REPORT SANmZED **-~d** UJAdJJv **T~xttD** DETERMINATION

OF THE PARTITION COEFFICIENT

{N

· OCTANOL/WATER)

OF 1,1,3,3·TETRAMETHYLBUTYL PEROXYPIVALA
TE -------·--; NOTOX Proje

1 , (' NOTOX Substanc

. t _ **-----~** · P age 1 of 1 7
-

## 1 , 1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE

**r~** NOTOX Project

, ___ _ ) CONFIDENTIALITY STATEMENT This report contains the unpublished results of research sponsored
by AKZO 

## Nobel Chemicals

B
. V
. . Reproduction

, issue or disclosure
to third parties 

## in any form is not permitted without prior written authorization from

t he 

## sponsor.



# c 3 **~T** 

# !11A - Page 2
-1,1,3,3·TETRAMETHYLBUTYL PEROXYPIVALATE

/-- **--~** NOTOX Pro

j e(t

) **~~~/** STATEMENT
OF GLP COMPLIANCE NOTOX B. V., 's ·Herto9enbosch,
The Netherlands The study described

in this report has been correctly reported and was conducted

in compliance with the most recent edition of: The OECD Principles
of Good Laborqtory Practice which are essentially

in conformity with: The United States Food and Drug Administration. Title 21 Code of Federal Regulations Part 58. The United States Environmental Protection Agency (FIFRA). Title 40 Code of Federal Regulations Part 160. The United States Environmental Protection Agency (TSCA). Tit1e·40 Code of Federal Regulations Part 792. Study Director Ir. M.J.C. Brekelmans \ **Date~** '-..

Page 3 Management J.A.M.W. van Helvoirt H ead-<J.f-Gh"'m.i.§..t

ry **Oat~"**'1,1,3,3-TETRAMETHYLBUTYLPEROXYPIVALATENOTOXProject\QUALITYASSURANCESTATEMENTNOTOXB.V.,'s-Hertogenbosch,TheNetherlandsThisreportwasauditedbytheNOTOXQualityAssuranceUnittoensurethatthemethodsandresultsaccuratelyreflecttherawdata.ThedatesofQualityAssuranceinspectionsandauditsaregivenbelow.Duringtheon-siteinspectionsproceduresapplicabletothistypeofstudywereinspected.DATESOFQAUINSPECTIONS/AUDITSon-siteinspection(s)17July1997protocolinspection(s)22May1997reportaudit(s)11August1997HeadofQualityAssuranceC.J.M.itchellB.Sc.--\Date\/)-·Page4-REPORTINGDATES17July199722May199711August19971,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE SUMMARY The determination

of the partition coefficient (n-octanol/water)

was based on the following guidelines: OECD Guidelines for Testing of Chemicals, Guideline No. **117~** 'Partition coefficient (n-octanol/water) High Performance Liquid Chromatography (HPLC) method' (adopted March 30, 1989). EEC directive 92/69 EEC, Part A, Methods for the determination.

of physico- chemical properties, A.8: 'Partition coefficient',
EEC Publication
no. L383, December 1992. In the HPLC chromatograms

of 1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE using spectrophotometric detection
at 205 nm, 4 peaks were observed and therefore

4 P 0 w values were determined. For the major component
of 1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE,
the Pow value was 8.08

' 104 (log P 0 w **~** 4.9). For the impurities, Pow values were 1.29

• 105 (log **Pow~** 5.1), 1.49

• 10 5 (log **Pow~** 5.2) and 1.45

• 106 (log P 0 w **~** 6.2). The temperature

of the mobile phase was between 24.2 and 24.4 'C during the measurements.
Page 5 -1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE

NOT OX Pro **je~t '--~**  PREFACE Sponsor Study Monitor Testing Facility Study Director Study plan TEST SUBSTANCE Identification Chemical name CAS No. Description Batch Purity Test substance storage Stability under storage conditions Expiry date
AKZO Nobel Chemicals B.V. P.O. Box '247 3800 AE AMERSFOORT **~~** (

) **NiiTOXB:-·v-:----~--/** Hambakenwetering

3 5231 DO 's-Hertogenbosch **T-lw--N.e~.** r· **~~** ----

_/ Sta;T23JU]y1997-

- Comp
1 eted: 23 Ju.l y 1997 1,1,3,3-tetramethylbutyl peroxypiva1ate Propaneperoxic acid, 2,2-dimethy1-,1,1,3,3- tetramethy1buty1 ester (9CI) 22288-41-4 **ColorJrlA~A** 1inuid **Csee-GeF1:4·f-ivat:-~**

of Analysis In freezer
in the dark Stable Not indicated The sponsor is responsible for the completeness and GLP Compliance
of all test substance data.

- Page 6 -1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE PURPOSE ANO PRINCIPLE
**~-~** NOTOX Proje:Ct **'-.._~** The purpose of the study was to determine the partition coefficient

of the test substance between n-octanol and water. The partition coefficient (n-octanol/water)

(P 0 w) is defined as the ratio of the equilibrium concentrations

in a two phase system consisting
of n·octanol and water. Four different methods are available for the determination

of the partition coefficient. The principle
of each method is described below: 1. Flask-shaking method Dissolved test substance
is mixed thoroughly with n-octano1 and water. After phase separation, the concentration

of the test substance

in both phases is determined
by a suitable analytical method. 2. HPLC-method A solution of the test substance
is injected onto
a reversed-phase HPLC-column. The log P 0 w·value is calculated from the retention time of the test substance. 3. Estimation from the water solubility and n-octanol solubility The solubility
of the test substance

in n-octanol and in water is determined. The quotient of the n-octanol- and water solubility

is a rough estimation
of the partition coefficient. 4. Calculation from the structural formula Calculation methods are based on the theoretical fragmentation

of the molecule into suitable substructures for which reliable log P w increments are known. The log P 0 is obtained
by summing the fragment values and the correction terms for intramolecular interactions. Lists of fragment constants and correction terms are available. Based on information supplied
by the sponsor, the logPow of 1,1,3,3- TETRAMETHYLBUTYL PEROXYP!VALATE
was expected
to be between

0 and 6. Therefore, the HPLC method was chosen for the determination

of the partition coefficient (n-octanol/water). Page 7

·1,1,3,3-TETRAMETHYLBUTYLPEROXYPIVALATENOTOXGUIDELINESThestudyproceduredescribedinthisreportwasbasedonthefollowingguidelines:OrganizationforEconomicCo-operationandDevelopment(OECD},OECDGuidelineforTestingofChemicals,GuidelineNo.117:'Partitioncoefficient(n-octanol/water}HighPerformanceLiquidChromatography(HPLC}method'(adoptedMarch30,1989}.EuropeanEconomicCommunity(EEC],EECdirective92/69EEC,PartA,Methodsforthedeterminationofphysico-chemicalproperties,A.8:'Partitioncoefficient',EECPublicationno.L383,December1992.ARCHIVINGNOTOXB.V.willarchivethefollowingdataforatleast10years:protocol,report,testsubstancereferencesampleandrawdata.-Page81,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE TEST SYSTEM AND RATIONALE A High Performance Liquid Chromatograph with
a spectrophotometric detector. The stationary phase was bonded silica

c 18
. The mobile phase was 70/30 {v/v) Acetonitrile/water.

See also the section 'HPLC conditions'. The test system is recognized
by the international guidelines (OECD, EEC). REFERENCE SUBSTANCES/HPLC METHOD Six chemicals for which log P 0 w has been reported were used to calibrate the elution time in units of log Pow· The mixture of reference substances consisted of: Reference substance nitrobenzene (z.s., Merck) toluene (p.a., Merck} bromobenzene (99%, GC, Aldrich Chemie Co., Inc, USA) 1,4·dichlorobenzene {z.s., Merck} 1,2,4-trichlorobenzene (LiChrosolv, Merck} 2,4-DDT {99%, HPLC, Riedel de Haen, Seelze, Germany} 1 log Paw values according
to the OECD guideline. - Page
9

log Pow' -"-"---/ 1.9 2.7 3.0 3.4 4.2 6.2

# 1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE PERFORMANCE

OF THE HPLC METHOD Preparation

of the solutions

NOTOX 

# Proj~~ **~·** 

# Solutions
of the reference substances were prepared
in methanol(HPLCgrade, Labscan Limited Co,, Dublin, Ireland) at concentrations between

1 and 2 gil. A mixture of the reference substances was prepared
by adding 50 **~1** 

# of each solution
to a volumetric flask. Subsequently, this flask was made up with mobile phase to a final vo1ume of 10.0 m1. For the determination

of
t 0 

# (retention time of the unretarded component), a solution of formamide (p.a., Merck, Darmstadt, Germany)
in methanol was prepared at a concentration

of 1.5 gil and thereafter diluted
10 times with mobile phase, A stock solution of 1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE
was prepared by dissolving 58,5 mg test substance

in 50.0 m1 acetonitrile. The stock solution was diluted 25 times with mobile phase, resulting

in the test solution. The corresponding blank solution was prepared
by dissolving acetonitrile

25 times with mobile phase. Performance

of the test The solutions were injected
in the following sequence: the mixture of reference substances, mobile phase, the formamide solution, the test solution (in duplicate), the corresponding blank solution, the mixture of reference substances and the formamide solution. The chromatograms were recorded for approximately

15 minutes. Temperature

of measurement The temperature

of the mobile phase was recorded intermittently during the measurements.

Page 101,1,3,3·TETRAMETHYLBUTYL PEROXYPIVALATE
NOTOX **Proje~~** HPLC CONDITIONS The conditions used for the High Performance Liquid Chromatographic method are described below: Column Mobile phase Flow Detection Injection volume DATA HANDLING LiChrospher 100 RP·18; 125 x 4 (I.D.) mm; d **~** 5 **~m** (Merck, Darmstadt, Germany) 78/30 (v/v) acetonitrile (HPLC-grade, Labscan Limited Co., Dublin, Ireland)/Milli·Q water (Millipore Corp., Bedford, MA, USA) 1 ml/min UV at **~=** 205 nm for the test substance and and the blank solution and **A~** 210 nm for the references and formamide. 100 **~1** for the test substance and the blank solution and 10 **~1** for the references and formamide. 1. The capacity factor (k') is calculated from the retention
of the substance concerned (tr) and the unretarded component

(t 0 ): k' = (tr

· t 0 )/t 0 2. Using the capacity factors of the reference substances,

a plot of log P 0 w (x-value) versus log k' (y-value) was constructed, using linear regression analysis. The log Pow value of each component
of the test substance was calculated from the following equation: log **k'~** a'log Pow+
b Using high performance liquid chromatography, especially large log P 0 w values (between

0 and 6) can be accurately determined from the liquid chromatographic retention data. Page 11

# 1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE RESULTS Representative HPLC chromatograms

of the test solution, the corresponding blank solution, the mixture of reference substances, and the formamide solution are shown in Figures

1 to 4. In the HPLG chromatograms

of **1,1,~ETRAMETHYLBUTYL** 

# PEROXYPIVALATE, using detect ion at 205 nm, one large **a~** 

# ...,sma 11 peaks were observed.
It 

# was assumed that the large peak derives rrom the major component
of test substance whereas the small peaks derives from impurities. The

t 0



# was determined
to be 0.96 

# minutes
as a 

# mean value of both measurements (i.e 0.96 and 0.96 minutes). The mean values of the retention times, k' values, log k' values, log 

# P 0 

# w and 

# P 0 

# w values are summarized

in Table 1. Table

1 

# Results of the test. Substance

tr


# '

' k



# ' 

# log
k 

# log Pow nitrobenzene

1. 73 

# 0.802 -0.096 1.9 toluene

2.65 1.760 0.246 2.7 bromobenzene

2.85

1. 

# 969 0.294 3.0 1,4-dichlorobenzene

3.50 2.641 0.422 3.4 1,2,4-trichlorobenzene

5.06 4.268 0.630 4.2 2,4-DDT
12,50 12.016 1.080 6.2 test substance lr2.eA.L_U.major component} 6.78 

# 6.057



# - 0.782 **-~** 

# 4.9 :_ -"- 

# - -



# - ·---· --



# - 

# --- - 

### ~

**-~**



## ---- 

# (

Pow 

# 1--~. oa 

# xlO~

, 

# ----------~



# - **----•~-~---· -~~---""•~·~--~** 

A= 

### Mean value of the retention times of both chromatograms. 

## Interpolated from the regression line:
y • 

## 0.263x

· 0.510 {r•0.991, n•6) Note: The calculations were performed using not rounded values. 

# The temperature

of the mobile phase was 24.2-24.4'C during the measurements.

Page 12/--------- 1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE
NOTOX Projec(_ __

) !HWHEL
R (

component

1 Figure

1 HPLC chromatogram

of a 46.8 mg/1 1,1,3,3-TETRAMETHYLBUTYL- PEROXYPIVALATE solution recorded at **~=** 205 nm. Note: The peaks between

0 and 6 minutes were also observed
in the chromatograms

of the blank {see Figure 2). Therefore
it was assumed that these peaks did not derive from the test substance. Page 131,1,3,3-TETRAMETHYLBUTYLPEROXYPIVALATEFigure2HPLCchromatogramofablankrecordedat205nm.Page141,1,3,3·TETRAMETHYLBUTYL PEROXYPIVALATE 1 **~** nitrobenzene 2 **~** toluene 3 **~** bromobenzene

6 4 = 1,4-dichlorobenzene 5 **~** 1,2,4-trichlorobenzene 6 = 2,4-DDT Figure

3 HPLC chromatogram

of the mixture of reference substances

in mobile phase recorded
at A= 210 nm. Page 15

2 3

11,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE CHAMHEL

R Figure

4 HPLC chromatogram

of a 0.15 g/1 formamide solution
in mobile phase recorded
at **).~** 210 nm. Page 1647= AKZONOBEL '!CS·331 ··:i

**~** . • ...

. •. Priidu"Ct liamo i'rada:;nilme **Satehhiil~ber** •
:

• .; • 'I

/--------....... NOTOX **Proje~~** Certificate of Analysis Centro/ Research : 1,1 ,3,3 .- tetramethylbutyl peroxy pivalale C : !rlg6nox.A21L.__

) **·-=-~~~~·,.,.."'**

· Page 17