Rosin, reaction products with acrylic acid

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:

26 Jul - 23 Aug 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 F (Ready Biodegradability: Manometric Respirometry Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Rosin, reaction products with acrylic acid

EC Number:

280-192-2

EC Name:

Rosin, reaction products with acrylic acid

Cas Number:

83137-13-7

Molecular formula:

N.A. - UVCB substance

IUPAC Name:

Rosin, reaction products with acrylic acid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 646729
- Expiration date of the lot/batch: 19 Nov 2018
- Certificate of Analysis date: 1 Feb 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: No special requirements
- Stability under test conditions: Yes

ThOD of test material was calculated to be 2.60 mg O2/mg test item

Study design

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

- Source of inoculum/activated sludge: Joint Abbotsford Mission Environmental Systems wastewater treatment plant, Abbotsford, BC.
- Laboratory culture: Sludge was washed twice by centrifugation 10 min at 2500 RPM, 4 °C and resuspension in mineral medium.
- Storage conditions: The washed, resuspended sludge was in stored in a foil-wrapped 4-L flask with approximately equal headspace and stirred at test temperature for five days.
- Preparation of inoculum for exposure: pH adjustment (pH 4.9 to pH 6.1)
- Concentration of sludge: ca. 4 mg/mL

Duration of test (contact time):

28 d

Initial test substance concentration

Initial test substance concentration 1

Initial conc.:

ca. 35 mg/L

Based on:

test mat.

Initial test substance concentration 2

Initial conc.:

ca. 91 mg/L

Based on:

ThOD

Parameter followed for biodegradation estimation

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

TEST CONDITIONS
- Composition of medium: Mineral medium per OECD 301F
- Additional substrate: none
- Solubilising agent: None. Test substance was weighed and added directly to test vessels. Larger fragments of material were avoided, but no effort was made to reduce particle size before addition.
- Test temperature: 19 - 21 °C
- pH: 7.2 - 7.8
- pH adjusted: no
- Suspended solids concentration: 20 mg/L
- Continuous darkness: yes

TEST SYSTEM
- Culturing apparatus: 2-L flask sealed with a rubber bottle stopper connected to a manometric respirometer and a controlled inlet source of ultra-high purity grade oxygen. Each bottle contained a CO2 trap suspended from the top of the bottle stopper. Total fill volume was 1.5 liters.
- Number of culture flasks/concentration: two
- Method used to create aerobic conditions: stirring and controlled introduction of oxygen gas
- Measuring equipment: online manometer
- Test performed in open system: no
- Details of trap for CO2 and volatile organics if used: small scintillation vial with ~4.0 g of potassium hydroxide pellets

CONTROL AND BLANK SYSTEM
- Inoculum blank: duplicate
- Abiotic sterile control: single flask as per test substance, with addition of mercuric chloride (50 mg/L final) as biocide
- Toxicity control: single flask with both test substance (ca. 35 mg/L) and reference substance (60 mg/L)

Reference substance

Reference substance:

benzoic acid, sodium salt

Remarks:

60 mg/L

Results and discussion

% Degradation

Key result

Parameter:

% degradation (O2 consumption)

Value:

>= 17 - <= 33

Sampling time:

28 d

Remarks on result:

other: values for duplicate flasks. Mean, 25% degradation

Details on results:

Detailed results on oxygen utilization of inoculum blanks are provided in Table 1. Oxygen utilization is within range of guideline validity criteria.

Detailed results on oxygen utilization of test substance are provided in Table 2 and are graphically depicted in the illustration

Detailed results on oxygen utilization by the reference substance and in the toxicity control are provided in Table 3. The toxicity control showed 40% degradation by day 8, indicating no toxicity by the test substance.

Detailed results on oxygen utilization in the abiotic control are provided in Table 4. The abiotic control showed negative oxygen utilization throughout the experiment. While the reason for this is unclear, the results show that the test substance does not exert an abiotic demand for oxygen.

BOD5 / COD results

Results with reference substance:

46% degradation by day 3, 64% biodegradation by day 5 (see Table 3).

Any other information on results incl. tables

Table 1. Oxygen use in inoculum blanks

Day	Blank 1 oxygen use (mg)	Blank 2 oxygen use (mg)	Mean blank oxygen use (mg)
0	-0.2	-0.4	-0.3
1	-4.1	-11.6	-7.8
2	-3.3	-12.5	-7.9
3	0.6	-7.8	-3.6
4	9.1	1.0	5.0
5	8.0	-1.4	3.3
6	5.7	-2.6	1.5
7	3.3	-4.8	-0.7
8	-2.6	-8.4	-5.5
9	-4.2	-9.5	-6.8
10	-3.2	-8.2	-5.7
11	1.9	-5.3	-1.7
12	1.7	-5.6	-1.9
13	3.1	-4.4	-0.6
14	3.3	-6.5	-1.6
15	4.7	-5.8	-0.5
16	3.3	-6.0	-1.3
17	2.9	-5.0	-1.0
18	4.1	-3.9	0.1
19	4.6	1.4	3.0
20	5.5	2.1	3.8
21	9.6	4.3	6.9
22	10.8	0.3	5.5
23	10.6	-1.1	4.7
24	10.6	0.3	5.4
25	13.4	3.0	8.2
26	11.5	0.6	6.0
27	4.9	-3.4	0.7
28	9.7	-0.6	4.5

Table 2. Raw and blank-corrected oxygen use, BOD and percent biodegradation in test samples

Day	Raw Test 1 oxygen use (mg)	Raw Test 2 oxygen use (mg)	Corrected Test 1 oxygen use (mg)	Corrected Test 2 oxygen use (mg)	BOD Test 1 (mg O2/L)	BOD Test 2 (mg O2/L)	% Biodeg Test 1	% Biodeg Test 2	% Biodeg Mean
0	-0.2	-0.3	0.1	0.0	0.066	0.000	0%	0%	0%
1	-13.5	0.1	-5.6	7.9	-3.757	5.286	-4%	6%	1%
2	-10.8	-1.2	-2.9	6.7	-1.928	4.455	-2%	5%	1%
3	-0.5	8.4	3.1	12.0	2.061	7.979	2%	9%	6%
4	7.9	18.8	2.8	13.7	1.895	9.143	2%	10%	6%
5	7.8	18.0	4.5	14.7	2.992	9.775	3%	11%	7%
6	8.2	18.1	6.6	16.5	4.422	11.005	5%	12%	8%
7	8.3	20.7	9.0	21.5	6.018	14.330	7%	16%	11%
8	7.4	19.9	12.9	25.4	8.578	16.956	9%	19%	14%
9	7.2	19.4	14.0	26.3	9.342	17.521	10%	19%	15%
10	10.3	22.5	16.0	28.2	10.639	18.818	12%	21%	16%
11	13.1	24.8	14.8	26.5	9.841	17.688	11%	19%	15%
12	14.0	28.5	15.9	30.5	10.606	20.314	12%	22%	17%
13	16.0	31.0	16.6	31.7	11.071	21.112	12%	23%	18%
14	16.2	32.5	17.8	34.1	11.836	22.741	13%	25%	19%
15	16.4	34.2	16.9	34.8	11.271	23.173	12%	25%	19%
16	16.0	33.8	17.3	35.2	11.537	23.439	13%	26%	19%
17	17.4	35.6	18.4	36.7	12.268	24.437	13%	27%	20%
18	18.6	37.0	18.5	36.9	12.302	24.603	14%	27%	20%
19	19.3	38.3	16.3	35.3	10.839	23.539	12%	26%	19%
20	20.5	39.5	16.8	35.7	11.171	23.805	12%	26%	19%
21	23.4	43.1	16.5	36.2	11.005	24.104	12%	26%	19%
22	27.8	46.8	22.3	41.2	14.862	27.496	16%	30%	23%
23	27.5	48.2	22.8	43.4	15.194	28.958	17%	32%	24%
24	27.8	71.1	22.4	65.7	14.928	43.787	16%	48%	32%
25	30.7	52.8	22.5	44.6	15.028	29.723	17%	33%	25%
26	28.7	50.7	22.7	44.6	15.128	29.756	17%	33%	25%
27	25.1	47.2	24.4	46.4	16.258	30.953	18%	34%	26%
28	27.7	50.3	23.2	45.7	15.460	30.488	17%	33%	25%

Table 3. Raw and blank-corrected oxygen use, BOD and percent biodegradation in a reference and toxicity controls

Day	Raw Reference oxygen use (mg)	Corrected Ref oxygen use (mg)	BOD Ref (mg O2/L)	% Biodeg Ref	Raw Tox control oxygen use (mg)	Corrected Tox oxygen use (mg)	BOD Tox (mg O2/L)	% Biodeg Tox
0	-0.2	0.1	0.066	0%	-0.2	0.1	0.066	0%
1	-3.5	4.3	2.893	3%	13.1	20.9	13.391	7%
2	60.8	68.7	45.815	46%	53.0	60.8	40.562	21%
3	75.6	79.2	52.797	53%	65.4	69.0	46.014	24%
4	94.6	89.5	59.679	60%	88.8	83.7	55.822	29%
5	99.5	96.3	64.167	64%	100.4	97.1	64.766	34%
6	100.2	98.7	65.796	66%	105.4	103.9	69.254	36%
7	99.7	100.5	66.993	67%	109.0	109.8	73.177	38%
8	98.2	103.7	69.154	69%	107.9	113.4	75.604	40%
9	97.2	104.1	69.387	69%	108.5	115.4	76.901	40%
10	99.7	105.4	70.285	70%	111.5	117.2	78.131	41%
11	103.5	105.2	70.152	70%	112.7	114.4	76.269	40%
12	103.5	105.5	70.318	70%	111.9	113.9	75.904	40%
13	105.1	105.8	70.518	71%	112.9	113.6	75.704	40%
14	104.3	105.9	70.617	71%	112.7	114.3	76.203	40%
15	105.1	105.7	70.451	71%	112.9	113.5	75.638	40%
16	103.4	104.8	69.853	70%	111.2	112.6	75.039	39%
17	104.1	105.2	70.119	70%	112.8	113.9	75.904	40%
18	105.3	105.2	70.152	70%	113.2	113.1	75.405	39%
19	105.9	102.9	68.622	69%	112.4	109.4	72.945	38%
20	106.9	103.1	68.755	69%	113.6	109.8	73.211	38%
21	109.5	102.6	68.390	68%	114.5	107.6	71.714	38%
22	112.0	106.5	70.983	71%	115.0	109.5	72.978	38%
23	110.7	106.0	70.651	71%	113.0	108.3	72.180	38%
24	111.9	106.5	70.983	71%	115.2	109.8	73.177	38%
25	113.6	105.4	70.285	70%	114.4	106.2	70.817	37%
26	111.3	105.3	70.185	70%	111.3	105.3	70.185	37%
27	107.8	107.1	71.382	71%	108.5	107.8	71.847	38%
28	110.1	105.6	70.385	70%	110.5	106.0	70.651	37%

Table 4.  Raw and blank-corrected oxygen use, BOD and percent biodegradation in abiotic controls

Day	Raw Abiotic control oxygen use (mg)	Corrected Abiotic oxygen use (mg)	BOD Abiotic (mg O2/L)	% Biodeg Abiotic
0	-0.1	0.2	0.133	0%
1	-11.0	-3.1	-2.095	-2%
2	-18.9	-11.0	-7.314	-8%
3	-19.5	-16.0	-10.639	-12%
4	-16.1	-21.1	-14.064	-15%
5	-18.9	-22.1	-14.762	-16%
6	-22.2	-23.8	-15.859	-17%
7	-23.2	-22.5	-14.995	-16%
8	-28.0	-22.5	-15.028	-17%
9	-28.7	-21.9	-14.596	-16%
10	-28.4	-22.7	-15.161	-17%
11	-25.7	-24.0	-16.025	-18%
12	-26.8	-24.9	-16.590	-18%
13	-25.6	-25.0	-16.657	-18%
14	-26.6	-25.0	-16.690	-18%
15	-16.2	-15.6	-10.406	-11%
16	-17.0	-15.6	-10.406	-11%
17	-15.9	-14.8	-9.874	-11%
18	-15.4	-15.5	-10.307	-11%
19	-14.8	-17.8	-11.836	-13%
20	-13.5	-17.3	-11.504	-13%
21	-16.2	-23.1	-15.394	-17%
22	-8.5	-14.0	-9.342	-10%
23	-9.3	-14.0	-9.342	-10%
24	-8.4	-13.8	-9.210	-10%
25	-6.4	-14.6	-9.708	-11%
26	-9.0	-15.0	-10.007	-11%
27	-12.3	-13.0	-8.678	-10%
28	-10.1	-14.6	-9.741	-11%

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

difference in test substance degradation <20% (16%), ref substance degradation >60% by day 14 (day 5), toxicity control degradation >25% by day 14 (40%), oxygen uptake by inoculum blanks <60 mg/L in 28 days (mean, 4.5 mg/1.5 L), pH within range 6-8.5.

Interpretation of results:

not readily biodegradable

Conclusions:

In duplicate flasks, MTDID 28640 was degraded 17% and 33% (mean, 25%) in an OECD 301F test. MTDID 28640 was not inhibitory to biodegradation.

Executive summary:

Degradability of MTDID 28640 was assessed according to OECD 301F. Duplicate flasks were used for blanks and the test substance, with single flasks for reference substance (sodium benzoate), toxicity control, and abiotic (mercuric chloride-killed) samples. The reference substance was degraded adequately by day 5. The test substance was biodegraded 17% and 33% (average, 25%) by day 28. The toxicity control was biodegraded 40% in the same time period. The test substance is not readily biodegradable but is not inhibitory to biodegradation.

The test was conducted according to internationally accept guidelines under GLP criteria. It is reliable without restrictions and is suitable for Risk Assessment, Classification & Labelling, and PBT Analysis.