Oleic acid, compound with (Z)-octadec-9-enylamine (1:1)

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:

28 Nov 2017 - 04 Jan 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

July 1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Hessisches Ministerium für Umwelt, Klimaschutz, Landwirtschaft und Verbraucherschutz, Wiesbaden, Germany

Test material

Test material information

Constituent 1

Reference substance name:

Oleic acid, compound with (Z)-octadec-9-enylamine (1:1)

EC Number:

248-813-1

EC Name:

Oleic acid, compound with (Z)-octadec-9-enylamine (1:1)

Cas Number:

28065-97-6

Molecular formula:

C34H69NO2 C36H67NO2 C36H69NO2 C36H71NO2 C36H73NO2

IUPAC Name:

oleic acid, compound with (Z)-octadec-9-enylamine (1:1)

Study design

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, domestic, non-adapted

Details on inoculum:

- Source of inoculum/activated sludge: activated sludge from sewage plant at Taunusstein-Bleidenstadt, Germany
- Storage conditions: sludge was aerated for 1d with CO2-free compressed air (use of a 'CO2 scrubbing apparatus' described in the OECD Test Guideline 301 B)
- Storage length: 1d
- Pretreatment: The activated sludge was washed twice with mineral nutrient solution. After resolution with mineral nutrient medium the sludge was aerated by means of compressed humidified air for about 4 h. Before use as inoculum for the CO2-Evolution-Test the sludge was homogenised in a "Waring Blender" at low speed for 2 min. The filtrate was used as inoculum. Within the study the test solutions were stirred by means of magnetic stirrers in order to distribute the test item (or control item) and oxygen in a way to dissolve them at the maximum solubility.
- Water filtered: yes
- Type and size of filter used, if any: activated sludge was filtered through a cotton filter previously carefully rinsed with deionised water. The filtrate was used as inoculum (1% of the final volume of the test solution) on the same day of preparation.

Duration of test (contact time):

28 d

Initial test substance concentration

Initial test substance concentration 1

Initial conc.:

47.144 other: mg/3.5 L

Based on:

test mat.

Initial test substance concentration 2

Initial conc.:

ca. 10 mg/L

Based on:

TOC

Initial test substance concentration 3

Initial conc.:

2 852 other: mg CO2/g test item

Based on:

ThCO2

Parameter followed for biodegradation estimation

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

TEST CONDITIONS
- Composition of medium: according to the guideline
- Solubilising agent: dichloromethane solvent was evaporated passively under a hood
- Test temperature: 20.2 to 22.3 °C
- Aeration of dilution water: The aeration rate of the test system was controlled at 4 L/h being assured by precise flowmeters

TEST SYSTEM
- Culturing apparatus: 5-L amber carboys closed (fill volume: 3500 mL) with stoppers with tubing for gas inlet and outlet (gas exit line)
- Number of culture flasks/concentration: 2 (per test item) and 1 reactor (for toxicity control, blank and reference, respectively)
- Method used to create aerobic conditions: Before starting the test, the mineral nutrient solution with inoculum but without any test component were aerated for 1d with CO2-free compressed air, in order to purge the system of carbon dioxide. Hereafter the test was started by addition of the test item. The gas outlet (exit air line) was connected to at least three CO2 absorber bottles filled with 100 mL 0.025 N Ba(OH)2 (connection in series), and bubbling of CO2 free compressed air through the solution was continued. Periodically the CO2 absorber nearest the test vessel was removed for analysis, and a new CO2 absorber was connected farest from the carboy. Precipitation of BaCO3 in the second CO2 trap indicated that the absorber bottle nearest the test vessel had to be changed and analysed for CO2.
- Measuring equipment: Degradation was calculated as a percentage of the theoretical CO2 (%TCO2) that should have been produced from the organic matter of the test item by complete combustion. The CO2 being generated was calculated to the nearest 0.01 mg and biodegradation values were rounded up to the nearest full percent (Schott Instruments, Titronic universal Ser.-No. 00693538).
- Test performed in open system: yes
- Details of trap for CO2 and volatile organics if used: Resting Ba(OH)2 (0.025 N) in the CO2 trap removed from the trap system was titrated with HCl (0.05 N) using phenolphthalein as an indicator. Each millilitre HCl of difference between the blank control and the test series with the test item titrated corresponds to 1.1 mg of CO2 produced. For this purpose values of titration for the 'Blank' were subtracted from that of the test unit with the test item.

SAMPLING
- Sampling frequency: t2d, t5d, t9d, t13d, t19d, t23d, t28d, and t29d
- Sampling method: At t28 d 1 mL conc. HCl was given into the test solutions in order to stop the test and to make the dissolved CO2 or inorganic carbon volatile and to purge the system of CO2

CONTROL AND BLANK SYSTEM
- Inoculum blank: yes, 2 reactors
- Toxicity control: yes, 1 reactor
- Reference control: yes, 1 reactors

Reference substance

Reference substance:

benzoic acid, sodium salt

Results and discussion

% Degradation

Parameter:

% degradation (CO2 evolution)

Value:

79

Sampling time:

28 d

Remarks on result:

other: mean of two replicates

BOD5 / COD results

Results with reference substance:

The biodegradation of the reference compound reached the pass level of 75% ThCO2 by day 9.

Any other information on results incl. tables

Table 1: 1st Test Solution, Test Item

Time [d]	Concentration: 47.144 mg / 3.5 L  ThCO2: 134.5 mg CO2/ 3.5 L
	mg CO2 produced in the Test solution, cumulative	% TCO2 (= % Degradation) ***
2	3.18	2
5	23.63	18
9	76.14	57
13	84.82	63
19	92.87	69
23	100.77	75
28	107.31	80 ****
29	112.13	83 *****

*TOC: Total Organic Carbon

**ThCO2: Theoretical amount of CO2 that can be generated by the test item

***% TCO2: Percentage of total CO2 the test item has generated in relation to the ThCO2

****The test solution was stopped by the addition of 1 mL conc. HCl;

*****The difference between this titration and that before is due to the liberation of CO2 after acidification of the test solution.

 

Table 2: 2nd Test Solution, Test Item

Time [d]	Concentration: 47.144 mg / 3.5 L  ThCO2: 134.5mg CO2/ 3.5 L
	mg CO2 produced in the Test solution, cumulative	% TCO2 (= % Degradation) ***
2	2.58	2
5	25.03	19
9	71.16	53
13	80.53	60
19	88.74	66
23	92.97	69
28	96.47	72 ****
29	101.42	75 *****

*TOC: Total Organic Carbon

**ThCO2: Theoretical amount of CO2 that can be generated by the test item

***% TCO2: Percentage of total CO2 the test item has generated in relation to the ThCO2

****The test solution was stopped by the addition of 1 mL conc. HCl;

*****The difference between this titration and that before is due to the liberation of CO2 after acidification of the test solution.

 

Table 3: Results of Sodium Benzoate

Time [d]	Concentration: 126.6 mg / 3.5 L ThCO2: 269.0 mg CO2/ 3.5 L
	mg CO2 produced in the Test solution, cumulative	% TCO2 (= % Degradation)
2	73.04	27
5	141.66	53
9	201.77	75
13	215.00	80
19	221.80	82
23	227.51	85
28	231.41	86 ****
29	237.07	88 *****

*TOC: Total Organic Carbon

**ThCO2: Theoretical amount of CO2 that can be generated by the test item

***% TCO2: Percentage of total CO2 the test item has generated in relation to the ThCO2

****The test solution was stopped by the addition of 1 mL conc. HCl;

*****The difference between this titration and that before is due to the liberation of CO2 after acidification of the test solution.

Table 4 Toxicity Control - Oleylamine Oleate and Sodium Benzoate

Time [d]	Concentration: 126.4 mg Sodium Benzoate +47.144 mg Test Item ThCO2: 403.1 mg CO2/ 3.5 L
	mg CO2 produced in the Test solution, cumulative	% TCO2 (= % Degradation)
2	67.53	17
5	150.74	37
9	267.15	66
13	283.95	70
19	305.74	76
23	323.30	80
28	333.95	83 ****
29	347.89	86 *****

*TOC: Total Organic Carbon

**ThCO2: Theoretical amount of CO2 that can be generated by the test item

***% TCO2: Percentage of total CO2 the test item has generated in relation to the ThCO2

****The test solution was stopped by the addition of 1 mL conc. HCl;

*****The difference between this titration and that before is due to the liberation of CO2 after acidification of the test solution.

Table 5: Validity criteria

Criterion from the guideline	Outcome	Validity criterion fulfilled
Difference of extremes of replicate values of the removal of the test chemical at the plateau, at the end of the test or at the end of the 10-d window, as appropriate, is less than 20%.	<20%	Yes
Percentage degradation of the reference compound has reached the pass levels by day 14.	70%	Yes
The toxicity control should degrade to at least 35% (based on DOC) or at least 25% (based on ThOD or ThCO2) within 14 d.	70%	Yes
The total CO2 evolution in the inoculum blank at the end of the test should not normally exceed 40 mg/L medium.	<40 mg/L	Yes

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”

Interpretation of results:

readily biodegradable

Conclusions:

Calculated from the organic carbon content of the test item and the measured CO2 generation, 83% of the theoretical CO2 (ThCO2) has been generated by the test item within 28d of test period in the first culture with 47.144 mg of test item. 75.4% of the theoretical CO2 (ThCO2) has been generated by the test item within 28d in the second culture with 47.144 mg of test item. The calculated mean degradation value of the test item was 79%. Even the “10-days-window” was met though it is not considered for the kind of test item tested. Thus, the test item should be regarded as “readily biodegradable". The control item sodium benzoate was degraded 88%, and the conditions of the "10-days-window" were met within 9 days (75% degradation). The total CO2-evolution of the Blank was 93.4 mg CO2. Thus the test is regarded to be valid. The results obtained with the toxicity control indicate that there was no toxicity of the test item towards microorganisms at the concentration used within the test as a degradation value of >30% (here: 86%) was achieved in total within that treatment.