Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Environmental fate & pathways

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:
18 April 1985 - 16 May 1985
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1985
Report date:
1985

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
EPA OPPTS 835.3100 (Aerobic Aquatic Biodegradation)
Version / remarks:
EPA Chemical Fate Test Guidelines, publ. No. EPA 560/6-83-003, Method CG-2000 (Aerobic Aquatic Biodegradation) 1983.
Deviations:
not specified
Principles of method if other than guideline:
For a period of about one month, beginning on or about 02 October 1984, the test material was stored in an outside storage area (i.e. not temperature controlled or monitored). The samples for study use were taken after this time. It was determined that the storage of MRD-83-303 in this area would not have caused any adverse effects on the study in question.
GLP compliance:
yes
Remarks:
Good Laboratory Practice Regulations set forth in 40 CFR Part 792.

Test material

Constituent 1
Chemical structure
Reference substance name:
Isooctyl acetate
EC Number:
250-701-2
EC Name:
Isooctyl acetate
Cas Number:
31565-19-2
Molecular formula:
C10H20O2
IUPAC Name:
Acetic acid, isooctyl ester
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Batch II

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: For a period of about one month, beginning on or about 02-Oct-84, the test material was stored in an outside storage area (i.e. not temperature controlled or monitored). The samples for study use were taken after this time. It was determined that the storage of MRD-83-303 in this area would not have caused any adverse effects on the study in question.
- Stability under test conditions: The stability of the test material with regard to aquatic biodegradation is the subject of this report.
- Solubility and stability of the test substance in the solvent/vehicle:
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:
- Preliminary purification step (if any):
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid:

FORM AS APPLIED IN THE TEST (if different from that of starting material): colorless liquid (not different)

Study design

Oxygen conditions:
aerobic
Inoculum or test system:
other: natural sediment (freshwater) and activated sludge
Details on inoculum:
- Source of inoculum/activated sludge (e.g. location, sampling depth, contamination history, procedure): pond sediment: dredge sample from Colonial Park, East Millstone, New Jersey.; activated sludge: fresh from the Raritan Valley-Somerset Sewage Authority, Bridgewater, New Jersey.
- Method of cultivation: Acclimation Phase (April 4, 1985 to April 18, 1985): The acclimation medium was prepared on April 4, 1985 by adding the following to a 2L Erlenmeyer flask: 1 mL each of mineral salt, Solutions I, II, and III, 10.1562 g of sediment, 100 mL of activated sludge and 4. 900 mL of glass distilled water. The mixture was homogenized in a Waring blender for 5 minutes and then filtered through glass wool, and 25.1 mg casamino acids and 25.5 mg yeast extract were added to the filtrate. Nine µL of test substance (specific gravity = 0.87) were added to the acclimation medium on days 0, 7, and 11. The acclimation medium was shaken gently on the gyrorotatory shaker for 13 days in the dark. The Erlenmeyer flask was stoppered with gauze which was removed only at times of test material addition.
- Storage conditions: dark
- Storage length: 13 days
- Preparation of inoculum for exposure: On day 13 (17-Apr-85) the contents of the acclimation medium flask were poured into an Imhoff settling cone and the contents were allowed to settle overnight. The cone was covered with a gauze pad. The
resulting supernatant served as the biodegradation phase test systems. 100 mL of adapted inoculum and 1 mL of each mineral salt solution were also added to each test system.
Duration of test (contact time):
28 d
Initial test substance concentration
Initial conc.:
>= 8.5 - <= 8.53 mg/L
Based on:
test mat.
Parameter followed for biodegradation estimationopen allclose all
Parameter followed for biodegradation estimation:
CO2 evolution
Parameter followed for biodegradation estimation:
other: TC removal
Details on study design:
TEST CONDITIONS
- Composition of medium: 1 mL each of mineral salt Solutions, 10.1562 g of sediment, 100 mL of activated sludge and 900 mL of glass distilled water (acclimation medium).
- Mineral Salt Solutions: I: NH4CI (35 g/L), KNO3 (15 g/L), K2HPO4 x 3H2O (76 g/L), NaH2P04 x H20 (26 g/L); II: KCl (10 g/L), MgS04 (20 g/L), FeS04 x 7H20 (1 g/L); III: CaCl2 (5 g/L), ZnCl2 (0.05 g/L), MnCl2 x 4H20 (0.5 g/L), CuCI2 x 2H2O (0.07 g/L), CoCl2 x 6H20 (0.003 g/L), H3BO3 (0.001 g /L), MoO3 (0.0004 g/L)
- Test temperature: 20.5 - 24 °C
- pH adjusted: no
- Continuous darkness: yes

TEST SYSTEM
- Culturing apparatus: 2 L Gledhill flasks
- Number of culture flasks/concentration: 3 replicates
- Measuring equipment: As more carbonate is formed, the trap solution becomes more acidic so that less HCI titrant is required to reach the equivalence point.
- Test performed in closed vessels: The eight flasks were sealed and placed on the gyrorotatory shaker.
- Details of trap for CO2 and volatile organics if used: Upon addition of the above, 10 mL of 0.2 N sodium hydroxide (NaOH) were added to the center reservoir of each flask.


SAMPLING
- Sampling frequency: Day 5, 8, 12, 15, 19, 22, 26.
- Sampling method: The flasks were removed from the shaker and the spent NaOH was emptied from the reservoirs, combined with 2x10 mL rinsings of distilled water into 125 mL Erlenmeyer flasks. A few drops of phenolphthalein indicator were added to the spent solutions and they were titrated against 0.1 N (approximately) hydrochloric acid (HCI) to a clear endpoint. The amount of HCI required to reach the endpoint was recorded. Ten mL of fresh 0.2N NaOH were added to each reservoir and the flask contents were sparged with air through the sidearms for 5 minutes. The flasks were resealed with rubber septums and they were returned to the gyrorotatory shaker and agitated in the dark.
Day 28: Ten mL TC sample aliquots were removed from the flask contents and analyzed for total carbon (TC). The flask contents were acidified to a pH of less than 3 with concentrated H2SO4 and agitated on the shaker for four more hours. The flasks were then removed from the shaker and the reservoir contents were treated as on day 5, 8, 12, ... etc.

CONTROL AND BLANK SYSTEM
- Inoculum blank: 3 replicates
- Toxicity control: Positive control Phthalic acid: 101.6 mg/L and negative Control HgCl2: 52 g/L
Reference substance
Reference substance:
other: phthalic acid

Results and discussion

Preliminary study:
no
% Degradationopen allclose all
Key result
Parameter:
% degradation (CO2 evolution)
Value:
64.9
Sampling time:
28 d
Parameter:
other: TC removal
Value:
98.7
Sampling time:
28 d
Details on results:
CO2 evolution rate for test material mineralization was found to be 1.9x10^-2/day which corresponds to a biodegradation half-life (t^1/2) of 36.4 days, or slightly greater than one month.

BOD5 / COD results

Results with reference substance:
Loss of phthalic acid from the positive control flask: 59.9 % based on CO2 evolution; 101.1 % based on TC removal (28 days). These results are consistently high enough to indicate that a viable bacterial population was present in the inoculum.
Likewise, the data for the negative control flask indicates that the presence of HgCl2 inhibited degradation.

Any other information on results incl. tables

Table 1: Amount (mg) of Carbon evolved as Carbon Dioxide (1)

Test system

Day 5

Day 8

Day 12

Day 15

Day 19

Day 22

Day 26

Day 28

Mean – Test (2)

1.15

0.15

0.27

0.11

0.36

0.18

0.24

0.10

Cumulative Test

1.15

1.30

1.57

1.68

2.04

2.22

2.46

2.56

Positive Control – Phthalic acid

2.19

0.14

0.15

 - (3)

 -

0.07

0.10

0.04

Cumulative Positive Control

2.19

2.33

2.48

2.48

2.48

2.55

2.65

2.69

Negative Control – HgCl2 inhibited

-        

-        

-        

-        

-        

0.01

 -

 -

Cumulative negative Control

-        

-        

-        

-        

-        

0.01

0.01

0.01

Temperature °C

24.0

22.0

21.5

20.5

24.0

22.5

22.5

22.0

(1) Calculated using the mean daily control values (Appendix II)

(2) Mean from Test A, B, and C daily values

(3) CO2 evolved was less than the mean daily control values

Table 2: Total carbon, total carbon dioxide evolution, percent material degradation, and percent TC removal

Flask Solutions

TC day 1 (mg/L)

TC day 28 (mg/L)

Cumulative carbon evolved (mg) as CO2 day 28 (Table 1)

Percent material degraded day 28

Percent TC removal day 28

Mean – Test

5.225

1.336

2.56

64.9 %

98.7 %

Positive Control – Phthalic acid

5.781

1.234

2.69

59.9 %

101.1 %

Negative Control – HgCl2 inhibited

5.393

7.011

0.01

0.2 %

- 36.7 % (a)

Mean Control

1.202

1.283

 -

 -

 -

(a) The questionable negative value occurred because TC day 28 > day 0 for the negative control.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Interpretation of results:
other: ultimately biodegradable
Conclusions:
The substabce is considered as biodegradable according to Guideline EPA OPPTS 835.3100 (Aerobic Aquatic Biodegradation). Based on the CO2 evolution, the substance can be considered as readily biodegradable according to OECD criteria (OECD 301B).
Executive summary:

Based on the test results (64.9 % based on CO2 evolution) for ultimate biodegradation, the substance was determined to be (readily) biodegradable after 28 d.