REAKTIV-GELB F-68 072 FW

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

biodegradation in water: inherent biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 April 2006 to 09 June 2006

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

GLP compliance:

yes (incl. QA statement)

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

Type: mixed population of aquatic microorganisms (activated sludge)
Origin: Aeration tank of sewage treatment plant Leverkusen-Burrig
Date of collection: 2006-04-24
Pretreatment: activated sludge is washed twice with mineral medium; separation of the sludge by centrifugation
Concentration of activated sludge in reaction mixture: 0.4 g ss/l

Duration of test (contact time):

28 d

Initial conc.:

100 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

The following type of flasks were used
Test suspension (2 parallels)
A measured volume of mineral medium containing a known concentration of the test item (as the nominal sole source of organic carbon) and inoculum.

Procedure control (2 parallels)
In order to check the procedure, the reference chemical sodium benzoate is tested by setting up 2 flasks in parallel to the normal tests runs.

Inoculum blank (2 parallels)
A measured volume of mineral medium containing only inoculum.

Toxicity control (1 parallel)
Toxic effects of the test item are determined in a toxicity control flask (containing test item reference compound and inoculum).

Each flask is inoculated with 0.4 g ss per litre medium. The mixtures are aerated in the dark at 20 - 25°C.

Degradation is followed by DOC analysis at frequent intervals over a 28 day period. The degree of biodegradation is calculated by expressing the concentration of DOC removed (corrected for that in the blank inoculum control) as a percentage of the concentration initially present.

Reference substance:

benzoic acid, sodium salt

Preliminary study:

Not applicable

Test performance:

All validity criteria of the test were met.

Parameter:

% degradation (DOC removal)

Value:

6

Sampling time:

28 d

Details on results:

0 % degradation after 1 day.
3 % degradation after 7 days.
2 % degradation after 14 days.
0 % degradation after 21 days.
4% degradation after 27 days.
6 % degradation after 28 days.

Results with reference substance:

The reference compound showed 100 % degradation after 14 days

Study data – DOC determination:

	Flask no.	DOC after n days (mg/l)
		3h	1d	7d	14d	21d	27d	28d
Test item	5	98.90	105.2	100.9	103.0	111.2	98.50	97.19
	6	101.2	108.1	100.7	103.5	112.7	102.5	100.6
Blank	1	4.359	4.870	8.370	9.477	9.411	8.714	8.199
	2	4.771	4.863	8.416	10.27	9.992	9.352	9.085
Mean 1 + 2		4.565	4.867	8.393	9.874	9.702	9.033	8.642
Reference compound	3	98.97	28.20	6.086	10.38	9.768	9.352	9.194
	4	99.51	28.47	5.708	10.26	9.804	9.354	8.922
Toxicity control	7	195.7	130.2	100.3	103.5	111.4	100.2	99.57

 

Comments:After 0h 96.91 mg/l DOC were measured in the flask with test item before addition of innoculum.

 

The pH value in all Flasks at all sampling times varied between 7.0 and 8.0 and could always be satisfactory adjusted to a pH of 7.4 ± 0.2.

 

Degradation

 

Test item	% degradation after n days
Flask no	3h	1d	7d	14d	21d	27d	28d
5	0	0	2	1	0	5	6
6	0	0	4	3	0	3	5
Mean	0	0	3	2	0	4	6

 

Reference compound	% degradation after n day
Flask no	3h	1d	7d	14d	21d	27d	28d
3	0	75	100	99	100	100	99
4	0	75	100	100	100	100	100
Mean	0	75	100	100	100	100	100

 

Toxicity control	% degradation after n days
Flask no	3h	1d	7d	14d	21d	27d	28d
7	0	34	52	51	47	52	52

 

Comments:The used concentrations of the test item did not show toxic effects to bacteria.

Validity criteria fulfilled:

yes

Interpretation of results:

not inherently biodegradable

Conclusions:

Within 28 days, a degradation of 6 % was determined for Reaktiv Gelb F68072 FW.

Executive summary:

A study was performed to assess the inherent biodegradability of Reaktiv Gelb F68072 FW.

The study was conducted in accordance with the Draft updated OECD Guideline for Testing of Chemicals 302 B - Inherent Biodegradability. Study conducted in compliance with the Principles of Good Laboratory Practice (GLP). 

 

A solution of Reaktiv Gelb F68072 FW in a mineral medium was inoculated with activated sludge and incubated under aerobic conditions. Degradation was followed by DOC determinations at 0h, 3h, Id, 7 d, 14 d, 21 d, 27 d and 28 d.

 

Reaktiv Gelb F68072 FW showed:

  

0 % degradation after 1 day.

3 % degradation after 7 days.

2 % degradation after 14 days.

0 % degradation after 21 days.

4 % degradation after 27 days.

6 % degradation after 28 days.

The reference compound showed 100 % degradation after 14 days.

 

Thus, the test item is not inherently biodegradable.

Reference 2

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29. June to 24. August 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: ISO 11734

GLP compliance:

not specified

Oxygen conditions:

anaerobic

Inoculum or test system:

anaerobic bacteria

Details on inoculum:

- Source of inoculum: Municipal sewage Kriftel
- Storage conditions: at 35°C +/- 2°C thermostat Incubator, the temperature has remained during the test in range of 33.0°C- 37.0°C
- Storage length: 7 days
- Preparation of inoculum for exposure: bacterial biomass anaerobic digester of a semi-continuous operation, through a 0.5 mm sieve to separate coarse particles, washed, resuspended to the DOC value of the class does not exceed 10 mg/L (C)
100 mL/L of anaerobic inoculum suspension with a solids content of 32.4 g/L, corresponding to 3.24 The final volume in the dry mass of the audit method.
- Pretreatment:the inoculum was 7 days for depletion of carbon in the incubator
mixed at 35 ± 2 °C
- Type and size of filter used, if any: 0.5 mm

Duration of test (contact time):

ca. 56 d

Initial conc.:

113 mg/L

Based on:

DOC

Initial conc.:

500 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

CO2 evolution

Parameter followed for biodegradation estimation:

CH4 evolution

Details on study design:

TEST CONDITIONS
-Test vessel: 1000 mL glass tank with magnetic stirrer
- Composition of medium: ISO 11734 Punkt 6.2 Testmedium
- Additional substrate:
- Test temperature: 33 - 37 °C
- pH: 7.2
- pH adjusted: yes
- Suspended solids concentration: 100 mL/L
- Stock solution: 2000 mg/L; 2000 mg/800 mL weighed in highly purified water, measured pH 4.3; adjusted to pH 7.2, stirred for 2 hours at 22°C, measured pH 7.2, filled up to 1000 mL and used as stock solution.

TEST SYSTEM
- Culturing apparatus:
- Number of culture flasks/concentration: 3
- Method used to create aerobic conditions:
- Method used to create anaerobic conditions:
- Measuring equipment:
- Test performed in open system:
- Details of trap for CO2 and volatile organics if used: graduated gas mouse
- Other:

SAMPLING
- Sampling frequency:
- Sampling method:
- Sterility check if applicable:
- Sample storage before analysis:
- Other:

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control:
- Toxicity control:
- Other:

STATISTICAL METHODS:

Reference substance:

benzoic acid, sodium salt

Parameter:

% degradation (CO2 evolution)

Value:

< 20

Sampling time:

56 d

Parameter:

% degradation (CH4 evolution)

Value:

< 20

Sampling time:

56 d

Results with reference substance:

> 60% gas formation

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

A significant anaerobic biodegradation was not observed. The formation of gas within 56 days remained below 20% of the theoretical maximum.

Executive summary:

A significant anaerobic biodegradation was not observed. The formation of gas within 56 days remained below 20% of the theoretical maximum.

Reference 3

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1999-03-24 to 1999-04-21

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.4-E (Determination of the "Ready" Biodegradability - Closed Bottle Test)

Deviations:

no

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

other: mixed population of aquatic microrganisms (activated sludge)

Details on inoculum:

Inoculum
source of sewage effluent: secondary effluent of a domestic sewage treatment plant (Kläranlage Odenthal);
Treatment given: separation of coarse particles by filtration
Concentration of effluents in reaction mixture: 5 mL/L (SOP 2030-6600103-99 D)
Preconditioning: aeration 5 days

Duration of test (contact time):

28 d

Initial conc.:

8 mg/L

Based on:

test mat.

Initial conc.:

2.9 mg/L

Based on:

other: Reference Substance: Sodium bensoate, 99% Purity

Parameter followed for biodegradation estimation:

O2 consumption

Remarks:

During the study period the biodegradation of the test substance is determined on the basis of the reduction of dissolved oxygen.

Details on study design:

TEST CONDITIONS

- Solubilising agent (type and concentration if used): no data
- Test temperature: 20 ± 1°C
- pH adjusted: yes
- other: Improved type for a volatile substance used (no further details given)
CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Toxicity control: yes (Sodium benzoate)

TEST SYSTEM

- Measuring equipment: According to SOP 2030-0000104-98 D
- Test performed in closed vessels

INOCULUM:
Source of sewage effluent: secondary effluent of a domestic sewage treatment plant (Kläranlage Odenthal)
Treatment given: Separation of coarse particles by filtration
Concentration of effluents in reaction mixture: 5 mL/L
-Preconditioning: aeration for 5 days.
Study condition:
Duration: 28 days


SAMPLING (- Sampling frequency - Sampling method- Sample storage before analysis : according to the guideline) as well as SOP 2030-6600401-96 D

REFERENCE SUBSTANCE: sodium benzoate
PURITY: 99%
Initial concentration: 2.9 mg/L reference substance

Reference substance:

benzoic acid, sodium salt

Test performance:

Test substance is suspended in a mineral medium, inoculated with a mixed population of aquatic microorganisms and incubated for 28 days under aerobic conditions in the dark at 20 ± 1°C

Parameter:

other: dissolved oxygen (DO) consumption

Value:

0

Sampling time:

28 d

Details on results:

Points of degradation plot (test substance): 1 % degradation after 7 d 0 % degradation after 14 d 1 % degradation after 21 d 0 % degradation after 28 d
The test results indicate no hazard potential.

Results with reference substance:

Points of degradation plot (reference substance): 70 % degradation after 7 d 78 % degradation after 14 d 85 % degradation after 21 d 84 % degradation after 28 d
see more details below.

 

 

Results-DO depletion: % Degradation

 

 

Test substance Test concentration: 8.4 mg/L ThoD: 839 mg/g COD---mg/g	DO depletion after n days                                 [mg/L]
No.1: (mt0-mtx)-(mb0-mbx)	7 d 0.04	0.00	0.07	0.00
No.2: (mt0-mtx)-(mb0-mbx)	0.00	0.00	0.07	0.00
D1: % Degradation	1	0	1	0
D2: % Degradation	0	0	1	0
D, Mean	0	0	1	0

 

Reference substance Test concentration: 2.9 mg/L ThoD: 1665 mg/g COD---mg/g	DO depletion after n days                                              [mg/L]
No.1: (mt0-mtx)-(mb0-mbx)	7d 3.35	14d 3.74	21d 4.08	28d 4.03
No.2: (mt0-mtx)-(mb0-mbx)	3.40	3.76	4,13	4.06
D1: % Degradation	69	77	84	83
D2: % Degradation	70	78	86	84
D, Mean	70	78	85	84

 

*Toxicity control	DO depletion after n days                                              [mg/L]
No.1: (mt0-mtx)-(mb0-mbx)	7d 3.29	14d 3.70	21d 4.08	28d 4.03
No.2: (mt0-mtx)-(mb0-mbx)	3.34	3.82	4.13	4.10
D1: % Degradation	28	31	34	34
D2: % Degradation	28	32	35	35
D, Mean	28	32	35	35

 

*In the toxicity control 8.4 mg/L test substance and 2.9 mg/L reference substance were used. The ThOD values respectively COD values of these substances are shown in the table above.

 

 

Dissolved oxygen(mg O2/l, mean values)

 

No. of days	Blank inoculum (bottles 1-10)	Refernce substance (bottles 11-20)	Test substance (bottles 41-50)	Toxicity Control (bottles 51-60)
0	9.42	9.41	9.36	9.35
7	9.15	5.76	9.08	5.76
14	8.96	5.2	8.94	5.13
21	8.98	4.86	8.85	4.8
28	8.71	4.65	8.8	4.57

 

Oxygen depletionafter 28 d (mg/l, mean values):

 

Inoculum blank	No data
Test substance	0
Reference substance	84
Toxicity control	35

 

Result: Within the test period of 28 days, a degradation of 0 % was determinded.

Biodegradability:

0 % (28 d)

Method: Modified OECD Screening Test

The test results indicate no hazard potential.

The test substance is not readily biodegradable; so longer resident times of the substance in surface water are to be expected. However on the basis of the ecotoxicological test results there is no hazard potential.

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

The test results indicate no hazard potential.
The test substance is not readily biodegradable.

Executive summary:

Result: Within the test period of 28 days, a degradation of 0 % was determinded. The test substance has to be classified as “Not Readily Biodegradable”

Biodegradability = 0 %

The test results indicate no hazard potential.

Reference 4

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 2017-11-19 to 2018-08-30

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 A (Ready Biodegradability: DOC Die Away Test)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: MEP, P.R. China, the Guideline for the Testing of Chemicals, Degradation and Accumulation (the Second Edition), No. 301A "DOC Die-Away Test"

Version / remarks:

2013.9

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Oxygen conditions:

aerobic

Inoculum or test system:

sewage, predominantly domestic (adaptation not specified)

Details on inoculum:

A fresh sample of activated sludge was collected from a sewage treatment plant on November 22, 2017 as recommended by the OECD Guidelines for the Testing of Chemicals. The sample of activated sludge was collected from the aeration tank of Shenyang North Sewage Treatment Plant (No.258, Kunshan Xilu Huanggu District Shenyang City) prior to the test. It is a properly working treatment plant receiving predominantly domestic sewage.

Duration of test (contact time):

28 d

Initial conc.:

20 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

TEST CONDITIONS
- Test temperature: 22+/-2
- pH: 7 - 8
- Aeration of dilution water: yes
- Suspended solids concentration: 30 mg/L
- Continuous darkness: no, diffuse light

TEST SYSTEM
- Number of culture flasks/concentration: 2

SAMPLING
- Sampling frequency: day 0, 2, 4, 7, 10, 14, 19, 23, 28

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control: yes
- Toxicity control: yes
- Other: positive control

Reference substance:

benzoic acid, sodium salt

Test performance:

The validity criteria were met.

Key result

Parameter:

% degradation (DOC removal)

Value:

< 3

Sampling time:

28 d

Details on results:

The substance reached a biodegradation level of ca. 2.8% (DOC removal) after 28 d in an OECD 301A guideline study. Based on these results, the test item is not readily biodegradable according to OECD criteria under the test conditions.

Results with reference substance:

The percentage degradation of reference item attained 98.7% after 2 days and 100.2% at the 14th day, which exceeded the 70% pass level. The test was considered valid.

In the toxicity control containing test item, reference item and inoculums, the percentage degradationbased on total DOC attained47.6%after 2 days and 48.7%at the 14^th day, which exceeded 35%,indicating that the test item can be assumed to be not inhibitory to activated sludge under the conditions of the test.

In the adsorption control containing test item and sterilising agent, the percentage removal of test item was 2.8% at the 28 day, the test item can be assumed to be not adsorbed by activated sludge particles.

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

The substance reached a biodegradation level of ca. 2.8% (DOC removal) after 28 d in an OECD 301A guideline study. Based on these results, the test item is not readily biodegradable according to OECD criteria under the test conditions.

Description of key information

No studies on the "Biodegradation in water: ready biodegradability" are available for the target substance in itself. Nevertheless, studies for this endpoint have been conducted with structural analogue substances. All three structural analogues were classified as 'not readily biodegradable' under the conditions of the respective study. Additionally, a study was performed to assess the inherent biodegradability of Reactive Yellow F68072. The results of the study showed that the test substance is not inherently biodegradable.

Summarising, Reactive Yellow F68072 is neither considered inherently nor readily biodegradable.

Key value for chemical safety assessment

Biodegradation in water:

not biodegradable

Additional information

No studies on the "Biodegradation in water: ready biodegradability" are available for the target substance in itself. Nevertheless, studies for this endpoint have been conducted with structural analogue substances and showed that these substances are not readily biodegradable.

A study was performed to assess the inherent biodegradability of Reactive Yellow F68072. The study was conducted in accordance with the Draft updated OECD Guideline for Testing of Chemicals 302 B - Inherent Biodegradability. Study conducted in compliance with the Principles of Good Laboratory Practice (GLP). A solution of Reaktiv Gelb F68072 FW in a mineral medium was inoculated with activated sludge and incubated under aerobic conditions. Degradation was followed by DOC determinations at 0h, 3h, Id, 7 d, 14 d, 21 d, 27 d and 28 d.

Reactive Yellow F68072 showed:

0 % degradation after 1 day.

3 % degradation after 7 days.

2 % degradation after 14 days.

0 % degradation after 21 days.

4 % degradation after 27 days.

6 % degradation after 28 days.

Thus, the test substance is not inherently biodegradable.