks over the 28-day exposure period. Therefore, FAT 40'549/A was found to be practically non-biodegradable (2.7 %) under the test conditions over 28 days.

The inherent biodegradability of FAT 40'549/A was investigated in the "28-Day Zahn-Wellens / EM PA Test". The mean concentration of DOC (Dissolved Organic Carbon) in the test flasks containing the test article varied from 57.0 mg/l to 61.9 mg/l over the exposure period of 28 days and was higher in comparison to the initial mean DOC concentration of 56.2 mg/l measured on day 0 after 3 hours of exposure. Practically no DOC-removal was observed during the first three hours of exposure. Consequently, FAT 40'549/A was found to be non-biodegradable over the 28-day test period.

In conclusion, FAT 40'549/A was non-biodegradable.