Trisodium 2,4-diamino-3,5-bis-[4-(2-sulfonatoethoxy)sulfonyl)phenylazo]benzenesulfonate

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:

Dec. 05, 1996 to Jan. 29, 1997

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:

other: EEC Directive 92/69 EEC, Annex No. L 383A, C.4-D for testing chemicals (29,12,92)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Swiss GLP

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: WP2/96
- Expiration date of the lot/batch: 11/2004

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Solubility of the test substance in the solvent/vehicle: ca. 50 g/l [in water]

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

Strain / Inoculation: Mixture of polyvalent bacteria collected on December 10,1996 from the aeration tank of a domestic sewage treatment plant, PRO RHENO Basel communal.
Sludge pretreatment: The sludge was washed 4 times with tap water and the last time with test medium. An amount corresponding to 3 g dry material per liter (±10 %) was mixed with test medium and then aerated until use. pH value: 7.2
Bacteria concentration: 30 mg/l suspended solids

Duration of test (contact time):

28 d

Initial conc.:

100 mg/L

Based on:

test mat.

Initial conc.:

97 mg/L

Based on:

ThOD/L

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

Stock solutions: 2.50 g/500 ml reference substance in distilled water.
Test concentrations: 25.0, 24.9, 25.1, and 25.0 mg test substance in Flask 1, 2, 3 and 4 respectively, 100 mg/l of the reference (20 ml stock solution in 1000 ml test medium)
Temperature: 21 – 21.5 °C
Lighting: indirect daylight
The pH value of all incubation flasks were measured at the beginning and at the end of the test.

Reference substance:

benzoic acid, sodium salt

Remarks:

Sodium benzoate

Preliminary study:

no data

Test performance:

Tested chemicals were inoculated in the testing vessels with microorganisms. During the test period, the Biochemical Oxygen Demand was measured continuously by means of a BOD- meter. Biodegradability was calculated on the basis of the theoretical oxygen demand (ThOD) or of the COD if the ThOD could not be calculated.

Parameter:

% degradation (O2 consumption)

Value:

0

Sampling time:

28 d

Details on results:

The biodegradation of the toxicity control corrected by the blank control and measured as BOD versus ThOD was 63 % after 14 days, 66 % after 28 days.
The abiotic degradation measured as BOD was 0 % after 28 days.

Parameter:

BOD5

Value:

10 mg O2/g test mat.

Results with reference substance:

The biodegradation of the reference substance corrected by the blank control and measured as BOD versus ThOD was 85 % after 14 days, 89 % after 28 days.

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

FAT 45176/A was not readily biodegradable.

Executive summary:

The ready biodegradability of FAT 45176/A was determined in a 28 days manometric respirometry test according to OECD 301F and EU C.4-D.The test substance and the reference substance were tested in a concentration of about 100 mg/l, corresponding to 97 mg ThOD/l00 mg and 167 mg ThOD/l00 mg, respectively.

Strain / Inoculation used was mixture of polyvalent bacteria collected on December 10, 1996 from the aeration tank of a domestic sewage treatment plant, PRO RHENO Basel communal.

The sludge was washed 4 times with tap water and the last time with test medium. An amount corresponding to 3 g dry material per liter (±10%) was mixed with test medium and then aerated until use. The pH value was 7.2 and bacteria concentration was 30 mg/l suspended solids.

The mean value of the biodegradation of FAT 45176/A corrected by the blank control and measured as BOD versus ThOD was 0 % after 28 days. While biodegradation of the reference substance after 28 days 89 % and the biochemical oxygen demand (BOD₅) after 5 days was 10 mg 0₂/g

On the basis of the present study FAT 45176/A was not readily biodegradable.

Reference 2

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Mar. 17, 1998 to Jul. 21, 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: "Testing Methods for New Chemical Substances" (July 13, 1974, Kanpogyo No.5, Planning and Coordination Bureau, Environment Agency, Yakuhatu No.615, Pharmaceutical Affairs Bureau, Ministry of Health and Welfare, and 49 Kikyoku No. 3 92, Japan

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: WP2/96

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room Temp.
- Stability under test conditions: stable under the storage conditions
- Solubility of the test substance in the solvent/vehicle: Water ^ 100 g/L

Oxygen conditions:

aerobic

Inoculum or test system:

mixture of sewage, soil and natural water

Details on inoculum:

On-site sludge sampling was carried out at the 10 locations in Japan.
Sludge sampling method: (1) City sewage: Return sludges from sewage plants were collected. (2) Rivers, lake and sea: Surface water and surface soil which was in contact with the atmosphere were collected.
Mixing of fresh and old activated sludge: The filtrate (5 L) of the supernatant of the activated sludge cultivated about 3 months was mixed all together with the mixed filtrate (5 L) of the supernatant of a newly collected sludge (10 sources). The mixed filtrate (10 L) was aerated sufficiently through prefiltered open-air, after the pH value of the mixture was adjusted to 7.0 ±1.0.

Duration of test (contact time):

28 d

Initial conc.:

100 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Parameter followed for biodegradation estimation:

test mat. analysis

Details on study design:

Cultivation temperature: 25 ± 1 °C
After the termination of the cultivation, the test solution (water + test substance), the test solutions (sludge + test substance) and the test solution (control blank) were pretreated for high performance liquid chromatography (HPLC) analysis.

Reference substance:

aniline

Preliminary study:

no data

Test performance:

no data

Parameter:

% degradation (O2 consumption)

Value:

0

Sampling time:

28 d

Parameter:

% degradation (test mat. analysis)

Remarks:

By HPLC

Value:

0

Sampling time:

28 d

Results with reference substance:

28 d: 74.0 %

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

FAT 45176/A was not readily biodegradable by microorganisms.

Executive summary:

Biodegradation test of FAT 45'176/A by microorganisms was carried out according to OECD 301C (Modified MTTI Test). Conditions of cultivation used was

 

(1) Concentration of test substance 100 mg/L

(2) Concentration of activated sludge 30 mg/L (as the concentration of suspended solid)

(3) Volume of test solution 300 mL

(4) Cultivation temperature 25 ± 1 °C

(5) Cultivation duration 28 days

 

On-site sludge sampling was carried out at 0 locations in Japan. Sludge sampling method used was City sewage (Return sludges from sewage plants were collected) and Rivers, lake and sea Surface water and surface soil which was in contact with the atmosphere were collected.

 

The measurement of biochemical oxygen demand (BOD) by means of a closed system oxygen consumption measuring apparatus while determination of test substance by means of high performance liquid chromatography (HPLC).

 

Based on the study results, the percentage biodegradation by BOD and HPLC was ‘0 %’. Main component in the test substance changed to components contained in the test substance, but the test substance was not biodegraded by microorganisms under the present test conditions.

In conclusion, FAT 45176/A was not readily biodegradable by microorganisms.

Description of key information

Reactive Orange 136 was not readily 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 45176/A was determined in a 28 days manometric respirometry test according to OECD 301F and EU C.4-D.The test substance and the reference substance were tested in a concentration of about 100 mg/l, corresponding to 97 mg ThOD/l00 mg and 167 mg ThOD/l00 mg, respectively.

Strain / Inoculation used was mixture of polyvalent bacteria collected on December 10, 1996 from the aeration tank of a domestic sewage treatment plant, PRO RHENO Basel communal. The sludge was washed 4 times with tap water and the last time with test medium. An amount corresponding to 3 g dry material per liter (±10%) was mixed with test medium and then aerated until use. The pH value was 7.2 and bacteria concentration was 30 mg/l suspended solids.

The mean value of the biodegradation of FAT 45176/A corrected by the blank control and measured as BOD versus ThOD was 0 % after 28 days. While biodegradation of the reference substance after 28 days 89 % and the biochemical oxygen demand (BOD₅) after 5 days was 10 mg 0₂/g

On the basis of the present study FAT 45176/A was not readily biodegradable.

In another key study (1998), biodegradation test of FAT 45'176/A by microorganisms was carried out according to OECD 301C (Modified MTTI Test). Conditions of cultivation used was

(1) Concentration of test substance 100 mg/L

(2) Concentration of activated sludge 30 mg/L (as the concentration of suspended solid)

(3) Volume of test solution 300 mL

(4) Cultivation temperature 25 ± 1 °C

(5) Cultivation duration 28 days

 

On-site sludge sampling was carried out at 0 locations in Japan. Sludge sampling method used was City sewage (Return sludges from sewage plants were collected) and Rivers, lake and sea Surface water and surface soil which was in contact with the atmosphere were collected.

The measurement of biochemical oxygen demand (BOD) by means of a closed system oxygen consumption measuring apparatus while determination of test substance by means of high performance liquid chromatography (HPLC).

 

Based on the study results, the percentage biodegradation by BOD and HPLC was ‘0 %’. Main component in the test substance changed to components contained in the test substance, but the test substance was not biodegraded by microorganisms under the present test conditions.

In supporting studies, the BOD₅ of FAT 45176/A was 14 mg O₂/g. The COD of FAT 45176/A was 869 mg O₂/g. The ratio of BOD₅/COD is 1.6 %, which means the test substance is not readily biodegradable.

In another supporting study, FAT 45'176/A was investigated for its inherent biodegradability in the 28-Day "Zahn-Wellens/ EMPA Test". The mean concentration of DOC (Dissolved Organic Carbon) in the test flasks containing the test article slightly decreased from initially 76.3 mg/l to 62.7 mg/l over the exposure period of 28 days. Expressed as percentage DOC removal, a mean biodegradation of 18 % was noted by the end of the test (day-28). Therefore, FAT 45'176/A was found to be slightly biodegradable over the 28-day exposure period under the present test conditions.

From the results above, it can be concluded thatFAT 45176/A was not readily biodegradable.