Reaction products of tetrazotized 5-amino-2-[(E)-2-(4-amino-2-sulfophenyl)ethenyl]benzenesulfonic acid with 6-amino-4-hydroxynaphthalene-2-sulfonic acid and 4-amino-5-hydroxynaphthalene-2,7-disulfonic acid in 2-[bis(2-hydroxyethyl)amino]ethanol

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The read-across approach should be used for data gap filling of Bayscript Blaukomponente TEA using environmental fate, ecotoxicity and human health toxicity studies of Bayscript Blaukomponente MDA. This approach is justified by structural similarity, i.e. a nearly identical composition, between source and target substance. Since the same method of manufacturing is used for Bayscript Blaukomponente MDA and Bayscript Blaukomponente TEA the composition of these UVCB substances is nearly identical. Bayscript Blaukomponente MDA (source) and Bayscript Blaukomponente TEA (target) exhibit three main organic salt constituents. Structures are mainly differing in the functional groups of the cationic amines MDA and TEA used as counterions.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Bayscript Blaukomponente TEA is a multi-constituent substance, consisting of three main constituents. The cationic counterion of Bayscript Blaukomponente TEA is 2,2'2" nitrilotris[ethanol] (TEA) (CAS no. 105-59-9).
Bayscript Blaukomponente TEA is manufactured in water and is handled and used as an aqueous solution during its lifetime. It is manufactured as a 34 % solution and in the further handling diluted by mixing with other aqueous colourant solutions to give aquatic inks. The solid substance has only been isolated for REACH registration purposes.

Typical concentrations depend on each registrant’s company specific legal entity composition. No impurity relevant for classification and labelling was identified for the composition.

Bayscript Blaukomponente MDA is a multi-constituent substance, consisting of the same three main constituents as Bayscript Blaukomponente TEA. The counterion of Bayscript Blaukomponente MDA is 2,2'-(methylimino)diethanol (MDA) (CAS no. 102-71-6).
Bayscript Blaukomponente MDA is manufactured in water and is handled and used as an aqueous solution during its lifetime. It is manufactured as a 41 % solution and in the further handling diluted by mixing with other aqueous colourant solutions to give aquatic inks. The aqueous solution has been registered under REACH since it was not possible at the time of registration to extract the solid from the aqueous solution without decomposition.

The concentration ranges for each of the three constituents calculated for the water-free source substance are shown in chapter AE A.2. Typical concentrations depend on each registrant’s company specific legal entity composition. No impurity relevant for classification and labelling was identified for the composition.


3. ANALOGUE APPROACH JUSTIFICATION
The UVCB substances Bayscript Blaukomponente MDA (source) and Bayscript Blaukomponente TEA (target) mainly consist each of three identical organic constituents. The structural formula of these constituents is an estimation as acid form due to the analytical results, other isomers with same molecular weight are possible. At the end of the production process the acid form is neutralized in aqueous solution to an ammonium salt with 2,2'-(methylimino)diethanol (for Bayscript Blaukomponente MDA) and 2,2'2" nitrilotris[ethanol] (for Bayscript Blaukomponente TEA), respectively. The constituents of source and target mainly differ in 2,2'-(methylimino)diethanol (MDA) (CAS no. 102-71-6) and 2,2'2" nitrilotris[ethanol] (TEA) (CAS no. 105-59-9) used as cationic counterion.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

assessment report

Parameter:

% degradation (O2 consumption)

Value:

29

Sampling time:

7 d

Parameter:

% degradation (O2 consumption)

Value:

99

Sampling time:

14 d

Parameter:

% degradation (O2 consumption)

Value:

100

Sampling time:

21 d

Parameter:

% degradation (O2 consumption)

Value:

100

Sampling time:

28 d

Details on results:

Biodegradation started on day 6 with 20% degradation and reached 100% on day 15.

Parameter:

COD

Value:

0.728 mg O2/g test mat.

Results with reference substance:

The reference compound sodium benzoate showed 88 % degradation after 14 days.

Validity criteria fulfilled:

yes

Remarks:

Ready biodegradability of reference compound ≥ 60 percent within 14 days. -In the toxicity control degradation rates > 25 % within 14 days. -Replicates difference< 20% . - Oxygen uptake of blank inoculum=< 60mg/l. -No pH influence

Interpretation of results:

readily biodegradable

Conclusions:

A test on ready biodegradability was conducted according to OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test). After 28 days showed 100 % degradation fulfilling the 10 day window.

Executive summary:

This study was designed to assess the ready biodegradability of BAYSCRIPT Blaukomponente (non-preserved, filtrated). It was conducted in accordance with the Council Regulation (EC) No 440/2008, Method C.4-D “Manometric Respirometry Test“(2008). This test method is in all essential parts identical with OECD Guideline 301 F (adopted July 1992). A suspension of 100 mg/L active ingredient of the test test item (= 283.7 mg/L test item) in a mineral medium, equalling to 50-100 mg ThOD or COD/Litre as the nominal sole source of organic carbon, was stirred in a closed flask and inoculated at a constant temperature (22 ± 1 °C) for up to 28 days under aerobic conditions in the dark. The consumption of oxygen (BOD) was determined by measuring the drop in pressure in the automated respirometer flasks. Evolved carbon dioxide was absorbed in sodium hydroxide. The amount of oxygen taken up by the test item (corrected for uptake by blank inoculum, run in parallel) was expressed as a percentage of chemical oxygen demand (COD). The endogenous activity of the inoculum was checked running parallel blanks with inoculum but without test item. A reference compound (sodium benzoate) was run in parallel to check the operation of the procedures. A toxicity control (test item and reference compound mixed) was not run in parallel, because the chosen concentration of the test item was not inhibitory to microorganisms. Degradation was followed by the determination of oxygen uptake and measurements were taken at frequent intervals to allow the identification of the beginning and end of biodegradation and the slope of the biodegradation curve. The test lasted for 28 days. Because of the nature of biodegradation and of the mixed bacterial populations used as inoculum, determinations of test item and inoculum blank were carried out in triplicate and of reference compound in duplicate. The oxygen uptake was calculated from the readings taken at regular and frequent intervals, using the method given by the manufacturer of the equipment. At the end of incubation, the pH was measured in the flasks. The degradation rate of the test item was set in relation to its experimentally determined COD. As the COD implicitly covers the oxygen demand for the nitrification process, there was no need to take additional correction measures into account. The 28-day degradation was 100 % and for the positive control (with reference substance) the biodegrdation was 88 %. Therefore, the substance is considered to be “Readily Biodegradable, fulfilling the 10 day window“.

Reference 2

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

- Type: mixed population of aquatic microorganisms (activated sludge)
- Origin: aeration tank of a wastewater plant treating predominantly domestic sewage (Wupper area water authority, WWTP Odenthal)
- Date of collection: 2012-08-22
- Concentration of inoculum: 30 mg/L suspended solids
Pre-treatment of the inoculum:
- The sludge was washed twice by adding mineral medium and centrifuging for 10 min at 2000 rpm and 20 °C and decanting off the supernatant.
- An aliquot of the wet sludge was dried in order to determine the wet weight / dry weight ratio of the sludge and to prepare a stock suspension (activated sludge) of 3 g dw/L.
- The calculated amount of sludge, needed to achieve 300 mL of this stock suspension, was dissolved in mineral medium and then filled up to a defined end volume.
- Before use, the inoculum was stored for one day at room temperature under continuous shaking with aeration.

Duration of test (contact time):

28 d

Initial conc.:

100 mg/L

Based on:

test mat.

Details on study design:

A suspension of 100 mg/L active ingredient of the test test item (= 283.7 mg/L test item) in a mineral medium, equalling to 50-100 mg ThOD or COD/Litre as the nominal sole source of organic carbon, was stirred in a closed flask and inoculated at a constant temperature (22 ± 1 °C) for up to 28 days under aerobic conditions in the dark.
The consumption of oxygen (BOD) was determined by measuring the drop in pressure in the automated respirometer flasks. Evolved carbon dioxide was absorbed in sodium hydroxide. The amount of oxygen taken up by the test item (corrected for uptake by blank inoculum, run in parallel) was expressed as a percentage of chemical oxygen demand (COD).
The endogenous activity of the inoculum was checked running parallel blanks with inoculum but without test item. A reference compound (sodium benzoate) was run in parallel to check the operation of the procedures.
A toxicity control (test item and reference compound mixed) was not run in parallel, because the chosen concentration of the test item was not inhibitory to microorganisms.
Degradation was followed by the determination of oxygen uptake and measurements were taken at frequent intervals to allow the identification of the beginning and end of biodegradation and the slope of the biodegradation curve.
The degradation rate of the test item was set in relation to its experimentally determined COD. As the COD implicitly covers the oxygen demand for the nitrification process, there was no need to take additional correction measures into account.
Exposure conditions:
Test volume : 250 mL
Test apparatus : OxiTopControl System (WTW)
Mixing : 1 magnetic stirrer per test vessel
Incubation time : 28 days
Incubation temperature : 22 ± 1 °C

Reference substance:

benzoic acid, sodium salt

Remarks:

Purity : 99.7 %

Parameter:

% degradation (O2 consumption)

Value:

29

Sampling time:

7 d

Parameter:

% degradation (O2 consumption)

Value:

99

Sampling time:

14 d

Parameter:

% degradation (O2 consumption)

Value:

100

Sampling time:

21 d

Parameter:

% degradation (O2 consumption)

Value:

100

Sampling time:

28 d

Details on results:

Biodegradation started on day 6 with 20% degradation and reached 100% on day 15.

Parameter:

COD

Value:

0.728 mg O2/g test mat.

Results with reference substance:

The reference compound sodium benzoate showed 88 % degradation after 14 days.

Validity criteria fulfilled:

yes

Remarks:

Ready biodegradability of reference compound ≥ 60 percent within 14 days. -In the toxicity control degradation rates > 25 % within 14 days. -Replicates difference< 20% . - Oxygen uptake of blank inoculum=< 60mg/l. -No pH influence

Interpretation of results:

readily biodegradable

Conclusions:

A test on ready biodegradability was conducted according to OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test). After 28 days showed 100 % degradation fulfilling the 10 day window.

Executive summary:

This study was designed to assess the ready biodegradability of BAYSCRIPT Blaukomponente (non-preserved, filtrated). It was conducted in accordance with the Council Regulation (EC) No 440/2008, Method C.4-D “Manometric Respirometry Test“(2008). This test method is in all essential parts identical with OECD Guideline 301 F (adopted July 1992). A suspension of 100 mg/L active ingredient of the test test item (= 283.7 mg/L test item) in a mineral medium, equalling to 50-100 mg ThOD or COD/Litre as the nominal sole source of organic carbon, was stirred in a closed flask and inoculated at a constant temperature (22 ± 1 °C) for up to 28 days under aerobic conditions in the dark. The consumption of oxygen (BOD) was determined by measuring the drop in pressure in the automated respirometer flasks. Evolved carbon dioxide was absorbed in sodium hydroxide. The amount of oxygen taken up by the test item (corrected for uptake by blank inoculum, run in parallel) was expressed as a percentage of chemical oxygen demand (COD). The endogenous activity of the inoculum was checked running parallel blanks with inoculum but without test item. A reference compound (sodium benzoate) was run in parallel to check the operation of the procedures. A toxicity control (test item and reference compound mixed) was not run in parallel, because the chosen concentration of the test item was not inhibitory to microorganisms. Degradation was followed by the determination of oxygen uptake and measurements were taken at frequent intervals to allow the identification of the beginning and end of biodegradation and the slope of the biodegradation curve. The test lasted for 28 days. Because of the nature of biodegradation and of the mixed bacterial populations used as inoculum, determinations of test item and inoculum blank were carried out in triplicate and of reference compound in duplicate. The oxygen uptake was calculated from the readings taken at regular and frequent intervals, using the method given by the manufacturer of the equipment. At the end of incubation, the pH was measured in the flasks. The degradation rate of the test item was set in relation to its experimentally determined COD. As the COD implicitly covers the oxygen demand for the nitrification process, there was no need to take additional correction measures into account. The 28-day degradation was 100 % and for the positive control (with reference substance) the biodegrdation was 88 %. Therefore, the substance is considered to be “Readily Biodegradable, fulfilling the 10 day window“.

Description of key information

This study was designed to assess the ready biodegradability of BAYSCRIPT Blaukomponente (non-preserved, filtrated). It was conducted in accordance with the Council Regulation (EC) No 440/2008, Method C.4-D “Manometric Respirometry Test“(2008). This test method is in all essential parts identical with OECD Guideline 301 F (adopted July 1992). A suspension of 100 mg/L active ingredient of the test test item (= 283.7 mg/L test item) in a mineral medium, equalling to 50-100 mg ThOD or COD/Litre as the nominal sole source of organic carbon, was stirred in a closed flask and inoculated at a constant temperature (22 ± 1 °C) for up to 28 days under aerobic conditions in the dark. The consumption of oxygen (BOD) was determined by measuring the drop in pressure in the automated respirometer flasks. Evolved carbon dioxide was absorbed in sodium hydroxide. The amount of oxygen taken up by the test item (corrected for uptake by blank inoculum, run in parallel) was expressed as a percentage of chemical oxygen demand (COD). The endogenous activity of the inoculum was checked running parallel blanks with inoculum but without test item. A reference compound (sodium benzoate) was run in parallel to check the operation of the procedures. A toxicity control (test item and reference compound mixed) was not run in parallel, because the chosen concentration of the test item was not inhibitory to microorganisms. Degradation was followed by the determination of oxygen uptake and measurements were taken at frequent intervals to allow the identification of the beginning and end of biodegradation and the slope of the biodegradation curve. The test lasted for 28 days. Because of the nature of biodegradation and of the mixed bacterial populations used as inoculum, determinations of test item and inoculum blank were carried out in triplicate and of reference compound in duplicate. The oxygen uptake was calculated from the readings taken at regular and frequent intervals, using the method given by the manufacturer of the equipment. At the end of incubation, the pH was measured in the flasks. The degradation rate of the test item was set in relation to its experimentally determined COD. As the COD implicitly covers the oxygen demand for the nitrification process, there was no need to take additional correction measures into account. The 28-day degradation was 100 % and for the positive control (with reference substance) the biodegrdation was 88 %. Therefore, the substance is considered to be “Readily Biodegradable, fulfilling the 10 day window“.

Key value for chemical safety assessment

Biodegradation in water:

readily biodegradable