Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

other information

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

other: REACH Practical guide 6: How to report read-across and categories

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

other: REACH TGD, Chapter R.6: QSARs and grouping of chemicals

Deviations:

not specified

GLP compliance:

no

Reference 2

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

disregarded due to major methodological deficiencies

Study period:

2004/05/05 to 2004/06/02

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: Study performed according to OECD 301D and GLP requirements. But since the test has been done on 1,3-Propanediamine,N-tallow-,diacetate with solvant (butylglycol), it is not possible to take into account this study in our dossier.

Qualifier:

according to guideline

Guideline:

EU Method C.4-E (Determination of the "Ready" Biodegradability - Closed Bottle Test)

Deviations:

yes

Remarks:

The percentage degradation of the reference item didn't reach 70% by 14 days (52%)

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

yes

Remarks:

The percentage degradation of the reference item didn't reach 70% by 14 days (52%)

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, non-adapted

Details on inoculum:

The inoculum was from the activated sludge of the biological wastewater treatment plant in Abidos (64). The activated sludge obtained 1 week before seeding the flasks, was diluted to 400 mg/L of equivalent dry material, and stirred. Before seeding the flasks, the diluted activated sludge was filtered to 11 µm. The test medium was inoculated with 7.8 mL/L of filtered sludge.

Details on study design:

The parent solution of teh item (60mg/L) was added to silica gel that was already in the flask. Flasks are then ultrasonicated during 5 min and then kept under impregnation during 2hours.

Reference substance:

other: sodium benzoate

Parameter:

% degradation (O2 consumption)

Value:

58

Sampling time:

28 d

Validity criteria fulfilled:

no

Remarks:

see details in conclusions

Conclusions:

The following validity criteria were fulfilled:
- the difference of extremes of replicate values of the removal of test chemical at the plateau, at the end of the test, or at the end of the 10d-window, was less than 20%
- the oxygen consumption was < 1.5mg/L at 28d. The residual oxygen concentration in the test series was > 0.5mg/L
- in the toxicity control flasks, containing both the reference item and the test substance, the percentage biodegradation reached 38% after 28d. Since this value was higher than 25%, the test item is considered as not inhibitory for inoculum.
But the percentage degradation of the reference item didn't reach 70% by 14 days (52%)

Executive summary:

Under the test conditions, the percentage of biodegradation of Dinoramac S36 reached 58% at the end of the test (28 days). According to OECD guideline 301, the test item can nearly be considered as readily biodegradable. The test item did not inhibit the micoorganisms as shown by the toxicity control flask. The validity criteria were fulfilled except the degradation for the reference substance that didn't reach 70% after 14 days.

Reference 3

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1990-01-24 to 1990-02-28

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Experiment performed under GLP, according guidelines with acceptable deviations. All quality criteria were met.

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

acceptable deviations (see details below)

Principles of method if other than guideline:

The test has been performed according to a slightly modified test guideline:
- Instead of an effluent/extract/medium, activated sludge was used as an inoculum (see details in the Study Design section);
- Ammonium chloride was omitted from the medium to prevent nitrification;
- As the test substance is a poorly soluble substance in water, the test substance was first dissolved in dichloromethane (0.56 mL) then added to 2g silica gel (100-200 mesh) weighed in a glass petri dish. The solvent was allowed to evaporate by placing the petri dish in a vnetilated hood for 3 hours, and the entire contents were then transferred to the BOD bottle. Although no additional oxygen consumption is expected controls with silica gel were carried out as well.

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

Secondary activated sludge was obtained from the RZWI Nieuwgraaf in Duiven (1990-01-24). The RZWI Nieuwgraaf is an activated sludge plant treating predominantly domestic wastewater.

Instead of an effluent/extract/medium, activated sludge was used as an inoculum. The inoculum was taken from an activated sludge plant. The sludge was preconditioned to reduce the endogenous respiration rates. To precondition, the sludge (200 mg dry weight/L) was aerated for a period of seven days. The sludge was diluted to a concentration in the BOD bottles of 2 mg DW/L.

Duration of test (contact time):

28 d

Initial conc.:

2 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

Test bottles: the test was performed in 280 mL BOD (Biological Oxygen Demand) bottles.

Stock solutions: The sodium acetate was added to the bottles using a stock solution of 1 g/L. For the application of the test substance on silica gel, a stock solution of 1 g/L on dichloromethane was used.

The biochemical oxygen demand (BOD) of the test substance and sodium acetate were calculated from the oxygen concentrations in the bottles with and without test substance but with silica gel and the bottles with and without sodium acetate but without silica gel.

Reference substance:

other: sodium acetate

Test performance:

The dissolved oxygen concentrations were determined electrochemically using a oxygen electrode and meter.

Parameter:

% degradation (O2 consumption)

Value:

20

Sampling time:

5 d

Parameter:

% degradation (O2 consumption)

Value:

50

Sampling time:

15 d

Parameter:

% degradation (O2 consumption)

Value:

76

Sampling time:

28 d

Details on results:

The test substance was biodegraded in the closed bottle test and thus is classified as readily biodegradable. The addition of silica gel in the closed bottle test reduced the concentration of the test substance. Due to this reduction, the biodegradation was probably not influenced by the test substance.

Results with reference substance:

Percentages biodegradation of sodium acetate in the closed bottle test:
73% after 5 days
88% after 15 days
90% after 28 days

Validity criteria fulfilled:

yes

Remarks:

see details in the conclusions section

Interpretation of results:

readily biodegradable, but failing 10-day window

Conclusions:

The test substance was biodegraded 76% after 28 days and thus can be classified as readily biodegradable. Over 60% biodegradation was not achieved between day 5 and day 15. The test substance is a mixture and this is a surfactant, it can be degraded sequentially by a consortium of microorganisms (Van Ginkel, 1996). The degradation curve will therefore be the sum of a number of growth curves. The 10-day time window criterion was developed on the assumption that a testsubstance is degraded according to a “standard” growth curve in ready biodegradability tests. The time-window should be ignored as a pass fail criterion because of these scientific reasons.

The validity of the test is shown by the oxygen consumption in the control bottle with sodium acetate and an endogenous respiration of 0.6 mg/L. The pH pf the medium at the end of the test period was 7.4.

Executive summary:

A ready biodegradability test was performed according to OECD guideline 301D and GLP requirements in order to allow the measurement of the biodegradability of 1,3-Propanediamine,N-tallow-,diacetate in an aerobic aqueous medium. The test substance was biodegraded to 76% after 28 days. The validity criteria were fulfilled. The test substance can be considered as readily biodegradable.

Reference 4

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Study period:

1990/01/24 - 1990/02/28

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Test performed according guidelines with acceptable deviations. Temperature was not mentioned in the rapport. Test was performed according the guidelines and therefore the temperature should be 22 +/- 2 °C. No certificate of analysis present but purity of substance is mentioned in report.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

deviations are acceptable

Principles of method if other than guideline:

Instead of an effluent/extract mixture, activated sludge was used as an inoculum.
(as described by: J. Blok, chemosphere 14, 1985, 1805-1820; EEC update of test methods C3 to C10 of Annex V DocXI/501/89).
The inoculum was taken from an activated sludge plant treating predominantly domestic waste water. The sludge was preconditioned to reduce endogenous respiration rates. To this end, the sludge (200 mg dry wt/liter) was aerated for a period of 7 days. The sludge was diluted to a concentration in the biochemical oxygen demand (BOD) bottles of 2 mg dry wt/liter.

Ammonium chloride was omitted from the medium to prevent nitrification

The test compound is a poorly soluble substance in water and therefore the test substance was first dissolved in dichloromethane. The test substance in dichloromethane was added to silica gel weighed in a glass petri dish. The solvent was allowed to evaporate by placing the petri dish in a ventilated hood, and the entire contents were then transferred to the BOD bottles. Although no additional oxygen consumption was expected, controls with silica gel were carried out as well.
This technique was described in: N. Nyholm, C. Seiero: “Biodegradability testing of poorly soluble compounds by means of manometric respirometry”. ISO/TC 147SC5/WG4 (1989)

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): activated sludge plant RWZI Nieuwgraaf in Duiven, treating predominantly domestic wastewater
- Preparation of inoculum for exposure: 200 mg/l DW/l was aerated for one week to reduce the endogenous respiration
- Sludge concentration in BOD bottels: 2 mg/l

Duration of test (contact time):

28 d

Initial conc.:

2 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

TEST CONDITIONS
- Composition of medium: 8.5 mg KH2PO4, 21.75 mg K2HPO4, 33.4 mg Na2HPO4.2H2O, 22.5 mg MgSO4.7H2O, 27.5 mg CaCl2, 0.25 mg FeCl3.6H2O.
- Solubilising agent (type and concentration if used): Dichloromethane (p.a. quality; concentration: pure)
- Test temperature: 22 +/-2°C
- pH: 7.4
- pH adjusted: no
- Aeration of mineral medium: yes
- Suspended solids concentration: sludge DW in BOD bottles = 2 mg/l
- Continuous darkness: yes

TEST SYSTEM
- Culturing apparatus: incubator
- Number of culture flasks/concentration:
8 control flasks without test material and without inoculum (abiotic control)
8 control flasks without test material and with inoculum (inoculum blank)
8 test flasks with test material on silica gel and with inoculum( Test)
8 test flasks with sodium acetate and with inoculum (Reference control)
8 control flasks without test material but with inoculum and evaporated silica gel (inoculum blank)
- Method used to create aerobic conditions: aeration with pressured air
- Measuring equipment: oxygen meter

SAMPLING
- Sampling frequency: day 0, 5, 15, 28
- Sampling method: BOD bottles are discarded after the oxygen measurement


CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control: yes (non-sterile)
- Toxicity control: no
- Reference control: yes
- Other: inoculum blank with silica gel = with evaporated silica gel (Dichloremethane without test substance is adde to silica gel and is then evaporated)

CALCULATIONS

oxygen consumption test compound (BOD) = average oxygen concentration test compound with silica - averag oxygen concentration inoculum blank with silica
Oxygen consumption reference (BOD) = average oxygen concentration reference compound - average concentration inoculum blank

Biodegradation % = BOD (mg/l) / THOD or COD (mg/l)
Biodegradation % sodium acetate is calculated with THOD
Biodegradation % Duomeen T = calculated with COD

Parameter:

% degradation (O2 consumption)

Value:

62

Sampling time:

28 d

Validity criteria fulfilled:

yes

Remarks:

endogenous respiration < 1,5 mg/l = 0.6 mg/l; residual oxgyen concentration at day 28 > 0.5 mg/l; 88% biodegradation acetate day 15 (therefore expected to pass 60% at day 14); differences replicates < 20% at end of test.

Interpretation of results:

readily biodegradable

Conclusions:

The biodegradation percentage of N-tallow-diaminopropane is 62% at day 28. The test was performed according the guidelines with acceptable deviations. The validity criteria of the test were fulfilled.
N-tallow-diaminopropane should therefore be classified as readily biodegradable

Executive summary:

In order to assess ecotoxicological effects of chemicals, a ready biodegradability test is performed which allows the measurement of the biodegradability in an aerobic aqueous medium. N-tallow-diaminopropane is biodegraded (62%) in the closed tottle test (OECD guideline 301D) perfomed in confirmity with the OECD principles of Good I.atoratory Practices (GLP). Therefore, N-tallow-diaminopropane should be classified as readily biodegradable. In this test, the presence of silica gel in the bottles reduced the concentration of N-tallow-diaminopropane in the medium so that the toxicity of the test substance does not influence the biodegradation.

Reference 5

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

supporting study

Study period:

13-03-2008 17-04-2008

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Experiment performed under GLP, according guidelines with acceptable deviations. All quality criteria were met

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

acceptable deviations

Principles of method if other than guideline:

Ammonium chloride was omitted from the medium to prevent nitrification

GLP compliance:

yes (incl. QA statement)

Oxygen conditions:

aerobic

Inoculum or test system:

natural water

Details on inoculum:

River water was sampled at 13-03-2008 from the river Rhine near Heveadorp, The Netherlands. The river water was preconditioned to reduce the endogenous respiration rates (one week in total). Small particles were removed by sedimentation after 1 day aeration.

Duration of test (contact time):

28 d

Initial conc.:

3 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

TEST CONDITIONS
- Composition of medium: The nutrient medium of the closed bottle test contained per litre of deionized water: 8.5 mg KH2PO4, 21.75 mg K2HPO4, 33.4 mg Na2HPO4.2H2O, 22.5 mg MgSO4.7H2O, 27.5 mg CaCl2, 0.25 mg FeCl3.6H2O.
- The closed bottles were filled with river water and medium at a ratio of 1:1.
- Test temperature: 22-23°C
- pH: 7.3 - 7.0
- pH adjusted: no
- Aeration of mineral medium / river water: yes
- Continuous darkness: yes


TEST SYSTEM
- Culturing apparatus: incubator
- Number of culture flasks/concentration:
10 bottles containing only inoculum = inoculum blank
10 bottles containing only inoculum + 1 g silica gel/bottle = inoculum blank (silica)
10 bottles containing test substance , inoculum and 1 g silica gel / bottle =test
6 bottles containing sodium acetate and inoculum = reference control
- Method used to create aerobic conditions: aeration with pressured air
- Measuring equipment: oxygen meter


SAMPLING
- Sampling frequency: day 0, 7, 14, 21 ,28, 42
- Sampling method: the bottles from day 0, 7, 14 , 21 and 28 were discarded after the oxygen measurement (reference control is measured on day 0,7 and 14) (two duplicate bottle for each measurement)


CONTROL AND BLANK SYSTEM
- Inoculum blank: yes
- Abiotic sterile control: no
- Toxicity control: no
- Reference control: yes



CALCULATIONS:
THOD N-oleyl-1,3-diaminopropane = 3.0 mg/mg
THOD sodiumacetate = 0.8 mg/mg

Oxygen consumption (mg/l) (BOD) = mean oxygen concentration (mg/l) inoculum blank - mean oxygen concentration (mg/l) test (or reference)
inoculum blank with silica gel is used for calculations of test
inoculum blank is used for calculations of referecence control

Biodegradation (%) = BOD/THOD *100

Parameter:

% degradation (O2 consumption)

Value:

66

Sampling time:

28 d

Remarks on result:

other: Over 60% biodegradation was not achieved within a period of 10 days immediately following the attainment of 10% biodegradation.

Validity criteria fulfilled:

yes

Remarks:

endogenous respiration of 1.0 mg/l; total mineralization of reference compound 81% at day 14; >0.5 mg/l O2 in all the bottles during the test; differences duplicates <20% at end test

Interpretation of results:

readily biodegradable

Conclusions:

N-oleyl-1,3-diaminopropane is biodegraded 66% at day 28 in the Closed Bottle test and should therefore be classified as readily biodegradable. Over 60% biodegradation was not achieved within a period of 10 days immediately following the attainment of 10% biodegradation. N-oleyl-1,3-diaminopropane is a mixture of alkyl-1,3-diaminopropanes and primary alkylamines. Moreover surfactants such as alkyl-1,3-diaminopropanes are degraded sequentially by a consortium of microorganisms (vanGinkel 1996). The degradation curve will therefore be the sum of a number of growth
curves. The 10-day time window criterion was developed on the assumption that a test substance is degraded according to a “standard” growth curve in ready biodegradability tests. The time-window should be ignored as a pass fail criterion because of these scientific
reasons.

Executive summary:

In order to assess the biotic degradation, a ready biodegradability test was performed which allows the biodegradability to be measured in an aerobic aqueous medium. The ready biodegradability was determined in the Closed Bottle test performed according to slightly modified OECD, EU and ISO Test Guidelines, and in compliance with the OECD principles of Good Laboratory Practice. N-oleyl-1,3-diaminopropane in the presence of silica gel did not cause a reduction in the endogenous respiration. The test substance is therefore considered to be non-inhibitory to the inoculum. N-oleyl-1,3-diaminopropane was biodegraded 66% at day 28 in the Closed Bottle test. Hence this substance should be classified as readily biodegradable. The test is valid as shown by an endogenous respiration of 1.0 mg/L and by the total mineralization of the reference compound, sodium acetate. Sodium acetate was degraded 81% of its theoretical oxygen demand after 14 days. Finally, the most important criterion was met by oxygen concentrations >0.5 mg/L in all bottles during the test period.

Description of key information

Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates is considered as readily biodegradable. 

Key value for chemical safety assessment

Biodegradation in water:

readily biodegradable

Type of water:

freshwater

Additional information

A ready biodegradability test was performed according to OECD guideline 301D and GLP requirements in order to allow the measurement of the biodegradability of Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, diacetates in an aerobic aqueous medium (Van Ginkel, 1990). The test substance was biodegraded to 76% after 28 days. The validity criteria were fulfilled. The test substance can be considered as readily biodegradable.

 

Similar biodegradability tests have been performed for N-C16-18-alkyl-(evennumbered)C18 unsaturated) propane-1,3-diamine and (Z)-N-9-octadecenylpropane-1,3-diamine. Similarly to other diamines and the diamine acetate, these substance were shown to be readily biodegradable indicating that the presence of acetate has no influence on the degradability of the diamine acetate. For more details, see the read-across justification document attached.