Trichloro(N,N-dimethyloctylamine)boron

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:

key study

Study period:

The study was conducted between 03 November 2014 and 01 December 2014.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not applicable

Oxygen conditions:

aerobic

Inoculum or test system:

other: Mixture of sewage and natural water

Details on inoculum:

A mixed population of active sewage sludge micro-organisms were obtained between
22 September 2014 and 25 September 2014 from ten different sampling sites around the UK.

The sampling sites were as follows:

a) Domestic sewage plants (x3)
i) Liverpool
ii) Loughborough, Leicestershire
iii) Gloucester

b) Industrial sewage plant (x1)
i) Derby

c) Freshwater samples (x3)
i) Leeds and Liverpool Canal, Liverpool
ii) River Derwent, Belper, Derbyshire
iii) River Severn, Gloucester

d) Lake Water (x1)
i) Allestree Lake, Derby

e) Sea Water Samples (x2)
i) Huttoft, Eastern Coast
ii) Hightown, North Western Coast
The sample types and volumes sampled from each site were as follows:

a) Sewage Plants: 1 liter of return sludge at each sewage disposal plant.
b) Freshwater, lake and sea: 1 liter of surface water and 1 liter of surface soil on the bank/beach which is in contact with the atmosphere.

The samples obtained from the sampling sites were mixed thoroughly and the mixture allowed to settle. The floating foreign matter was removed and the supernatant filtered through a coarse filter paper. The filtrate (4 liters) was then mixed with 2 liters of synthetic sewage and transferred to a culture vessel. The pH of the culture mixture was determined to be 7.0. The culture mixture was constantly aerated via a narrow bore glass pipette at a temperature of approximately 25 °C.

The culture was allowed to settle daily for approximately 30 minutes and approximately 1/3 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.

The sludge was found to have active microflora including a variety of protozoa, including ciliates, flagellates and a large population of motile bacteria. The sludge formed cloudy flocs when on aeration.

Duration of test (contact time):

28 d

Initial conc.:

30 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

Mineral Medium
The mineral medium used in this study was that recommended in the OECD Guidelines.

Procedure
Preparation of Seeded Dilution Water
Determination of the suspended solids level of the active sludge culture was carried out according to the method given in the Japanese Industrial Standard JISK 0102-1981. The suspended solids (ss) concentration of the culture was determined to be 1300 mg ss/L. Seeded dilution water was prepared by addition of 615 mL of the active sludge culture to a final volume of 800 mL of mineral medium to give a suspended solids level of 1000 mg ss/L and maintained on aeration at approximately 25 °C for 5 days prior to use in the study in order to reduce the basal BOD of the seeded dilution water. On subsequent dilution in the preparation of the test series, suspended solids levels of 30 mg ss/L and 100 mg ss/L were obtained for the reference and test item preparations respectively.

Experimental Preparation
Test Item
The test item was dispersed directly in mineral medium. Prior to weighing the test item was warmed to approximately 80 ºC in order to ensure homogenenity.

An amount of test item (15 mg) was dispersed in mineral medium (400 mL) with the aid of high shear mixing (approximately 7500 rpm, 5 minutes) prior to the addition of inoculum (50 mL) and water to a final volume of 500 mL to give the test concentration of 30 mg/L.

Additional vessels were prepared as above using deionized water (500 mL) without the addition of inoculum.

A test concentration of 30 mg/L was selected for use in the study following the recommendations of the Test Guideline.

Preparation of Test System
The following test concentrations were prepared in 500 mL glass bottles:

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 of 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 450 mL of mineral medium and 50 mL of seeded dilution water of suspended solids level of 1000 mg ss/L 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 500 mL of deionized water.


On Day 0, one of each inoculated mineral medium at 30 and 100 mg ss/L, one procedure control, one test item in inoculated mineral medium, and one test item in deionized water vessel were sampled for pH determination.

All remaining inoculum control, procedure control and test item vessels were placed in the CES Multi-Channel Aerobic Respirometer.

The system consists of a sample flask sealed by a sensor head/CO2 trap immersed in a temperature controlled water bath. The samples were stirred for the duration of the study with a magnetically coupled stirrer.
As biodegradation progresses, the micro-organisms convert oxygen to carbon dioxide which is absorbed in the ethanolamine (50% v/v) CO2 trap causing a net reduction in gas pressure within the sample flask. The pressure reduction triggers the electrolytic process, generating oxygen and restoring the pressure in the sample flask. The magnitude of the electrolyzing current and the duration of the current is proportional to the amount of oxygen supplied to the micro-organisms. The data generated from the respirometer's own battery backed memory was collected on the hard disk drive of a non-dedicated computer.

The test was conducted in diffuse light at a temperature of 25 ºC.

On Day 28, one of each inoculated mineral medium at 30 and 100 mg ss/L, one procedure control, three test item in inoculated mineral medium, and one test item in deionized water vessel that were considered to have given the most consistent BOD values over the study period were selected for pH determinations.

The remaining vessels which were not sampled were discarded and not reported. Additional replicate vessels were prepared and incubated in order that in the event of a leak in the test system a replicate vessel could be discarded without jeopardizing the integrity of the study.

Evaluations
Observations
The appearance of the test dispersions was recorded on Days 0, 14 and 28.

Physico-chemical Measurements
The temperature of the water bath was recorded daily.

pH Measurements
The pH of the relevant inoculum control, test item and procedure control vessels was recorded on Days 0 and 28 using a Hach HQ40d Flexi handheld meter.

Data Evaluation
Theoretical Oxygen Demand
The Theoretical Oxygen Demand (ThOD) for a compound CcHhClclNnPpSsOoNana was calculated using the following equation:

ThOD (NO3) = (16 (2c + ½ (h-cl) + 5/2n + 5/2p + 3s +½na – o)) / molecular weight

Percentage degradation
The percentage degradation in terms of oxygen consumption was calculated using the following equation:

% degradation = ((BOD – B) / ThOD) x 100
Where:

BOD = Biological Oxygen Demand of the test or standard material.
B = Oxygen consumption in basal mineral medium to which inoculum is added (control).
ThOD = Theoretical Oxygen Demand to completely oxidize the reference item.

Reference substance:

aniline

Remarks:

test concentration of 100 mg/L

Parameter:

% degradation (O2 consumption)

Value:

42

Sampling time:

28 d

Details on results:

The BOD of the inoculated mineral medium (control) at 30 mg ss/L was 52.02 mg O2/L at the end of the test and therefore satisfied the validation criterion given in the OECD Guidelines. Although the control BOD was in excess of 30 mg O2/L at the end of the test, this is considered not to affect the integrity of the study given that the upper level of 60 mg O2/L given in the OECD Guidelines was not exceeded and that all other validation criteria were satisfied.

The pH of the inoculated test item vessels at the end of the study ranged between 7.9 and 8.0 and therefore satisfied the validation criterion given in the OECD Guidelines.

Percentage Degradation Values
The test item attained 38%, 48% and 40% degradation with a mean of 42% degradation calculated from oxygen consumption values after 28 days.

Parameter:

BOD5

Value:

52.02 mg O2/g test mat.

Results with reference substance:

Aniline attained 76% degradation after 28 days calculated from oxygen consumption values. 

Validation Criteria

Aniline attained biodegradation rates of 66% after 7 days and 74% after 14 days thereby confirming the suitability of the inoculum and culture conditions.

Calculation Table for the Degree of Degradation by BOD

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 (%)
Aniline Plus Inoculum (30 mg ss/L)	309	221.50	66	263.26	74	287.72	76	-
Control (30 mg ss/L)	-	18.46	-	33.24	-	52.02	-
Test item Plus Inoculum (100 mg ss/L)                        R1	56.7	65.80	-4	106.24	20	145.20	38	42
Test item Plus Inoculum (100 mg ss/L)                       R2	56.7	65.06	-5	106.32	20	150.50	48
Test item Plus Inoculum (100 mg ss/L)                       R3	56.7	64.02	-7	105.58	19	146.06	40
Test item in Deionized Water	-		-		-		-	-
Control (100 mg ss/L)	-	68.14	-	94.88	-	123.54	-

R₁– R₃= Replicates 1 to 3

Test concentration of 100 mg/L and 30 mg ss/L was added to the procedure control vessel

Test concentration of 30 mg/L and 100 mg ss/L was added to each test item vessel

Negative degradation values are due to measured BOD values being less than control values

 

 

Validity criteria fulfilled:

yes

Interpretation of results:

inherently biodegradable

Conclusions:

Under study conditions, the test substance can be considered as inherently biodegradable.

Executive summary:

A study was performed to assess the inherent biodegradability of the trichloro(N,N-dimethyloctylamine) boron in an aerobic aqueous medium according to OECD Guideline 301C , in compliance with GLP. The test substance 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 and incubated in diffuse light at 25 °C for 28 days. The degradation of the test substance 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 substance attained 38%, 48% and 40% degradation with a mean of 42% degradation calculated from oxygen consumption values after 28 days. Under study conditions, the test substance can be considered as inherently biodegradable (Harlan, 2015).

Reference 2

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13 March 1996 to 11 April 1996

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No GLP methodology followed.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.4-C (Determination of the "Ready" Biodegradability - Carbon Dioxide Evolution Test)

Deviations:

no

GLP compliance:

no

Oxygen conditions:

aerobic

Inoculum or test system:

other: Activated sludge freshly obtained from a municipal sewage treatment plant.

Details on inoculum:

Activated sludge freshly obtained from a municipal sewage treatment plant: "Waterschap de Maaskant", s-Hertogenbosch, the Netherlands.

The sludge were kept under continuous aeration until further treatment. The concentration of suspended solids was 5.7 g/L in the concentrated
sludge. This was outside the protocolled range (3-5 g/L). However, this was found to have had no effect on the outcome of the study. Before use, the sludge was allowed to settle for at least 30 min and the liquid decanted for use as inoculum at the amount of 10 ml/l of mineral medium.

Reason for selection: The test has been accepted internationally (EEC, OECD) for determining the "ready biodegradability of the test substances
under aerobic conditions.

Duration of test (contact time):

28 d

Initial conc.:

28.5 mg/L

Based on:

test mat.

Remarks:

57 mg was dissolved in 2 L medium

Parameter followed for biodegradation estimation:

CO2 evolution

Reference substance:

other: Sodium acetate

Parameter:

% degradation (CO2 evolution)

Value:

> 10

Sampling time:

28 d

Remarks on result:

other: Bottle A

Parameter:

% degradation (CO2 evolution)

Value:

25

Sampling time:

28 d

Remarks on result:

other: Bottle B

Results with reference substance:

The positive control substance was degraded 72% in 14 days and 81% dagradation was reached at the end of the test period.

Theorical CO2 production:

The total carbon content (TC) of  was determined to be 42.2%. The theorical CO2 production (ThCO2) of TK 12146, was calculated to be 1.547 mg CO2/mg.

The concentration was 57.0 (A) and 57.2 mg (B) TK 12146 in 2 litres test medium. Hence, the theorical CO2 production following complete degradation was 88.2 mg per 2 litres for A and 88.5 mg for B.

The positive control contained 81.6 mg sodium acetate (ThCO2=1.073 mg CO2/mg) resulting in a theorical CO2 production following complete degradation of 87.6 mg per 2 litres.

 

Biodegradation:

The relative degradation values calculated from the measurments performed during the test period revealed no significant degradation of test substance, in bottle A (significant:>10%) and 25% biodegradation in bottle B.

 

In the toxicity control more than 25% degradation occured in 14 days (based on ThCO2). Therefore, the test substance was assumed to be not inhibitory.

 

Monitoring of temperature and pH.

The temperature recorded during the study varied between 20 and 23°C, except for a few hours on day 22, when a deviation of 2°C from the temperature range mentioned in the protocol was noted, but was considered not to have affected the integrity of this study.

The pH values of the different test media were:

 

	Just before the start of the test	On day 28
Blank control (A)	7.45	7.81
Blank control (B)	7.48	7.93
Positive control	7.49	8.17
TK 12146 (A)	7.48	7.63
TK 12146 (B)	7.49	7.60
Toxicity control	7.53	8.01

 

Acceptability of the test:

The positive control substance was degraded 72% in 14 days and 81% degradation was reached at the end of the test period.

The total CO2 release in the blank reached a total value of 13 mg CO2 per 2 litres of medium.

The differnce of duplicate values for %-degradation of TK 12146, was always less than 20.

In the toxicty control TK 12146 was found to have no inhibiting effect on microbial activity.

 

TK was not readily biodegradable under the conditions of the modified Sturm test presently performed.

Validity criteria fulfilled:

yes

Interpretation of results:

under test conditions no biodegradation observed

Conclusions:

Under the study conditions, the trichloro(N,N-dimethyloctylamine)boron was considered as not readily biodegradable.

Executive summary:

A study was conducted to determine the biodegradation in water of the test substance,trichloro(N,N-dimethyloctylamine)boron according to OECD Guideline 301 B and EU Method C.4-C in CO2 evolution test (Modified Sturm test) without GLP. Trichloro(N,N-dimethyloctylamine) boron was tested at approximate concentration of 57 mg per 2 liters, corresponding to ca. 12 mg TC/L. The Total Carbon content (TC) of test substance was determined to be 42.2%. The Theorical CO2 production (ThCO2) of test substance was calculated to be 1.547 mg CO2/mg. The relative degradation values calculated from the measurements performed during the test period revealed no significant degradation of test substance, in bottle A (SIGNIFICANT >10%), and 25% degradation in bottle B. In the control toxicity of test substance was found to have no inhibiting effect on microbial activity. Since all acceptability criteria prescribed by protocol were met, this study was considered to be valid. Under the study conditions, the trichloro(N,N-dimethyloctylamine)boron was considered as not readily biodegradable (Desmares-Koopmans, 1996).

Key value for chemical safety assessment

Biodegradation in water:

not biodegradable

Additional information

A study was conducted to determine the biodegradation in water of the test substance according to OECD Guideline 301 B and EU Method C.4-C in CO2 evolution test (Modified Sturm test) without GLP. Trichloro(N,N-dimethyloctylamine) boron was tested at approximate concentration of 57 mg per 2 liters, corresponding to ca. 12 mg TC/L. The Total Carbon content (TC) of test substance was determined to be 42.2%. The Theorical CO2 production (ThCO2) of test substance was calculated to be 1.547 mg CO2/mg. The relative degradation values calculated from the measurements performed during the test period revealed no significant degradation of test substance, in bottle A (10%), and 25% degradation in bottle B. In the control toxicity of test substance was found to have no inhibiting effect on microbial activity. Since all acceptability criteria prescribed by protocol were met, this study was considered to be valid. Under the study conditions, the trichloro(N,N-dimethyloctylamine)boron was considered as not readily biodegradable (Desmares-Koopmans, 1996).

A study was performed to assess the inherent biodegradability of the trichloro(N,N-dimethyloctylamine) boron in an aerobic aqueous medium according to OECD Guideline 301C , in compliance with GLP. The test substance 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 and incubated in diffuse light at 25 °C for 28 days. The degradation of the test substance 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 substance attained 38%, 48% and 40% degradation with a mean of 42% degradation calculated from oxygen consumption values after 28 days. Under study conditions, the test substance can be considered as inherently biodegradable (Harlan, 2015).