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REACH

L-Aspartic acid, N,N'-1,2-ethanediylbis-, sodium salt (1:3)

EC number: 416-530-4 | CAS number: 178949-82-1

Life Cycle description
Uses advised against

Environmental fate & pathways

Adsorption / desorption

Currently viewing:

Administrative data

Link to relevant study record(s)

Reference 1

Endpoint:

adsorption / desorption: screening

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

02.04.93 - 07.04.93

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Principles of method if other than guideline:

Adsorption coefficients were determined for the partitioning of the test substance at two different concentrations between an aqueous phase and a known mass of activated sewage sludge either pre-exposed to the test substance (adapted) or not (nonadapted). Inactivation (sterilisation) of incubations containing adapted sludge was attempted using sodium azide (Na3N) to prevent biodegradative losses of the test substance during equilibration. However, Na3N was not an effective sterilant with respect to microorganisms capable of degrading the test substance and as such biodegradative losses of the test substance from these incubations resulted in low recoveries and inaccurate determinations of adsorption coefficients. The ineffectiveness of Na3N to inactivate the adapted sludge was assessed in a parallel study in which biodegradative mineralisation of radiolabelled test substance was quantified by liquid scintillation counting (LSC) of evolved 14C-CO2 trapped in ethanolamine.

Adapted and nonadapted sludge which had been incubated with the test substance during the adsorption tests was subsequently used to determine coefficents of desorption.

Liquid-solid partitioning in adsorption and desorption tests was assessed using a mixture of radiolabelled and non-radiolabelled test substance with quantification of test substance concentrations of both the liquid phase and sludge suspended solid phase by LSC.

A preliminary study (equivalent to test controls containing no sludge) concluded adsorption of the test compound to labware (e.g. glass vessels used for incubations, pipettes etc) was insignificant.

Methodological weaknesses arise from the following:
- no report of tests undertaken to establish equilibration time between test substance and sludge
- no report of pH measurment or control
- no report of blank tests (i.e. sludge + aqueous phase only)
- no report of control of ionic strength of aqueous phase
- no report of temperature control

GLP compliance:

yes

Type of method:

batch equilibrium method

Media:

sewage sludge

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
no data

Radiolabelling:

yes

Test temperature:

no data

Details on study design: HPLC method:

not applicable

Analytical monitoring:

yes

Details on sampling:

Adsorption tests:

1. Centrifugation of 100 mL test substance/sludge after 3 h incubation.
2. 2 x 2 mL supernatent samples taken by pipette and added to 17 mL Lumagel (scintillation cocktail) for liquid scintillation counting (LSC).
3. Supernatent discarded and sludge solids resuspended in 100 mL water.
4. 2 x 1 mL suspended sludge solids samples taken by pipette and added to 9 mL deionised water and 10 mL Lumagel for LSC analysis.

Desorption tests:

5. Centrifugation of 100 mL water/sludge after 3 hrs incubation.
6. 2x 2 mL supernatant samples taken by pipette and added to 17 mL Lumagel (scintillation fluid) for LSC.
7. Supernatent discarded and sludge solids resuspended in 100 mL water.
8. 2 x 1 mL suspended sludge solids samples taken by pipette and added to 9 mL deionised water and 10 mL Lumagel for LSC analysis.

Details on matrix:

- Type of sludge: activated domestic sewage sludge either pre-exposed or non-exposed to the test substance.
- Source of sludge: adapted sludge: obtained from previous CAS- test ECM ETS 465/03; non-adapted sludge: obtained from Bocholt Waste Water Treatement Works, Belgium.
- Pretreatment: none reported

PREPARATION OF SLUDGE SOLIDS
- Separation from wastewater: no data
- Further preparation procedures: no data
- Details of lyophilization: not performed
- Details of desiccation: not performed

Details on test conditions:

PRELIMINARY STUDY

To assess losses of test substance to test vessels (glass centrifuge tube) and pipette tips:

Preliminary vessels 1 and 2:

100 mL of 0.1 mg/L non-radiolabelled test substance
+ 0.050 mL radiolabelled test substance stock solution

The test container was rinsed with 3 x 10 mL of methanol to give a total rinse volume of 30 mL from which 10 mL was analysed by LSC.
The pipette tip was rinsed with 3 x 1 mL methanol and the 3 mL total rinse volume was analysed by LSC.

TEST CONDITIONS
- Buffer: none
- pH: not reported
- Suspended solids concentration: determined at 105 oC: adapted sludge = 1.89 g/L and non-adapted sludge = 2.35 g/L; determined at 550 oC: adapted sludge = 1.53 g/L and non-adapted sludge = 1.92 g/L

TEST SYSTEM
- Type, size and further details on reaction vessel: 200 mL glass centrifuge tube
- Water filtered (i.e. yes/no; type of size of filter used, if any): no data
- Amount of sludge and water per treatment: 100 mL
- Measuring equipment: LSC
- Test performed in closed vessels due to significant volatility of test substance: no data
- Test performed in open system: no data
- Method of agitation: continuous aeration
- Method of preparation of test solution:

The test solutions were prepared by mixing the appropriate volumes of activated sludge, radiolabelled test substance, stock solution of non-labelled test substance and sodium azlde solution (20 mg/mL) as per the following:

vessels 1 and 2:

100 mL non-adapted activated sludge
+ 0.050 mL radiolabelled test substance stock solution
+ 0.1 mL non-radiolabelled test sustance stock solution

vessels 3 and 4:

100 ml non-adapted activated sludge
+ 0.050 mL radiolabelled test susbtance stock solution
+ 1.0 mL stock solution of the non-labelled test substance stock solution

vessels 5 and 6:

100 ml adapted activated sludge
+ 0.050 mL radiolabelled test substance stock solution
+ 0.1 mL non-radiolabelled test substance stock solution
+ 1 mL sodium azide solution

vessels 7 and 8:

100 mL activated sludge
+ 0.050 mL labeled test matertal
+1.000 mL stock solution of the non-labeled test matertal
+ 1 mL sodium azide solution

- Are the residues from the adsorption phase used for desorption: yes. After centrifugation and sampling, supernatants from adsorption tests were discarded and vessels refilled with 100 mL water. After 3 hours incubation, desorption coefficients were determined by analysis of aqueous and soilid phases for test substance.

Computational methods:

Sorption Coefficient
Kd = Concentration of test material on solids/Concentration of test materials in water = (DPM on solids/DPM in water) x (Wt of water/Wt of solids)

Key result

Sample No.:

Type:

Value:

388 dimensionless

% Org. carbon:

100

Sample No.:

Type:

Value:

455 dimensionless

% Org. carbon:

100

Sample No.:

Type:

Value:

372 dimensionless

% Org. carbon:

100

Sample No.:

Type:

Value:

295 dimensionless

% Org. carbon:

100

Details on results (HPLC method):

not applicable

Adsorption and desorption constants:

Adsorption coefficients (Kd abs)

Vessels 1 & 2 (0.1 mg/L test substance + non-adapted sludge): mean Kd abs = 40

Vessels 3 & 4 (1.0 mg/L test substance + non-adapted sludge): mean Kd abs = 37

Vessels 5 & 6 (0.1 mg/L test substance + adapted sludge): mean Kd abs = 71 ((low mass balance (see Table 2) indicates biodegradative loss of test substance and inaccurate adsorption coefficient)

Vessels 7 & 8 (1.0 mg/L test substance + adapted sludge): mean Kd abs = 90 (low mass balance (see Table 2) indicates biodegradative loss of test substance and inaccurate adsorption coefficient)

Desorption coefficients (Kd des)

Vessels 1 & 2 (0.1 mg/L test substance + non-adapted sludge): mean Kd des = 421

Vessels 3 & 4 (1.0 mg/L test substance + non-adapted sludge): mean Kd des = 334

Vessels 5 & 6 (0.1 mg/L test substance + adapted sludge): mean Kd des = 1034 ((low mass balance (see Table 2) indicates biodegradative loss of test substance giving inaccurate desorption coefficient)

Vessels 7 & 8 (1.0 mg/L test substance + adapted sludge): mean Kd des = 1298 (low mass balance (see Table 2) indicates biodegradative loss of test substance giving inaccurate desorption coefficient)

Also See Table 2

Recovery of test material:

See Table 2

Concentration of test substance at end of adsorption equilibration period:

See Table 2

Concentration of test substance at end of desorption equilibration period:

See Table 2

Transformation products:

not specified

Details on results (Batch equilibrium method):

- Preliminary tests assessed adsorption of the test substance onto labware to be insignificant (see Table 1).
- Losses of test substance from vessels 5 to 8 in which the test substance was incubated with adapted sludge were attributed to biodegradation. The adsorption coefficients determined for these incubations were therefore unreliable (see Table 2).

Table 1:

		Preliminary vessel 1	Preliminary vessel 2
test substance concentration (mg/L)		0.1	0.1
t₀spike (dpm)		214382	214382
supernatant	dpm	218370	208677
supernatant	% of t₀	101.86	97.34
test vessel rinses	dpm	1035	695
test vessel rinses	% of t₀	0.48	0.32
pipette tip rinses	dpm	35	22
pipette tip rinses	% of t₀	0.02	0.01
% recovery		102	98

Table 2: LSC measurements for the adsorption/desorption test and calculations of the Kd values.

Vessel #	1	2	3	4	5	6	7	8
t₀-spike (dpm)	214382	214382	214382	214382	214382	214382	214382	214382
ADSORPTION
dpm on solids (dpm/g)	79894	82011	74603	75661	71489	75745	80000	81702
dpm In water (dpm/g)	1953	2055	1978	2086	1262	894	1146	740
Kd abs	41	40	38	36	57	85	70	110
% recovery	98	103	99	104	67	50	62	43
DESORPTION
dpm on solids (dpm/g)	46561	48677	37566	35450	47660	62979	50638	63830
dpm In water (dpm/g)	120	107	101	120	63	48	43	45
Kd des	388	455	372	295	757	1312	1178	1418
% recovery	138	128	122	131	104	no	86	102

NOTE: Adsorption coefficents (Kd abs) and desorption coefficients (Kd des) for vessels 5 to 8 are not reliable due to biodegradative losses of the test compound.

Validity criteria fulfilled:

not applicable

Conclusions:

In a GLP study, adsorption of trisodium EDDS to activated sludge was considered minimal.

Executive summary:

In a GLP study, adsorption (Kd abs) and desorption (Kd des) coefficients were determined (using the batch equilibrium method) for incubations of two different concentrations (0.1 and 1.0 mg/L) of trisodium EDDS with activated domestic sewage sludge that had either been pre-exposed (adapted) to the test substance or not (non-adapted). The addition of sodium azide to incubations with adapted sludge was ineffective in preventing the biodegradation of the test substance and therefore Kd abs and Kd des values derived from these tests were considered unreliable. Incubations of the test substance at concentrations of 0.1 and 1.0 mg/L with non-adapted sludge yielded K abs values of 40 and 37, respectively. Corresponding Kd des values of 421 and 334 were determined using the sludge residues from these adsorption tests. A 14C-radiolabelled test substance tracer was used to assess partitioning between the aqueous and suspended solid phases of the sludge. A preliminary study determined the adsorption of test substance, trisodium EDDS, to labware (i.e. glass test vessel and pipette tips) to be insignificant.

From the Kd abs and Kd des values derived, it was concluded that adsorption of trisodium -EDDS to activated sewage sludge was minimal.

Reference 2

Endpoint:

adsorption / desorption: screening

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

From May 24, to June 01, 1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 106 (Adsorption - Desorption Using a Batch Equilibrium Method)

Deviations:

yes

Remarks:

See section 'Any other information on materials and methods'

GLP compliance:

yes (incl. QA statement)

Type of method:

batch equilibrium method

Media:

soil

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
No data

Radiolabelling:

Test temperature:

Room temperature

Details on study design: HPLC method:

Not applicable

Analytical monitoring:

yes

Details on sampling:

Concentrations of test sample: 25.1 mg/L in 0.01 M CaCl2

Details on matrix:

Soil I (Samples #1 and #2):
COLLECTION AND STORAGE
- Geographic location: Wageningen, The Netherlands.
- Collection procedures: no data
- Sampling depth (cm): no data
- Storage conditions: no data
- Storage length: no data
- Soil preparation (e.g.: 2 mm sieved; air dried etc.): 2 mm sieved and air dried

PROPERTIES
- Soil texture
- % sand: 81.3
- % silt: 11.1
- % clay: 6.0
- Horizon: 0-30 cm
- Soil taxonomic classification: strong silty sand
- Soil classification system: no data
- Soil series: no data
- Soil order: no data
- pH: 4.0
- Dry weight content (%) - 99.6
- Organic carbon (%): 1.4
- CEC (meq/100 g): 4.7
- Carbonate as CaCO3: 0.2 %
- Insoluble carbonates (%): no data
- Extractable Cations (Ca, Mg, Na, K, H) (MEQ/100 g): Ca: 1.33; Mg: 0.17; Na: <0.01; K: 0.23; H: 5.5
- Special chemical/mineralogical features: no data
- Clay fraction mineralogy: no data
- Moisture at 1/3 atm (%): no data
- Bulk density (g/cm3): no data
- Biomass (e.g. in mg microbial C/100 mg, CFU or other): no data

Soil II (Samples #3 and #4):
COLLECTION AND STORAGE
- Geographic location: Kloosterburen, The Netherlands.
Collection procedures: no data
- Sampling depth (cm): no data
- Storage conditions: no data
- Storage length: no data
- Soil preparation (e.g.: 2 mm sieved; air dried etc.): 2 mm sieved and air dried

PROPERTIES
- Soil texture
- % sand: 41.7
- % silt: 39.8
- % clay: 13.6
- Horizon: 0-30 cm
- Soil taxonomic classification: strong sandy loam
- Soil classification system: no data
- Soil series: no data
- Soil order: no data
- pH: 7.5
- Dry weight content (%) - 99.9
- Organic carbon (%): 1.8
- CEC (meq/100 g): 10.9
- Carbonate as CaCO3: 3.1 %
- Insoluble carbonates (%): no data
- Extractable Cations (Ca, Mg, Na, K, H) (MEQ/100 g): Ca: 9.06; Mg: 1.0; Na: 0.15; K: 0.96; H: 0.6
- Special chemical/mineralogical features: no data
- Clay fraction mineralogy: no data
- Moisture at 1/3 atm (%): no data
- Bulk density (g/cm3): no data
- Biomass (e.g. in mg microbial C/100 mg, CFU or other): no data

Soil III (Samples #5 and #6):
COLLECTION AND STORAGE
- Geographic location: Beek, The Netherlands.
- Collection procedures: no data
- Sampling depth (cm): no data
- Storage conditions: no data
- Storage length: no data
- Soil preparation (e.g.: 2 mm sieved; air dried etc.): 2 mm sieved and air dried

PROPERTIES
- Soil texture
- % sand: 11.9
- % silt: 74.7
- % clay: 12.1
- Horizon: 0-30 cm
- Soil taxonomic classification: weak sandy loam
- Soil classification system: no data
- Soil series: no data
- Soil order: no data
- pH: 6.6
- Dry weight content (%) - 99.9
- Organic carbon (%): 1.1
- CEC (meq/100 g): 7.5
- Carbonate as CaCO3: 0.2%
- Insoluble carbonates (%): no data
- Extractable Cations (Ca, Mg, Na, K, H) (MEQ/100 g): Ca: 5.93; Mg: 0.61; Na: 0.22; K: 0.17; H: 1.0
- Special chemical/mineralogical features: no data
- Clay fraction mineralogy: no data
- Moisture at 1/3 atm (%): no data
- bulk density (g/cm3): no data
- Biomass (e.g. in mg microbial C/100 mg, CFU or other): no data

Details on test conditions:

TEST CONDITIONS
Three soil types were tested (Soil I, Soil II and Soil III). per soil type, triplicate 5 g samples of soil were pre-equilibrated/washed with 10 mL of distilled water in 50 mL glass vials for 5 days at room temperature with continuous agitation by tumbling. Thereafter, vials were centrifuged for 10 minutes at 170 g and 20oC and most of the supernatants removed. Adsorption/desorption screening was performed on the soil precipitates at room temperature. A 16 h equilibration time was used in adsorption and desorption testing. Soil residues used in adsorption testing were used.

TEST SYSTEM
- Buffer: no buffers used
- pH: pH of the test soils was measured prior to equilibration with test substance (method of pH measurment not reported). pH of test vessels (soil + test substance solution) was not measured.
- Suspended solids concentration: 5 g air dried soil

Adsorption screening test:
- Preparation of test solution: test substance dissolved in 0.01 M CaCl2
- Reaction vessel: 50 mL glass vial
- Water filtered: no data
- Amount of soil/water per treatment: 5 g soil (air-dried weight) per 25 mL test substance solution (25.1 test substance in 0.01 M CaCl2)
- Soil/solution ratio: 1:5
- Reaction vessels/concentration: per soil type (i.e. soil I, II and III) duplicate vessels contained 5 g soil (air-dried weight) and 25 mL of 25.1 mg/L test substance in 0.01 M CaCl2. Also per soil type a singular blank contained 5 g soil (air-dried weight) and 25 mL of 0.01 M CaCl2. A singular control contained 25 mL of 25.1 mg/L test substance in 0.01 M CaCl2 only to assess loss of the test substance due to adsorption to the interior of the vessel.
- Measuring equipment: for the liquid phase (i.e. test substance and CaCl2 solutions) quantities were determined gravimetrically. pH measurement prior to and after equilibration of test substance with soil was not reported.
- Test performed in closed vessels for agitation by continuous tumbling.
- Equilibration time: 16 h
- Cetrifugation: test vessels centrifuged for 10 minutes at 170 x g and 20oC for separation of soil and solution phases.
- Supernatants analysed by HPLC for test substance concentration.
- Soil residues from adsorption tests were used for desorption tests.

Desorption screening test:
- Reaction vessel: same 50 mL glass vial used in adsorption tests.
- Amount of soil/water per treatment: same 5 g soil (air-dried weight) used in adsorption tests per 25 mL 0.01 M CalCl2.
- Soil/solution ratio: 1:5
- Extractions: two sequential extractions
- Cetrifugation: test vessels centrifuged for 10 minutes at 170 x g and 20 oC for separation of soil and solution phases.
- Supernatents analysed by HPLC for test substance concentration.

Sample No.:

Duration:

16 h

Initial conc. measured:

130.97 mg/kg soil d.w.

Sample No.:

Duration:

16 h

Initial conc. measured:

135.84 mg/kg soil d.w.

Sample No.:

Duration:

16 h

Initial conc. measured:

142.47 mg/kg soil d.w.

Sample No.:

Duration:

16 h

Initial conc. measured:

142.82 mg/kg soil d.w.

Sample No.:

Duration:

16 h

Initial conc. measured:

135.39 mg/kg soil d.w.

Sample No.:

Duration:

16 h

Initial conc. measured:

137.05 mg/kg soil d.w.

Sample no.:

Duration:

32 h