2-Propenoic acid, 2-chloro-, 3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl ester

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

biodegradation in water: inherent biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

March 2019

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

The study was conducted in accordance OECD 302C guideline with one deviation, under GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 302 C (Inherent Biodegradability: Modified MITI Test (II))

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Identification: C6SFCLA
- batch: 6SFCT83018
- Expiration date of the lot/batch: 31 Marc 2020
-Purity: 98.1%


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark

Oxygen conditions:

aerobic

Inoculum or test system:

mixture of sewage, soil and natural water

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure):
a) Domestic sewage plants
i) Liverpool
ii) Loughborough, Leicestershire iii) Gloucester
b) Industrial sewage plant
i) Derby
c) Freshwater samples
i) Leeds and Liverpool Canal, Liverpool ii) RiverDerwent,Belper,Derbyshire iii) River Severn, Gloucester
d) Lake Water (x1)
i) Allestree Lake, Derby
e) Sea Water Samples
i) Huttoft, Eastern Coast
ii) Hightown, North Western Coast
- Laboratory culture: No information
- Method of cultivation: Not relevant
- Storage conditions: constantly aerated at a temperature of approximately 25 °C and shielded from light.
- Storage length: 2 days
- Preparation of inoculum for exposure: The culture was allowed to settle daily for approximately 30 minutes and approximately one third of the volume of the supernatant removed. An equal volume of 0.1% synthetic sewage was added and the aeration re-started again. Synthetic sewage was prepared by dissolving glucose, peptone and monopotassium phosphate in deionized water at a concentration of 0.1% w/v. The pH of the synthetic sewage and culture was adjusted daily to within the range pH 7.0 ±1.0 with sodium hydroxide or phosphoric acid.
- Concentration of sludge: The suspended solids (ss) concentration of the culture was determined to be 2500 mg ss/L
- Water filtered: yes
- Type and size of filter used, if any: coarse

Duration of test (contact time):

ca. 28 d

Initial conc.:

ca. 30 mg/L

Based on:

act. ingr.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

TEST CONDITIONS
- Composition of medium:
olution a
pH = 7.2
Solution b
Solution c
Solution d
pH = 7.2
Envigo Study Number: VQ33YJ
Mineral Medium
KH2PO4 K2HPO4 Na2HPO4.12H2O NH4Cl
CaCl2 MgSO4.7H2O FeCl3.6H2O
8.50 g/L 21.75 g/L 44.60 g/L 1.70 g/L
27.50 g/L 22.50 g/L 0.25 g/L
To 1 liter (final volume) of purified water* was added the following volumes of
solutions a - d:
3 mL of Solution a
3 mL of Solution b 3 mL of Solution c 3 mL of Solution d
- Additional substrate: inoculum as previously described
- Solubilising agent (type and concentration if used): none
- Test temperature: between 23 and 26 °C.
- pH: pH 7.0 ±1.0
- pH adjusted: yes
- CEC (meq/100 g): not reported
- Aeration of dilution water: not reported
- Suspended solids concentration: 2500 mg ss/L
- Continuous darkness: yes

TEST SYSTEM
- Culturing apparatus: 500 mL glass bottles
- Number of culture flasks/concentration: 1. Three replicate bottles containing inoculated mineral medium to act as the inoculum control for the test item vessels. These bottles contained 50 mL of seeded dilution water of suspended solids level of 1000 mg ss/L and 450 mL of mineral medium to give a final suspended solids level of 100 mg ss/L.
2. Three replicate bottles containing inoculated mineral medium to act as the inoculum control for the procedure control vessels. These bottles contained 15 mL of seeded dilution water of suspended solids level of 1000 mg ss/L and 485 mL of mineral medium to give a final suspended solids level of 30 mg ss/L.
3. Three replicate bottles containing inoculated mineral medium and the reference item, aniline, at a concentration of 100 mg/L to act as the procedure control. These bottles were prepared by diluting a 50 mL aliquot of a 1000 mg/L stock solution of aniline with 435 mL of mineral medium and 15 mL of seeded dilution water with a suspended solids level of 1000 mg ss/L to give a test concentration of 100 mg/L, and a final suspended solids concentration of 30 mg ss/L.
4. Five replicate bottles containing inoculated mineral medium and the test item at a concentration of 30 mg/L. These bottles were prepared by dispersing an amount of test item (15 mg) with 400 mL of mineral medium with the aid of high shear mixing (approximately 7500 rpm, 15 minutes) prior to the addition of 50 mL of seeded dilution water with a suspended solids level of 1000 mg ss/L and further mineral medium (50 mL) to give a test concentration of 30 mg/L, and a final suspended solids concentration of 100 mg ss/L.
5. Three replicate bottles containing the test item in deionized water alone at a concentration of 30 mg/L. These bottles were prepared by dispersing an amount of test item (15 mg) with 400 mL of deionized water with the aid of high shear mixing (approximately 7500 rpm, 15 minutes) prior to adjusting to a final volume of 500 mL with deionized water.
- Method used to create aerobic conditions: not reported
- Method used to create anaerobic conditions: not relevnt
- Measuring equipment: The DOC analyses were carried out using a Shimadzu TOC V-CPH TOC Analyzer
- Test performed in closed vessels due to significant volatility of test substance: The system consists of a sample flask sealed by a sensor head/CO2 trap immersed in a temperature controlled water bath
- Test performed in open system: Not relevant
- Details of trap for CO2 and volatile organics if used: ethanolamine (50% v/v)


SAMPLING
- Sampling frequency: 0,7,14,21 anms 28 days
- Sampling method: not reported
- Sterility check if applicable: not applicable
- Sample storage before analysis: not applicable


CONTROL AND BLANK SYSTEM
- Inoculum blank: Yes; containing inoculum only as previously described.
- Abiotic sterile control: no
- Toxicity control: not relevant

STATISTICAL METHODS: The results of statistical analysis using Student’s t-test on the Biological Oxygen Demand values showed significant differences (P> 0.05) between the control and test item vessels

Parameter:

% degradation (O2 consumption)

Value:

ca. 0

Sampling time:

7 d

Parameter:

% degradation (O2 consumption)

Value:

ca. 0

Sampling time:

14 d

Parameter:

% degradation (O2 consumption)

Value:

ca. 0

Sampling time:

21 d

Key result

Value:

ca. 0

Sampling time:

28 d

Remarks on result:

other:

Remarks:

The test item attained 0% degradation calculated from oxygen consumption values after 28 days.

Sample Description	ThOD (mg O2/L)	Day 7		Day 14		Day 28
		BOD (mg O2/L)	Degradation (%)	BOD (mg O2/L)	Degradation (%)	BOD (mg O2/L)	Degradation (%)	Mean Degradation (%)
Inoculum Control (30 mg ss/L)	309	24.24	-	40.44	-	59.90	-	-
Procedure Control Plus Inoculum (30 mg ss/L)		202.30	58	244.28	66	282.76	72	-
Inoculum Control (100 mg ss/L)		49.56	-	87.84	-	121.54	-
Test item Plus Inoculum (100 mg ss/L)R1	17.1	31.82	0*	64.06	0*	90.26	0*	0*
Test item Plus Inoculum (100 mg ss/L)R2	17.1	31.98	0*	63.76	0*	90.54	0*
Test item Plus Inoculum (100 mg ss/L)R3	17.1	30.74	0*	66.30	0*	97.84	0*
Test item in Deionized Water	-	3.38	0*	7.58	0*	13.08	-	-

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

The test item cannot be considered to be inherently biodegradable under the strict terms and conditions of OECD Guideline No. 302C.

Executive summary:

A study was performed to assess the inherent biodegradability of the test item in an aerobic aqueous medium. The methods followed are designed to be compatible with the OECD Guidelines for Testing of Chemicals (1981) No. 302C Inherent Biodegradability; Modified MITI (II) Test and in the MITI Gazette, 19 July 1974 under paragraph 7-1: Method for Testing the Biodegradability of Chemical Substances by Micro-organisms.

The test item was prepared as an aqueous dispersion at a concentration of 30 mg/L, inoculated with micro-organisms from a laboratory culture originating from 10 different sites throughout the UK (3 Domestic sewage plants, 1 industrial sewage plant, 3 freshwater samoles, 1 lake water and 2 sea water samples) and incubated in diffuse light at temperatures of between 23 and 26°C for 28 days. The degradation of the test item was assessed by the measurements of oxygen consumption on Days 0 and 28. Control solutions with inoculum and the reference item, aniline, were used for validation purposes.

Aniline attained biodegradation rates of 58% after 7 days and 66% after 14 days thereby confirming the suitability of the inoculum and culture conditions (Validity Criteria: If the precentage degradation of aniline calculated from the oxygen consumption does not exceed 40% after 7 days , and 65% after 14 days , the test is regarded as invalid).

The test item attained 0% degradation calculated from oxygen consumption values after 28 days.

Reference 2

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

7 Oct - 7 Dec 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 D (Ready Biodegradability: Closed Bottle Test)

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): secondary effluent from a domestic sewage treatment plant, Naruse Clean Center, Machida; collected October 8, 2010
- Storage conditions: aerobic conditions
- Pretreatment: The inoculum was filtered using a No. 5A filter to remove the suspended matter.
- Concentration of sludge: not reported
- Water filtered: yes
- Type and size of filter used, if any: No. 5A

Duration of test (contact time):

28 d

Initial conc.:

2.98 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

TEST CONDITIONS
- Composition of medium:
Liquid A: Aqueous solution containing 2.175% K2HPO4, 0.85% KH2PO4, 3.34% Na2HPO4*2H2O and 0.05% NH4Cl
Liquid B: Aqueous solution containing 2.75% CaCl2
Liquid C: Aqueous solution containing 2.25% MgSO4*7H2O
Liquid D: Aqueous solution containing 0.025% FeCl3*6H2O
Purified water, Grade 4 (Japanese Industry Standards (JIS) K0557) was used. Aqueous solution containing 0.1% each of liquids A, B, C and D.
- Test temperature: 20 ± 1 °C
- pH: 6.8 - 7.8
- Concentration of inoculum in test vessels: 50 µL/L
- Continuous darkness: yes

TEST SYSTEM
- Culturing apparatus: BOD bottles
- Number of culture flasks/concentration: 2
- Method used to create aerobic conditions: mineral medium saturated with oxygen
- Measuring equipment: BOD bottles

SAMPLING
- Sampling frequency: days 0 and 28

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes, 1 replicate
- Abiotic sterile control: yes, 1 replicate
- Toxicity control: none

Reference substance:

benzoic acid, sodium salt

Parameter:

% degradation (O2 consumption)

Remarks:

BOD

Value:

4

Sampling time:

28 d

Parameter:

% degradation (test mat. analysis)

Remarks:

GC

Value:

0

Sampling time:

28 d

Details on results:

- Transformation products: The amounts of C6 Alcohol were less than 0.01 mg in the test suspensions and abiotic controls. The formation rates of C6 Alcohol were calculated to be all less than 1% in test suspensions and abiotic controls. The amounts of alpha-Chloroacrylic acid were all less than 0.01 mg in test suspension and abiotic controls. The formation rates of alpha-Chloroacrylic acid were calculated to be all less than 2% in test suspensions and abiotic controls.

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Description of key information

4%  biodegradation after 28 days (BOD, OECD 301 D)

Key value for chemical safety assessment

Biodegradation in water:

under test conditions no biodegradation observed

Type of water:

freshwater

Additional information

Key Study

Two studies are available on the biodegradability of 3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctyl 2-chloropropenoic acid ester (CAS 96383-55-0). The first study was conducted as closed bottle test according to OECD guideline 301 D and GLP (Makido, 2010). Secondary effluent from a domestic sewage treatment plant was used as inoculum and exposed to the test substance at a concentration of 2.98 mg/L. Less than 10% biodegradation (4% after 28 days) was observed based on BOD and residual test material, respectively. Consequently, the test substance is not considered readily biodegradable.

Supporting Study

A study was performed to assess the inherent biodegradability of the test item in an aerobic aqueous medium. The methods followed are designed to be compatible with the OECD Guidelines for Testing of Chemicals (1981) No. 302C Inherent Biodegradability; Modified MITI (II) Test and in the MITI Gazette, 19 July 1974 under paragraph 7-1: Method for Testing the Biodegradability of Chemical Substances by Micro-organisms.

The test item was prepared as an aqueous dispersion at a concentration of 30 mg/L, inoculated with micro-organisms from a laboratory culture originating from 10 different sites throughout the UK(3 Domestic sewage plants, 1 industrial sewage plant, 3 freshwater samples, 1 lake water and 2 sea water samples)and incubated in diffuse light at temperatures of between 23 and 26°C for 28 days. The degradation of the test item was assessed by the measurements of oxygen consumption on Days 0 and 28. Control solutions with inoculum and the reference item, aniline, were used for validation purposes.

The test item attained 0% degradation calculated from oxygen consumption values after 28 days.

The results obtained from the OECD 302C, Inherent Biodegradability are in agreement with the results from the OECD 301D study (the substance is not ready biodegradable). It is likely the study was conducted to see if any further degradation than that seen in the OECD 301D, would be seen when using a mixture of naive bacterial cultures and those subject to industrial waste. However, as the result was similar to that of the 301D it is assumed measuring the residual amounts of test item as prescribed by the 302C was unnecessary. Furthermore, it should be noted that the substance has vapour pressure of 4.9 Pa at 20 °C and a low water solubility, making the substance highly volatile. The OECD 302C guideline states that the method is not applicable for volatile substances. During the test no measurements or adaptations were taken to address the volatility of the substance. The results do, however, agree with the OECD 301D study showing the limited degradability of the substance despite the use of varying bacterial cultures. The OECD 302C study further supports the key study indicating the substance is not degradable.