trisodium 3-[(E)-2-(4-{[4-chloro-6-({2-[(4-{[4-(ethenesulfonyl)phenyl]amino}-6-fluoro-1,3,5-triazin-2-yl)amino]propyl}amino)-1,3,5-triazin-2-yl]amino}-5-sulfonaphthalen-1-yl)diazen-1-yl]naphthalene-1,5-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:

10 July 2006 to 02 October 2006

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:

not specified

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

other: DIN 38414-59.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40826/A
Batch no.: TZ 5604 BOP 01/06
Expiration date: February 01, 2011
Purity: Content of organic part (Na-salt): approx. 78 %; Oligomers: 13 %; Main component: approx. 48 %
Solubility in water: Approx. >50 g/L at room temperature
Stability in water: Max. 7 days at room temperature
pH: 7.6 (1 g/L)
Aggregate state/physical form at room temperature: Solid (orange powder)
Storage conditions: At room temperature at about 20 °C, away from direct sunlight
Specific instructions: Store in desiccator

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, adapted

Details on inoculum:

The study was performed with aerobic activated sludge from a wastewater treatment plant (ARA Ergolz II, Füllinsdorf, Switzerland) treating predominantly domestic wastewater. The sludge was washed twice with tap water by centrifugation and the supernatant liquid phase was decanted. A homogenized aliquot of the final sludge suspension was weighed, thereafter dried and the ratio of wet to dry weight was calculated. Based on this ratio, calculated amounts of wet sludge were suspended in test water to obtain a concentration equivalent to 4 g (±10 %) dry material per liter. During holding, the sludge was aerated at room temperature until use. Prior to use, the sludge was diluted with test water to a concentration of 1 g per liter (dry weight basis). This diluted activated sludge was used as inoculum to give a final concentration of 30 mg dry material per liter.

Duration of test (contact time):

28 d

Initial conc.:

102 mg/L

Based on:

ThOD/L

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

- Apparatus: The test flasks (500-mL Erlenmeyer flasks, labeled with all necessary information to ensure unmistakable identification) were incubated under continuous stirring in a SAPROMAT D12 (Voith GmbH, Heidenheim, Germany). Oxygen consumption was recorded manually by taking a daily reading at least each working day.
- Electro-chemical analysis process: The biodegradation process consumes the dissolved oxygen in the liquid and generates CO2. The CO2 is adsorbed by soda lime and the total pressure decreases in the airtight test flasks. The pressure drop 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 a copper sulfate solution.
- Test duration: 28 days
- Light conditions: Darkness
- Test temperature: 22 °C, maintained with a built-in thermostat and checked once per week.
- pH: Prior to test start, the pH was measured in each test flask before the addition of the activated sludge inoculum. At the end of incubation the pH was measured again in each test flask.

Reference substance:

benzoic acid, sodium salt

Parameter:

% degradation (O2 consumption)

Value:

0

Sampling time:

28 d

Details on results:

The percent biodegradation of the test item was calculated based on the theoretical oxygen demand of 1.13 mg 02/mg test item without nitrification (ThODNH4), and 1.90 mg O2/mg test item with nitrification (ThODNO3). The biochemical oxygen demand (BOD) of the test item FAT 40826/A in the test media was slightly higher than in the inoculum controls. At the end of the 28-day exposure period, the mean biodegradation of FAT 40826/A amounted to 8 % without nitrification or 5 % with nitrification, respectively. FAT 40826/A was not biodegradable under the test conditions within 28 days.

Parameter:

COD

Value:

0.82 mg O2/g test mat.

Parameter:

BOD5

Value:

0 mg O2/g test mat.

Results with reference substance:

91% after 28 days

Abiotic control:

no degradation of the test item under the conditions of the test within 28 days.

 

Toxicity control:

the course of biodegradation over the 28-day exposure period was similar to the two controls containing only the reference item. Within 14 days of exposure, biodegradation amounted to 60 % and 47 % based on the THOD-NH4 and THOD-NO3, respectively. Thus, according to guidelines, the test item had no inhibitory effect on activated sludge microorganisms at the tested concentration of 102 mg/L.

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

FAT 40826/A was found not to be biodegradable under the conditions of the test within 28 days.

Executive summary:

FAT 40826/A was investigated for its ready biodegradability in a manometric respirometry test over 28 days according to EU Commission Directive 92/69 EEC, C.4-D (1992) and OECD Guideline for Testing of Chemicals No. 301 F (1992). The biochemical oxygen demand (BOD) of the test item FAT 40826/A in the test media was slightly higher than in the inoculum controls. However, at the end of the 28-day exposure period, the mean biodegradation of FAT 40826/A was <10 %. Consequently, FAT 40826/A was not biodegradable under the test conditions within 28 days. No degradation of the test item occurred in the abiotic control under the test conditions within 28 days. In the toxicity control, containing both FAT 40826/A and the reference item sodium benzoate, no inhibitory effect on the biodégradation of the reference item was determined. Thus, FAT 40826/A had no inhibitory effect on the activity of activated sludge microorganisms at the tested concentration of 102 mg/L. In the procedure controls, the reference item sodium benzoate was degraded by an average of 88 % by Exposure Day 14, and by an average of 91 % by the end of the test (Exposure Day 28), thus confirming suitability of the activated sludge. The BOD5 (biochemical oxygen demand after five days) was recorded based on the EU Commission Directive 92/69 EEC, C.5, Degradation: Biochemical Oxygen Demand, 1992. After the test duration of five days, no oxygen consumption was determined in the two flasks containing the test item FAT 40826/A. Thus, the BOD5 of test item was 0 mg O2/mg test item. The COD (chemical oxygen demand) of the test item FAT 40826/A was determined according to the EU Commission Directive 92/69 EEC, C.6, following DIN 38414-S9. The COD of test item was 0.82 mg O2/mg test item. Consequnetly, the BOD5/COD quotient cannot be calculated and is close to zero. Based on the study results, FAT 40826/A was not found as readily biodegradable.

Reference 2

Endpoint:

biodegradation in water: inherent biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

28 August 2006 to 28 November 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 302 C (Inherent Biodegradability: Modified MITI Test (II))

Deviations:

yes

Remarks:

See Material and methods

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40826/A
Batch no.: TZ 5604 BOP 01/06
Expiration date: February 01, 2011
Purity: Content of organic part (Na-salt): approx. 78 %; Oligomers: 13 %; Main component: approx. 48 %
Solubility in water: Approx. >50 g/L at room temperature
Stability in water: Max. 7 days at room temperature
pH: 7.6 (1 g/L)
Aggregate state/physical form at room temperature: Solid (orange powder)
Storage conditions: At room temperature at about 20 °C, away from direct sunlight
Specific instructions: Store in desiccator

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, adapted

Details on inoculum:

The study was performed with aerobic activated sludge from a wastewater treatment plant (ARA Ergolz II, Füllinsdorf, Switzerland) treating predominantly domestic wastewater. The sludge was washed twice with tap water by centrifugation and the supernatant liquid phase was decanted. A homogenized aliquot of the final sludge suspension was weighed, thereafter dried and the ratio of wet to dry weight was calculated. Based on this ratio, calculated amounts of wet sludge were suspended in test water to obtain a concentration equivalent to 4 g dry material per liter (±10 %). During holding, the sludge was aerated at room temperature until use. Prior to use, the sludge was first thoroughly mixed and then diluted with test water to a concentration of 1 g per liter (dry weight basis). Based on the determined dry weight of this diluted activated sludge, defined amounts were added to test water to obtain a final concentration of 100 mg dry material per liter

Duration of test (contact time):

28 d

Initial conc.:

50 mg/L

Based on:

ThOD/L

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

- Apparatus: The test flasks (500-mL Erlenmeyer flasks, labeled with the necessary information to ensure unmistakable identification) were incubated under continuous stirring in a SAPROMAT 012 (Votth GmbH, Heidenheim, Germany). Oxygen consumption was recorded manually by taking a daily reading at least on each working day.
- Electrochemical analysis process: The biodegradation process consumes the dissolved oxygen in the liquid and generates CO2 The CO, is adsorbed by soda lime and the total pressure decreases in the airtight test flasks. The pressure drop 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 a copper sulfate solution.
- Test duration: 28 days
- Light conditions: Darkness
- Test temperature: 22 °C, maintained with a built-in thermostat and checked once per week.
- pH: Prior to test start, the pH was measured in each test flask before the addition of the activated sludge inoculum (Table 3). At the end of incubation, the pH was measured again in each test flask.

Reference substance:

benzoic acid, sodium salt

Parameter:

% degradation (O2 consumption)

Value:

0

Sampling time:

28 d

Details on results:

The percent biodegradation of the test ttem was calculated based on the theoretical oxygen demand of 1.13 mg O2/mg test item without nitrification (ThODNH4) and 1.90 mg O2/mg test item with nitrification (ThODNO3). The biochemical oxygen demand (BOD) of the test item FAT 40826/A in the test media was in the range of the inoculum controls throughout the study period of 28 days.
Consequently, FAT 40826/A was not biodegradable under the test conditions wtthin 28 days

Results with reference substance:

The percent biodegradation of the reference item sodium benzoate was calculated based on the theoretical oxygen demand of 1.67 mg O2,/mg. In the procedure controls, the reference item (sodium benzoate) was degraded by an average of 65 % and 78 % by Exposure Days 7 and 14, respectively, thus confirming suitability of the activated sludge. At the end of the test (Day 28), the reference item was degraded by an average of 86 %.

No significant degradation of the test item occurred in the abiotic control under the test conditions within 28 days. In the toxicity control, containing both FAT 40826/A and the reference item sodium benzoate, FAT 40826/A had no inhibitory effect on the activity of activated sludge microorganisms at the tested concentration of 50 mg/L. In the procedure controls, the reference item (sodium benzoate) was degraded by an average of 65 % and 78 % by Exposure Days 7 and 14, respectively, thus confirming suitability of the activated sludge. At the end of the test (Day 28), the reference item was degraded by an average of 86 %.

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

FAT 40826/A was not inherently biodegradable under the test conditions wtthin 28 days.

Executive summary:

In a GLP-compliant study, FAT 40826/A was investigated for its inherent biodegradability in a manometric respirometry test over 28 days based on the OECD Guideline for Testing of Chemicals No. 302 C (1981). The following modifications were made: activated sludge from one source was used, the activated sludge was not fed during the holding period of maximum seven days, the test was run at 22 °C, the test water composition was slightly changed, and only the biological oxygen demand (BOD) was monitored, no test item specific analysis was performed. The percent biodegradation of the test item was calculated based on the theoretical oxygen demand of 1.13 mg O2/mg test item without nitrification (ThOD_NH4) and 1.90 mg O2/mg test item with nitrification (ThOD_NO3). The biochemical oxygen demand (BOD) of the test item FAT 40826/A in the test media was in the range of the inoculum controls. Consequently, FAT 40826/A was not biodegradable under the test conditions within 28 days. No significant degradation of the test item occurred in the abiotic control under the test conditions within 28 days. In the toxicity control, containing both the test substance and the reference item sodium benzoate, the test substance had no inhibitory effect on the activity of activated sludge microorganisms at the tested concentration of 50 mg/L. In the procedure controls, the reference item (sodium benzoate) was degraded by an average of 65 % and 78 % by Exposure Days 7 and 14, respectively, thus confirming suitability of the activated sludge. At the end of the test (Day 28), the reference item was degraded by an average of 86 %. Based on the study results, FAT 40826/A was not inherently biodegradable under the test conditions within 28 days.

Description of key information

The test substance is neither inherently nor readily biodegradable.

Key value for chemical safety assessment

Biodegradation in water:

not biodegradable

Type of water:

freshwater

Additional information

FAT 40826/A was investigated for its ready biodegradability in a manometric respirometry test over 28 days according to EU Commission Directive 92/69 EEC, C.4-D (1992) and OECD Guideline for Testing of Chemicals No. 301 F (1992). The biochemical oxygen demand (BOD) of the test item FAT 40826/A in the test media was slightly higher than in the inoculum controls. However, at the end of the 28-day exposure period, the mean biodégradation of FAT 40826/A was <10 %. Consequently, FAT 40826/A was not biodegradable under the test conditions within 28 days. No degradation of the test item occurred in the abiotic control under the test conditions within 28 days. The BOD5 (biochemical oxygen demand after five days) was recorded based on the EU Commission Directive 92/69 EEC, C.5, Degradation: Biochemical Oxygen Demand, 1992. After the test duration of five days, no oxygen consumption was determined in the two flasks containing the test item FAT 40826/A. Thus, the BOD5 of test item was 0 mg O2/mg test item. The COD (chemical oxygen demand) of the test item FAT 40826/A was determined according to the EU Commission Directive 92/69 EEC, C.6, following DIN 38414-S9. The COD of test item was 0.82 mg O2/mg test item. Based on the study results, FAT 40826/A was not found as readily biodegradable. In another a GLP-compliant study, FAT 40826/A was investigated for its inherent biodegradability in a manometric respirometry test over 28 days based on the OECD Guideline for Testing of Chemicals No. 302 C (1981). The following modifications were made: activated sludge from one source was used, the activated sludge was not fed during the holding period of maximum seven days, the test was run at 22 °C, the test water composition was slightly changed, and only the biological oxygen demand (BOD) was monitored, no test item specific analysis was performed. The percent biodegradation of the test item was calculated based on the theoretical oxygen demand of 1.13 mg O2/mg test item without nitrification (ThOD_NH4) and 1.90 mg O2/mg test item with nitrification (ThOD_NO3). The biochemical oxygen demand (BOD) of the test item FAT 40826/A in the test media was in the range of the inoculum controls. Consequently, FAT 40826/A was not biodegradable under the test conditions within 28 days. Based on the study results, FAT 40826/A was not inherently biodegradable under the test conditions within 28 days.