fatty acid chloride, C12, reaction product with sodium glutamate and sodium hydroxide

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

11.93 - 12.93

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 E (Ready biodegradability: Modified OECD Screening Test)

Deviations:

not specified

GLP compliance:

no

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, non-adapted

Details on inoculum:

STP Niederrad

Duration of test (contact time):

28 d

Initial conc.:

>= 5 - <= 40 mg/L

Based on:

DOC

Parameter:

% degradation (DOC removal)

Value:

88

Sampling time:

28 d

Validity criteria fulfilled:

not specified

Interpretation of results:

readily biodegradable

Conclusions:

The test item is readily biodegradable in 10 -d window 86 % and after 28 d (88%)

Executive summary:

The test item is readily biodegradable in 10 -d window 86 % and after 28 d (88%)

Reference 2

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Non-GLP guideline study with acceptable restrictions (test performance is briefly reported)

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 E (Ready biodegradability: Modified OECD Screening Test)

GLP compliance:

no

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

- - Source of inoculum/activated sludge: not specified

Duration of test (contact time):

28 d

Initial conc.:

>= 5 - <= 40 mg/L

Based on:

DOC

Parameter followed for biodegradation estimation:

DOC removal

Details on study design:

TEST CONDITIONS
- Composition of medium: anorganic nutrient solution containing micronutrients and essential vitamines
- Test temperature: 20 - 25 °C
- Continuous darkness: yes

TEST SYSTEM
- Other: The test solution was placed on a mechanical shaker

SAMPLING
- Sampling frequency: Samples were taken after 7, 14, 21, 27 and 28 days.

CONTROL AND BLANK SYSTEM
- Inoculum blank: Yes, a trial without the test or reference substance was conducted

Reference substance:

benzoic acid, sodium salt

Parameter:

% degradation (DOC removal)

Value:

94.4

Sampling time:

28 d

Details on results:

The test substance reached a biodegradation of 94.4% within a test period of 28 days. The 10 day-window was met. Thus, the test substance is readily biodegradable according to OECD criteria.

Results with reference substance:

The reference substance was biodegraded to 97.9% within 28 days.

Table 1: Biodegradation of reference and test substance in % DOC removal throughout the test period of 28 days

Days	Reference substance	Test substance
7	93.0	81.4
14	97.8	90.5
21	97.2	93.0
27	97.8	95.5
28	97.9	94.4

Validity criteria fulfilled:

not specified

Interpretation of results:

readily biodegradable

Reference 3

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

10.2004 - 11-2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, non-adapted

Initial conc.:

50 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

CO2 evolution

Reference substance:

acetic acid, sodium salt

Parameter:

% degradation (CO2 evolution)

Value:

100

Sampling time:

21 d

Results with reference substance:

82 % after 14 days and 100 % after 28 days

Validity criteria fulfilled:

yes

Interpretation of results:

readily biodegradable

Conclusions:

The test item reached the 10 % level (beginning of biodegradation) after an adaptation phase of 4 days. The biodegradation was rapid and reached the pass level of 60 % already on day 10. The biodegradation came to a maximum of 100 % after 21 days.

Executive summary:

The ready biodegradability was determined with a non adapted activated sludge for the test item over a period of 28 days in the Modified Sturm Test. The test item was tested with a concentration of 50 mg/L in duplicates, corresponding to a carbon content (TOC) of 12.4 mgC/L in the test vessels. The biodegradation of the test item was followed by titrimetric analyses of the quantity of CO2 produced by the respiration of bacteria. The degradation was finished on day 28 by acidification; the last titration was made on day 29. The percentage Co2 production was calculated in relation to the theoretical CO2 (ThCo2) of the test item. The biodegradation was calculated for each titration time.

To check the activity of the test system sodium acetate was used as functional control. The percentage degradation of the functional control reached the pass level of 60 % after 8 days. In the toxicity control containing both test and reference item a biodegradation rate of 82 % occurred within 14 days and came to a maximum of 100 % after 28 days. The test item in the toxicity control did not inhibit the biodegradation of the reference item.

The test item reached the 10 % level (beginning of biodegradation) after an adaptation phase of 4 days. The biodegradation was rapid and reached the pass level of 60 % already on day 10. The biodegradation came to a maximum of 100 % after 21 days.

Reference 4

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2021-11-18 to 2021-12-17

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

Test item fatty acid chloride, C12, reaction product with sodium glutamate and sodium hydroxide
Batch number LAB AA101.20 Grobgut
Purity 100 % (UVCB)
Chemical composition Disodium 2- (dodecanoylamino)pentanedioate (main component)
Water solubility 7.8 g/L
pH 9.9 at concentration of: 7800 [mg/L]
TOC* 46.0 %
Appearance colorless powder (solid)
Expiry date 2023-08-23
Recommended storage Room temperature

* The TOC was measured at the test facility with a carbon analyser.

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, non-adapted

Details on inoculum:

- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure):
Municipal sewage treatment plant, D-31137 Hildesheim, Germany
- Receipt: 2021-11-15
- Pretreatment/Concentration of sludge:
The activated sludge was washed twice with chlorine free tap water. After the second washing the settled sludge was resuspended in mineral salts medium and was maintained in an aerobic condition by aeration with CO2 free air for two days before test start. Further treatment see section ‘preparation of the test vessels’. 4.27 mL/ L of this mixture were used to initiate inoculation (25.0 mg/L dw).

Duration of test (contact time):

28 d

Initial conc.:

23 other: mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

TEST CONDITIONS
- Composition of medium: Mineral salts medium acc. to OECD 301 B / CO2 Evolution Test
- Test temperature: Nominal 22 ± 2 °C, actually measured 21.0 – 23.0 °C
- Dispersion treatment: Continuous stirring
- Aeration: 30 - 100 mL/min
- Photoperiod: Low light conditions (brown glass bottles)

TEST SYSTEM
- Culturing apparatus: 5000 mL brown glass flasks
- Number of culture flasks/concentration: 1 for the reference item, 1 for toxicity control (test and reference item), 2 for the control, 2 for the test item
- Method used to create aerobic conditions: Aeration with 30 - 100 mL/min
- Measuring equipment: Visual check of aeration twice per day
- Details of trap for CO2 and volatile organics if used:
CO2 absorption vessels were connected to the air outlets of the incubation vessels via a series of 3 gas wash bottles, each containing 100 mL of a 0.0125 mol/L Ba(OH)2 solution.

- Course of the study:
The concentration of the test item and the theoretical CO2 production (ThCO2) were calculated based on the carbon content.

The following incubation vessels will be prepared:
- two for the inoculum control (C1, C2)
- one for the functional control (R1)
- two for the test item concentration (P1, P2)
- one for the toxicity control (T1)

The necessary amounts of ultrapure water, mineral salts medium and inoculum were placed in each incubation vessel. The vessels were aerated for 24 h with CO2 free air. After 24 h the CO2 absorption vessels were connected to the air outlets of the incubation vessels via a series of 3 gas wash bottles, each containing 100 mL of a 0.0125 mol/L Ba(OH)2 solution.

The test and reference item were weighed out. The test item was
weighed out into small beakers. A defined amount of ultrapure
water will be added to the test item. The test item dispersions and
the reference item will be transferred to the respective incubation
vessels with ultrapure water. The vessels were made up to 3 L
with ultrapure water and connected to the system for the
production of CO2 free air.


On day 28, 1 mL 37 % HCl was added to each of the vessels. Aeration was continued for further 24 h and the quantity of CO2 released was determined.



SAMPLING
- Sampling frequency:
Back titration of the residual Ba(OH)2 with 0.05 N HCL was carried out three times a week during the first ten days and thereafter twice weekly.
- Sampling method:
For each titration the first gas wash bottle was removed and a new bottle was connected to the last one.

CONTROL AND BLANK SYSTEM
- Inoculum blank: Test medium without test and/or reference item
- Abiotic sterile control: No
- Toxicity control: Test item and reference item in test concentration


STATISTICAL METHODS:
- The theoretical production of carbon dioxide (ThCO2) of the test item and functional control was calculated by the carbon content (1) and the molecular formula (2), respectively.

ThCO2 [mgCO2/mg] = 3.67 * TOC [mgC/mg test item] (1)

ThCO2 [mgCO2/mg] = (C-Atoms *molecular weight of CO2)/molecular weight of reference item) (2)


- The produced CO2 was calculated by: 1 mL HCl (c = 0.05 mol/L) = 1.1 mg CO2

- The net amount of CO2 produced was calculated by correcting the results of the test item and functional control for endogenous CO2 production of the inoculum controls.

- The biodegradation was calculated from the ratio theoretical CO2 production to net CO2 production:

Degradation [%] = (net CO2 * 100)/(THCO2 [mg CO2/3L])

Reference substance:

benzoic acid, sodium salt

Key result

Parameter:

% degradation (CO2 evolution)

Value:

64

Sampling time:

28 d

Details on results:

Based on the carbon content a ThCO2 of 1.69 mg CO2/mg test item was calculated. A test concentration of 23 mg/L, corresponding to a carbon content of 10.6 mg C/L in the test vessels was selected.

The adaptation phase of the functional control changed within 4 days into the degradation phase (degradation > 10 %). The course of the degradation was fast and the functional control reached the pass level of 60 % within 8 days and a biodegradation of 80% on day 28. The validity criterion degradation > 60 % after 14 days was fulfilled.

In the toxicity control containing both test item and reference item a biodegradation of 61 % was determined within 14 days, with 71 % after 28 days. The biodegradation of the reference item was not inhibited by the test item in the toxicity control.

The biodegradation of the test item is shown in comparison to the readily degradable functional control and the toxicity control. The 1st test item replicate reached the 10 % level (beginning of biodegradation) within 8 days and slightly missed the 60% pass-level at test end. The 2nd test item replicate reached the 10 % level (beginning of biodegradation) within 4 days and the 60 % pass level within 11 days. The mean biodegradation on day 28 was 64 %.

In the inoculum control the total CO2 production was 48.1 mg CO2/L after 28 days.

Results with reference substance:

In the toxicity control containing both test item and reference item a biodegradation of 61 % was determined within 14 days, with 71 % after 28 days. The biodegradation of the reference item was not inhibited by the test item in the toxicity control.

Validity criteria fulfilled:

yes

Interpretation of results:

readily biodegradable, but failing 10-day window

Conclusions:

Under the test conditions the test item is classified as readily biodegradable with a percentage biodegradation of 64 % within the 28-day period of the study.

Executive summary:

The ready biodegradability of the test item fatty acid chloride, C12, reaction product with sodium glutamate and sodium hydroxide (batch no.: LAB AA101.20 Grobgut) was determined with a non-adapted activated sludge over a test period of 28 days in the Modified Sturm Test. The study was conducted from 2021-11-18 to 2021-12-17 according to OECD 301 B at the test facility. The test item was tested at a concentration of 23 mg/L with 2 replicates corresponding to a carbon content (TOC) of 10.6 mg C/L in the test vessels. The test vessels were incubated at low light conditions and at a temperature of 22 ± 2 °C.

The biodegradation of the test item was followed by titrimetric analysis of the quantity of CO2 produced by the respiration of bacteria. The degradation was stopped on day 28 by acidification of the test solutions. The last titration was made on day 29 after residual CO2 had been purged from the test solutions over a period of 24 hours. The percentage CO2 production was calculated in relation to the theoretical CO2 production (ThCO2) of the test item. The biodegradation was calculated for each titration time.

To check the activity of the test system sodium benzoate was used as functional control. The percentage degradation of the functional control reached the pass level of 60 % within 8 days and a biodegradation of 80 % on day 28.

In the toxicity control containing both test and reference item a biodegradation of 61 % was determined within 14 days, with 71 % after 28 days. The biodegradation of the reference item was not inhibited by the test item in the toxicity control.

The biodegradation of the test item is shown in comparison to the readily degradable functional control and the toxicity control. The 1st test item replicate reached the 10 % level (beginning of biodegradation) within 8 days and slightly missed the 60% pass-level at test end. The 2nd test item replicate reached the 10 % level (beginning of biodegradation) within 4 days and the 60 % pass level within 11 days. The mean biodegradation on day 28 was 64%.

Under the test conditions the test item is classified as readily biodegradable with a percentage biodegradation of 64 % within the 28-day period of the study.

Biodegradation of the Test Item in Comparison to the Functional Control and Toxicity Control

	Biodegradation [%]
	Study Day [d]
	6	14	21	28
Test Item, 1^stReplicate	6	35	52	58
Test Item, 2^ndReplicate	47	64	68	69
Functional Control	48	76	80	80
Toxicity Control Test item + Reference item	24	61	66	71

Description of key information

In a key study the test item is classified as readily biodegradable with a percentage biodegradation of 64 % within the 28-day period of the study.

In three supporting studies (structural analogue or surrogate) ready biodegradability could be shown in the 10 -d windows and after the 28 day period.

In the supporting study with N-Lauroyl glutamic acid, sodium salt in aqueous solution a biodegradation of 100% was achieved after 28 days.

Key value for chemical safety assessment

Biodegradation in water:

readily biodegradable

Type of water:

freshwater