Reaction mass of lithium sodium hydrogen 4-amino-6-({5-[(5-chloro-2,6-difluoropyrimidin-4-yl)amino]-2-sulfonatophenyl}diazenyl)-5-hydroxy-3-[(4-{[2-(sulfonatooxy)ethyl]sulfonyl}phenyl)diazenyl]naphthalene-2,7-disulfonate and lithium sodium hydrogen 4-amino-6-({5-[(5-chloro-2,6-difluoropyrimidin-4-yl)amino]-2-sulfonatophenyl}diazenyl)-5-hydroxy-3-{[4-(vinylsulfonyl)phenyl]diazenyl}naphthalene-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:

1986

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)

Deviations:

yes

Remarks:

reduced duration of test

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

reduced duration of test

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

sewage, predominantly domestic (adaptation not specified)

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): aeration tank of a wastewater plant treating predominantly domestic sewage (Wupperverband); date of collection: 1986-08-18
- Pretreatment: none

Duration of test (contact time):

5 d

Parameter followed for biodegradation estimation:

DOC removal

Reference substance:

aniline

Key result

Parameter:

% degradation (DOC removal)

Value:

0

Sampling time:

5 d

Details on results:

0% degradation after 5 d

Because of the structure and intended use of this substance as a stable colouring agent, it is not anticiapted to show any appreciable degradation in an aerobic test for ready biodegradability. This fact has been confirmed even at a reduced duration test.

Results with reference substance:

Degradation of reference substance: 77% degradation after 5 d

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

Under the study conditions, the test substance was considered to be not readily biodegradable.

Executive summary:

A study was conducted to determine the ready biodegradability of the test substance according to OECD Guideline 301D and EU Method C.6. Sludge from a predominantly domestic wastewater treatment plant was exposed to the test substance for a reduced duration of only 5 d. Degradation of the test substance was monitored in terms of DOC removal. The test set-up also included a reference substance aniline. Within the test period, no degradation of the test substance was observed. The biodegradation of the reference substance was equivalent to 77%. Under the study conditions, the test substance was considered to be not readily biodegradable (Kanne, 1986).

Reference 2

Endpoint:

biodegradation in water: inherent biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May 30, 2001 to June 27, 2001

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: Modified EU Method C.4-D

Version / remarks:

an adapted inoculum was used to assess the inherent biodegradability

Deviations:

not applicable

Principles of method if other than guideline:

An adapted inoculum was used in this test in order to determine the inherent biodegradability of the test substance.

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, adapted

Details on inoculum:

- Type: Mixed population of aquatic microorganisms (activated sludge)
- Origin: Aeration tank of sewage treatment plant Leverkusen Bürrig
- Date of collection: 2001-04-30
- Adaptation of the activated sludge to the test substance: 100 mg/L test substance and activated sludge (1 g/L suspended solids) were incubated at room temperature for 4 weeks in a flask with permanent aeration
- Concentration of inoculum: 0.30 g ss/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:

(The consumption of oxygen was determined by measuring the quantity of oxygen (produced electrolytically) required to maintain constant gas volume in the respirometer flask.)

Details on study design:

PRETREATMENT OF THE TEST SUBSTANCE
- 25 mg test substance was weighed out on aluminium foil. This substance inclusive of the aluminium foil was added to the test vessels to give a test substance concentration of 100 mg/L.

EXPOSURE CONDITIONS
- Test volume: 250 mL
- Test apparatus: Voith Sapromat
- Mixing: One magnetic stirrer per test vessel
- Incubation time: 28 d
- Incubation temperature: 20±1°C

TREATMENT
- Control: 3 replicates
- Test substance: 3 replicates
- Reference substance: 3 replicates
- Toxicity Control: 2 replicates

CHEMICAL ANALYSIS
- Nitrate-N/Nitrite-N method: Determination of nitrite nitrogen and nitrate nitrogen and the sum of both by flow analysis (CFA and FIA) and spectrometric detection
- Standard: EN ISO 13395
- Test apparatus: Continuous Flow Analyser SKALAR SAN Plus System

Reference substance:

benzoic acid, sodium salt

Remarks:

(Purity: 99%)

Key result

Parameter:

% degradation (O2 consumption)

Value:

ca. 2

Sampling time:

28 d

Details on results:

pH of the test vessels: 7.6

Degradation of test substance:
0% degradation after 2 d
1% degradation after 6 d
1% degradation after 8 d
2% degradation after 12 d
1% degradation after 14 d
2% degradation after 16 d
2% degradation after 20 d
2% degradation after 22 d
2% degradation after 26 d
2% degradation after 28 d

Results with reference substance:

Degradation of test substance:
45% degradation after 2 d
76% degradation after 6 d
82% degradation after 8 d
89% degradation after 12 d
91% degradation after 14 d
92% degradation after 16 d
93% degradation after 20 d
94% degradation after 22 d
95% degradation after 26 d
95% degradation after 28 d

- The used concentrations of the test substance did not show toxic effects to bacteria

- No oxygen consumption by nitrification was observed in the flask with test substance at the end of the test.

Validity criteria fulfilled:

yes

Interpretation of results:

not inherently biodegradable

Conclusions:

Under the study conditions, the test substance was considered to be ‘not inherently biodegradable’.

Executive summary:

A study was conducted to assess the inherent biodegradability of the test substance according to a modified EU Method C.4-D, in compliance with GLP. Pre-adapted activated sludge was exposed to 100 mg/L of the test substance in a closed flask at a constant temperature (20±1°C) for 28 d. Degradation was assessed by the determination of Biochemical Oxygen Demand (BOD) on Days 2, 6, 8, 12, 14, 16, 20, 22, 26 and 28. The test set-up also included a reference substance (benzoic acid, sodium salt). The test substance degradation was equivalent to 2% by Day 28. The reference substance showed 91% degradation by Day 14. Under the study conditions, the test substance was not considered to be inherently biodegradable (Müller, 2001).

Key value for chemical safety assessment

Biodegradation in water:

under test conditions no biodegradation observed

Additional information

A study was conducted to determine the ready biodegradability of the test substance according to OECD Guideline 301D and EU Method C.6. Sludge from a predominantly domestic wastewater treatment plant was exposed to the test substance for a reduced duration of only 5 d. Degradation of the test substance was monitored in terms of DOC removal. The test set-up also included a reference substance aniline. Within the test period, no degradation of the test substance was observed. The biodegradation of the reference substance was equivalent to 77%. Under the study conditions, the test substance was considered to be not readily biodegradable (Kanne, 1986).

A study was conducted to assess the inherent biodegradability of the test substance according to a modified EU Method C.4-D, in compliance with GLP. Pre-adapted activated sludge was exposed to100 mg/L of the test substance in a closed flask at a constant temperature (20±1°C) for 28 d. Degradation was assessed by the determination of Biochemical Oxygen Demand (BOD) on Days 2, 6, 8, 12, 14, 16, 20, 22, 26 and 28. The test set-up also included a reference substance (benzoic acid, sodium salt). The test substance degradation was equivalent to 2% by Day 28. The reference substance showed 91% degradation by Day 14. Under the study conditions, the test substance was not considered to be inherently biodegradable (Müller, 2001).