Reactive Orange 143

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:

15 March 2012 to 24 May 2012

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:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Name: FAT 40854/A TE
Batch: No.: TZ 5719 / BOP 02-11
Certificate of Analysis Date: November 10, 2011
Purity: Total Inorganic Impurities: 39.49 %
Total Organic Constituents: 60.51 %
Chemical Oxygen Demand: 0.83 mg oxygen per mg test item
Physical Appearance / Colour: Solid (Powder) / reddish-brown (according to the sponsor)
Expiry Date: April 01, 2016
Storage: At room temperature (20 ± 5 °C), under dark and dry conditions

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic (adaptation not specified)

Details on inoculum:

Aerobic activated sludge, microorganisms from a domestic waste water treatment plant was supplied by the sewage plant Darmstadt, Germany.

Duration of test (contact time):

28 d

Initial conc.:

32 g/L

Parameter followed for biodegradation estimation:

O2 consumption

Reference substance:

other: sodium benzoate

Parameter:

% degradation (O2 consumption)

Sampling time:

28 d

Remarks on result:

other: The mean biodegradation after 28 days of FAT 40854/A TE was -12% (ThODNH4) and (ThODNO3)

Results with reference substance:

The reference item sodium benzoate was sufficiently degraded to 80 % after 14 days and to 83 % after 28 days of incubation, thus confirming the suitability of the aerobic activated sludge inoculum used.

Biodegradation of Test Item

Percentage Biodegradation: The occurrence of nitrification was considered but not experimentally confirmed. The mean percentage biodegradation at the end of the 28-day exposure period was -12 % (ThODNH4) and (ThODNO3). The 10-day window was not passed. The degradation rate of FAT 40854/A did not reach 60 % within the 10-day window and after 28 days of incubation. According to the test guideline, FAT 40854/A is considered not to be readily biodegradable.

 

Biodegradation of Reference Item Sodium Benzoate

Percentage Biodegradation: The reference item sodium benzoate was sufficiently degraded to 80 % after 14 days and to 83 % after 28 days of incubation. The percentage biodegradation of the reference item confirms the suitability of the used aerobic activated sludge inoculum.

 

Biodegradation in the Toxicity Control

Percentage Biodegradation: In the toxicity control containing both the test item and reference 56 % biodegradation was noted within 14 days and 56 % biodegradation was determined after 28 days of incubation (ThODNH4) and (ThODNO3).

Conclusion: According to the test guidelines the test item can be assumed to be not inhibitory on the aerobic activated sludge microorganisms because degradation was >25 % within 14 days.

 

Abiotic Control

Oxygen Demand: The oxygen demand in the abiotic control was zero. No correction of the test item degradation rates had to be done.

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

FAT 40854/A is considered not to be readily biodegradable.

Executive summary:

In GLP-compliant study, ready biodegradability of FAT 40854/A in a Manometric Respirometry Test according to OECD 301 F and EU Method C.4-D. Test Species used was aerobic activated sludge (micro-organisms from a domestic wastewater treatment plant) was supplied by the sewage works of Darmstadt, Germany. The test item FAT 40854/A was investigated for its ready biodegradability in a manometric respirometry test over a period of 28 days. The biodegradation was followed by the oxygen uptake of the micro-organisms during exposure. As a reference item sodium benzoate was tested simultaneously under the same conditions as the test item and functioned as a procedure control. Degradation rate of test item calculated by the oxygen consumption of the aerobic activated sludge microorganisms after 28 days of incubation. Test item loading rate was 103 mg/L corresponding to an oxygen demand of about 85 mg/L (ThODNH4) and 85 mg/L (ThODNO3). The mean biodegradation after 28 days of FAT 40854/A was -12 % (ThODNH4) and (ThODNO3); the 10 day window criterion was not passed. The reference item sodium benzoate was sufficiently degraded to 80 % after 14 days and to 83 % after 28 days of incubation, thus confirming the suitability of the aerobic activated sludge inoculum used. Biodegradation of the toxicity control containing both, the test item and the reference item sodium benzoate, 56 % biodegradation was noted within 14 days and 56 % biodegradation after 28 days of incubation (ThODNH4) and (ThODNO3). Thus, the test item can be assumed to be not inhibitory to the aerobic activated sludge microorganisms. Based on the test results, FAT 40854/A is considered not to be readily biodegradable.

Reference 2

Endpoint:

biodegradation in water: inherent biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

31 January 2012 to 28 February 2012

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)

Version / remarks:

OECD 302B

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: ISO 9888

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identification FAT 40854/A TE
Description: Reddish-brown powder (determined at NOTOX)
Batch TZ 5719 / BOP 02-11
Content: 46.2 % (4 main constituents)
Test substance storage: At room temperature in the dark
Stability under storage conditions: Stable
Expiry date: 01 April 2016

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

The sludge was coarsely sieved and washed with mineral medium. The batch of sludge was used within 6 hours of sampling and aerated until required. A small amount of the sludge was weighed and dried at ca. 105 °C. From this result the amount of suspended solids (SS) in the final test medium was calculated (3.0 g/l).

Duration of test (contact time):

28 d

Initial conc.:

241.2 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

TEST CONDITIONS
- Composition of medium: 1 litre mineral medium contains: 10 ml of solution (A), 1 ml of solutions (B) to (D) and Milli-RO water
Stock solutions of mineral components
A) 8.50 g KH2PO4; 21.75 g K2HPO4; 67.20 g Na2HPO4.12H2O; 0.50 g NH4Cl; dissolved in Milli-Q water and made up to 1 litre, pH 7.4 ± 0.2
B) 22.50 g MgSO4.7H2O dissolved in Milli-Q water and made up to 1 litre.
C) 36.40 g CaCl2.2H2O dissolved in Milli-Q water and made up to 1 litre.
D) 0.25 g FeCl3.6H2O dissolved in Milli-Q water and made up to 1 litre.

- Test temperature: between 20 and 23 °C.

- pH: of the different test solutions during the test period ranged between 6.5 and 7.7, except for the reference substance on day 7, 17 and 21. On day 7 the pH was 8.2 and adjusted to 7.7 and on day 17 and 21 the pH was 6.2 and adjusted to 7.0 (using 1 N H2SO4).

- Oxygen concentrations: between 7.8 and 9.1 mg/l.

- Aeration of dilution water: Not before the test, the test is aerated continuously.

- Final concentration suspended solids in the test mixtures: 0.75 g SS/L.

- The ratio of inoculum and FAT 40854/A or reference substance (as DOC) was 3:1.

- Continuous darkness: yes

Reference substance:

aniline

Preliminary study:

In a pre-test, to determine the concentration to be tested in the main study, a DOC concentration was measured in a solution containing 0.5008 g FAT 40854/A per litre. Hence, the DOC concentration was 131.7 mg/L.

Test performance:

Preparation of test vessels

1) Test substance mixture (1 vessel)
- 500 ml mineral medium
- 500 ml activated sludge (final concentration in test mixture: 0.75 g SS/L)
- 380 ml stock solution of FAT 40854/A. The stock solution was prepared by dissolving 2.4276 g FAT 40854/A in 500 mL mineral medium.
- Made up to 2 litres with mineral medium.
- The DOC of the test solution of 1.2138 g FAT 40854/A was 241.2 mg/L (t=0h).
- The expected ratio of inoculum and FAT 40854/A (as DOC) was 3:1.

2) Blank control mixture (1 vessel)
- 500 mL mineral medium
- 500 mL activated sludge (final concentration in test mixture: 0.75 g SS/L)
- Made up to 2 litres with mineral medium.

3) Reference substance mixture (1 vessel)
- 500 mL mineral medium
- 500 mL activated sludge (final concentration in test mixture: 0.75 g SS/L)
- 712.6 mg aniline.
- Made up to 2 litres with mineral medium.
- The DOC of this solution of 356.3 mg/L was 273.7 mg/L (t=0h).
- The ratio of inoculum and aniline (as DOC) was 3:1.

Sampling and DOC analysis:
Prior to sampling the volume of the vessels was checked for evaporation and if necessary Milli-Q water was added. The medium was mixed thoroughly and a sample of ca. 100 mL was taken. This sample was passed through a rough paper filter (S&S 604), and thereafter through a 0.45 µm filter (S&S FP 030/0.45). At least, the first 5 mL was discarded. Duplicate samples of ca. 40 mL were then taken from the filtrate. All samples not analysed freshly were stored in a deep-freeze until analysis.

Frequency of sampling:
- At the start of the test (0 h) and 3h after addition of the test substance in order to estimate any adsorption of FAT 40854/A by the activated sludge;
- Day 1, 3, 7, 14, 21, 27 and 28.

Sample pretreatment:
- Day 1, 3, 7, 14, 21, 27 and 28: Not applicable, samples were analysed on the day of sampling.
- Day 14, reserve samples: Samples were stored in a freezer until analysis. Frozen samples were defrosted at room temperature.

Parameter:

% degradation (DOC removal)

Value:

12

Sampling time:

28 d

Details on results:

The results obtained during the test period showed no significant DOC removal in the first 3 hours of incubation in either of the test vessels indicating no or only limited adsorption to sludge during this period. The biodegradation values calculated from the DOC measurements performed during the 28-day test period revealed a maximum of 12 % biodegradation of FAT 40854/A. Thus, the biodegradation values did not reveal more than 20 % biodegradation of FAT 40854/A during the test period.

Results with reference substance:

The removal of aniline was at least 70 % within 14 days (98 %).

Validity criteria fulfilled:

yes

Interpretation of results:

not inherently biodegradable

Conclusions:

FAT 40854/A was not inherently biodegradable under the conditions of the 'Zahn-Wellens / EMPA Test' presently performed.

Executive summary:

In a GLP-compliant study, the inherent biodegradability of FAT 40854/A was investigated in a 'Zahn-Wellens / EMPA Test' according to EU method C.9 and OECD 302B. At the start of the study the Dissolved Organic Carbon (DOC) concentration of the test solution containing FAT 40854/A (1.2138 g/l) was 241.2 mg/l and the DOC concentration of the reference solution containing Aniline (356.3 mg/l) was 273.7 mg/l. The ratio of inoculum and FAT 40854/A /aniline (as DOC) was 3:1.

The results obtained during the test period showed no significant DOC removal in the first 3 hours of incubation in any of the test vessels indicating no or only limited adsorption to sludge during this period. The biodegradation values calculated from the DOC measurements performed during the 28-day test period revealed a maximum of 12 % biodegradation of FAT 40854/A. Thus, the biodegradation values did not reveal more than 20 % biodegradation of FAT 40854/A during the test period. Since all acceptability criteria prescribed by the protocol were met, this study was considered to be valid. In conclusion, FAT 40854/A was not inherently biodegradable.

Description of key information

The substance is neither inherently biodegradable nor ready biodegradable.

Key value for chemical safety assessment

Biodegradation in water:

not biodegradable

Type of water:

freshwater

Additional information

In aGLP-compliant study, ready biodegradability of FAT 40854/A was investigated in a Manometric Respirometry Test according to OECD 301 F and EU Method C.4-D using aerobic activated sludge over a period of 28 days. The mean biodegradation after 28 days of FAT 40854/A was -12 % (ThODNH4) and (ThODNO3); the 10-day window criterion was not passed. The reference item sodium benzoate was sufficiently degraded to 80 % after 14 days and to 83 % after 28 days of incubation, thus confirming the suitability of the aerobic activated sludge inoculum used. Biodegradation of the toxicity control containing both, the test item and the reference item sodium benzoate, 56 % biodegradation was noted within 14 days and 56 % biodegradation after 28 days of incubation (ThODNH4) and (ThODNO3). Thus, the test item can be assumed to be not inhibitory to the aerobic activated sludge microorganisms. Based on the test results, FAT 40854/A is considered not to be readily biodegradable. In another study, the inherent biodegradability of FAT 40854/A was investigated in a 'Zahn-Wellens / EMPA Test' according to EU method C.9, OECD 302B. The results obtained during the test period showed no significant DOC removal in the first 3 hours of incubation in any of the test vessels indicating no or only limited adsorption to sludge during this period. The biodegradation values calculated from the DOC measurements performed during the 28-day test period revealed a maximum of 12 % biodegradation of FAT 40854/A . Thus, the biodegradation values did not reveal more than 20 % biodegradation of FAT 40854/A during the test period. In conclusion, FAT 40854/A was not inherently biodegradable.