Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3-sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

Biodegradation in water: screening tests

Administrative data

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: see 'Remark'

Remarks:

The reliability is rated 2 because the study followed a standard guidelin of reference (OCDE 301B), which describes a procedure designed to evaluate this endpoint. The results were reviewed for reliability and assessed as valid. However the study was not conducted under GLP condition.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 B (Ready Biodegradability: CO2 Evolution Test)

Deviations:

no

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3- sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

IUPAC Name:

Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3- sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

Constituent 2

Reference substance name:

Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3-sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

EC Number:

700-610-3

Molecular formula:

UVCB substance - not applicable

IUPAC Name:

Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3-sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

Details on test material:

- Name of test material (as cited in study report): Sepisol Fast Blue 2BR
- Substance type: organic salt
- Appareance/physical state: dark blue powder
- Analytical purity: 90 - 95% w/w
- Lot/batch No.: 905536
- Stability under test conditions: stable under test conditions
- Storage condition of test material:room temperature

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES
- Water solubility (under test conditions): 73 mg/L

Study design

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic (adaptation not specified)

Duration of test (contact time):

28

Initial test substance concentration

Initial test substance concentration 1

Initial conc.:

15.4

Based on:

test mat.

Initial test substance concentration 2

Initial conc.:

10

Based on:

other: TOC (Total Oragnic Carbon)

Parameter followed for biodegradation estimation

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

TEST CONDITIONS
- Test temperature: 20°C +/- 2°C

TEST SYSTEM
- Measuring equipment: The amount of CO2 produced is calculated by titrimetric dosage, after titration of the barium hydroxide asolution thanks to sulfochromic oxidation.
- Details of trap for CO2 and volatile organics if used: During microbial degradation, the CO2 formed is trapped/absorbed in external absorption bottles containing a solution of barium hydroxide.

Reference substance

Reference substance:

acetic acid, sodium salt

Results and discussion

Test performance:

Validity of test
- 22 mg/800 mL of CO2 produced by the "blank control" at the end of the study (< 40 mg/L)
- Biodegradation of 75% (> 60%) for the reference substance (sodium acetate) at the 14 th day
=> Test validated

Toxicity:
- Combined degradation of 17% following a 14 days period for the reference and test substances, indicating a toxic effect to the microbial population in the inoculum.

% Degradation

Parameter:

% degradation (CO2 evolution)

Value:

38

Sampling time:

28

Remarks on result:

other: (38% degradation reach within 21 d)

BOD5 / COD results

Results with reference substance:

75% of biodegradation at the 14th day > than 60%

Any other information on results incl. tables

The following results are presented as the mean of the two assays (test material and blank control):

CO2 produced (in mg)

	Number of days
	1	2	6	9	12	14	16	19	21	23	28	28*
Na acetate Control	13.3	6.6	8.5	3.8	11.9	4.6	6.6	0.7	0.2	4.6	4.84	4.18
Blue 2BR 10 mg/L of TOC	2.2	1.65	1.8	2.4	11.6	1.3	3.0	0.4	1.3	1.9	3.1	2.5
Inhibition Control	2.2	2.86	13.6	2.6	2.1	1.6	3.0	1.3	0.2	2.8	4.4	2.5
Blank control	2.2	1.65	1.8	2.4	2.1	1.3	3.0	0.3	0.0	1.9	3.1	2.5

CO2 produced (in mg) following "removal" of the "blank control" values:

 

	Number of days
Corrected CO2	1	2	6	9	12	14	16	19	21	23	28	28*
Na acetate Control	11.1	5.0	6.7	1.4	9.8	3.3	3.6	0.4	0.2	2.8	1.8	1.7
Blue 2BR 10 mg/L of TOC	0.0	0.0	0.0	0.0	9.5	0.0	0.0	0.1	1.3	0.0	0.0	0.0
Inhibition Control	0.0	1.2	11.8	0.2	0.0	0.3	0.0	1.0	0.2	1.0	1.3	0.0

Degradation percentage comparing ThCO2 :

 

			Number of days
	0	2		4	6	9	12	14	16	19	21	23	28	28*
Reference substance	0%	22%		32%	46%	48%	68%	75%	82%	83%	83%	89%	92%	95%
Blue 2BR 10 mg/L of TOC	0%	0%		0%	0%	0%	33%	33%	33%	33%	38%	38%	38%	38%
Inhibition Control	0%	0%		2%	16%	17%	17%	17%	17%	19%	19%	20%	22%	22%

 

* after adding hydrochloric acid to each test vessel to drive off the carbon dioxide present in the test suspensions (left as carbonates and bicarbonates).

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Interpretation of results:

not inherently biodegradable

Conclusions:

Under the test condition, the biodegradation percentage of the test material (10 mg/L of TOC) has been assessed as 38% after a 28 days period of incubation.
According to the Guideline OECD 301, the degradation has failed to reach the 60% threshold so the test material is not considered to be readily
biodegradable.

Executive summary:

Test item, Sepisol Fast Blue 2BR, was tested using the OECD method 301B (CO2 evolution test).

Activated sludge from domestic wastewater treatment plant was used. 15.4 mg/L of the test material (corresponding to a TOC of 10 mg/L) was added to the mineral medium. The theoric CO2 quantity (ThCO2) was calculated to be 29 mg.

No organic solvent were used to facilitate the dispersion of the test material. Each test vessel was connected to a series of 4 absorption bottles, 3 of them containing 4 g/L of barium hydroxyde solution and one containing a sodium hydroxide solution.

The test was started by bubbling CO2 -free air through the suspensions.

CO2 evolution from the test suspension. Inoculum blanks and the reference substance (the positive control) were followed in parallel.

The test included:

- A blank control

- A positive control = sodium acetate

Abiotic and toxicitiy checks were included.

Analytical method: for measurement of evolved C02, the barium hydroxide absorber closesy to the test vessel was disconneted and titrate thanks to sulfochromic oxidation (NF ISO 14235).

Under the test condition, the biodegradation percentage of the test material (10 mg/L of TOC) has been determined as 38 % after a 28 days period of incubation.The % of biodegradation of the reference substance, sodium acetate, is greater than 60 % (75%) at the 14th days.

The biodegradation percentage of the inhibition control has only reached 17% after 14 hours. The standard mentions a 25 % threshold for non-toxic effect. As a result the test item could have a toxic effect in regards of the microorganisms used. The essay have not been reconducted.

According the guideline OCDE 301, the degradation has failed to reach the 60 % threshold and the test item is not considered to be readily biodegradable.