5,7-Di-t-butyl-3-[3,5-dimethyl-4-[(1,3,7,9-tetra-t-butyl-5-methyl-5H-benzo[d][1,3,2]benzodioxaphosphocin-11-yl)oxy]phenyl]-3H-benzofuran-2-one

Biodegradation in water: screening tests

Administrative data

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-05-08 to 2017-06-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

5,7-Di-t-butyl-3-[3,5-dimethyl-4-[(1,3,7,9-tetra-t-butyl-5-methyl-5H-benzo[d][1,3,2]benzodioxaphosphocin-11-yl)oxy]phenyl]-3H-benzofuran-2-one

EC Number:

817-187-7

Cas Number:

1803088-15-4

Molecular formula:

C54 H73 O5 P

IUPAC Name:

5,7-Di-t-butyl-3-[3,5-dimethyl-4-[(1,3,7,9-tetra-t-butyl-5-methyl-5H-benzo[d][1,3,2]benzodioxaphosphocin-11-yl)oxy]phenyl]-3H-benzofuran-2-one

Details on test material:

- State of aggregation: Solid / white
- Storage Condition: room temperature under light exclusion

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: GSID 1212253-1 OP3

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: test substance was weighed and placed in a culture bottle and then a fixed amount of mineral medium or purified water was added. It was confirmed that the pH of the test solution was 7.0.

Study design

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

Standard activated sludge, purchased from the Chemicals Evaluation and Research Institute (3-2-7 Miyanojin, Kurume, Fukuoka 839-0801, Japan), on April 19, 2017

Duration of test (contact time):

28 d

Initial test substance concentration

Initial conc.:

100 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation

Parameter followed for biodegradation estimation 1

Parameter followed for biodegradation estimation:

O2 consumption

Parameter followed for biodegradation estimation 2

Parameter followed for biodegradation estimation:

test mat. analysis

Details on study design:

TEST CONDITIONS
- Composition of medium: Mineral medium was prepared by adding each 6 mL of stock solution A, B, C and D to 1976 mL of purified water (which was adapted to quality of Japanese Pharmacopeia) and adjusted the pH to 7.0.
Stock solutions for mineral medium
Solution A:
Potassium dihydrogen orthophosphate, KH2PO4 0.85 g
Dipotassium hydrogen orthophosphate, K2HPO4 2.175g
Disodium hydrogen orthophosphate dodecahydrate, Na2HP04• 12H2O, 4.46 g
Ammonium chloride, NH4CI, 0.17 g
Dissolve in water and make up to 100 mL. The pH value of the solution should be 7.2.
Solution B
Magnesium sulphate heptahydrate, MgS04 • 7H2O, 2.25 g
Dissolve in water and make up to 100 mL.
Solution C
Calcium chloride anhydrous, CaCl2, 2.75 g
Dissolve in water and make up to 100 mL.
Solution D
Iron(III) chloride hexahydrate, FeCl3• 6H20, 0.025 g
Dissolve in water and make up to 100 mL. The pH of the solution should be 7.2.

- Test temperature: 25 ± 1°C

TEST SYSTEM
- Culturing apparatus: culture bottle in oxygen consumption analyzer for 28 days
- Number of culture flasks/concentration: 3
- Measuring equipment: oxygen consumption was continuously measured by using a Coulometer (OM-3001 A, Ohkura Electric Co., Ltd.)

CONTROL AND BLANK SYSTEM
- Inoculum blank: 1 bottle, Mineral medium 290 mL + Inoculum 900 mg/L solution 10 mL
- Abiotic control: Test substance (30.00 mg) + Purified water 300 mL
- Activity control: Aniline 29.5 µL (30.12 mg) + mineral medium 290 mL + Inoculum 900 mg/L solution 10 mL

Reference substance

Reference substance:

aniline

Results and discussion

% Degradation

% Degradation 1

Key result

Parameter:

% degradation (O2 consumption)

Value:

1

Sampling time:

28 d

% Degradation 2

Key result

Parameter:

% degradation (test mat. analysis)

Remarks:

calculated by HPLC analysis

Value:

0

Sampling time:

28 d

Details on results:

The biodegradability of the test substance after 28 days as calculated from the BOD curve were 0%, -1% and 0%, respectively, and the mean biodegradability of test substance was 1%.
The biodegradability of aniline was 61 % after 7 days and 65% after 14 days, and the oxygen consumption curve was normal. Thus, that the degradability activity of the active sludge was normal. The difference of maximum value of the biodegradability and minimum value after 28 days became 1%, and it was under 20%. Therefore, the test was valid.

On the test solution after 28 days culture, the residual concentration of the test substance was measured by the HPLC method. In the abiotic control (test substance + water), the test substance was detected at 101% relative to the amount supplied and in the test suspensions (test substance + sludge) bottles, detected at 101%, 101% and 101%.

BOD5 / COD results

Results with reference substance:

The biodegradability of aniline was 61 % after 7 days and 65% after 14 days, and the oxygen consumption curve was normal.

Any other information on results incl. tables

The test was considered valid because:

- the degradation of the reference item at day 7 was >= 40 %.

- the degradation of the reference item at day 14 was >= 65 %.

- The difference of the maximum value of the biodegradability and the minimum value after 28 days became 1%, and it was under 20%.

Table 1: Biodegradability calculated from measured oxygen consumption (moc)

Groups	Theoretical Oxygen Demand (mg)	day 7		day 14		day 21		day 28
		moc [mg]	Degradability(%)	moc [mg]	Degradability (%)	moc [mg]	Degradability (%)	moc [mg]	Degradability (%)
Test suspensionbottle1	81.8	3.0	-1	5.6	0	6.0	0	6.0	0
Test suspension bottle2	81.8	2.5	-1	5.4	0	5.6	0	5.7	-1
Test suspension bottle3	81.8	2.9	-1	5.6	0	6.0	0	6.1	0
Activity control (Aniline)	90.6	58.5	61	64.4	65	66.7	67	69.3	69
Inoculum blank	—	3.5	-	5.2	-	6.0	-	6.4	-
Abiotic control	—	0.0	-	0.0	-	0.0	-	0.0	-

 

Table 2: Residues of the test substance determined by HPLC

Groups	Amount supplied (mg)	Peak areas	Concentrations calculated by the 2-point calibration (mg/L)	Residual amount (mg)	Ratio relative to the amount supplied (%)
Test suspension bottle 1	30.00	166465	149.4532	30.38"	101
Test suspension bottle 2	30.00	166373	149.3706	30.36	101
Test suspension bottle 3	30.00	165974	149.0113	30.29	101
Inoculum blank	-	N.D. *	-	-	-
Abiotic control	30.00	165975	149.0116	30.44	101

*Quantification limit: 0.3760 mg/L(1000 as the peak area) or less was indicted as N.D.

   

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed