6,6',6''-(1,3,5-triazine-2,4,6-triyltriimino)trihexanoic acid, compound with 2,2',2''-nitrilotriethanol (1:3)

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:

1982-11-25 to 1982-12-16

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Non GLP but guideline study. The percent biodegradability was determined based on DOC measurements of flask 1 (flask 2 cracked during test).

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 E (Ready biodegradability: Modified OECD Screening Test)

Deviations:

yes

Remarks:

Measurements of DOC removal were not performed in duplicate. One test flask cracked during the test.

GLP compliance:

no

Oxygen conditions:

aerobic

Inoculum or test system:

mixture of sewage, soil and natural water

Details on inoculum:

- Source of inoculum/activated sludge: Mixture of polyvalent bacteria
a) Secondary effluent from the sewage treatment plant of Rheinfelden, Switzerland.
b) In the river Rhine (Basle city, Switzerland)
c) From a suspension of garden soil from Vouvry, Switzerland

Duration of test (contact time):

ca. 21 d

Initial conc.:

ca. 19.3 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

TEST CONDITIONS
- Composition of medium: according to specifications of the OECD method No. 301 E
- Test temperature: 23±2 °C
- pH: no data
- pH adjusted: no data
- Continuous darkness: no, indirect daylight

TEST SYSTEM
- Culturing apparatus: 2000 mL erlenmeyer flasks
- Number of culture flasks: 2
- DOC measurement: weekly

CONTROL AND BLANK SYSTEM
- Inoculum blank: Yes
- Positive control: Yes

Reference substance:

benzoic acid, sodium salt

Parameter:

% degradation (DOC removal)

Value:

ca. 76

Sampling time:

21 d

Details on results:

Biodegradation measured as mg DOC/L and corrected by the blank control was calculated as 76 % in 21 days.

Results with reference substance:

Biodegradation corrected by the blank control and measured as DOC (mg/L) was 90 % in 7 days.

Reference 2

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1983-11-23 to 1983-12-21

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Non GLP study. Deviations from the updated OECD-guideline (adopted 1992): Filter was 0.45 instead of 0.20 micrometer.

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 E (Ready biodegradability: Modified OECD Screening Test)

Deviations:

yes

Remarks:

Non GLP study. Deviations from the updated OECD-guideline (adopted 1992): Filter was 0.45 instead of 0.20 micrometer.

GLP compliance:

no

Oxygen conditions:

aerobic

Inoculum or test system:

mixture of sewage, soil and natural water

Details on inoculum:

- Source of inoculum/activated sludge:
Mixture of polyvalent bacteria collected on 23/11/1983
a) Secondary effluent from the sewage treatment plant of CH~4310 Rheinfelden
b) In the river Rhine" (Basle-city)
c) From a suspension of garden soil from CH-1896 Vouvry

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
- Composition of medium: in accordance to the guideline
- Test temperature: 23 +/- 2 °C
- Continuous darkness: indirect daylight

TEST SYSTEM
- Culturing apparatus: 2000ml erlenmeyer flasks
- Number of culture flasks/concentration: duplicates

CONTROL AND BLANK SYSTEM
- Inoculum blank: one blank control

Reference substance:

aniline

Parameter:

% degradation (DOC removal)

Value:

68

Sampling time:

28 d

Details on results:

Biodegradation measured as mg DOC/L and corrected by the blank control was calculated as 68 % in 28 days.

Results with reference substance:

Biodegradation corrected by the blank control and measured as DOC (mg/L) is 86 % in 14 days.

After 28 days 64 -73% removal of DOC was observed.

Days	mean DOC removal minus blanks
7	32
14	30
21	66
27	68
28	68

Reference 3

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Non-GLP study, only pre-test

Qualifier:

according to guideline

Guideline:

other: MITI guideline

GLP compliance:

no

Oxygen conditions:

aerobic

Inoculum or test system:

other: standard sludge/water

Duration of test (contact time):

28 d

Initial conc.:

100 mg/L

Based on:

test mat.

Initial conc.:

30 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

TOC removal

Parameter followed for biodegradation estimation:

test mat. analysis

Details on study design:

two experimental set-ups were chosen:
A 100mg/L test material and 30mg/L active sludge
B 30mg/L test material and 100mg/L active sludge

Test performance:

After pretreatlng cultured solution, HPLC analysis was carried out for organic solvent layer.

Parameter:

% degradation (TOC removal)

Value:

64 - 73

Sampling time:

28 d

Remarks on result:

other: 100 mg/L test mat

Parameter:

% degradation (test mat. analysis)

Value:

91 - 100

Sampling time:

28 d

Remarks on result:

other: 100 mg/L test mat

Parameter:

% degradation (TOC removal)

Value:

67

Sampling time:

28 d

Remarks on result:

other: 30mg/L test mat.

Parameter:

% degradation (test mat. analysis)

Value:

100

Sampling time:

28 d

Remarks on result:

other: 30 mg/L test mat

Details on results:

The test substance completely biodegrades within 28 days and generates multi-component-residue via degradation-intermediate.
Identification of the residues was not possible however, melamine formation could be excluded.

Reference 4

Endpoint:

biodegradation in water: screening tests

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Basic data given

Qualifier:

according to guideline

Guideline:

other: OECD 306 Biodegradability in seawater

GLP compliance:

no

Oxygen conditions:

aerobic

Inoculum or test system:

other: seawater

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): expected non-polluted location (90 m depth in the Trondheimsfjord, Norway)
- Storage conditions: stored in the dark in 10-l tanks at 20 ± 2 °C for degradation of inherent organic materials (aging).
- Storage length: 2 to 5 days
- Preparation of inoculum for exposure: aerated for 20 min, seawater fortified with nutrient solution enhancing bacterial growth
- Water filtered: yes
- Type and size of filter used, if any: 50 µm

Duration of test (contact time):

28 d

Initial conc.:

2 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

TEST CONDITIONS
- Composition of medium: according to guideline
- Additional substrate: no
- Solubilising agent (type and concentration if used): not applicable
- Test temperature: 20±2 °C
- pH: no data
- pH adjusted: no data
- CEC (meq/100 g): no data
- Aeration of dilution water: no data
- Suspended solids concentration: no data
- Continuous darkness: yes
- Other: test substance diluted in "normal" seawater, which hda been aged for 3 to 5 days, essential nutrients for the bacteria added to the seawater

TEST SYSTEM
- Culturing apparatus: bottles with stoppers (no air space)
- Number of culture flasks/concentration: 2
- Method used to create aerobic conditions: seawtaer aerated for 20 min prior to exposure
- Measuring equipment: "dissolved oxygen meter" (YSI Instruments, YSI Inc., Yellow Springs, OH, USA)
- Test performed in closed vessels due to significant volatility of test substance: not specified
- Test performed in open system: no

SAMPLING
- Sampling frequency: after 0, 5, 15 and 28 days

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control: no
- Toxicity control: no
- Other: reference (aniline)

Reference substance:

aniline

Parameter:

% degradation (O2 consumption)

Value:

19.6

Sampling time:

28 d

Interpretation of results:

other: The substance is poorly degradabale in seawater.

Reference 5

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study has been performed according to a precursor of the OECD Modified Screening Test

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 301 E (Ready biodegradability: Modified OECD Screening Test)

Version / remarks:

precursor of OECD TG 301 E

Principles of method if other than guideline:

Modified OECD Screening Test performed according to:
OECD (1976). Proposed Method for the Determination of the Biodegradability of Surfactants Used in Synthetic Detergents. Paris.
Umweltbundesamt (1978). OECD Chemicals Testing Programme.Expert Group C "Persistence" (Degradation/Accumulation) Draft Working Papers, March 31, 1978

GLP compliance:

no

Oxygen conditions:

aerobic

Inoculum or test system:

other: wwtp effluent (non-adapted)

Duration of test (contact time):

19 d

Initial conc.:

20 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

TEST CONDITIONS
- Composition of medium: medium with a trace metal solution and an essential vitamin solution; ammonium concentration tripled compared to OECD procedure for an optimal C:N:P ratio
- Additional substrate: no
- Solubilising agent (type and concentration if used): no
- Test temperature: ambient

Parameter:

% degradation (DOC removal)

Value:

96

Sampling time:

19 d

Interpretation of results:

readily biodegradable

Reference 6

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study well documented, meets generally accepted scientific principles, acceptable for assessment. Key study because biodegradation under environmental conditions tested.

Qualifier:

no guideline followed

Principles of method if other than guideline:

CO2 evolution test

GLP compliance:

not specified

Oxygen conditions:

aerobic

Inoculum or test system:

other: Mixed liquor activated sludge (secondary effluent)

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): West Bay County WWTP, Bay City, Michigan, treating primarily municipal sewage up to 4 mio gallons per day

Duration of test (contact time):

6 d

Initial conc.:

> 600 - < 5 700 µg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

TEST CONDITIONS
- Composition of medium: minimal salts basal medium according to Stainer et al. (1966; J Gen Microbiol 43: 159-271)
- Additional substrate: no
- Solubilising agent (type and concentration if used): no
- Test temperature: 25 °C
- pH:
- pH adjusted: yes/no
- CEC (meq/100 g):
- Aeration of dilution water:
- Suspended solids concentration:
- Continuous darkness: yes
- Other:

TEST SYSTEM
- Culturing apparatus: 500-mL polypropylene Erlenmeyer flasks, sealed with rubber stoppers and fitted with CO2 traps
- Number of culture flasks/concentration: 2
- Method used to create aerobic conditions:
- Method used to create anaerobic conditions:
- Measuring equipment:
- Test performed in closed vessels due to significant volatility of test substance:
- Test performed in open system:
- Details of trap for CO2 and volatile organics if used: glass tubular CO2 traps (4 mL 1N KOH)
- Other: test material amended with 14C-TEA to yield 0.7 µCi/mL

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

CONTROL AND BLANK SYSTEM
- Inoculum blank:
- Abiotic sterile control: yes, via addition of formaldehyde (final concentration: 2.8%)
- Toxicity control:
- Other: performed in duplicate

STATISTICAL METHODS:

Parameter:

% degradation (test mat. analysis)

Value:

ca. 100

Sampling time:

5 d

Remarks on result:

other: Primary biodegradation of 5.7 mg/l TEA in a test with 164 mg/l municipal activated sludge

Parameter:

% degradation (test mat. analysis)

Value:

ca. 100

Sampling time:

1 d

Remarks on result:

other: Primary biodegradation of 0.6 mg/l TEA in a test with 164 mg/l municipal activated sludge

Details on results:

The rate constants in all test batches for degradation and mineralization are >0.359. According to GHS legislation, the substance is rapidly biodegradable. Therefore it can be assumed that the substance is also readily biodegradable.

Biodegradation (activated sludge):

Activated sludge (mg/L MLSS)	Test item concentration (mg/L)	Biodegradation rate constant (/d)	Biodegradation half-life (d)	CO2/ThCO2 (%)
164	0.6	6.86	0.1	56+-10
164	5.7	1.36	0.51	54+-3
818	0.6	30	0.02	55+-1
818	5.7	6.7	0.01	52+-0

 

Mineralisation (activated sludge):

Activated sludge (mg/L MLSS)	Test item concentration (mg/L)	Mineralisation rate constant (/d)	Mineralisation half-life (d)
164	0.6	1.92	0.36
164	5.7	1.04	0.67
818	0.6	2.86	0.24
818	5.7	1.69	0.41

Validity criteria fulfilled:

yes

Interpretation of results:

other: rapidly biodegradable according to GHS (readily biodegradability is therefore assumed)

Description of key information

The compound consists of a triazine part and an amine part. Under environmental conditions it is present in its dissociated form. An OECD 301 E study is available for the compound itself. Additionally, separate assessments for biodegradation were conducted for the triazine and the amine part, respectively.  In conclusion, neither the triazine nor the amine part of the compound can be regarded as persistent or even very persistent in the environment. The compound is readily biodegradable (by OECD criteria).

Additional information

The compound consists of a triazine part and an amine part. Under environmental conditions it is present in its dissociated form. An OECD 301 E study is available for the compound itself. Additionally, separate assessments for biodegradation were conducted for the triazine and the amine part, respectively.

CAS 80584-92-5

The ready biodegradability of the test compound was assessed in a test following OECD guideline No. 301E (1981). The relative biodegradation values calculated from the measurements performed during the test period revealed 76 % biodegradation of the test compound.

In conclusion, the test compound is classified as readily biodegradable.

CAS 80584 -91 -4

The ready biodegradability of the triazine part was assessed in a modified OECD screening test in accordance to OECD guideline 301 E. The test item was exposed to the inoculum for 28 days. After exposure period a DOC removal of 68 % was detected. Therefore, the test item can be considered to be biodegradable but not readily biodegradable.

Furthermore, supporting data on biodegradation are available. The biodegradability of the test item was investigated under different sludge / test item ratios. TOC removal and removal of test item with specific analysis after 28 days exposure was investigated. The results indicate complete primary degradation after 28days and 68% biodegradability based on TOC removal after 28 days.

It can be concluded that the triazine part of the compound is not persistent or very persistent and at least inherently biodegradable not fulfilling specific criteria.

CAS 102 -71 -6

As key study regarding the biodegradation of the amine part a test performed by West et al. (1996) is identified. In a test with activated sludge biodegradation half-lifes between 0.01 and 0.51 days and mineralization half-lifes between 0.24 and 0.67 days were found, depending on the initial concentration of the test substance (164 and 818 mg/L) and the concentration of the activated sludge (0.6 and 5.7 mg/L MLSS).

These results indicate, that the compound is readily biodegradable. A ready test performed by Gerike (1978) according to a precursor of the Modified OECD Screeening Test (OECD TG 301 E) using non-adapted effluent from a waste water treatment plant as inoculum supports these results.

While the substance is readily biodegradable in freshwater systems, it is not readily biodegradable in seawater as concluded from an OECD 306 test published by Eide-Haugmo et al. (2009/2012) and Brakstad et al. (2012). The compound is not readily biodegradable in marine systems (19.6% ThBOD).

In conclusion, neither the triazine nor the amine part of the compound can be regarded as persistent or even very persistent in the environment.