main component 1 (isomer 1): 2-{6-fluoro-4-[3-(2,5-disulfo-phenylazo)-4-hydroxy-2-sulfonapht-7-ylamino]-1,3,5-triazin-2-ylamino}-3-{6-fluoro-4-[3-(1,5-disulfonaphth-2-ylazo)-4-hydroxy-2-sulfonaphth-7-ylamino]-1,3,5-triazin-2-ylamino}-propane sodium salt;main component 1 (isomer 2): 2-{6-fluoro-4-[3-(2,5-disulfo-phenylazo)-4-hydroxy-2-sulfonaphth-7-ylamino]-1,3,5-triazin-2-ylamino}-3-{6-fluoro-4-[3-(2,5-disulfo-phenylazo)-4-hydroxy-2-sulfonaphth-7-ylamino]-1,3,5-triazin-2-ylamino}-propane sodium salt;main component 2: 2,3-bis-{6-fluoro-4-[3-(2,5-disulfo-phenylazo)-4-hydroxy-2-sulfonaphth-7-ylamino]-1,3,5-triazin-2-ylamino}-propane sodium salt;main component 3: 2,3-bis-{6-fluoro-4-[3-(1,5-disulfonaphth-2-ylazo)-4-hydroxy-2-sulfonaphth-7-ylamino]-1,3,5-triazin-2-ylamino}-propane sodium salt

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

biodegradation in water: ready biodegradability

Remarks:

OECD 301 F

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04 October 1996 to 17 December 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:

yes

Remarks:

DOC Determination: only one sample from each flask for DOC-determination instead of the measurement in duplicate.

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 38409, H-41, December 1980: Bestimmung des chemischen Sauerstoffbedarfs (CSB) im Bereich uber 15 mg/L

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40557/A
Batch-no.: PV1
Purity: 65 %
Appearance: Solid powder, red-brown
Solubility (g/l): >100
Stability in water: at least 48 h
Expiration date: September, 2002
Storage conditions: At room temperature

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

- Source: Domestic waste water treatment plant (ARA Ergolz Il, Fullinsdorf/Switzerland)
- Conditioning: The sludge used was washed three times with tap water and amounts corresponding to 4 g dry material per litre (±10 %) were mixed with Sorensen buffer solution (pH 7) and then aerated until use five days later. The sludge was 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:

act. ingr.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

- Biochemical Oxygen Demand: The test flasks were incubated on a SAPROMAT D12 (Voith GmbH) equipped with an evaluation program in the darkness at 22 °C.
- Chemical Oxygen Demand: A predetermined amount of the test article, dispersed in water, was oxidized by mercury sulfate containing potassium dichromate in a strong sulfuric acid medium with silver sulfate as a catalyst under reflux of 2 hours. The residual dichromate was determined by titration with standardized ferrous ammonium sulfate.

Reference substance:

aniline

Parameter:

% degradation (O2 consumption)

Value:

0.8

Sampling time:

28 d

Results with reference substance:

The biodegradation of the reference substance:
- after 14 days: 77.4 %
- after 28 days: 82.4 %

The biochemical oxygen demand (BOD) in test flasks 2 and 3, containing FAT 40557/A at a concentration of 100 mg/I with respect to active ingredient, was 0 and 2 mg O2/1 (corrected for the mean BOD of the control flasks), respectively by the end of the 28-day test period. Expressed as percentage biodegradation, 0 and 1.6 % were noted by the end of the 28-day test period, resulting in a mean of 0.8 %. Therefore, FAT 40'557/A was found to be nonbiodegradable under the present test conditions.

 The mean biochemical oxygen demand (BOD) of the test article after five days of exposure, corrected for the mean BOD of the control flasks, was 0 mg O2/1; therefore, the ratio BODs : COD = 0 : 80.2.

Chemical oxygen demand (COD):

The COD of the test substance was determined to be 80.2 mg O2/100 mg. The COD of the positive controls was determined to be 198.6 mg O2/L on average and no more than 10% of the oxidizing agent was used up in the blank controls.

Abiotic control:

The BOD of the Non inoculated flask containing the test substance and sterile test medium was 0 mg O2/L on exposure day 28. therefore, no abiotic degradation occurred throughout the 28-day exposure period.

Toxicity control:

The BOD of the toxicity control containing both test substance and the reference compound aniline showed a similar course of biodegradation over the 28-day exposure period as the two procedure controls containing the reference compound aniline only.

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

FAT 40557/A is not readily biodegradable.

Executive summary:

In a GLP-compliant study, the ready biodegradability of the test substance was determined in a 28 days Manometric Respirometry Test according to OECD guideline 301F. Biochemical oxygen demands of 0 and 2 mg O2/L were noted for the test article in the two test flasks at the end of the 28-day exposure period. Therefore, FAT 40557/A was found to be nonbiodegradable (mean of 0.8 %) under the test conditions over 28 days. The reference compound Aniline was biodegraded by an average of 77.4 % on exposure day 14, and reached an average biodegradation rate of 82.4 % by the end of the test (day 28). In the toxicity control, containing both FAT 40557/A and the reference compound Aniline, no inhibitory effect on the microorganisms was observed. Based on the test results, FAT 40557/A is not readily biodegradable. The test substance induced no inhibition of the activity of the microorganisms.

Reference 2

Endpoint:

biodegradation in water: inherent biodegradability

Remarks:

OECD guideline 302 B

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 October 1996 to 20 February 1997

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)

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identity: FAT 40557/A
Batch-no.: PV1
Purity: 65 %
Appearance: Solid powder, red-brown
Solubility (g/l): >100
Stability in Water: At least 48 hours
Expiration date: 30 September, 2002
Storage conditions: At room temperature at about 20 °C, away from direct sunlight.

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic (adaptation not specified)

Details on inoculum:

- Source: ARA Ergolz II, Füllinsdorf / Switzerland
- Conditioning: The activated sludge was centrifuged, the supernatant liquid phase decanted, the solid material resuspended in tap water and again centrifuged. This procedure was repeated twice. An aliquot of the final sludge suspension was weighed, dried and the ratio of wet sludge to dry sludge calculated. the amount of wet sludge corresponding to 4g ±10 % dry weight basis was added per one litre of Soerensen buffer (pH 7) and kept aerated until use six days later.
- An amount of sludge from a domestic waste water treatment plant, corresponding to 0.2g 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:

DOC

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

- The study was run under diffuse illumination.
- Temperature: 22.5-23°C
- Oxygen concentration: 8.0-8.7 mg O2 per litre.
- pH: 7.0-7.6

Reference substance:

aniline

Parameter:

% degradation (DOC removal)

Value:

>= 1 - <= 7

Sampling time:

28 d

Details on results:

During the exposure period of 28 days, the mean DOC concentration very slightly decreased from 65.9 mg/L (starting value after 3 hours of exposure) to 64.2-62.9 mg/L. No DOC removal was observed during the first three hours of exposure.

Results with reference substance:

Initial DOC concentration of aniline rapidly decreased within 7 days (97 %). After 28 days aniline was completely biodegraded.

Validity criteria fulfilled:

yes

Interpretation of results:

not inherently biodegradable

Conclusions:

The test substance was found to be practically non-biodegradable.

Executive summary:

The inherent biodegradability of the test substance was determined in a 28 days Zahn-Wellens/ EMPA Test according to the OECD Guideline 302B and GLP compliant. The test substance was tested at a concentrations of 200 mg/L DOC. No adsorption occurred on the activated sludge. The biodegradation of the test substance ranged between 1 and 7 % up to 28 days. Therefore, it was concluded that the test substance was practically non-biodegradable.

Description of key information

The test substance is not readily biodegradable.

Key value for chemical safety assessment

Biodegradation in water:

not biodegradable

Type of water:

freshwater

Additional information

The ready biodegradability of the test substance was determined in a GLP-compliant study according to OECD Guideline 301F (RCC, 1996). Biodegradation of the test substance was determined to be 0.8 % after 28 days and therefore it was concluded that the test substance was not readily biodegradable. 

The inherent biodegradability of the test substance was determined in a GLP-compliant test according to OECD Guideline 302B (RCC, 1997). The biodegradation of the test substance ranged between 1 and 7 %. In another supporting inherent biodegradability study 0 % of biodegradation was seen in OECD guideline 302 B.

The biological oxygen demand (BOD5) was examined in a GLP-compliant study according to ISO 5815 (corresponding to EC method C.5) (CG 1996). The BOD5 of the test substance was found to be 0 mg O2/L. The chemical oxygen demand (COD) was examined in a GLP-compliant study according to DIN 38409- H 41-1 (corresponding to EC method C.6. (CG 1996). The COD of the test substance is 798 mg O2/L. The BOD5/COD ratio supports the conclusion that the test substance is not readily biodegradable.