N-ethyl-N-[2-[1-(2-methylpropoxy)ethoxy]ethyl]-4-(phenylazo)aniline

Biodegradation in water and sediment: simulation tests

Link to relevant study record(s)

Reference

Endpoint:

biodegradation in water: sediment simulation testing

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20/10/2017 to 23/03/2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Guideline study, conducted to GLP and current methodology.

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 308 (Aerobic and Anaerobic Transformation in Aquatic Sediment Systems)

Deviations:

yes

Remarks:

See "Any other information" for details

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

No further details specified in the study report.

Radiolabelling:

no

Oxygen conditions:

aerobic/anaerobic

Inoculum or test system:

natural water / sediment: freshwater

Details on source and properties of surface water:

Water/sediment system with sandy sediment
This surface water comes from the river “La Véore”. “La Véore” is a river in the south-east of France, a tributary of the Rhone on the left bank. It flows entirely in the department of Drôme, in region Auvergne-Rhône-Alpes.
The sampling was carried out on the commune of “Livron sur Rhone” (GPS coordinates 44.810454, 4.820417).
Water and sediment from the river “La Véore” were sampled on 16 October 2017. Water was sampled from the river. Oxygen content, pH and water temperature were measured at sampling. The water was clear and non-turbid.
Till arrival to the laboratory, the water and sediment (water-logged) were stored refrigerated under aerobic conditions (open lid) in the dark, prior to use.

Water/sediment system with silty sediment
This surface water comes from the lone “La Morte”, dead arm of the Rhone. “La Morte” is located in Saint-Benoit, a commune of the department of Ain, in France.
The sampling was carried out on the commune of Saint Benoit (GPS coordinates 45.701794, 5.553743).
Water and sediment from the river “La Morte” were sampled on 12 October 2017. Water was sampled from the river. Oxygen content, pH and water temperature were measured at sampling. The water was clear, non-turbid with a pale brown colour.
Till arrival to the laboratory, the water and sediment (water-logged) were stored refrigerated under aerobic conditions (open lid) in the dark, prior to use.
Till arrival to the laboratory, the water and sediment (water-logged) were stored refrigerated under anaerobic conditions (closed) in the dark, prior to use.

Details on source and properties of sediment:

Water/sediment system with sandy sediment
Four freshly sampled water/sediment systems were used in this study. Sediment samples were taken from the entire 5 to 10 cm upper layer of the sediment for aerobic test and under for the anaerobic test. Associated water was collected from the same location as the sediment.
Sediment sampling was performed by RRCo under non-GLP condition.
Sub-samples of the sediment of each system were shipped to La Drôme-Laboratoire, for the determination of particles size distribution and total organic carbon in the sediments.
The four sediments were significantly different based on the texture and percentage organic carbon of the sediment. The difference between the organic carbon contents of the two sediments was at least 2%. The difference in [clay + silt] content of the two sediments was at least 20%.
Concerning water, origin, temperature, pH and O2 concentration were recorded, and for sediment, origin, depth of layer and observation (colour/smell) were registered.
After collection, the sediments were separated from the water by filtration and the sediment wet-sieved to a 2 mm sieve using excess location water that were then discarded. Sediments and water were stored together, water-logged (6-10cm water layer), in the dark at 4°C ± 2 for a maximum of 4 weeks.
A period of acclimation for the water-sediment system took place prior to the beginning of the test, in order to reach stable physicochemical conditions. Therefore, incubation flasks (the same ones used for the test) were filled with a mix of sediment and water in the desired proportion. Water-sediment systems were acclimatized for 1 week under test conditions (darkness, 12°C ± 2). During equilibration, dissolved oxygen, pH, and conductivity were determined each day for each system. The temperature in the temperature controlled room was monitored continuously. Measurements made during this period are detailed in the result part of this report.
Prior to start of the incubation period, the sediment layer of 8 flasks of each system were shipped to SADEF Laboratory, for the determination of the microbial biomass using the fumigation-extraction method (non-GLP).
Water and sediment from the river “La Véore” were sampled on 16 October 2017.
The aerobic sediment was sampled from the top 5 cm and was brown. The sediment was then sieved with a 2 mm sieve mesh. The sediment was kept covered with water and bring to the laboratory on the sampling day.
Till arrival to the laboratory, the water and sediment (water-logged) were stored refrigerated under aerobic conditions (open lid) in the dark, prior to use.
The anaerobic sediment was sampled with a corer at a depth of 10 to 30 cm and was brown.
The sediment was then sieved with a 2 mm sieve mesh. The sediment was kept covered with water and bring to the laboratory on the sampling day.
Till arrival to the laboratory, the water and sediment (water-logged) were stored refrigerated under anaerobic conditions (closed) in the dark, prior to use.

Water/sediment system with silty sediment
This surface water comes from the lone “La Morte”, dead arm of the Rhone. “La Morte” is located in Saint-Benoit, a commune of the department of Ain, in France.
The sampling was carried out on the commune of Saint Benoit (GPS coordinates 45.701794, 5.553743).
Water and sediment from the river “La Morte” were sampled on 12 October 2017. Water was sampled from the river. Oxygen content, pH and water temperature were measured at sampling. The water was clear, non-turbid with a pale brown colour.
The aerobic sediment was sampled from the top 5 cm and was light brown. The sediment was then sieved with a 2 mm sieve mesh. The sediment was kept covered with water and shipped to the laboratory on the sampling day.
Till arrival to the laboratory, the water and sediment (water-logged) were stored refrigerated under aerobic conditions (open lid) in the dark, prior to use.
The anaerobic sediment was sampled with a corer at a depth of 10 to 30 cm and was brown.
The sediment was then sieved with a 2 mm sieve mesh. The sediment was kept covered with water and bring to the laboratory on the sampling day.
Till arrival to the laboratory, the water and sediment (water-logged) were stored refrigerated under anaerobic conditions (closed) in the dark, prior to use.

Details on inoculum:

Not required

Duration of test (contact time):

100 d

Initial conc.:

10 µg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

test mat. analysis

Details on study design:

The test was performed in incubators containing the water-sediment system with a volume ratio from 3:1 to 4:1. For the sandy system, about 170 g of wet sediments and 300 ml of water and for the silt system about 150 g of wet sediments and 300 ml of water were weighed into 500 ml amber glass jars. Approximately 2 cm of sediment and a water layer of approximately 6 cm were obtained at the end of equilibration.
A stock solution of the test item (C.I. Yellow 124) was prepared in acetone at a concentration of 1 g.L-1 and mixed with glass beads. The solvent was eliminated with rotary evaporator and the glass beads were rinsed with water. Then, the test solution was prepared by agitation of the glass beads in water during 24 hours. This solution was analysed and according to the measured concentration, the test system was spiked with this solution to obtain a test item quantity of approximately 10 μg in the test system.
Two flasks with test item and one control were prepared, for each sampling time (i.e. 14 flasks containing the test item and 7 control flasks).
The test item was applied as an aqueous solution into the water phase of the incubation flasks, previously acclimatised. The aqueous phase was gently mixed in order to ensure homogeneity, disturbing the sediment as little as possible. An abiotic control was performed with water system sterilized.
The frequency of atmospheric air renewal of the headspace of the flasks in the aerobic test was to two times a week in order to compensate the oxygen consumption by the biomass. The ingoing air was allowed to bubble gently through the upper part of the water layer – in order to not disturb the sediment layer – before leaving the metabolism flasks.
A trap with ethylene glycol was used for the trapping of volatile products.
The test was performed under the darkness and at 12°C ± 2.
7 samplings have been performed during the test (0, 3, 7, 14, 23, 51 days and final time). For each sampling, two flasks containing the test item and a control flask were sacrificed. Abiotic control were sampled three times (14 days, 51 days and at final time).
At the final time, the sediment of 4 dedicated flasks were shipped to SADEF laboratory for the determination of microbial biomass.
The test item and identifiable transformation product (with known analytical standards) were measured for each sampling in the aqueous phase and in the sediments.
Mineralization and mass balance have not been performed because the molecules were not radiolabelled.
The water layer was carefully separated from the sediment. Dissolved oxygen, pH, and redox potential were determined at each sampling interval.
The sediment layer was mixed. Approximately 5 g of the sediment layer was weighed for determination of pH and redox potential. About 3.5 g of the “La Veore” sediment and 7 g of the “La morte” Sediment were weighed for determination of test item and transformation products in each sediment.
The water and sediment layer were analysed for the presence of C.I. Yellow 124 at each sampling interval.

Reference substance:

other: Solvent Yellow 124; CAS No. 34432-92-3

Test performance:

Measurements of microbial biomass were performed by Sadef laboratory.
At the beginning of the incubation period, microbial biomass was determined to be 33.2 mg C/kg dry sample for “La Véore” aerobic system, 15.0 mg C/kg dry sample for “La Véore” anaerobic system, 49,8 mg C/kg dry sample for “La Morte” aerobic system and 49,8 mg C/kg dry sample for “La Morte” anaerobic system.
The results indicate sufficiently viable conditions for both water/sediment systems.

Key result

Remarks on result:

other: No degradation compounds was detected

Key result

Compartment:

natural water / sediment: freshwater

DT50:

11 d

Type:

(pseudo-)first order (= half-life)

Temp.:

12.9 °C

Remarks on result:

other: "La Véore" aerobic

Key result

Compartment:

natural water / sediment: freshwater

DT50:

4.1 d

Type:

(pseudo-)first order (= half-life)

Temp.:

12.9 °C

Remarks on result:

other: "La Morte" aerobic

Key result

Compartment:

natural water / sediment: freshwater

DT50:

28.4 d

Type:

(pseudo-)first order (= half-life)

Temp.:

12.9 °C

Remarks on result:

other: "La Véore" anaerobic

Key result

Compartment:

natural water / sediment: freshwater

DT50:

2.8 d

Type:

(pseudo-)first order (= half-life)

Temp.:

12.9 °C

Remarks on result:

other: "La Morte" anaerobic

Transformation products:

not measured

Volatile metabolites:

no

Details on results:

Parameters measured during incubation period
Temperature
During the incubation period, the temperature was within the range 11.4 – 14.4°C.

Distribution of C.I. Yellow 124
“La Véore” aerobic system
At the start of incubation, 8.3-7.6 μg (corresponding to 80% of the applied dose) of the 10 μg the nominally applied of C.I. Yellow 124 were recovered in the layer of water in both systems. After 3 days of incubation, the amount of C.I. Yellow 124 decreased to 3.7-3.1 μg (corresponding to 34% of the applied dose) in the water layer and increased to 4.9-6.4 μg (corresponding to 57% of the applied dose) in the sediment layer. Subsequently, C.I. Yellow 124 further decreased in the water layer to approximately 0 μg (LOQ < 0.2 μg of C.I. Yellow 124 / L) and decreased in the sediment layer to approximately 0.2 μg (0.18 – 0.19 μg (corresponding to 2 % of the applied dose) at the end of the incubation period.
In both total systems, the initial amount of detected C.I. Yellow 124 was 9.2 – 8.9 μg (corresponding to 91% of the applied dose). During the whole incubation period a decrease of approximately 9 μg was observed.

“La Morte” aerobic system
At the start of incubation, 5.5-12.1 μg (corresponding to 88% of the applied dose) of the 10 μg the nominally applied of C.I. Yellow 124 were recovered in the layer of water in both systems. After 3 days of incubation, the amount of C.I. Yellow 124 decreased to 1.9-1.7 μg (corresponding to 18% of the applied dose) in the water layer and increased to 3.8-4.1 μg (corresponding to 39% of the applied dose) in the sediment layer. Subsequently, C.I. Yellow 124 further decreased in the water layer to approximately 0 μg (LOQ < 0.2 μg of C.I. Yellow 124 / L) and decreased in the sediment layer to approximately 0.3 μg (0.33 – 0.19 μg (corresponding to 3 % of the applied dose) at the end of the incubation period.
In both total systems, the initial amount of detected C.I. Yellow 124 was 7.3 – 14.3 μg (corresponding to 108% of the applied dose). During the whole incubation period a decrease of approximately 10 μg was observed.

“La Veore” anaerobic system
At the start of incubation, 6.2-6.1 μg (corresponding to 62% of the applied dose) of the 10 μg the nominally applied of C.I. Yellow 124 were recovered in the layer of water in both systems. After 7 days of incubation, the amount of C.I. Yellow 124 decreased to 0.9-1.2 μg (corresponding to 10% of the applied dose) in the water layer and increased to 3.4-2.5 μg (corresponding to 40% of the applied dose) in the sediment layer. Subsequently, C.I. Yellow 124 further decreased in the water layer to approximately 0 μg (LOQ < 0.2 μg of C.I. Yellow 124 / L) and decreased in the sediment layer to approximately 0.2 μg (0.22 – 0.25 μg (corresponding to 2 % of the applied dose) at the end of the incubation period.
In both total systems, the initial amount of detected C.I. Yellow 124 was 7.0 – 8.6 μg (corresponding to 78% of the applied dose). During the whole incubation period a decrease of approximately 7.5 μg was observed.

“La Morte” anaerobic system
At the start of incubation, 8.6-4.8 μg (corresponding to 67% of the applied dose) of the 10 μg the nominally applied of C.I. Yellow 124 were recovered in the layer of water in both systems. After 3 days of incubation, the amount of C.I. Yellow 124 decreased to 0.9-1.0 μg (corresponding to 9% of the applied dose) in the water layer and increased to 4.0-2.7 μg (corresponding to 34% of the applied dose) in the sediment layer. Subsequently, C.I. Yellow 124 further decreased in the water layer to approximately 0 μg (LOQ < 0.2 μg of C.I. Yellow 124 / L) and decreased in the sediment layer to approximately 0 μg at the end of the incubation period.
In both total systems, the initial amount of detected C.I. Yellow 124 was 10.9 – 5.5 μg (corresponding to 82% of the applied dose). During the whole incubation period a decrease of approximately 8.2 μg was observed.

Carbon in NaOH traps
In carbon traps, 493 mg.L-1 of inorganic carbon was found in the control, 489 and 499 mg.L-1 in the both samples of “La Véore” system. For “La Morte” system, 507 mg.L-1 of inorganic carbon was found in the control, 517 mg.L-1 in the test 1 and 513.5 mg.L-1 in the test 2. No significant production of inorganic carbon was highlighted in test system compared to the control.

Analysis of metabolites in Q-TOF
Sediment and aqueous phase samples were analysed for the presence of metabolites using a high resolution LC-MS.
A visual examination of data was firstly done by overlaying control and test samples. Then a subtraction chromatogram was generated using Metabolite Detect software from Bruker by the subtraction of control to test sample.
No metabolite was highlighted.
Finally, a list of expected metabolite structures was created using literature or Metabolite Predict software from Bruker with phase I metabolism pathways and were screened in the samples.
No expected metabolite was highlighted.

DT50 and DT90 determination
The DT50 and DT90 calculations for the decrease of C.I. Yellow 124 in the test systems are based on the individual quantities measured.
The half-life calculated for the water layer is not a degradation DT50 but a dissipation half-life because transfer from water to sediment is included in the dissipation process.
For the sediment, degradation was assessed from the maximum onwards at a time ( 3 days for “La morte” anaerobic sediment, 7 days for “la morte” aerobic sediment, 14 days for “La veore” aerobic and anaerobic sediment) where C.I. Yellow 124 is present in the water layer at a concentration less than 10% of applied quantity so that transfer from water to sediment is reduced.
The SFO model fitted for water, sediment and total system for both test systems. The DFOP model, FOMC model and HS model fitted for several conditions.
Considering the c2 for the different models, the results with the “best” model were used in the exposure/persistence assessments.

Distribution of C.I. Yellow 124

 

La Véore” aerobic system

Water samples:

	Date		Time (days)	Control	Test 1	Test 2	Control	Test 1	Test 2	Control	Test 1	Test 2
				Concentration (ug/L)			Water volume (mL)			Quantity (µg)			Mean quantity (µg)
Initial time		12/12/2017	0	<LOQ	26.821	25.453	309.580	309.710	299.330	<LOQ	8.31	7.62	7.96
Sampling 1		15/12/2017	3	<LOQ	12.421	10.463	307.800	299.950	299.890	<LOQ	3.73	3.14	3.43
Sampling 2		19/12/2017	7	0.042	4.632	5.116	302.200	302.840	304.170	<LOQ	1.40	1.56	1.48
Sampling 3		26/12/2017	14	<LOQ	1.937	1.789	298.650	297.540	297.680	<LOQ	0.58	0.53	0.55
Sampling 4		04/01/2018	23	<LOQ	0.926	0.800	301.570	296.450	300.080	<LOQ	0.27	0.24	0.26
Sampling 5		01/02/2018	51	<LOQ	0.021	0.063	304.290	308.190	296.520	<LOQ	0.01	0.02	0.01
Sampling 6		22/03/2018	100	<LOQ	0.042	0.021	304.450	298.640	302.520	<LOQ	0.01	0.01	0.01
Abiotique (Sampling 4)		26/12/2017	14		0.168			299.22			0.05		0.05
Abiotique (Sampling 5)		01/02/2018	51		0.105			282.260			0.03		0.03
Abiotique (Sampling 6)		22/03/2018	100		0.147			281.600			0.04		0.04

Sediment samples

Time (days)	Control	Test 1	Test 2	Control	Test 1	Test 2	Control	Test 1	Test 2	Control	Test 1	Test 2
	Sample weighing (g)			Quantity (ug)			sediment weight (g)			Quantity in sediment (µg)			Mean quantity (µg)
0	3.6	3.6	3.5	< LOQ	0.0204	0.031	152.3	150.4	149.5	< LOQ	0.87	1.31	1.09
3	3.5	3.5	3.6	< LOQ	0.1156	0.1502	150.0	151.7	153.7	< LOQ	4.92	6.42	5.67
7	3.5	3.5	3.5	< LOQ	0.1372	0.0978	151.9	151.6	153.6	< LOQ	5.93	4.24	5.09
14	3.6	3.7	3.7	< LOQ	0.054	0.0716	151.7	153.8	160.3	< LOQ	2.31	2.98	2.65
23	3.5	3.5	3.5	< LOQ	0.054	0.0408	157.3	154.7	153.3	< LOQ	2.44	1.78	2.11
51	3.5	3.5	3.6	< LOQ	0.0154	0.0164	154.7	151.7	153.9	< LOQ	0.68	0.70	0.69
100	3.6	3.5	3.6	< LOQ	0.004	0.004	159.4	150.8	158.1	< LOQ	0.18	0.19	0.18
14		3.6			0.0944			153.6			4.05		4.05
51		3.6			0.0778			162.3			3.56		3.56
100		3.6			0.0874			153.3			3.74		3.74

Total system:

				Water			Sediment			Total system
	Date		Time (days)	Control	Test 1	Test 2	Control	Test 1	Test 2	Test 1	Test 2
				Quantity in water (µg)			Quantity in sediment (µg)			Quantity (µg)		Mean quantity (µg)
Initial time		12/12/2017	0	<LOQ	8.31	7.62	< LOQ	0.87	1.31	9.18	8.93	9.05
Sampling 1		15/12/2017	3	<LOQ	3.73	3.14	< LOQ	4.92	6.42	8.65	9.56	9.10
Sampling 2		19/12/2017	7	<LOQ	1.40	1.56	< LOQ	5.93	4.24	7.33	5.80	6.57
Sampling 3		26/12/2017	14	<LOQ	0.58	0.53	< LOQ	2.31	2.98	2.88	3.52	3.20
Sampling 4		04/01/2018	23	<LOQ	0.27	0.24	< LOQ	2.44	1.78	2.71	2.02	2.37
Sampling 5		01/02/2018	51	<LOQ	0.01	0.02	< LOQ	0.68	0.70	0.69	0.72	0.71
Sampling 6		22/03/2018	100	<LOQ	0.01	0.01	< LOQ	0.18	0.19	0.19	0.19	0.19
Abiotique (Sampling 4)		26/12/2017	14		0.05			4.05		4.10		4.10
Abiotique (Sampling 5)		01/02/2018	51		0.03			3.56		3.59		3.59
Abiotique (Sampling 6)		22/03/2018	100		0.04			3.74		3.78		3.78

 

Time (days)	% of applied quantity of test item
	water	sediment	Water + sediment
0	80%	11%	91%
3	34%	57%	91%
7	15%	51%	66%
14	6%	26%	32%
23	3%	21%	24%
51	0%	7%	7%
100	0%	2%	2%

 

“La Morte” aerobic system

Water samples:

	Date		Time (days)	Control	Test 1	Test 2	Control	Test 1	Test 2	Control	Test 1	Test 2
				Concentration (ug/L)			Water volume (mL)			Quantity (µg)			Mean quantity (µg)
Initial time		12/12/2017	0	<LOQ	17.621	39.032	308.600	314.880	308.840	<LOQ	5.55	12.05	8.80
Sampling 1		15/12/2017	3	<LOQ	6.000	5.811	299.730	308.400	298.720	<LOQ	1.85	1.74	1.79
Sampling 2		19/12/2017	7	0.042	2.084	2.653	305.170	296.530	304.460	<LOQ	0.62	0.81	0.71
Sampling 3		26/12/2017	14	<LOQ	0.463	0.800	301.300	300.500	300.620	<LOQ	0.14	0.24	0.19
Sampling 4		04/01/2018	23	<LOQ	0.232	0.189	301.870	292.510	297.070	<LOQ	0.07	0.06	0.06
Sampling 5		01/02/2018	51	<LOQ	0.000	0.000	276.190	273.580	274.440	<LOQ	0.00	0.00	0.00
Sampling 6		22/03/2018	100	<LOQ	0.042	0.021	279.030	258.230	259.440	<LOQ	0.01	0.01	0.01
Abiotique (Sampling 4)		26/12/2017	14		0.021			263.400			0.01		< LOQ
Abiotique (Sampling 5)		01/02/2018	51		0.021			276.990			0.01		< LOQ
Abiotique (Sampling 6)		22/03/2018	100		0.021			263.500			0.01		< LOQ

Sediment samples:

Time (days)	Control	Test 1	Test 2	Control	Test 1	Test 2	Control	Test 1	Test 2	Control	Test 1	Test 2
	Sample weighing (g)			Concentration (ug/L)			sediment weight (g)			Quantity in sediment (µg)			Mean quantity (µg)
0	7.0	7.0	7.0	< LOQ	0.0732	0.0868	168.9	176.4	173.8	< LOQ	1.78	2.20	1.99
3	7.0	6.9	7.0	< LOQ	0.1458	0.1652	181.3	172.1	178.1	< LOQ	3.78	4.10	3.94
7	7.0	6.9	7.0	< LOQ	0.0928	0.2144	178.4	176.0	181.1	< LOQ	2.36	5.44	3.90
14	7.0	7.2	6.9	< LOQ	0.0844	0.0764	185.7	181.0	171.7	< LOQ	2.23	1.92	2.08
23	7.0	7.0	6.9	< LOQ	0.0554	0.0774	183.1	182.6	178.9	< LOQ	1.45	2.03	1.74
51	6.9	7.0	7.0	< LOQ	0.0242	0.033	200.8	205.7	198.9	< LOQ	0.70	0.97	0.84
100	7.0	7.0	7.0	< LOQ	0.011	0.006	205.8	214.1	209.0	< LOQ	0.32	0.18	0.25
14		7.0			0.0144			189.5			0.39		0.39
51		7.0			0.0072			177.1			0.18		0.18
100		7.0			0.0098			186.6			0.26		0.26

Total system:

				Water			Sediment			Total system
	Date		Time (days)	Control	Test 1	Test 2	Control	Test 1	Test 2	Test 1	Test 2
				Quantity in water (µg)			Quantity in sediment (µg)			Quantity (µg)		Mean quantity (µg)
Initial time		12/12/2017	0	<LOQ	5.55	12.05	< LOQ	1.78	2.20	7.32	14.25	10.79
Sampling 1		15/12/2017	3	<LOQ	1.85	1.74	< LOQ	3.78	4.10	5.64	5.83	5.73
Sampling 2		19/12/2017	7	<LOQ	0.62	0.81	< LOQ	2.36	5.44	2.98	6.24	4.61
Sampling 3		26/12/2017	14	<LOQ	0.14	0.24	< LOQ	2.23	1.92	2.37	2.16	2.27
Sampling 4		04/01/2018	23	<LOQ	0.07	0.06	< LOQ	1.45	2.03	1.52	2.09	1.80
Sampling 5		01/02/2018	51	<LOQ	0.00	0.00	< LOQ	0.70	0.97	0.70	0.97	0.84
Sampling 6		22/03/2018	100	<LOQ	0.01	0.01	< LOQ	0.32	0.18	0.33	0.19	0.26
Abiotique (Sampling 4)		26/12/2017	14		0.01			0.39		0.40		0.40
Abiotique (Sampling 5)		01/02/2018	51		0.01			0.18		0.19		0.19
Abiotique (Sampling 6)		22/03/2018	100		0.01			0.26		0.27		0.27

 

Time (days)	% of applied quantity of test item
	water	sediment	Water + sediment
0	88%	20%	108%
3	18%	39%	57%
7	7%	39%	46%
14	2%	21%	23%
23	1%	17%	18%
51	0%	8%	8%
100	0%	3%	3%

 

“La Veore” anaerobic system

Water samples:

	Date		Time (days)	Control	Test 1	Test 2	Control	Test 1	Test 2	Control	Test 1	Test 2
				Concentration (ug/L)			Water volume (mL)			Quantity (µg)			Mean quantity (µg)
Initial time		12/12/2017	0	<LOQ	20.126	20.400	291.650	307.070	300.590	<LOQ	6.18	6.13	6.16
Sampling 1		15/12/2017	3	<LOQ	5.768	16.968	292.320	301.520	304.540	<LOQ	1.74	5.17	3.45
Sampling 2		19/12/2017	7	0.042	2.947	3.768	296.720	305.870	310.380	<LOQ	0.90	1.17	1.04
Sampling 3		26/12/2017	14	<LOQ	1.137	1.516	306.760	308.710	301.430	<LOQ	0.35	0.46	0.40
Sampling 4		04/01/2018	23	<LOQ	1.600	1.200	297.010	311.760	314.050	<LOQ	0.50	0.38	0.44
Sampling 5		01/02/2018	51	<LOQ	0.126	0.063	302.540	303.450	297.970	<LOQ	0.04	0.02	0.03
Sampling 6		22/03/2018	100	<LOQ	0.126	0.063	298.740	304.790	307.660	<LOQ	0.04	0.02	0.03
Abiotique (Sampling 4)		26/12/2017	14		0.211			326.880			0.07		0.07
Abiotique (Sampling 5)		01/02/2018	51		0.126			243.460			0.03		0.03
Abiotique (Sampling 6)		22/03/2018	100		0.147			307.890			0.05		0.05

Sediment samples:

Time (days)	Control	Test 1	Test 2	Control	Test 1	Test 2	Control	Test 1	Test 2	Control	Test 1	Test 2
	Sample weighing (g)			Concentration (ug/L)			sediment weight (g)			Quantity in sediment (µg)			Mean quantity (µg)
0	3.5	3.5	3.5	< LOQ	0.019	0.057	155.3	154.3	152.3	< LOQ	0.84	2.50	1.67
3	3.5	3.5	3.5	< LOQ	0.0844	0.3056	154.4	151.7	156.0	< LOQ	3.69	13.15	8.42
7	3.5	3.5	3.5	< LOQ	0.0786	0.0572	152.0	154.5	152.4	< LOQ	3.39	2.51	2.95
14	4.0	4.1	4.1	< LOQ	0.1468	0.1674	153.9	155.3	151.2	< LOQ	5.63	6.42	6.03
23	3.6	3.5	3.5	< LOQ	0.0948	0.1134	156.9	147.8	152.7	< LOQ	4.17	4.77	4.47
51	3.5	3.5	3.5	< LOQ	0.0394	0.0202	153.8	153.1	153.6	< LOQ	1.71	0.87	1.29
100	3.6	3.6	3.6	< LOQ	0.004	0.005	154.3	155.1	151.7	< LOQ	0.18	0.23	0.21
14		3.6			0.06			154.5			2.55		2.55
51		3.5			0.051			161.1			2.33		2.33
100		3.6			0.0328			159.4			1.46		1.46

Total system:

				Water			Sediment			Total system
	Date		Time (days)	Control	Test 1	Test 2	Control	Test 1	Test 2	Test 1	Test 2
				Quantity in water (µg)			Quantity in sediment (µg)			Quantity (µg)		Mean quantity (µg)
Initial time		12/12/2017	0	<LOQ	6.18	6.13	< LOQ	0.84	2.50	7.02	8.63	7.82
Sampling 1		15/12/2017	3	<LOQ	1.74	5.17	< LOQ	3.69	13.15	5.43	18.32	11.87
Sampling 2		19/12/2017	7	<LOQ	0.90	1.17	< LOQ	3.39	2.51	4.30	3.68	3.99
Sampling 3		26/12/2017	14	<LOQ	0.35	0.46	< LOQ	5.63	6.42	5.98	6.88	6.43
Sampling 4		04/01/2018	23	<LOQ	0.50	0.38	< LOQ	4.17	4.77	4.67	5.14	4.91
Sampling 5		01/02/2018	51	<LOQ	0.04	0.02	< LOQ	1.71	0.87	1.75	0.89	1.32
Sampling 6		22/03/2018	100	<LOQ	0.04	0.02	< LOQ	0.18	0.23	0.22	0.25	0.24
Abiotique (Sampling 4)		26/12/2017	14		0.07			2.55		2.62		2.62
Abiotique (Sampling 5)		01/02/2018	51		0.03			2.33		2.36		2.36
Abiotique (Sampling 6)		22/03/2018	100		0.05			1.46		1.51		1.51

 

Time (days)	% of applied quantity of test item
	water	sediment	Water + sediment
0	62%	17%	78%
3	35%	84%	119%
7	10%	30%	40%
14	4%	60%	64%
23	4%	45%	49%
51	0%	13%	13%
100	0%	2%	2%

 

“La Morte” anaerobic system

Water samples:

	Date		Time (days)	Control	Test 1	Test 2	Control	Test 1	Test 2	Control	Test 1	Test 2
				Concentration (ug/L)			Water volume (mL)			Quantity (µg)			Mean quantity (µg)
Initial time		12/12/2017	0	<LOQ	28.505	16.463	294.640	303.910	291.180	<LOQ	8.66	4.79	6.73
Sampling 1		15/12/2017	3	<LOQ	3.137	3.116	289.870	301.220	304.300	<LOQ	0.94	0.95	0.95
Sampling 2		19/12/2017	7	0.042	1.853	1.768	291.850	306.750	300.200	<LOQ	0.57	0.53	0.55
Sampling 3		26/12/2017	14	<LOQ	0.168	0.463	306.140	306.730	305.580	<LOQ	0.05	0.14	0.10
Sampling 4		04/01/2018	23	<LOQ	0.021	0.021	305.820	302.270	307.010	<LOQ	0.01	0.01	0.01
Sampling 5		01/02/2018	51	<LOQ	0.000	0.000	295.220	301.510	305.000	<LOQ	0.00	0.00	0.00
Sampling 6		22/03/2018	100	<LOQ	0.126	0.063	298.580	304.050	306.560	<LOQ	0.04	0.02	0.03
Abiotique (Sampling 4)		26/12/2017	14		0.063			315.740			0.02		< LOQ
Abiotique (Sampling 5)		01/02/2018	51		0.021			245.720			0.01		< LOQ
Abiotique (Sampling 6)		22/03/2018	100		0.042			307.780			0.01		< LOQ

Sediment samples:

Time (days)	Control	Test 1	Test 2	Control	Test 1	Test 2	Control	Test 1	Test 2	Control	Test 1	Test 2
	Sample weighing (g)			Concentration (ug/L)			sediment weight (g)			Quantity in sediment (µg)			Mean quantity (µg)
0	7.0	7.0	7.0	< LOQ	0.0878	0.0276	178.2	174.9	176.7	< LOQ	2.25	0.69	1.47
3	6.9	7.0	7.0	< LOQ	0.149	0.1124	186.1	168.4	175.2	< LOQ	3.99	2.72	3.36
7	7.0	7.0	7.0	< LOQ	0.0276	0.0222	179.3	175.4	177.8	< LOQ	0.71	0.56	0.63
14	7.4	7.4	7.3	< LOQ	0.0218	0.0784	176.4	176.6	177.5	< LOQ	0.52	1.87	1.19
23	7.0	6.9	6.9	< LOQ	0.0092	0.0016	170.5	182.6	173.6	< LOQ	0.23	0.04	0.13
51	7.0	7.0	7.0	< LOQ	0.001	0.0012	181.2	177.9	177.1	< LOQ	0.03	0.03	0.03
100	7.0	7.0	6.9	< LOQ	0.000	0.000	180.1	172.8	171.9	< LOQ	0.00	0.00	0.00
14		7.4			0.06			169.5			1.38		1.38
51		6.9			0.0132			187.4			0.36		0.36
100		7.0			0.0358			170.0			0.88		0.88

Total system:

				Water			Sediment			Total system
	Date		Time (days)	Control	Test 1	Test 2	Control	Test 1	Test 2	Test 1	Test 2
				Quantity in water (µg)			Quantity in sediment (µg)			Quantity (µg)		Mean quantity (µg)
Initial time		12/12/2017	0	<LOQ	8.66	4.79	< LOQ	2.25	0.69	10.91	5.49	8.20
Sampling 1		15/12/2017	3	<LOQ	0.94	0.95	< LOQ	3.99	2.72	4.93	3.67	4.30
Sampling 2		19/12/2017	7	<LOQ	0.57	0.53	< LOQ	0.71	0.56	1.28	1.09	1.18
Sampling 3		26/12/2017	14	<LOQ	0.05	0.14	< LOQ	0.52	1.87	0.57	2.01	1.29
Sampling 4		04/01/2018	23	<LOQ	0.01	0.01	< LOQ	0.23	0.04	0.23	0.05	0.14
Sampling 5		01/02/2018	51	<LOQ	0.00	0.00	< LOQ	0.03	0.03	0.03	0.03	0.03
Sampling 6		22/03/2018	100	<LOQ	0.04	0.02	< LOQ	0.00	0.00	0.04	0.02	0.03
Abiotique (Sampling 4)		26/12/2017	14		0.02			1.38		1.40		1.40
Abiotique (Sampling 5)		01/02/2018	51		0.01			0.36		0.36		0.36
Abiotique (Sampling 6)		22/03/2018	100		0.01			0.88		0.89		0.89

 

Time (days)	% of applied quantity of test item
	water	sediment	Water + sediment
0	62%	17%	78%
3	35%	84%	119%
7	10%	30%	40%
14	4%	60%	64%
23	4%	45%	49%
51	0%	13%	13%
100	0%	2%	2%

 

“La véore” aerobic system

Test system	Compartment	Kinetics	Visual fit	c^2	DT50 (days)	DT90 (days)
« La Véore »	water	SFO	moderate	7.49	2.8	9.1
		FOMC	good	1.17	2.4	11.2
		DFOP	good	1.01	2.5	11.2
	sediment	SFO	moderate	4.61	20.6	68.3
	Total system	SFO	moderate	10.6	11.0	36.5
		HS	moderate	12.27	10.8	38.7

 

“La Morte” aerobic system

Test system	Compartment	Kinetics	Visual fit	c2	DT50 (days)	DT90 (days)
« La Morte »	water	SFO	moderate	9.5	1.4	4.6
		FOMC	good	1.94	1.0	5.7
	sediment	SFO	moderate	14.99	14.9	49.4
		FOMC	moderate	8.27	10.0	107.3
	Total system	SFO	moderate	16.85	6.1	20.3
		FOMC	moderate	7.18	4.1	40.9
		DFOP	moderate	8.34	3.7	30.9
		HS	moderate	8.37	3.3	30.1

 

“La Veore” anaerobic system

Test system	Compartment	Kinetics	Visual fit	c2	DT50 (days)	DT90 (days)
« La Morte »	water	SFO	moderate	10.39	3.2	10.5
		HS	moderate	9.15	3.1	10.3
	sediment	SFO	good	3.29	17.6	58.4
	Total system	SFO	moderate	21.03	28.4	94.2

 

“La Morte” anaerobic system

Test system	Compartment	Kinetics	Visual fit	c2	DT50 (days)	DT90 (days)
« La Morte »	water	SFO	moderate	9.5	1.4	4.6
		FOMC	good	1.94	1.0	5.7
	sediment	SFO	moderate	38.12	2.7	7.9
	Total system	SFO	moderate	15	3.1	10.3
		FOMC	moderate	14.55	2.8	12.4

Validity criteria fulfilled:

yes

Conclusions:

At the start of incubation, 6.2 - 8.8 μg (corresponding to 62 – 88 % of the applied dose) of the 10 μg the nominally applied of C.I. Yellow 124 were recovered in the layer of water in both systems.
Between 3 and 14 days of incubation, the amount of C.I. Yellow 124 decreased in the water layer and increased in the sediment layer.
Subsequently, C.I. Yellow 124 further decreased in the water layer to approximately 0 μg (LOQ < 0.021 μg of C.I. Yellow 124 / L) and decreased in the sediment layer to approximately 0.2 μg at the end of the incubation period.
In both total systems, the initial amount of detected C.I. Yellow 124 was 7.8-10.8 μg (corresponding to 78 – 108 % of the applied dose). During the whole incubation period a decrease of 7.6 – 10.6 μg was observed.
No metabolite was identified through this test. The decrease of C.I. Yellow 124 measured concentration could be due to mineralization (CO2 production) or important adsorption on sediment (Non-extractable (bound) residues.

Executive summary:

The objective of this study was to assess the aerobic transformation rate of the test item in aquatic sediment and the measurement of the distribution of the test substance and its transformation products between the two phases according to OECD guideline 308.

 

The test was performed in the dark in a water-sediment system. The test item was applied as an aqueous solution into the water phase of the test system. Concentration of the test item and the transformation products was measured regularly in samples taken from the water phase and the sediment. This allowed the measurement of the transformation rate of the test item in the sediment and in a water-sediment system, as well as its distribution between these two phases. The identification of transformation products was based on comparison with control.

 

The test was performed on two aerobic and anaerobic aquatic sediment for 100 days at the most.

 

Upon addition to the water layer, C.I. Yellow 124 partitioned between the water and sediment.

At the start of incubation, 6.2 - 8.8 μg (corresponding to 62 – 88 % of the applied dose) of the 10 μg the nominally applied of C.I. Yellow 124 were recovered in the layer of water in both systems.

Between 3 and 14 days of incubation, the amount of C.I. Yellow 124 decreased in the water layer and increased in the sediment layer.

Subsequently, C.I. Yellow 124 further decreased in the water layer to approximately 0 μg (LOQ < 0.021 μg of C.I. Yellow 124 / L) and decreased in the sediment layer to approximately 0.2 μg at the end of the incubation period.

In both total systems, the initial amount of detected C.I. Yellow 124 was 7.8-10.8 μg (corresponding to 78 – 108 % of the applied dose). During the whole incubation period a decrease of 7.6 – 10.6 μg was observed.

 

No metabolite was identified through this test. The decrease of C.I. Yellow 124 measured concentration could be due to mineralization (CO2 production) or important adsorption on sediment (Non-extractable (bound) residues).

 

The DT₅₀and DT₉₀ values of C.I. Yellow 124 in both water/sediment systems are shown in the table below.

Test system	Compartment	Kinetics	Visual fit	ꭓ2	DT50(days)	DT90(days)
“La Véore” aerobic	Water	DFOP	Good	1.01	2.5	11.2
	Sediment	SFO	Moderate	4.61	20.6	68.3
	Total system	SFO	Moderate	10.6	11.0	36.5
“La Morte” aerobic	Water	FOMC	Good	1.94	1.0	5.7
	Sediment	FOMC	Moderate	8.27	10.0	107.3
	Total system	FOMC	Moderate	7.18	4.1	40.9
“La Véore” anaerobic	Water	HS	Moderate	9.15	3.1	10.3
	Sediment	SFO	Good	3.29	17.6	58.4
	Total system	SFO	Moderate	21.03	28.4	94.2
“La Morte” anaerobic	Water	FOMC	Good	1.94	1.0	5.7
	Sediment	SFO	Moderate	38.12	2.7	7.9
	Total system	FOMC	Moderate	14.55	2.8	12.4

 

Description of key information

The DT₅₀and DT₉₀values of C.I. Yellow 124 in both water/sediment systems are shown in the table below.

Test system	Compartment	Kinetics	Visual fit	ꭓ2	DT50(days)	DT90(days)
“La Véore” aerobic	Water	DFOP	Good	1.01	2.5	11.2
	Sediment	SFO	Moderate	4.61	20.6	68.3
	Total system	SFO	Moderate	10.6	11.0	36.5
“La Morte” aerobic	Water	FOMC	Good	1.94	1.0	5.7
	Sediment	FOMC	Moderate	8.27	10.0	107.3
	Total system	FOMC	Moderate	7.18	4.1	40.9
“La Véore” anaerobic	Water	HS	Moderate	9.15	3.1	10.3
	Sediment	SFO	Good	3.29	17.6	58.4
	Total system	SFO	Moderate	21.03	28.4	94.2
“La Morte” anaerobic	Water	FOMC	Good	1.94	1.0	5.7
	Sediment	SFO	Moderate	38.12	2.7	7.9
	Total system	FOMC	Moderate	14.55	2.8	12.4

 

Key value for chemical safety assessment

Half-life in freshwater:

3.1 d

at the temperature of:

12 °C

Half-life in freshwater sediment:

20.6 d

at the temperature of:

12 °C

Additional information

The objective of this study was to assess the aerobic transformation rate of the test item in aquatic sediment and the measurement of the distribution of the test substance and its transformation products between the two phases according to OECD guideline 308.

The test was performed in the dark in a water-sediment system. The test item was applied as an aqueous solution into the water phase of the test system. Concentration of the test item and the transformation products was measured regularly in samples taken from the water phase and the sediment. This allowed the measurement of the transformation rate of the test item in the sediment and in a water-sediment system, as well as its distribution between these two phases. The identification of transformation products was based on comparison with control.

The test was performed on two aerobic and anaerobic aquatic sediment for 100 days at the most.

 

Upon addition to the water layer, C.I. Yellow 124 partitioned between the water and sediment.

At the start of incubation, 6.2 - 8.8 μg (corresponding to 62 – 88 % of the applied dose) of the 10 μg the nominally applied of C.I. Yellow 124 were recovered in the layer of water in both systems.

Between 3 and 14 days of incubation, the amount of C.I. Yellow 124 decreased in the water layer and increased in the sediment layer.

Subsequently, C.I. Yellow 124 further decreased in the water layer to approximately 0 μg (LOQ < 0.021 μg of C.I. Yellow 124 / L) and decreased in the sediment layer to approximately 0.2 μg at the end of the incubation period.

In both total systems, the initial amount of detected C.I. Yellow 124 was 7.8-10.8 μg (corresponding to 78 – 108 % of the applied dose). During the whole incubation period a decrease of 7.6 – 10.6 μg was observed.

 

No metabolite was identified through this test. The decrease of C.I. Yellow 124 measured concentration could be due to mineralization (CO2production) or important adsorption on sediment (Non-extractable (bound) residues).

 

The DT₅₀and DT₉₀values of C.I. Yellow 124 in both water/sediment systems are shown in the table below.

Test system	Compartment	Kinetics	Visual fit	ꭓ2	DT50(days)	DT90(days)
“La Véore” aerobic	Water	DFOP	Good	1.01	2.5	11.2
	Sediment	SFO	Moderate	4.61	20.6	68.3
	Total system	SFO	Moderate	10.6	11.0	36.5
“La Morte” aerobic	Water	FOMC	Good	1.94	1.0	5.7
	Sediment	FOMC	Moderate	8.27	10.0	107.3
	Total system	FOMC	Moderate	7.18	4.1	40.9
“La Véore” anaerobic	Water	HS	Moderate	9.15	3.1	10.3
	Sediment	SFO	Good	3.29	17.6	58.4
	Total system	SFO	Moderate	21.03	28.4	94.2
“La Morte” anaerobic	Water	FOMC	Good	1.94	1.0	5.7
	Sediment	SFO	Moderate	38.12	2.7	7.9
	Total system	FOMC	Moderate	14.55	2.8	12.4

 

[Type of water: freshwater]