Reaction mass of 1,2,2,6,6-pentamethylpiperidin-4-yl hexadecanoate and 1,2,2,6,6-pentamethylpiperidin-4-yl octadecanoate

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:

1st March - 29th March 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study conducted in accordance with official OECD Guidelines

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: March 31, 2011, No.0331-7, Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare; March 29 2011No.5, Manufacturing Industries Bureau, Ministry of Environment, Japan

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 1,2,2,6,6-pentamethylpiperidin-4-yl hexadecanoate and 1,2,2,6,6-pentamethylpiperidin-4-yl octadecanoate

EC Number:

700-792-4

Molecular formula:

C26H51NO2, C28H55NO2

IUPAC Name:

Reaction mass of 1,2,2,6,6-pentamethylpiperidin-4-yl hexadecanoate and 1,2,2,6,6-pentamethylpiperidin-4-yl octadecanoate

Test material form:

other: SOLID

Details on test material:

- Name of test material (as cited in study report): T-1640L
- Physical state: white to pale yellow solid
- Analytical purity: 98.74%
- Purity test date: 20 May 2011
- Lot/batch No.: 10121
- Expiration date of the lot/batch: 12 April 2013
- Stability under test conditions: stable
- Storage condition of test material: refrigerated in the dark under dry conditions

Study design

Oxygen conditions:

anaerobic

Inoculum or test system:

activated sludge, domestic, adapted

Details on inoculum:

On-site sludge sampling was carried out at 10 locations in Japan (samples were from surface water
and surface soil of rivers, lakes, and inland sea; or return sludges from sewage plants). Activated
sludge, which was prepared and controlled in this laboratory according to test method described in
Section 5 a), was used in this study (sampling period: December, 2011, initimion date ofuse: January
10,2012). The activated sludge, which was cultivated for 18.5 hours after the synthetic sewage was
added, was used. The synthetic sewage was prepared according to the following method; glucose,
peptone, and potassium dihydrogenphosphate were dissolved in purified water, and the pH of the
solution was adjusted to 7.0±1.0.

Duration of test (contact time):

ca. 28 d

Initial test substance concentration

Initial conc.:

ca. 100 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

Instruments and conditions ofcultivation
a) Instruments for cultivation
Closed system oxygen consumption measuring apparatus
Temperature controlled bath (containing a measuring unit)
AI-0001 (Asahi Techneion Co., Ltd.)
Data sampler OM7000A (Ohkura Electric Co., Ltd.)
Vessel Glass vessel
Absorbent for carbon dioxide
Soda lime No.1 (for absorption ofcarbon dioxide,
Wako Pure Chemical Industries, Ltd.)
b) Conditions ofcultivation
Cultivation temperature 25±1°C
Cultivation duration 28 days (under dark conditions)
Stirring method Each test solution was stirred by a stirrer.

Determination of test item
The test item was analyzed quantitatively with LC-MS.
The test item is a mixture of seven components: main components, two components; the other
components (hereinafter referred to as subcomponents), five components. The main components
and the subcomponents were analyzed quantitatively under respective analytical conditions. It
was difficult to perfonn quantitative analysis on the same chromatogram, because the content rates
ofthe subcomponents were extremely low compared with those ofmain components. The main
components of the test item was detected as two peaks and subcomponents of the test item was
detected as five peaks on the chromatogram, so these peaks were analyzed.
1) Determination method
The test item was determined with absolute calibration curve method using one concentration
ofstandard solution.
In order to confirm the validity of this determination method, the calibration curve was made
using three concentrations of standard solution, 0.300, 0.600, and 1.20 mg/L (see Figs. 2, 3).
As a result, the regression line of the calibration curve was a straight line from the origin,
therefore the validity was confirmed.

Analytical conditions
Instrument: Liquid chromatograph-mass spectrometer
HPLCsystem: Alliance2695 (Waters)
Mass spectrometer: ZQ2000 (Waters)

LC conditions
Column: L-column ODS (150mm x 2.1 mmJ.D., particle size 5 J.Ull, Chemicals Evaluation and Research Institute)
Column temp: 40°C
Eluent: A :Tetrahydrofuran I pentafluoropropionic acid (l00011 v/v)
B : Ultrapure water I pentafluoropropionic acid (l00011 v/v)
Gradient condition
Time (min) A(%) B(%)
0.0 40 60
10.0 80 20
15.0 80 20

Flow rate: O.2mL/min
Injection volume: Main components: 1 JlL
Sub components: 10 JlL
Mass conditions
Ionization mode: Electrospray ionization (ES!)
Detection ion: Positive
Detection mode: Selected ion monitoring (S1M)
Monitoring ion (m/z) MaincomPOnent: n-C1sH31 : 410.2 [M+Ht
n-C17H3S : 438.1 [M+Ht
Subcomponent: n-CnH23 : 354.1 [M+Ht
n-C13H27 : 382.1 [M+Ht
n-ClJI29: 396.1 [M+Ht
n-C1JI33: 424.2 [M+Ht
n-C19H39 : 466.2 [M+Ht
 (see Figs. 12, 13)
Ion source temp. 120°C
Desolvation temp. 350°C
Cone voltage Main component: 10 V
Subcomponent: 30 V

Reference substance

Reference substance:

aniline

Results and discussion

% Degradation

Parameter:

% degradation (O2 consumption)

Value:

ca. 54

Sampling time:

28 d

Remarks on result:

other: continuous measurement

Applicant's summary and conclusion

Conclusions:

Most of the linear-alkyl chain of the test item was biodegraded, and a part of the test item and 1,2,2,6,6-pentamethyl-4-piperidinol remained under the conditions of this study. It is considered that the residue of the test item will disappear and change ultimately to 1,2,2,6,6-pentamethyl-4-piperidinol.