Reaction mass of sodium amino-bis{[4-(ethenylsubstituted)phenyl]diazenyl}-hydroxynaphthalenesulfonate and polysodium amino-{[4-(ethenylsubstituted)phenyl]diazenyl}-{[4-(ethenylsubstituted)-2-sulfonatophenyl]diazenyl}-hydroxynaphthalenesulfonate and polysodium amino-bis{[4-(ethenylsubstituted)-2-sulfonatophenyl]diazenyl}-hydroxynaphthalenesulfonate

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:

2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

Guidelines followed

GLP compliance:

yes

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
None

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

Activated sludge from a biologic sewage treatment plant was used. The chosen plant treats mostly domestic sewage.
The sludge was taken from the activation basin of the ESN (Stadtentsorgung Neustadt) sewage treatment plant, Im Altenschemel, NW-Lachen-Speyerdorf.
Date of collection: 24. Jul. 2014, batch no: 20140724.
The sludge was filtrated through a clean cotton cloth, then washed with tap water twice, with test medium once and re-suspended in test medium. It was then aerated until use in the test. Determination of dry matter gave 4080 mg suspended solids/litre.

Duration of test (contact time):

ca. 28 d

Initial conc.:

ca. 103.8 mg/L

Based on:

other: Test item

Initial conc.:

ca. 40.8 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

The following instruments and devices were used in the performance of the study:
- Data logger for temperature, ebro
- Analytical scales Mettler Toledo XS DU 205
- Precision scales Mettler Toledo XS 6001S
- Adjustable pipettes with one-way tips Rainin®;
- Carbon analyser TOC multi N/C 2100S, Analytik Jena
- Glass measuring flasks
- pH-meter 3310 wtw
- Heating chamber Heraeus
- Orbital shaker GFL 3017
- Ultrasonic bath Sonorex RK 510H Bandelin
- Fridge

Reference substance:

aniline

Preliminary study:

None

Test performance:

None

Parameter:

% degradation (test mat. analysis)

Value:

51

Sampling time:

14 d

Remarks on result:

other: The total biodegradation in the toxicity control after 14 days was 51 % (criterion: > 35 %).

Details on results:

None

Results with reference substance:

None

Findings and results

DOC-Values

DOC Values in mg/L before addition of the test medium to an end volume of 250 mL

Day	Procedure Control 1	Procedure Control 2	Positive Control 1	Positive Control 2	Test Item
					Replicate 1	Replicate 2	Abiotic Control	Adsorption Control	Toxicity Control
0	0.79	1.16	67.48	77.03	37.1	40.1	47.66	48.51	103.12
Volume in mL	100	100	119	119	125	125	128	128	145
theoretical value	0.32	0.46	32.25	36.81	18.62	20.12	24.39	24.83	59.78
measured value	0.00	0.00	40.05	40.05	40.76	40.76	40.77	40.85	80.58

 

DOC Values in the test in mg/L

Day	Procedure Control 1	Procedure Control 2	PositiveControl 1	PositiveControl 2	Test Item
					Replicate 1	Replicate 2		Abiotic Control		Adsorption Control		Toxicity Control
0.125	1.86	1.45	42.07	42.68	22.47	24.10	26.93		27.72		59.52
4	0.90	0.73	1.35	1.62	24.67	24.13	30.19		29.79		24.17
11	1.27	1.11	1.20	1.70	24.15	24.54	29.57		30.75		29.16
18	0.22	0.11	1.55	0.67	25.95	27.93	29.81		28.65		28.97
28	1.66	1.22	2.56	1.24	27.22	30.05	32.74		34.80		31.39

The calculated values for the percentage degradation are presented in the following table (Degradation values in %):

Day	Positive Control 1	Positive Control 2	Test item
			Replicate 1	Replicate 2	Abiotic Control	Adsorption Control	Toxicity Control
4	98.7	98.0	-14.6	-3.9	-12.1	-11.2	59.6
11	100.0	98.8	-10.3	-4.0	-9.8	-13.4	51.7
18	96.6	98.8	-23.9	-23.7	-10.7	-9.3	50.2
28	97.2	100.5	-23.9	-27.5	-21.6	-28.0	48.2

 

The measured DOC concentrations at the end of the test in the test replicates and in the abiotic control were slightly higher than at the beginning. This might be caused by dissolution of undissolved test item during the test. Therefore, negative degradation values were calculated.

Validity

- The difference within replicates at the end of the test was 3.6 % (criterion: <= 20%).

- The positive control reached the pass level of 98 % on day 4 (criterion: > 70% within 14 days).

- The total biodegradation in the toxicity control after 14 days was 51 % (criterion: > 35 %).

  All the criteria were met to validate this study according to test guideline.

 

Validity criteria fulfilled:

yes

Interpretation of results:

other: not readily biodegradable

Conclusions:

The following data were determined for the test item FAT 40865/A TE:
10-day-window: not detected
Degradation at the end of 10-day-window: not detected
Degradation at the end of the test: 0 %

Therefore, the test item FAT 40865/A TE is not readily biodegradable following OECD test guideline No. 301A/EU method C.4-A.
The test item can be stated as “not toxic to the inoculum at a concentration of 103.6 mg/L.”

Executive summary:

The test item FAT 40865/A was investigated for its ready biodegradability over 28 days following DOC die-away test according to OECD Test Guideline for the Testing of Chemicals No. 301A and EU Method No. C.4-A.

Activated sludge was used as inoculum at a concentration of 25 mg dry matter/L.

Aniline was used as positive control and at the end of 4 days, 98 % degradation was recorded.

In the toxicity control, the biodegradation amounted to 51 % within 14 days. Therefore, according to the test guideline, the test item can be stated as “not toxic to the inoculum at a concentration of 103.6 mg/L.”

Reference 2

Endpoint:

biodegradation in water: inherent biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Guideline:

OECD Guideline 302 B (Inherent biodegradability: Zahn-Wellens/EMPA Test)

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

The composition of the medium is taken from OECD Guideline. As in EU-Method C.9 a different composition is given, this is stated as a deviation from the EU Method.

Principles of method if other than guideline:

None

GLP compliance:

yes

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
None

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

Activated sludge from a biologic sewage treatment plant was used. The chosen plant is treating mostly household sewage. The sludge was taken from the activation basin of the ESN (Stadtentsorgung Neustadt) sewage treatment plant, Im Altenschemel, NW-Lachen-Speyerdorf.
Date of collection: 07. Aug. 2014
The sludge was washed with tap water twice, then filtrated through a cloth, resuspended in test medium and then aerated.
The dry matter was determined as 4100 mg suspended solids/L.

Duration of test (contact time):

ca. 28 d

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

None

Reference substance:

aniline

Preliminary study:

None

Test performance:

None

Parameter:

% degradation (DOC removal)

Value:

8

Sampling time:

28 d

Details on results:

None

Results with reference substance:

None

DOC Values

The measured/recalculated DOC-values in mg/L at the beginning (before addition of the inoculum):

Day	Control		Positive Control		Test Item		Abiotic Control	Toxicity Control
	1	2	1	2	1	2
Measured Value	1.21	0.87	286.35	283.51	226.18	222.65	221.61	503.54
Calculated Start Conc.	0.81	0.58	190.77	188.88	150.79	148.43	147.79	335.81
Theoretical Start Conc.	0.00	0.00	200.00	200.00	200.27	200.27	200.13	399.73

 

Calculated start concentrations are based on DOC measurement of the test solutions with a volume of 1 L (before adding inoculum and filling up to the final volume of 1500 mL). Theoretical start concentrations are based on the organic carbon content of test item resp. positive control, based on TOC measurement of the respective stock solutions.

 

The measured DOC values (mg/L) in the course of the test are presented in the following table:

Day	Control		Positive control		Test Item		Abiotic	Toxicity
	1	2	1	2	1	2
0.125	2.04	1.10	194.19	196.69	171.45	166.95	167.21	360.27
5	4.76	4.74	6.88	6.31	172.70	185.98	182.90	170.60
12	4.46	4.87	4.55	4.03	183.34	172.94	188.27	187.27
19	6.24	5.93	7.00	5.84	137.14	140.80	187.39	180.26
27	5.84	3.59	4.25	5.91	147.93	132.69	192.38	118.70
28	4.32	3.66	5.75	3.72	161.67	155.20	186.63	152.32
29*	5.19	4.14	5.84	6.74	121.05	130.07	148.89	108.25

*repeated measurement on day 29

Calculation of biodegradation was based only on the content of dissolved organic carbon. Because of degradation in conjunction with precipitation and dissolution of test item during the running study, scattering DOC values and scattering degradation values were likely to occur.

These effects are confirmed by the measured DOC concentrations in the test replicates and the toxicity control on day 28. The DOC concentrations were much higher than on day 27. For verification, an additional measurement on day 29 was performed. The measured values on Day 29 were much lower than the previously measured values. The measured DOC values in the abiotic control were also significantly lower on day 29 compared to day 28.

 pH Values

pH was measured at each sampling time or once per week and, if necessary, adjusted to pH 7.0 ± 0.5. The values are presented in the following table:

Day	Control		Positive Control		Test Item		Abiotic Control	Toxicity Control
	1	2	1	2	1	2
0	7.6 to 7.3	7.6 to 7.4	7.7 to 7.3	7.7 to 7.2	7.6 to 7.4	7.6 to 7.3	7.1	7.6 to 7.2
5	6.9	6.9	7.8 to 7.0	7.8 to 7.1	7.2	7.2	7.2	7.8 to 7.1
12	6.7	6.8	7.1	7.1	7.2	7.2	7.1	7.3
19	6.6	6.7	5.1 to7.2	5.2 to 6.9	7.2	7.2	7.1	6.9
27	6.4 to 7.3	6.6	6.3 to 6.6	6.0 to 6.8	7.1	7.1	7.2	6.0 to 6.7

 

Validity criteria fulfilled:

yes

Interpretation of results:

not inherently biodegradable

Conclusions:

The test item can be considered as “not inherently biodegradable“.
The degree of biodegradation reached 8 % after 28 days.

Executive summary:

The Inherent biodegradability of FAT 40865/A was determined in a 28 day Zahn-Wellens test following OECD Test Guideline for the Testing of Chemicals No. 302B and EU Method No. C.9 at a nominal concentration of 200 mg organic carbon/L in test medium Aniline was used as positive control, and at the end of 5 days, 99 % degradation was recorded. Activated sludge from a domestic sewage treatment plant was used as inoculum (concentration 601 mg dry matter/L). To measure the degradation of test item and positive control, the content of dissolved organic carbon (DOC) in the test vessels was measured seven times during the test. All validity criteria were met. The per cent biodegradation of the test item was calculated based on dissolved organic carbon (DOC).

The following data could be determined for the test item FAT 40865/A TE:

Degradation at the end of the test: 8%

Lag-phase: Day 0 - 16

Degradation phase: Not determinable

Therefore, FAT 40865/A TE is stated as not inherently biodegradable.

Description of key information

FAT 40865/A TE is stated as not inherently biodegradable.

Additional information

The test item FAT 40865/A was investigated for its ready biodegradability over 28 days following DOC die-away test according to OECD Test Guideline for the Testing of Chemicals No. 301A and EU Method No. C.4-A. Activated sludge was used as inoculum at a concentration of 25 mg dry matter/L. Aniline was used as positive control and at the end of 4 days, 98 % degradation was recorded. In the toxicity control, the biodegradation amounted to 51 % within 14 days. Therefore, according to the test guideline, the test item can be stated as “not toxic to the inoculum at a concentration of 103.6 mg/L.

In another study, the Inherent biodegradability of FAT 40865/A was determined in a 28 day Zahn-Wellens test following OECD Test Guideline for the Testing of Chemicals No. 302B and EU Method No. C.9 at a nominal concentration of 200 mg organic carbon/L in test medium Aniline was used as positive control, and at the end of 5 days, 99 % degradation was recorded. Activated sludge from a domestic sewage treatment plant was used as inoculum (concentration 601 mg dry matter/L). To measure the degradation of test item and positive control, the content of dissolved organic carbon (DOC) in the test vessels was measured seven times during the test. All validity criteria were met. The percent biodegradation of the test item was calculated based on dissolved organic carbon (DOC).

The following data could be determined for the test item FAT 40865/A TE:

Degradation at the end of the test: 8%

Lag-phase: Day 0 - 16

Degradation phase: Not determinable

Therefore, FAT 40865/A TE is stated as not inherently biodegradable.