esterification product of 2-hydroxypropane-1,3-diyl-bis(2-methylprop-2-enoate) and 3-hydroxypropane-1,2-diyl-bis(2-methylprop-2-enoate) and diphosphorus pentoxide

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:

3 December 2001 - 28 January 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study under GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

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)

Test material

Test material information

Constituent 1

Reference substance name:

1224866-76-5

Cas Number:

1224866-76-5

IUPAC Name:

1224866-76-5

Details on test material:

- Name of test material (as cited in study report): Glypho
- Substance type: pure active substance
- Physical state: liquid
- Stability under test conditions: Stable
- Storage condition of test material: Refrigeration at ca. 4°C
- Other: bottles not to be filled > 90%, oxygen inhibits polymerisation

Study design

Oxygen conditions:

aerobic

Inoculum or test system:

sewage, predominantly domestic, non-adapted

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): ARA Ergolz II, Füllinsdorf, Switzerland
- Preparation of inoculum for exposure: The sludge was washed once with tap water by centrifugation and the supernatant liquid phase was decanted. A homogenized aliquot of the final sludge suspension was weighed, thereafter dried and the ratio of wet to dry weight was calculated. Based on this ratio, wet sludge were suspended in test water to obtain a concentration equivalent to 4 g (±10%) dry material per liter. During the holding period before use, the sludge was aerated at room temperature. Prior to use, the sludge was diluted with test water to a concentration of 1 g per liter (dry weight basis). This diluted activated sludge was added to test water to obtain a final concentration of 30 mg dry material per liter.
- Pretreatment: None
- Concentration of sludge: 30 mg/L dry weight basis in reactor
- Initial cell/biomass concentration: Not reported
- Water filtered: No

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:

O2 consumption

Details on study design:

TEST CONDITIONS
- Composition of medium:
Stock solution 1
KH2PO4 8.50 g/L
K2HPO4 21.75 g/L
Na2HPO4 ∙ 2H2O 33.40 g/L
NH4Cl 0.50 g/L
Stock solution 2
MgSO4 ∙ 7H2O 22.50 g/L
Stock solution 3
CaCl2 ∙ 2H2O 36.40 g/L
Stock solution 4
FeCl3 ∙ 6H2O 0.25 g/L

10 mL of stock solution No. 1 and 1 mL each of stock solution Nos. 2, 3 and 4 were combined and made up to 1000 mL with purified water. The pH was adjusted from 7.7 to 7.4 with a diluted hydrochloric acid solution. This mixture was used for all dilutions and preparation of activated sludge.

- Additional substrate: None
- Solubilising agent (type and concentration if used): None
- Test temperature: 22 °C
- pH: 7.2 - 7.4 at test initiation.
- pH adjusted: No
- Aeration of dilution water: Sealed bottles stirred in a SAPROMAT D12 (Voith GmbH, Heidenheim,
Germany)
- Suspended solids concentration: 30 mg/L sludge solids dry weight
- Continuous darkness: Yes

TEST SYSTEM
- Culturing apparatus: SAPROMAT D12 (Voith GmbH, Heidenheim, Germany), 500 mL reactor volume, 250 mL solution per reactor.
- Number of culture flasks/concentration: Two
- Method used to create aerobic conditions: Continuous stirring. Consumed oxygen replaced by electrolytic generation of a copper sulfate solution.
- Measuring equipment: SAPROMAT D12 (Voith GmbH, Heidenheim, Germany)
- Details of trap for CO2 and volatile organics if used: Soda lime, used to trap CO2

SAMPLING
- Sampling frequency: Once daily on working days, non-destructive testing
- Sampling method: Electrode-type manometer.

CONTROL AND BLANK SYSTEM
- Inoculum blank: Yes, in duplicate
- Abiotic sterile control: Yes, single flask inhibited with mercury dichloride
- Toxicity control: Yes, single flask

CONCENTRATIONS:
- Test substance flasks: 100 mg/L
- Reference substance flasks: 100 mg/L
- Abiotic control: 100 mg/L test substance, 10 mg/L HgCl2
- Toxicity control: 100 mg/L test substance plus 100 mg/L reference substance
- The test item was first weighed onto glass slides. The glass slides were then transferred to the designated test flasks and test water was added. No emulsifiers or solvents were used but ultrasound dispersion was employed for five minutes to obtain a suspension of the test item as homogeneous as possible. ThOD of test substance could not be calculated, but COD was measured according to the EU Commission Directive 92/69/EEC, C.6 following DIN 38414-S9. The COD determination was not GLP-compliant but was performed in compliance with DIN EN 45001 (General criteria for the operation of testing laboratories).

STATISTICAL METHODS:

BOD = [(mg O2 uptake/L by test and/or reference item) - (mg O2 uptake/L by inoculum control)] / [mg test and/or reference item per liter]

% biodegradation = [BOD (mg O2 /mg chemical)] / [COD or ThOD (mg O2 /mg chemical)]

where COD is used for test substance and ThOD is used for reference substance

Reference substance

Reference substance:

benzoic acid, sodium salt

Results and discussion

% Degradation

Parameter:

% degradation (O2 consumption)

Value:

82

Sampling time:

28 d

Remarks on result:

other: Biodegradation >10% on day 7, >60% on day 17

BOD5 / COD results

Results with reference substance:

The percent biodegradation of the reference item sodium benzoate was calculated based on cumulative oxygen demand (Table 1) and the theoretical oxygen demand of 1.67 mg O2/mg. In the procedure controls, the reference item was degraded by an average of 85% by Exposure Day 14, thus confirming suitability of the activated sludge.

In the toxicity control, the course of biodegradation over the 28-day exposure period was intermediate between the reference substance and the test substance (Figure 1). Within 14 days of exposure, biodegradation amounted to 64%. According to the test guidelines, the test substance had no inhibitory effect on activated sludge microorganisms at a concentration of 100 mg/L because biodegradation in the toxicity control was >25% within 14 days.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

% difference in extreme values <20% at end of test, % degradation of reference compound > 60% on day 14.

Interpretation of results:

readily biodegradable

Conclusions:

CAS# 1224866-76-5 is readily biodegradable

Executive summary:

CAS# 1224866-76-5 biodegradability was assessed in a respirometric assay conducted according to OECD301F. Because the test substance theoretical oxygen demand could not be calculated, chemical oxygen demand was measured and used to calculate percent biodegradation. Reference substance was sodium benzoate. Test substance biodegradation was >10% by day 7, >60% by day 17, and 82% on day 28. CAS# 1224866-76-5 is readily biodegradable.

The test was conducted in accordance with OECD guideline and compliant with GLP criteria. It is considered reliable without restrictions and is suitable for use in Risk Assessment, Classification & Labeling, and PBT Analysis.