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STUDY PLAN

D-erythro-hex-2-enonic acid, .gamma.-lactone, monosodium

salt [CAS RN 6381-77-7]:

Determination of Partition Coefficient (n-Octanol/Water)

Study Director: J A Walker

Test Facility: Harlan Laboratories Ltd.

Shardlow Business Park

Shardlow

Derbyshire

DE72 2GD

UK

Sponsor: Univar USA Inc.

17425 NE Union Hill Road

Redmond

WA 98052

UNITED STATES OF AMERICA

Study Monitor: Greenwich Chemical Consulting, Inc.

3300 PGA Boulevard, Suite 635

Palm Beach Gardens

FL 33410

UNITED STATES OF AMERICA

Harlan Study Number: 41401172
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CONTENTS

CONTENTS....................................................................................................................................2

SIGNATURES................................................................................................................................3

GOOD LABORATORY PRACTICE.............................................................................................4

GENERAL INFORMATION .........................................................................................................5

Proposed Schedule.................................................................................................................5

Distribution List.....................................................................................................................5

Study Plan Amendments and Deviations...............................................................................5

Quality Assurance..................................................................................................................5

Reporting ...............................................................................................................................5

Archiving ...............................................................................................................................6

1 INTRODUCTION AND PURPOSE.....................................................................................7

1.1 Guidelines / Regulations..........................................................................................7

2 TEST AND REFERENCE ITEMS .......................................................................................8

2.1 Test Item..................................................................................................................8

2.2 Reference Item.........................................................................................................8

3 TEST......................................................................................................................................9

3.1 Partition Coefficient (n-Octanol/Water)..................................................................9

3.1.1 Test Specific Guidelines / Regulations......................................................9

3.1.2 Test System ...............................................................................................9

3.1.3 Principle of the Test...................................................................................9

3.1.4 Performance of the Test.............................................................................9

3.1.5 Calculation...............................................................................................10
Harlan Study Number: 41401172 Study Plan Page 3

SIGNATURES

Study Director: J A Walker

..................................................

Date:

Test Facility

Management:

..................................................

Date:

For the Sponsor: John V. Massingdale

Greenwich Chemical Consulting, Inc.

.................................................

Date:
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GOOD LABORATORY PRACTICE

This study will be performed in compliance with UK GLP standards (Schedule 1, Good

Laboratory Practice Regulations 1999 (SI 1999/3106 as amended by SI 2004/0994)). These

regulations are in accordance with GLP standards published as OECD Principles on Good

Laboratory Practice as revised 1997 (ENV/MC/CHEM(98)17); and are in accordance with, and

implement, the requirements of Directives 2004/9/EC and 2004/10/EC.

These principles are compatible with Good Laboratory Practice regulations specified by

regulatory authorities throughout the European Community, the United States (EPA and FDA),

and Japan (MHLW, MAFF and METI).
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GENERAL INFORMATION

Proposed Schedule

Experimental Starting Date: 01 May 2014

Experimental Completion Date: 17 July 2014

Draft Report Date: 31 July 2014

Distribution List

The original study plan will be retained in the study file. Copies of the study plan along with any

amendments will be distributed as follows:

Study Monitor: John V. Massingdale

Greenwich Chemical Consulting, Inc.

Quality Assurance: Harlan Laboratories Ltd.

Study Plan Amendments and Deviations

Any intended change to the study plan will result in an amendment to study plan approved by the

Study Director and also signed by Test Facility Management and the Study Monitor.

Amendments will be distributed to all recipients of the study plan.

Deviations (unplanned changes) from the study plan will be documented and acknowledged by

the Study Director.

Quality Assurance

Quality Assurance will audit and inspect study-related procedures and will report any audit and

inspection results in writing to the Study Director and Test Facility Management.

Reporting

A GLP compliant report will be presented. A draft report will be sent to the Study Monitor for

review and comments before issue of the final report.
Harlan Study Number: 41401172 Study Plan Page 6

Archiving

Unless instructed otherwise by the Sponsor, the study plan, all raw data (paper and electronic)

and the final report will be retained in the Harlan Laboratories Ltd, Shardlow, UK archives for

five years after which instructions will be sought as to further retention or disposal. Further

retention or return of the data will be chargeable to the Sponsor.

No data will be discarded without contacting the Sponsor to obtain their written consent.
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1 INTRODUCTION AND PURPOSE

The purpose of this study is to evaluate the partition coefficient (n-octanol/water) of the test

item.

This study is being performed based on the U.S. Toxic Substances Control Act (TSCA)

Section 4 Test Rule and the regulations set forth under 40 CFR 799.5087.

1.1 Guidelines / Regulations

The test methods described are designed to be compatible with the procedures indicated by the

following internationally accepted guidelines and recommendations:

 OECD Guidelines for Testing of Chemicals.

Where testing is conducted to satisfy the requirements of specific guidelines and/or regulations,

this will be documented and reflected in the final report.
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2 TEST AND REFERENCE ITEMS

2.1 Test Item

The test item will be supplied by or on behalf of the Sponsor including the following information

if known: description, batch, purity, expiry / retest date, storage conditions and safety

precautions to be taken.

Information as provided by the Sponsor.

Identification: D-erythro-hex-2-enonic acid, .gamma.-lactone,

monosodium salt [CAS RN 6381-77-7]

2.2 Reference Item

Reference items, if used, will be identified in the individual test sections.
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3 TEST

3.1 Partition Coefficient (n-Octanol/Water)

The purpose of this test is to evaluate the partition coefficient (Pow) between n-octanol and water

for the test item.

As the test item is a salt, testing will be performed by the shake-flask method (as indicated by

test guidance).

3.1.1 Test Specific Guidelines / Regulations

The procedure outlined in this document has been designed to be compatible with the following

guidelines and recommendations:

 OECD 107, 27 July 1995

3.1.2 Test System

The test system consists of immiscible aqueous and n-octanol phases in the same vessel with a

defined amount of test item pre-dissolved in the appropriate phase. Partition of the test item

between both phases will be achieved by inversion of the mixture.

The justification for the selection of the test system and test method is that they are

recommended by international guidelines.

3.1.3 Principle of the Test

In principle, the log Pow can be determined by the shake flask method (log Pow between -2 and 4).

Limitations of the method are described in the relevant guidelines.

3.1.4 Performance of the Test

A preliminary estimate of the partition coefficient of the test item will be generated. This may

be obtained from the ratio of the approximate solubilities of the test item in n-octanol and water

(by visual assessment) or calculated using appropriate modeling software.

In order to determine a partition coefficient, equilibrium between all interacting components of

the system must be achieved, and the equilibrium concentrations of the items dissolved in the

two phases must be determined.
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Before the partition coefficient can be determined, the two individual phases will be

pre-saturated by shaking both n-octanol and water with a sufficient quantity of the opposite

phase. A stock solution of the test item will be prepared in either n-octanol saturated water or

water saturated n-octanol. The actual mass concentration of this stock solution will be

determined using a suitable analytical method.

Three ratios of the two phases will be carried out, each in duplicate, as detailed in the method

guidelines. Ratio selection will be based on the characteristics of the test item and analytical

methodology, but will contain both twice and half the n-octanol to water ratio of the middle

ratio.

The test solutions will be partitioned and phase separation performed by centrifugation before

analysis to determine the concentration of the test item in the initial stock solution and in both

phases. The total quantity of test item present in both phases will be calculated and compared

with the quantity of the test item originally introduced (derived from the analyzed stock solution

concentration).

3.1.5 Calculation

The partition coefficient (Pow) is defined as the ratio of the equilibrium concentrations (c) of a

dissolved test item in a two-phase system consisting of two largely immiscible solvents. The

partition coefficient therefore is the quotient of two concentrations (no dimension) and is usually

given in the form of its logarithm to base ten (log Pow).

All observations which may be used to interpret the results of the tests will be documented.

Where appropriate, mean values and standard deviation will be calculated and linear regression

analysis (least square method) will be applied.

Calculations performed will be presented in the study report.