Reaction mass of L-valine and ethanesulphonic acid and octadecan-1-ol and docosan-1-ol and eicosan-1-ol

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:

3/5/18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

no

Remarks:

ISO/IEC 17025:2005

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of L-valine and ethanesulphonic acid and octadecan-1-ol and docosan-1-ol and eicosan-1-ol

Cas Number:

2125692-22-8

Molecular formula:

C25-31H54-66NO5S

IUPAC Name:

Reaction mass of L-valine and ethanesulphonic acid and octadecan-1-ol and docosan-1-ol and eicosan-1-ol

Test material form:

solid

Study design

Oxygen conditions:

aerobic

Inoculum or test system:

activated sludge, non-adapted

Details on inoculum:

The Inoculum was collected from the Escatawpa, Mississippi POTW on March 16, 2018.
This inoculum was immediately taken to the lab, homogenized, then placed into a 6 L Erlenmeyer flask. A Teflon stir bar was then added to the flask. The inoculum was placed on a magnetic stir plate. A CO2-free aeration system, which uses a CO2 scrubber system consisting of NaOH, was used to purge the inoculum. The inoculum continued stirring and aerating, uninterrupted, throughout the 4 day conditioning period.

Duration of test (contact time):

28 d

Initial test substance concentration

Initial conc.:

64.1 other: %

Based on:

TOC

Parameter followed for biodegradation estimation

Parameter followed for biodegradation estimation:

CO2 evolution

Details on study design:

On March 20, 2018, a mineral stock solution was made up, as follows, according to
OECD method 301B specifications:
DI water: 38, 493 mL
Solution 1: 390 mL
Solution 2: 39 mL
Solution 3: 39 mL
Solution 4: 39 mL
For a total of: 39 L
Then, 2400 mL of the homogenized mineral stock solution was added to each 5 L reactor bottle. A Teflon stir bar was added to each reactor, which was then placed on a stir plate and connected to a CO2 scrubber system consisting of series of soda lime and 10N NaOH scrubbers. Air flow to the system was confirmed using a Restek ProFlow 6000
Flowmeter to ensure air flows were within the 30-100 mL/min range that is stated within the method. The remaining nutrient solution was connected to a CO2 scrubber system overnight. A Total Suspended Solids test was performed on the inoculum using a Hach Lange DR5000. The test was performed on a 1:10 dilution of inoculum to DI H20 in triplicate. The average TSS was calculated to be 2,110 mg/L. The 301B method requires 30 mg of TSS to be addedper liter of nutrient solution for a total of 3 L of nutrient-biomass solution. Therefore 43 mL of inoculum was added to each reactor bottle already containing the mineral medium.
The nutrient-biomass solution (2400 mL nutrient solution + 43 mL Inoculum) remained in the 5 L reactor bottles on a stir plate and hooked to the CO2 scrubber system for 24 hrs.
Total Organic Carbon Analysis On March 20, 2018, RespirTek, Inc. performed an analysis of the reference material to obtain a Total Organic Carbon (TOC) value.
On March 16, 2018, Galbraith Laboratories, Inc. performed an analysis of the test material to obtain a Total Organic Carbon (TOC) value.
It was noted on March 16, 2018, that Galbraith Laboratories received the sample identified incorrectly. The sample was labeled as DEN-TC1 and should have been labeled as INO-TC1. Galbraith Laboratories was notified of the error. On March 21, 2018, they re-issued the report with a note on the deviation. Using the percent TOC values, appropriate test chemical and positive control additions were made to obtain a final reactor TOC value of 10 mg/L for both the PC and TC. The total amount of product to be added to nutrient-inoculum solution was added to enough mineral stock solution (the remaining solution that scrubbed overnight) to obtain a final total reactor composition of 3 L.
 Brassicyl Valinate Esylate (INO-TC1): 46.8 mg INO-TC1 + 2400 mL CO2 Free
Mineral Stock Solution+ 43 mL biomass + 557 mL DI water.
 Sodium Benzoate (PC): 87 mL PC Sodium Benzoate Stock Solution + 2400 mL
CO2 Free Mineral Stock Solution + 43 mL biomass + 470 DI water.
 Blank (B): 2400 mL CO2 Free Mineral Stock Solution + 43 mL biomass + 557 mL DI water only. All reactors were delivered CO2-free air by passing compressed air through several soda lime and 10N NaOH scrubbers. The reactors were then continually stirred, kept in diffused light and allowed to vent into a three-series 0.05N NaOH scrubber solution. Each scrubber solution was analyzed for TIC (Total Inorganic Carbon) concentrations periodically throughout the extentof the test to determine concentrations of CO2 produced by each reactor. Scrubber solutions were periodically refreshed to ensure adequate absorption of CO2 was maintained. TIC analyses were performed on a Shimadzu TOC-V CSH instrument, which was calibrated prior to test initiation and periodically throughout the duration of the test. Test reactors were set up in duplicate for statistical validation of results, and a total of 9 sampling events was executed.

Reference substance

Reference substance:

benzoic acid, sodium salt

Results and discussion

% Degradation

Parameter:

% degradation (CO2 evolution)

Value:

84

Sampling time:

28 d

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Interpretation of results:

readily biodegradable

Conclusions:

Based on the testing conducted in accordance with the specified method above, test product, Brassicyl Valinate Esylate, achieved 84% biodegradation. The product met method requirements for a Ready Biodegradability classification.