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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: 41401172Harlan Study Number: 41401172 Study Plan Page 2



## CONTENTS CONTENTS ................................ ................................ ................................ ................................ .... 2 SIGNATURES ................................ ................................ ................................ ................................ 3 GOOD LABORATORY PRAC TICE ................................ ................................ ............................. 4 GENERAL INFORMATION ................................ ................................ ................................ ......... 5 Proposed Schedule ................................ ................................ ................................ ................. 5 Distribution Lis t ................................ ................................ ................................ ..................... 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 TE ST 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 ................................ ................................ ............................... 10Harlan 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:Harlan Study Number: 41401172 Study Plan Page 4



## GOOD LABORATORY PRAC TICE 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).Harlan Study Number: 41401172 Study Plan Page 5



## 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 s tudy plan will be retained in the study file. Copies of the study plan along with any amendments will be distributed as follows: S tudy 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 S tudy 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 pro cedures 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 S tudy 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.Harlan Study Number: 41401172 Study Plan Page 7



## 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.Harlan Study Number: 41401172 Study Plan Page 8



## 2 TEST AND REFERENCE ITEM S 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.Harlan Study Number: 41401172 Study Plan Page 9



## 3 TEST 3.1 Partition Coefficient (n - Octanol/Water) The purpose of this test is to evaluate the partition coefficient (P ow) 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 guideline s and recommendations :  OECD 1 07, 27 July 1995 3.1.2 Test System The test system consists of immiscible aqueous and n- octanol phase s 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 P ow can be determined by the shake flask method (log P ow between - 2 and 4) . Limitations of the method are describ ed 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.Harlan Study Number: 41401172 Study Plan Page 10

Before the partition coefficient can be determined, the two individual phase s 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 concentrati on 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 sto ck solution concentration). 3.1.5 Calculation The partition coefficient (P ow) 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 P ow). 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.