### November 5, 2010

Via Federal Express



### New Chemicals Program Off. ofPollution Prevention and Toxics Confidential Business Information Center [Rm. 6428] EPA EastBuilding Ronald Reagan Complex 1201 Constitution Ave., NW Washington, DC 20004-3302 

# Company Sanitized 

# P-11-083

CONTAINS NO CONFIDENTIAL BUSINESS INFORMATION



### Re: 

### PremanufactureNotice SubmissionunderTSCA Section5by [ ] 

### SubmissionReportNumber: CMTP101105088166808

Dear Sir/Madam: Enclosed you "vill fwd a Prcmanufacture Notice (PMN) submitted under Section S(a)(1) of the Toxic Substances Control Act. This PMN is being submitted to the {J.S. Environmental Protection Agency (EPA orthe Agency) onbehalfoff ]

## . This PMN is being submitted for the chemical substance identified below:U.S. Environmental Protection :\gency November 5, 2010 Page 2



### CA IndexName:

CAS Registry Number: Trade Name: TS Number: 

### TS-CMTP01 Submission Report Number: 

### CMTP101105088166808 CONTAINS NO CONFIDENTIAL BUSINESS INFORMATION



### User Fee PaymentNumbers: 

### Pay.govTrackingID 

### No.: 251QRS7P AgencyTrackingID

### No.: 74149916922
This PMN submission is made in accordance with the regulations given at 40 CFR 720 et seq. We request that the Agency begin review upon receipt. Enclosed with this letter are the confidential and sanitized versions of the PMN. Both of these versions have been created using the EPA's new e-PMN software. ] has also electronically remitted the PMN user fee of $2,500.00. The User Fee Payment Identity Numbers are given above. Finally, 

## f ) is claiming certain information in the PMN, including the chemical identity and company identity, as Confidential Business Information (CBI) in accordance with the provisions at 40 CFR 720.80. 

## *
Forall questions regarding the submission ofthis PMN, the technical contactis [ ] ofthe Firm. Heis reachable at [ 

## J. Sincerely,U.S. Env-ironmental Protection Agency November 5, 2010 Page 3



### cc:

Enclosures:


### • Conjidentialversion ofthe PMN 

### • Sanitizedversion ofthe PMN CONTAINS NO CONFIDENTIAL BUSINESS INFORMATION1111111111111111111 SANITIZED SUBMISSION PMN2010P1 

### PMN 

### Page 1
Form Approved. O.M.B. Nos. 2070-0012 and2070-0038 U.S. ENVIRONMENTAL PROTECTION AGENCY AGENCY USE ONLY

When completed send th1s form to !> tact Numoers: 202-564-8930/8940 :s ',--- Date of receipt: • ') "1



# 51110000063 P-11-063 Submission Re ort Number CMTP101105088166808 TS Number 196 Pay.gov 10: 251QRS7P; Agency 10: 74149916922 CMTP01 GENERAL INSTRUCTIONS • You must proVIde all information requested in this form to the extent that it 1s known to or reasonably ascertainable by you Make reasonable estimales if you do nol have actual data.

Before you complete this form. you should read the "Instructions Manual for Premanutacture Notification" (the Instructions Manual is available from the Toxic Substances Control Act (TSCA) Information Service by calling 202-554-1404, orfaxing 202-554-5603). • Ifa userfee has been remitted for this notice (40 CFR 700.45), indicate in the boxes above the TS-userfee identification number you have generated. Remember, your userfee ID number must also appear on your corresponding fee remittance. For mailing address information see the Help instructions 1n the e-PMN tool Part 1- GENERAL INFORMATION You must provide the currently correct Chemical Abstracts (CA) Name ofthe new chemical substance, even if you claim the identity as confidential. You may authorize another person to submit chemical identity information for you, butyour submission will not be complete and the review will not begin until EPA receives this information. A letter in support of your submission should reference your TS user fee identification number. For all Section 5 Notice submissions (paper or electronic) you must submit an original notice including all test data; if you claimed any information as confidential, an original sanitized copy must also be submitted. Part II- HUMAN EXPOSURE AND ENVIRONMENTAL RELEASE Ifthere are several manufacture, processing, or use operations to be described in Part II, sections A and B ofthis notice, reproduce the sections as needed. Part Ill- LIST OF ATTACHMENTS For paper submissions, attach additional sheets ifthere is not enough space to answer a question fully. Label each continuation sheet with the corresponding section heading. In Part Ill, list these attachments, any test data or other data and any optional information included in the notice. OPTIONAL INFORMATION You may include any information that you want EPA to consider in evaluating the new substance. On page 11 ofthis form, space has been provided for you to describe pollution prevention and recycling information you may have regarding the new substance. "Binding" boxes are included throughout this form for you to indicate yourwillingness to be bound to certain statements you make in this section, such as use, production volume, protective equipment
... The intention is to reduce delays that routinely accompany the development of consent orders or Significant New Use Rules. Checking a "binding" box in a PMN does not by itself prohibit the submitter from laterdeviating from the information (except chemical identity) reported in the form; however, in the case of exemption applications (such as TMEA, LVE, LOREX) certain information provided in such notifications is binding on the submitterwhen the Agency approves the exemption application, especially if the production volume "binding" box is chosen in a LVE. CONFIDENTIALITY CLAIMS You may claim any information in this notice as confidential. To assert a claim on the form, mark (X) the confidential box next to the information that you claim as confidential. To assert a claim in an attachment, circle or bracket the information you claim as confidential. If you claim information in the notices as confidential you must also provide a sanitized version ofthe notice (including attachments). For additional instructions on claiming information as confidential, read the Instructions Manual. EPA FORM 7710-25 (Rev. 6-09) TEST DATAAND OTHER DATA You are required to submit all test data in your possession or control and to provide a description of all other data known to or reasonably ascertainable by you, ifthese data are related to the health and environmental effects on the manufacture, processing, distribution in commerce, use, or disposal ofthe new chemical substance. Standard literature citations may be submitted for data in the open scientific literature. Complete test data (written in English), not summaries of data must be submitted if they do not appear in the open literature. You should clearly identify whether test data is on the substance oron an analog. Also, the chemical composition ofthe tested material should be characterized. Following are examples oftest data and other data. Data should be submitted according to the requirements of §720.50 ofthe Premanufacture Notification Rule (40 CFR Part 720). 

# D 0 

# D 

# 0 D 0 

# D D D D D D 

# D D Test Data (Check Below any included in this notice) Environmental fate data 

# D OtherData Health effects data 

# D Risk Assessments Environmental effects data 

# 0 Structure/activity relationships

Physical/Chemical Properties (A physical and chemical properties worksheet is located on the last page ofthis form.) Test data not in the possession or control ofthe submitter TYPE OF NOTICE (Check Only One) PMN (Premanufacture Notice) SNUN (Significant New Use Notice) TMEA (Test Marketing Exemption Application) LVE (Low Volume Exemption)@ 40 CFR 723.50(c)(1) LOREX (Low Release/Low Exposure Exemption) @ 40 CFR 723.50(c)(2) LVE Modification LOREX Modification Mock Submission Mark (X) if pending Letter of Support IS THIS A CONSOLIDATED PMN (Y/N)? #of chemicals or polymers (Prenotice Communication# required, enter#on p. 3). Mark (X) if any information in this notice is claimed as confidential. Replaces previous editions of EPA Form 7710-25 Page 1SANITIZED SUBMISSION 

### PMN Page 2 The public reporting and recordkeeping burden for this collection of information is estimated to average 93 hours per response. Send comments on the Agency's need for this information, the accuracy ofthe provided burden estimates, and any suggested methods for minimizing respondent burden, including through the use ofautomated collection techniques to the Director, Collection Strategies Division, U.S. Environmental Protection Agency (2822T), 1200 Pennsylvania Ave., NW, Washington, D.C. 20460. Include the OMB control number in any correspondence. Do not send the completed EPA Form 7710-25 to this address. 

### CERTIFICATION --A printed copy ofthis signature page, with original signature, must be submitted with CD or paper submission. I certify that to the best of my knowledge and belief: 1. The company named in Part I, section A, subsection 1 a ofthis notice form intends to

manufacture, import or process for a commercial purpose, other than in small quantities solely for research and development, the substance identified in Part I, Section B. 2. All information provided in this notice is complete and truthful as ofthe date of submission. 3. I am submitting with this notice all test data in my possession or control and a description ofall

other data known to or reasonably ascertainable by me as required by §720.50 ofthe Premanufacture Notification Rule. Additional Certification Statements: Ifyou are submitting a PMN, Intermediate PMN, Consolidated PMN, or SNUN, check the following userfee certification statement that applies: 

# D D The Company named in Part I, Section A has remitted the fee of$2500 specified in 40 CFR 700.45(b), or The Company named in Part I, Section A has remitted the fee of$1000 for an Intermediate PMN (defined @ 40 CFR 700.43) in accordance with 40 CFR 700.45(b), or The Company named in Part I Section A is a small business concern under 40 CFR 700.43 and has remitted a fee of$100 in accordance with 40 CFR 700.45(b). Ifyou are submitting a Low Volume Exemption (LVE) application in accordance with 40 CFR 723.50(c)(1) or a Low Release and Low Exposure Exemption (LoRex) application in accordance with 40 CFR 723.50(c)(2), check the following certification statements: 

# D The manufacturer submitting this notice intends to manufacture or import the new chemical substance for commercial purposes, other than in small quantities solely for research and development, under the terms of40 CFR 723.50. 

# D The manufacturer is familiar with the terms ofthis section and will comply with those terms; and 

# D The new chemical substance for which the notice is submitted meets all applicable exemption conditions. 

# D Ifthis application is for an LVE in accordance with 40 CFR 723.50(c)(1), the manufacturer intends to commence manufacture of the exempted substance for commercial purposes within 1year ofthe date ofthe expiration ofthe 30 day review period. The accuracy ofthe statements you make in this notice should reflect your best prediction ofthe anticipated facts regarding the chemical substance described herein. Any knowing and willful misrepresentation is subject to criminal penalty pursuant to 18 USC 1001. Signature and title of Authorized Official (Original Signature Required) Date Confidential

EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 2111111111111111111 SANITIZED SUBMISSION 

### PMNP 

## 3 liTTER Mark (X) the ··1... ' box next to any '" 'you claim as ...u1111ut:lltial 1a. 

### Pan•u•· Sul.uuuu• Notice (in U.S.) I.JO!)IIUCIIUdl Name ofAuthorized Official (first)XXX (last) XXX Position XXX Company XXX 

# 00 Mailing Address (number & street) XXX City XXX 

# I State 

# I Postal Code XXX email XXX 

### b. Agent (if A**~)** l,j!J,,..,.,,,
....... Name ofAuthorized Official (first>xxx (last) XXX Position XXX Company XXX 

# 00 Mailing Address (number & street) XXX City XXX 

# I State 

# I Postal Code XXX e-mail XXX Telephone XXX (include area code) c. JointSubr.nmn' (If 1.,\.lrlm.I.,II\!CII Ifyou are submitting this notice as part ofajointsubmission, mark (X) 

# I 

# [ J Name ofAuthorized Official (first) (last) Position Company Mailing Address (number &street) City 

# IState 

# IPostal Code e-mail Telephone (include area code) 

### 2. Technical (In U.S.) C• ·"· ·"· •VIIIIUCIIUCII Name ofAuthorized Official (first>xxx (last) XXX Position XXX Company XXX 

## lli] Mailing Address (number &street) XXX City XXX 

## I State 

# IPostal Code XXX e-mail XXX c'

## ~~~~d;·~~ea code) XXX Ifyou have had a prenotice communication (PC) concerning XXX Mark (X} if none \...Uiiuu"'"'"" 

### 3. this notice and EPA assigned a PC Numberto the notice, 

# D lli] enter the number. Ifyou.**pr~vio_usly** submitted an ....**~,.,,**'"""v' u"""""."'u"v' 1for the Mark (X) if none V\Jomu.,ntial XXX chemical substance covered bythis notice, enter the 

### 4. exemption number assigned by EPA. Ifyou previously 

# D lli] submitted a PMN for this substance enter the PMN number o.aajnnorl by EPA (i.e. withdrawn or Ifyou have submitted a notice of Bona fide intent to XXX Mark (X) if none vUr!UUCIIUCII 

### 5. manufacture or importforthe chemical substance covered 

# D bythis notice, enter the notice number assigned by EPA. 

### 6. Type of Notice- Mark (X) Manufacture Only 

# D Import Only 

## [X] 

# D 1. 2. 3. Both Binding Option 

# D Binding Option 

# D EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 771D-25 Page31111111111111111111

1. Class 1 or 2 chemical substances (for definitions of class 1 and class 2 substances, see the Instructions Manual) a. Class ofsubstance - Mark Class 1 

# D SANITIZED SUBMISSION

Class 2 CBI



# D D b. Chemical name (Currently correct Chemical Abstracts (CA) Name that is consistent with TSCA Inventory listings for similar 

# D substances. For Class 1 substances a CA Index Name must be provided. For Class 2 substances either a CA Index Name orCA Preferred Name must be which ever is based on current CA index nomenclature rules and

Method 1 (CAS Inventory Expert Service - a copy ofthe Identification report obtained from the CAS Inventory Expert EnterAttachment filename for Part I, Section B, 1. c. d. e.

Enter Attachment filename for Part I, Section B, 1. e.
EPA FORM 7710-25 (Rev. 6-09) provide a correct 

# D

Replaces previous editions of EPA Form 7710-25 Page 4111111111111111111111111111111111111111111111111111111111111 SANITIZED SUBMISSION PMN2010P4A 

### PMN Page 4a For a class 2 substance- (1) List the immediate precursor substances with their respective CAS Registry Numbers. (2) Describe the nature ofthe reaction or process. (3) Indicate the range of composition and the typical composition (where appropriate).
e. (1) List the immediate precursor substance names with their respective CAS Registry Numbers.

Enter Attachment filename for Part I, Section B, 1. e. (1) 

## I e. (2) Describe the nature ofthe reaction or process.

Enter Attachment filename for Part I, Section B, 1. e. (2) 

## I
e. (3) Indicate the range of composition and the typical composition (where appropriate).

Enter Attachment filename for Part I, Section B, 1. e. (3) 

## I Confidential 

# D

D D

D D



# D

EPA FORM 7710-25 (Rev. 6-09) Page 5**Repl~ces** previous editions of EPA Form 7710-25 -:**~** ,.;**~-** ,::J ,:_::1**-~** "·.1111111111111111111111111111111111111111111111111111111 SANITIZED SUBMISSION PMN2010P5 

### PMN Page 

## 5 Part I -- GENERAL INFORMATION - Continued Section B -- CHEMICAL IDENTITY INFORMATION -- Continued 2. Polymers (For a definition of polymer, see the Instructions Manual.) Confidential a. Indicate the number-average weight ofthe lowest molecular weight composition ofthe polymer you intend to manufacture. 

## [X] Indicate maximum weight percent of low molecular weight species (not including residual monomers, reactants, or solvents) below 500 and below 1,000 absolute molecular weight ofthat composition. Describe the methods of measurement or the basis for your estimates: GPC 

# D 

# IOther (Specify Below) 

# D 

# I Specify Other:

(i) lowest number average molecular (ii) maximum weight % below 500 molecular (iii) maximum weight % below 1000 molecular weight: weight: weight: XXX XXX XXX Enter Attachment filename for Part I, Section B, 2. a. 

## I 

# D b. You must make separate confidentiality claims for monomer or other reactant identity, composition information, and residual information. Mark (X) the "Confidential" box next to any item you claim as confidential

(1) - Provide the specific chemical name and CAS Registry Number (if a number exists) ofeach monomer or other reactant used in the manufacture ofthe polymer. (2) - Mark (X) this column ifentry in column (1) is confidential. (3) - Indicate the typical weight percent ofeach monomer or other reactant in the polymer. (4) - Choose "yes" from drop down menu ifyou want a monomer or other reactant used at two weight percent or less to be listed as part of the polymer description on the TSCA Chemical Substance Inventory. (5) - Mark (X) this column ifentries in columns (3) and (4) are confidential. (6) - Indicate the maximum weight percent ofeach monomer or other reactant that may be present as a residual in the polymer as manufactured for commercial purposes. (7) - Mark (X) this column ifentry in column (6) is confidential. Monomer or other reactant specific chemical name (1) XXX

CAS Registry Number (1) XXX XXX

CAS Registry Number (1) XXX XXX

CAS Registry Number (1) XXX XXX

CAS Registry Number (1) XXX XXX

CAS Registry Number (1) 

# IXXX Mark (X) this box ifthe data continues on the next page. EPA FORM 7710-25 (Rev. 6-09) Page 6 Typical Include in Max CBI composition identity CBI residual CBI (2) (3) (4) (5) (6) (7) X XXX X XXX X

X XXX X XXX X

X XXX X XXX X

X XXX X XXX X

X XXX X XXX X 

## IX] Replaces previous editions ofEPA Form 7710-25 :;t ...,.; '....., ·,:_; ::)r·SANITIZED SUBMISSION PMN2010P5-1 

### PMN Page 5 (1) Part I --GENERAL INFORMATION - Continued Section B -- CHEMICAL IDENTITY INFORMATION -- Continued 2. Polymers (For a definition ofpolymer, see the Instructions Manual.) Confidential a. Indicate the number-average weight ofthe lowest molecular weight composition ofthe polymeryou intend to manufacture. 

# D Indicate maximum weight percent of low molecular weight species (not including residual monomers, reactants, or solvents) below 500 and below 1,000 absolute molecular weight ofthat composition. Describe the methods of measurement orthe basis for your estimates: GPC 

# D I Other (Specify Below) 

# D 

# I Specify Other:

(i) lowest number average molecular (ii) maximum weight % below 500 molecular (iii) maximum weight % below 1000 molecular weight: weight: weight:

EnterAttachment filename for Part I, Section B. 2. a. 

# I 

# D b. You must make separate confidentiality claims for monomer or other reactant identity, composition information, and residual information. Mark (X) the "Confidential" box next to any item you claim as confidential

(1) - Provide the specific chemical name and CAS Registry Number (if a number exists) ofeach monomer or other reactant used in the manufacture ofthe polymer. (2) - Mark (X) this column ifentry in column (1) is confidential. (3) - Indicate the typical weight percent ofeach monomer or other reactant in the polymer. (4) - Choose "yes" from drop down menu ifyou want a monomer or other reactant used attwo weight percent or less to be listed as part of the polymer description on the TSCA Chemical Substance Inventory. (5) - Mark (X) this column ifentries in columns (3) and (4) are confidential. (6) - Indicate the maximum weight percent ofeach monomer or other reactant that may be present as a residual in the polymer as manufactured for commercial purposes. (7) - Mark (X) this column ifentry in column (6) is confidential. Monomer or other reactant specific chemical name Typical Include in Max CBI composition identity CBI residual CBI (1) (2) (3) (4) (5) (6) (7) XXX 

### X XXX 

### X XXX 

### X

CAS Registry Number (1) XXX XXX 

### X XXX 

### X XXX 

### X

CAS Registry Number (1) XXX XXX 

### X XXX 

### X XXX 

### X

CAS Registry Number (1) XXX

CAS Registry Number (1)

CAS Registry Number (1) Mark (X) this box ifthe data continues on the next page. 

# D
EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 71111111111111111111111111111111111111 1111111111111 SANITIZED SUBMISSION PMN2010P5A 

### PMN Page Sa c. Please identify which method you used to develop or obtain the specified chemical identity information reported in this notice (check one). Method 1 (CAS Inventory Expert Service

- a copy ofthe identification report obtained from CAS Inventory Expert Service must be submitted as an attachment to this notice) Enter Attachment filename for Part I, Section B, 2. c. IES Order Number 152304 Method 2 

# D (other source)
d. The currently correct Chemical Abstracts (CA) name for the polymer that is consistent with TSCA Inventory listings for similar polymers. XXX

CAS Registry Number (if a number already exists for the substance) XXX e. Provide a correct representative or partial chemical structure diagram, as complete as can be known, if one can be reasonably ascertained.

Enter Attachment filename for Part I, Section B, 2. e. 

# I CBI 

# D

D



# D
EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 8SANITIZED SUBMISSION 

### PMN Page 6 Part 

### I -- GENERAL INFORMATION -- Continued Section B -- CHEMICAL IDENTITY INFORMATION -- Continued 3. Impurities (a) - Identify each impurity that may be reasonably anticipated to be present in the chemical substance as manufactured for commercial purpose. Provide the CAS Registry Number if available. Ifthere are unidentified impurities, enter "unidentified." (b) - Estimate the maximum weight% of each impurity. Ifthere are unidentified impurities, estimate their total weight%. CAS Registry Maximum Conti- Impurity (a) Number Percent% (a) (b) dential

Mark (X) this box ifthe data continues on the next page. 

# D EnterAttachment filename for Part I, Section B, 3. 

## I 

# D 4. Synonyms- Enter any chemical synonyms for the new chemical identified in subsection 1 or 2. XXX 

## [Z]

Enter Attachment filename for Part I, Section B, 4. 

# I 

# D 5. Trade identification - List trade names for the new chemical substance identified in subsection 1 or 2. XXX 

# 00

Enter Attachment filename for Part I, Section B, 5. 

# I 

# D 6. Generic chemical name - Ifyou claim chemical identify as confidential, you must provide a generic name for your substance that reveals the specific chemical identity ofthe new chemical substance to the maximum extent possible. Refer to the TSCA Chemical Substance Inventory, 1985 Edition, Appendix B for guidance on developing generic names. Perfluoroalkyl acrylate copolymer

EnterAttachment filename for Part I, Section B, 6. 

## I 7. Byproducts - Describe any byproducts resulting from the manufacture, processing, use, or disposal ofthe newchemical substance. Provide the CAS Registry Number ifavailable. Byproduct (1) CAS Registry Number Conti-

.(2)- dential XXX XXX 

### X

Mark (X) this box ifthe data continues on the next page. 

# D

EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 911111111111111111111111111111111111111111 SANITIZED SUBMISSION PMN2010P7 

### PMN P

### age 

## 7 Part I -- GENERAL INFORMATION -- Continued Section C --PRODUCTION, IMPORT, AND USE INFORMATION: The information on this page refers to consolidated chemical number(s): 

# 01 02 03 04 Os Os Mark (X) the "Confidential" box next to any item you claim as confidential. 1. Production volume -- Estimate the maximum production volume during the first 12 months of production. Also estimate the maximum production volume for any consecutive 12-month period during the first three years of production. Estimates should be on 100% new chemical substance basis. For a Low Volume Exemption application, ifyou choose to have your notice reviewed at a lower production volume than 10,000 kg/yr, specify the volume and mark (x) in the binding box. Ifgranted, you are bound to this volume. Maximum first 12-month production (kg/yr) Maximum 12-month production (kg/yr) Confidential Binding Option

(100% newchemical substance basis) (100% new chemical substance basis) Mark (X) XXX XXX 

## [X] 

# 0 Enter Attachment filename for Part I, Section C, 1. 

# I CBI 

# 0 2. Use Information --You must make separate confidentiality claims for the description ofthe category of use, the percent ofproduction volume devoted to each category, the formulation ofthe newsubstance, and other use information. Mark (X) the "Confidential" Box nex1 to any item you claim as confidential. a. (1) --Describe each intended category of use ofthe new chemical substance by function and application. (2) --Mark (X) this column if entry column (1) is confidential business information (CBI). (3) --Indicate your willingness to have the information provided in column (1) binding. (4) --Estimate the percent oftotal production for the first three years devoted to each category of use. (5) --Mark (X) this column ifentry in column (4) is confidential business information (CBI). (6) --Estimate the percent ofthe newsubstance as formulated in mixtures, suspensions, emulsions, solutions, or gels as manufactured for commercial purposes at sites underyour control associated with each category of use. (7) --Mark (X) this column ifentry in column (6) is confidential business information (CBI). (8) --Indicate% of product volume expected for the listed "use" sectors. Mark more than one box ifappropriate. Mark (X) to indicate your willingness to have the use type provided in (8) binding. (9) --Mark-(X) this column ifentry{ies) in column (8)is (are)confidential business information (CBI). Category ofuse (1) Binding Prod %in % ofsubstance expected per use (by function and application i.e. a dispersive dye for CBI Option uction CBI Form- CBI (8) CBI finishing polyester fibers) Mark (X) % ulation Site- Con- Industrial Com- Binding (2) (3) (4) (5) (6) (7) limited sumer• mercial Option (9) Coating material for use in textiles and/or paper. XXX X XXX X XXX XXX XXX XXX 

### X

• Ifyou have identified a "consumer" use, please provide on a continuation sheet a detailed description ofthe use(s) ofthis chemical substance in consumer products. In addition include estimates ofthe concentration ofthe new chemical substance as expected in consumer products and describe the chemical reactions by which this substance loses its identity in the consumer product. Mark (X) this box ifthe data continues on the nex1 page. 

# 0 b. Generic use Ifyou claim any category of use description in subsection 2a as confidential, enter a generic description ofthat category. description Read the Instruction Manual for examples ofgeneric use descriptions. Not applicable.
EnterAttachment filename for Part I, Section C, 2. b. 

# I CBI 

# 0 3. Hazard Information -- Include in the notice a copy of reasonable facsimile ofany hazard warning statement, label, material safety

data sheet, orother information which will be provided to any person who is reasonably likely to be exposed to this substance Binding Option regarding protective equipment or practices for the safe handing, transport, use, or disposal ofthe new substance. List in part Ill Mark (X) hazard information you include. Mark (X) this box ifyou attach hazard information. 

## [XI 

# 0 EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 10SANITIZED SUBMISSION PMN2010P8 

### PMNP age 

### 8 Part II- HUMAN EXPOSURE AND ENVIRONMENTAL RELEASE SectionA - INDUSTRIALSITES CONTROLLED BYTHE SUBMITTER 

# IMark (X) the "Confidential" box nextto any item you claim as confidential The information on pages 8and Sa referto consolidated chemical number(s): 

# [ J 1 

# 02 03 04 5 

# Os Complete section A for each type ofmanufacture, processing, or use operation involving the new chemical substance at industrial sites you control. Importers do not have to complete this section for operations outside the U.S.; however, you may still have reporting requirements ifthere are further industrial processing or use operations after import. You must describe these operations. See instructions manual 1. Operation description Contia. Identity -- Enter the identity ofthe site atwhich the operation will occur. dential Name Site address (number and street) 

# 0 City County State ZIP code Ifthe same operation will occur at more than one site, enter the number ofsites. Identify the additional 

# 0 sites on a continuation sheet, and if any ofthe sites have significantly different production rates or 

## 0 operations, include all the information reguested in this section for those sites as attachments.**~** Mark (X) this box ifthe data continues on the next page. 

# 0 b. Type-- Manufacturing 

# 0 Processing 

# 0 Use 

# 0 0 Mark (X) c. Amount and Duration - Complete 1 or 2 as appropriate Contidential Maximum kg/batch (100% new chemical Hours/batch Batches/year 

# 0 1. Batch substance) Maximum kg/day Hours/day Days/year (1

00% newchemical substancel 

# 0 2. Continuous Mark (X) to indicate your willingness to d. Process description have your process description binding.**~** 

# ..

(1) Diagram the major unit operation steps and chemical conversions. Include interim storage and transport containers (specify- e.g. 5gallon pails, 55 gallon drum, rail car, tank truck, etc.). (2) Provide the identity, the approximate weight (by kg/day or kg/batch on a 100% new chemical substance basis), and entry point ofall starting materials and feedstocks (including reactants. solvents, catalysts, etc.), and ofall products, recycle streams, and wastes. Include cleaning chemicals (note frequency if not used daily or per batch.). (3) Identify by numberthe points of release, including small or intermittent releases, to the environment ofthe newchemical substance. If releasing to two media at the same step, assign asecond release numberfor the second medium. 

# D See Attachments #5 and #6.

EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 11

# 1~111~**Ill~ 1111111111~11111111111111111**  SANITIZED SUBMISSION PMN2010P8A 

### PMNPage 

## 8a Confidential Diagram ofthe major unit operation steps. 

# D

EnterAttachment filename for Part II, Section A, 1. d. 

# I 

# [ J

EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 12 

## ,.,SANITIZED SUBMISSION PMN2010P9 

### PMNP

## age 9 Part 11-- HUMAN EXPOSURE AND ENVIRONMENTAL RELEASE --Continued Section A - INDUSTRIAL SITES CONTROLLED BY THE SUBMITTER --Continued The Information on pages 9 and 9a referto consolidated chemical number(s): 

# [ J 1 

# l 2 

# 03 [ ]4 

# Ds Ds 2. Occupational Exposure-- You must make separate confidentiality claims for the description ofworker activity, physical form ofthe new chemical

substance, number ofworkers exposed, and duration ofactivity. Mark (X) the "Confidential" box nextto any item you claim as confidential.

(1) - Describe the activities (i.e. bag dumping, tote filling, unloading drums, sampling, cleaning, etc.) in which workers may be exposed to the substance. (2) Mark (X) this column ifentry in column (1) is confidential business information (CBI). (3) -- Describe any protective equipment and engineering controls used to protect workers. (4) and (6) -- Indicateyourwillingness to have the information provided in column (3) or (5) binding. (5) ·- Indicate the physical form(s) ofthe newchemical substance (e.g., solid: crystal. granule, powder, or dust) and % newchemical substance (if part ofa mixture) at the time of exposure. (7) -- Mark (X) this column ifentries in columns (3) and (5) are confidential business information (CBI). (8) -- Estimate the maximum number ofworkers involved in each activity for all sites combined. (9) Mark (X) this column ifentry in column (8) is confidential business information (CBI). (10) and (11) -- Estimate the maximum duration ofthe activity for anyworker in hours perday and days peryear. (12) -Mark (X) this column ifentries in columns (10) and (11) are confidential business information CBI). Worker activity Protective Equipment/ Binding Physical Binding ##of Maximum Duration (I.e., bag dumping, fiiUng CBI Option form(e) Option CBI Workers CBI CBI drums) Engineering Controls Mark (X) &%new Mark (X) Exposed eubstance Hrs!Day Days/Yr (12) (1) (2) (3) (4) (51 (6) (7) (8) {9) (10) (11)

Mark (X) this box ifthe data continues on the next page. 

### f 

## I Enter Attachment filename for Part II, Section A on the bottom ofpage 9a. 

# D
EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 13SANITIZED SUBMISSION 20 PMN 10P9A 

### PMNPage 

## 9a 3. Environmental Release and Disposal -- You must make separate confidentiality claims for the release number and the amount ofthe new chemical substance released and other release and disposal information. Mark (X) the "Confidential" box next to each item you claim as confidential. (1) -- Enter the number of each release point identified in the process description, part II, section A, subsection 1d(3). (2) -- Estimate the amount ofthe new substance released (a) directly to the environment or (b) into control technology (in kg/day or kg/batch). (3) Mark (X) this column ifentries in columns (1) and (2) are confidential business information (CBI). (4) -- Identify the media (stack air, fugitive air (optional-see Instruction Manual), surface water, on-site or off-site land or incineration, PO'TW, or other (specify)) to which the new substance will be released from that release point. (5) -- a. Describe control technology, if any, and control efficiencythat will be used to limit the release ofthe new substance to the environment. For releases disposed of on land, characterize the disposal method and state whether it is approved for disposal of RCRA hazardous waste. On a continuation sheet. for each site describe any additional disposal methods that will be used and whetherthe waste is subjectto secondary or tertiary on-site treatment. b. Estimate the amount released to the environment after control technology (in kg/day). (6) -- Mark (X) this column ifentries in columns (4) and (5) are confidential business information (CBI). (7) -- Identify the destination(s) of releases to water. Please supply NPDES (National Pollutant Discharge Elimination System) numbers for direct discharges or NPDES numbers ofthe PO'TW (Publicly Owned Treatment Works). Mark (X) ifthe PO'TW name or NPDES #is confidential business information (CBI). Release Amount ofNew Medium ofrelease Control technology and efficiency (you may wish to Number Substance Released CBI e.g. Stackair optionally attach efficiency data) CBI Binding (6) (1) (2a) (2b) (3) (4) (5a) Mark (5b) (X}

Mark (X) this box ifthe data continues on the next page. 

# D (7) Marl< (X) the destination(s of releases to water. NPDES# CBI



# D POTW--provide 

# D name(s) 

# D Navigable waterway- 

# D -provide name(s) 

# D Other--Specify 

# D EnterAttachment filename for Part II, Section A. 

# 1 

# D EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 14

# 11111~111~1111 SANITIZED SUBMISSION PMN2o1oP1o 

### PMN 

## Page 10 Part II- HUMAN EXPOSURE AND ENVIRONMENTAL RELEASE- Continued Section B -- INDUSTRIAL SITES CONTROLLED BY OTHERS The information on pages 10 and 10a referto consolidated chemical number(s): [ ] 

### 1 [ ] 2 [ ] 3 

# Ds Ds Complete section Bfortypical processing or use operations involving the new chemical substance at sites you do not control. Importers do not have to complete this section for operations outside the U.S.; however, you must report any processing or use activities after import. See the Instructions Manual. Complete a separate section Bforeach type ofprocessing, oruse operation involving the newchemicalsubstance. Ifthe same operation is performed at more than one site describe the typical operation common to these sites. Identify additional sites on acontinuation sheet. 1(a). Operation Description-- To claim information in this section as confidential, bracket (e.g.{}) the specific information that you claim as confidential. (1) Diagram the major unit operation steps and chemical conversions, including interim storage and transport containers (specify- e.g. 5gallon pails, 55 gallon drums, rail cars, tank trucks, etc). On the diagram. identify by fetter and briefly describe each worker activity. (2) •• Either in the diagram or in the text field 1 (b) below, provide the identity, the approximate weight (by kg/day or kg/batch, on an 100% new chemical substance basis), and entry point of all feedstocks (including reactants. solvents and catalysts, etc) and all products. recycle streams, and wastes. Include cleaning chemicals (note frequency ifnot used daily or per batch). (3) -- Either in the diagram or in the text field 1(b) below. identify by numberthe points of release, including small or intermittent releases, to the environment ofthe new chemical substance. (4) •• Please enterthe# ofsites (remember to identify the locations ofthese sites on acontinuation sheet):



## ...... 

# I Numberof Sites 

# I IConfidential

1(b). (Optional) This space is for atext description to clarify the diagram above. Confidential 

# D See Attachment #9.

EnterAttachment filename for Part II, Section Bonthe bottom of page 10a. 

# I 

# D

EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 15**-~** 

## •..

# Ill~**I~ Ill~** **11~1 ~~I** IIIII SANITIZED SUBMISSION PMN2010P10-1 

## Continuation Sheet ID

# IP10SB1 (a)(4)1 

# I Field

# I Part II, Section B, 1(a)(4). Operation Site Locations XXX

EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 16

# ~1111~111111111 1111 SANITIZED SUBMISSION PMN201OP10A 

## PMNPage 

## 10

## a 2. Worker Exposure/Environmental Release (1) - From the diagram above, provide the letterfor each worker activity. Complete 2-8 for each worker activity described. (2) -- Estimate the number ofworkers exposed for all sites combined. (4) Estimate the typical duration of exposure perworker in (a) hours per day and (b) days per year. (6) -- Describe physical form ofexposure and % new chemical substance (if in mixture), and any protective equipment and engineering controls, if any, used to protectworkers. (7) -- Estimate the percent ofthe new substance as formulated when packaged or used as afinal product. (9) -- From the process diagram above, enter the number ofeach release point Complete 9-13 for each release point identified. (10) -- Estimate the amount ofthe newsubstance released (a) directly to the environment or (b) into control technology to the environment (in kg/day or kg/batch). (12) Describe media of release i.e. stack air, fugitive air (optional-see Instructions Manual), surface water, on-site or off-site land or incineration, POTW, or other (specify) and control technology, ifany, that will be used to limit the release ofthe new substance to the environment. (14) -- Identify byproducts which may result from the operation. (3). 

### (5), (8). (11), (13) and (15) -- Mark 

### (X) this column if any ofthe proceeding entries are confidential business information (CBI). Letter #of Duration of Protective Equlp./Englneering Controls/Physical %new %in

of Workers CBI CBI CBI Activity Exposed Exposure Form substance Formulation

(1) (2) (3) (4a) (4b) (5) (6) (6) (7) (8) 

### XXX XXX 

### X XXX XXX 

### X XXX XXX XXX 

### X XXX XXX 

### X XXX XXX 

### X XXX XXX XXX 

### X XXX XXX 

### X XXX XXX 

### X XXX XXX XXX 

### X

Release Amountof New Substance Released CBI Media ofRelease &Control Technology CBI Number

(9) (108) (10b) (11) (12) (13) 

### XXX XXX XXX 

### X XXX 

### X XXX XXX XXX 

### X XXX 

### X XXX XXX XXX 

### X XXX 

### X

Mark 

### (X) this box ifthe data continues on the next page. 

# [ J <<' ' (14) Byproducts: 

# I XXX 

# I (15) CBI 

# 00 Enter Attachment filename for Part II, Section B. 

# D
EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 17**l~** k) •,.J _:;111111111 SANITIZED SUBMISSION PMN2010P11 

### PMN Page 11 

## OPTIONAL POLLUTION PREVENTION INFORMATION To claim information in the following section as confidential, bracket (e.g. 0) the specific information thatyou claim as confidentiaL In this section you may provide information not reported elsewhere in this form regarding your efforts to reduce or minimize potential risks associated with activities surrounding manufacturing, processing, use and disposal ofthe PMN substance. Please include new information pertinent to pollution prevention, including source reduction, recycling activities and safer processes or products available duetothe newchemical substance. Source reduction includes the reduction in the amount ortoxicity ofchemical wastes by technological modification. process and procedure modification, product reformulation, and/or raw materials substitution. Recycling refers to the reclamation of useful chemical components from wastes that would otherwise betreated or released as air emissions orwater discharges, or land disposal. Quantitative or qualitative descriptions of pollution prevention, source reduction and recycling should emphasize potential risk reduction in addition to compliance with existing regulatory requirements. The EPA is interested in the information to assess overall net reductions in toxicity or environmental releases and exposures. not the shifting of risks to other media (e.g.• air to water) or nonenvironmental areas (e.g., occupational or consumerexposure). To the extent known. information aboutthetechnology being replaced will assist EPA in its relative risk determination. In addition. information on the relative cost or performance characteristics ofthe PMN substance to potential alternatives may be provided. Describe the expected net benefits, such as

(1} an overall reduction in risk to human health or the environment; (2) a reduction in the generation ofwaste materials through recycling, source reduction or other means; (3) a reduction in the use ofhazardous starting materials, reagents, orfeedstocks; (4) a reduction in potential toxicity. human exposure and/or environmental release; or (5) the extent to which the newchemical substance may be a substitute for an existing substance that poses a greater overall risk to human health orthe environment. Information provided in thissection will be taken into consideration during the review ofthissubstance. See PMN Instructions Manual and Pollution Prevention Guidance manual for guidanceand examples. The newchemical substance is expected to offerthe following Pollution Prevention benefits: {XXX}

EnterAttachment filename for Pollution Prevention Page 11 

# I 

# D

EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 181111111111111111 SANITIZED SUBMISSION PMN2010P12 

### PMNP age 12 Part 

### Ill -- LIST OF ATTACHMENTS Attach continuation sheets for sections ofthe form, test data and other data (including physical/chemical properties and structure/activity information), and optional information after this page. Clearly identify the attachment and the section ofthe form to which it relates, ifappropriate. Numberconsecutively the pages ofany paper attachments. In the Number of Pages column below, enterthe inclusive page numbers of each attachment for paper submissions or enterthe total number of pages for each attachment for electronic submissions. Electronic attachments can be identified by filename. Mark (X) the "Confidential" box nextto any attachment name orfilename you claim as confidential. Read the Instructions Manual for guidance on howto claim any information in an attachment as confidential. You must include with the sanitized copy ofthe notice form a sanitized version ofany attachment in which you claim information as confidential. Nu Associated # Attachment Name Attachment Filename ofPag PMN Section CBI Number 001 Material Safety Data Sheet 7 002 Summary of Information on the New Chemical 1 Substance 003 CAS Inventory Expert Service Report and 4 PU, Sec.B. 2c. 

### IPt.l, Sec.B. 2e. Chemical Structure ofthe New Chemical 004 Impurities Analysis on the New Chemical 2 Pt.l, Sec.B, 3. Substance 005 Manufacturing and Processing Information on 2 the New Chemical Substance at Industrial Sites 006 Manufacturing and Processing Information on 1 the New Chemical Substance at Industrial Sites 007 Worker/Occupational Exposure Information on 1 the New Chemical Substance at Industrial Sites 008 Environmental Release and Disposal Information 1 on the New Chemical Substance at Industrial 009 Processing Information on the New Chemical 2 Pt.2, Sec.B, 1a. Substance at Industrial Sites Controlled by 010 Worker/Occupational Exposure and 1 Pt.2, Sec.B, End Environmental Release Information on the New 011 Relevant Data on Analogous Chemical 5 Substances 012 Physical and Chemical Properties Data on the 15 PU, Sec.B, 2a. 013 Health Effects Data on the NewChemical 19 Substance- Acute Oral Toxicity Test 014 Health Effects Data on the New Chemical 33 Substance- Skin Irritation Test 015 Health Effects Data on the NewChemical 32 016 Health Effects Data on the New Chemical 34 Substance- Ames (Bacterial Reverse Mutation)

Mark (X) this box ifthe data continues on the next page. 

# D EPA FORM 7710-25 (Rev. 6-09) Replaces previous editions of EPA Form 7710-25 Page 19

# lllllmllllllll SANITIZED SUBMISSION PMN2010P13 

### PMNP 

## 13 

## age PHYSICALAND CHEMICAL PROPERTIES WORKSHEET The information on this page refers to chemical number(s): 

# 01 [ J 2 

# l 

# J 3 

# 04 l 

# J 5 

# 06 To assist EPA's review ofphysical and chemical properties data, please complete the following worksheet for data you provide and include it in the 

# I notice. Identify the property measured, the value ofthe property, the units in which the property is measured (as necessary), and whether or not the property is claimed as confidential. Give the attachment number (found on page 12) in column (b). The physical state ofthe neat substance should be provided. These measured properties should be for the neat (100% pure) chemical substance. Properties that are measured for mixtures or formulations should be so noted(% PMN substance in_). You are not required to submit this worksheet; however, EPA strongly recommends that you do so, as it will simplify the review and ensure that confidential information is properly protected. You should submit this worksheet as a supplement to your submission of lest data. This worksheet is not a substitute for submission oftest data. Property Unit 

# I MarkX if Attachment Value Measured CBI Number or Estimate Mark (X) (a) Provided (b) (C) (MorE) (d) 

# 0 (solid) 

# I (liquid) (gas) Physical state of neat substance 

# 0 [X] 

# 0 Measured Vapor Pressure @ 

## oc 

# 0 Torr Temperature Density/relative density 

# 0 gfcm3 Solubility @ Temperature 

## ·c 

# 0 g/L Solvent Solubility in Water @ 

## ·c 

# D giL Temperature Melting Temperature 

# D •c Boiling I Sublimation Torr 

# 0 ·c temperature @ Spectra 

# 0 Dissociation constant 

# 0 Octanol/water partition coefficient 

# 0 Henry's Law constant 

# 0 Volatilization from water 

# D Volatilization from soil 

# 0 pH@ concentration 

# 0 Flammability 

# 0 Explodability 

# 0 Adsorption I Coefficient 

# 0
Particle Size Distribution 

# D ' Other- Specify 

# D

Page 20

### ATTACHMENT HEADER SHEET

Attachment Number 001 Attachment Name Material Safety Data Sheet 

### Associated PMN Section Number N/A
Does not contain CBI



### Report Number CMTP101105088166808TS-CMTP01 

### Attachment #1 MaterialSafetyDataSheet The following page provides a representative material safety data sheet (MSDS) on the flnal formulated productcontaining the new chemical substance. There are expected to bevarious formulations containing the new chemical substance. However, the MSDSs are all expected to be similar.

# [ 

# J MATERIAL SAFETY DATA SHEET 

# [ J Page 1 

# [ 

# J ISection 01 - Product Information Revision Date: 09/1 0/2010 Date of printing :09/10/2010

Identification ofthe company: 

# [ 

# J
[ Information ofthe substance/pjparation: Emergencytel. number: +1 800-424-9300 CHEMTREC Trade name: Material number: 

# [ Chemical family: Primary product use: 

# J Dispersion of perfluoroalkyl acrylic copolymerisate -preparation Intermediate product 

# ISection 02 - Composition information on hazardous ingredients Hazardous ingredients: Component CAS-no. (Trade secret no.} Dipropylene Glycol 25265-71-8 Ethoxylated Alcohols 

# ISection 03 - Hazards identification Expected Route ofentry: Inhalation: Skin contact: Eye contact: Ingestion: Skin absorption: Health effects ofexposure: Propanol, oxybis- ( 25265-71-8) no yes yes yes no Concentration
< 10% <5%

Dipropylene Glycol: LD50 (oral)(rat): 14.85 g/kg; LD50 (dermal){rat) : > 20 mllkg. Has a low order oftoxicity following acute ingestion, but little data are available. Based on animal data, it is estimated that the acute lethal dose in adults is greaterthan one pint. Liver and kidney damage were observed in animals. Minor corneal irritation was reported in animals. May cause mild skin irritation. Central nervous system depression was reported in dogs given 6 mllkg intravenously. No toxicity was noted when 5 mllkg was given orally. Extremely large amounts may produce liver and kidney injury. No untoward effects have been reported from the use ofdipropylene glycol nor would any be expected based on human experience.

# [ 

# J MATERIAL SAFETY DATA SHEET 

# [ J Page 2 

# [ 

# J

Ethoxylated Alcohol Revision Date: 09/10/2010 Date of printing :09/10/2010

Some ethoxylated alcohols have been known to cause eye and skin irritation. Ingestion may cause gastrointestinal irritation, diarrhea, nausea and vomiting. Known effects on other illnesses: No known effects on other illnesses for this product. listed carcinogen: lARC: NO NTP: NO OSHA: NO Other: No HMIS: Health: 1* Flammability: 1 Reactivity: 0 Personal protection: C 

# I Section 04 - Firstaid measures After inhalation: Get victim to fresh air. Give artificial respiration or oxygen if breathing has stopped. Get prompt medical attention. Do not give fluids ifvictim is unconscious. After contactwith skin: Wash thoroughly with soap and water for 15 minutes. If skin irritation occurs, seek medical attention. After contactwith eyes: Immediately flush eyes with running water for at least 

### 15 minutes, occasionally lifting the upper and lower eyelids. Seek prompt medical attention if redness or irritation occurs. After ingestion: Seek medical attention immediately. Advice to doctor ITreatment: None known. 

# I Section 05 - Firefighting measures Flashpoint: ca. 212 oF Hazardous combustion products: In case offires, hazardous combustion gases are formed: carbon oxides nitrogen oxides Hydrogen fluoride (HF) Extinguishing media:
Special fire fighting procedure: water sprayjet foam dry powder carbon dioxide Use NIOSH/MSHA approved self-contained breathing apparatus and other proper protective equipment where this chemical is involved in a fire. Use water spray to cool containers. Unusualfire and explosion hazards: Emits toxic fumes under fire conditions.

## MATERIAL SAFETY DATA SHEET 

# [ 

# J [ J Page 3 

# [ 

# J I Section 06 - Accidental release measures Steps to be taken in case of spill or leak: Revision Date: 09/10/2010 Date of printing :09/10/2010

Contain spill. Ensure adequate ventilation and wear appropriate personal protective equipment. Collect onto inert absorbent. Place in sealable container. Do not allow to contaminate water sources or sewers. 

# ISection 07 - Handling and storage Advice on safe handling: Avoid formation of aerosols. Avoid breathing vapors and contact with skin, eyes. and clothing. Further info on storage conditions: Store in original container. Store in a cool, dry, well-ventilated area. Keep container sealed when not in use. Keep from freezing. 

# I Section 08 - Exposure controls 

### Ipersonal protection Respiratory protection: If airborne concentrations pose a health hazard, become irritating or exceed recommended limits, use a NIOSH approved respirator in accordance with OSHA respiratory protection requirements under 29 CFR 1910.134 Hand protection: Butyl Rubber, PVC Or Neoprene. Eye protection: Safety glasses or chemical splash goggles. Other protective equipment: Clothing suitable to prevent skin contact. Advice on system design: Local ventilation recommended - mechanical ventilation may be used. IDLH: Not Determined 

# I Section 09 - Physical and chemical properties Form: Color: Odor: pH: Solubility in water: Density: Boiling point : Dispersion milky white to yellowish characteristic ca.4 miscible 1.1 g/cm3 ca. 212 °F

# MATERIAL SAFETY DATA SHEET 

# [ [ J Page 4 

# [ 

# J ISection 10 ·Stability and reactivitY Chemical stability: Stable. Hazardous Polymerization: Will not occur. Conditions to avoid: None known. Conditions to avoid: Strong oxidizing agents. 

# ISection 11 -Toxicological information Product information: LD50 > 2,000 mg/kg (rat) Revision Date: 09/10/2010 Date of printing :09/10/2010


# II Acute oral

## toxic~ Directive 1999/45/EC, Information based on the main component. Skin irritation: Eye irritation: non-irritant (rabbit) Method: 1999/45/EC non-irritant (rabbit eye) Method: 1999/45/EC 

# I Section 12 - Ecological information Product information: Fishtoxicity: Bacteria toxicity: Chemical oxygen demand (COD): Remarks: LC50 > 100 mg/1 (96 h, zebra fish (brachydanio rerio)) Method: evaluated IC50 > 100 mg/1 (activated sludge) Method: evaluated 550

- 600 mg/1 Method: estimated from the components No ecological testing was carried out on the preparation Ecological data are determined by analogy 

# ISection 13 - Disposal considerations Waste disposal information: Consult local, state, and federal regulations. RCRA hazardouswaste: No -- Not as sold. RCRA number: NONE 

# ISection 14- Transport information DOT lATA not restricted not restricted 

# J

## MATERIAL SAFETY DATA SHEET 

# [ 

# J [ J Page 5 

# J

IMDG not restricted 

# I Section 15- Regulatory information TSCA Status: Revision Date: 09/10/2010 Date ofprinting :09/10/2010

A component ofthis product is not listed on the TSCA Inventory. The component meets the definition of polymer as found in 40 CFR 723.250 and thereby is exempt from certain Premanufacture Notification requirements ofTSCA. SARA (section 311/312): Reactive hazard: no Pressure hazard: no Fire hazard: no Immediate/acute: yes Delayed/chronic: yes SARA 313 information: This product does notcontain any toxic chemical listed under Section 313 ofthe Emergency Planning and Community Right-To-Know Act of 1986. Clean Water Act: Contains no known priority pollutants at concentrations greater than 0.1%. Volatile organic compounds VOC: Remarks: Nottested 

### FDA: Permitted for Use per Section: 

# I Section 16 - Other information Other precautions: NONE This product is not registered with the FDA.

Avoid inhalation ofthis product in mist or aerosol forms. Label information: CAUTION! MAY CAUSE GASTROINTESTINAL DISTURBANCES IF INGESTED. REPEATED EXPOSURES MAY RESULT IN LIVER OR KIDNEY DAMAGE. Avoid breathing fumes, vapors, mists or spray. Avoid contact with skin and eyes. Use with adequate ventilation and/or respiratory protection. Wash thoroughly after handling. Keep container closed when not in use. Wear proper protective equipment when handling. Skin contact: wash thoroughly with soap and water for 15 minutes. If skin irritation occurs, seek medical attention. Wash contaminated clothing before reuse. Eye contact: flush with waterfor at least 15 minutes while holding eyelids open. Seek immediate medical attention. Ingestion: seek medical attention immediately. Inhalation: remove to

# MATERIAL SAFETY DATA SHEET 

# [ [ J Page 6 

# l J Revision Date: 09110/2010 Date of printing :09/10/2010

fresh air. Ifnot breathing, give artificial respiration. Ifbreathing is difficult, give oxygen. Seek medical attention if respiratory irritation continues. NFPA: Health: 1 Flammability: 1 Reactivity: 0 This information is supplied underthe OSHA Hazard Communication Standard, 29 CFR 1910.1200, and is offered in good faith based on data available to us that we believe to be true and accurate. The recommended industrial hygiene and safe handling procedures are believed to be generally applicable to the material. However, each user should review these recommendations in the specific context ofthe intended use and determine whether they are appropriate forthat use. No warranty, express or implied, is made regarding the accuracy ofthis data, the hazards connected with the use ofthe material, orthe results to be obtained from the use thereof. We assume no responsibility for damage or injury from the use ofthe product described herein. Data provided here are typical and not intended for use as product specifications. 

# J

### ATTACHMENT HEADER SHEET

Attachment Number 002 Attachment Name Summary of Information on the New Chemical Substance 

### Associated PMN Section Number N/A
Does not contain CBI



### Report Number CMTP101105088166808

### TS-CMTPOl 

### Attachment#2 Summaryof Infonnation on theNew ChemicalSubstance

I

'------------

### ATIACHMENT HEADER SHEET

Attachment Number003 Attachment Name CAS Inventory Expert Service Report and Chemical Structure ofthe New Chemical Substance 

### Associated PMN Section Number Pt.l, Sec.B, 2c. 

### I Pt.l, Sec.B, 2e.
Does not contain CBI



### Report Number CMTP101105088166808

### TS-CMTPOl 

### Attachment#3 Pt. 1, Sec. B,2 

### CASInvent01yExpertService Reportand ChemicalStructure of the New Chemical Substance

# Inventory

# ExpertService
Adivision oftheAmerican Chemical Society Phone: 800-631-1884, 614-447-3870 Fax: 614-447-3747 E-mail: answers@cas.org Web:

### www.cas.org/products/dientl



# [ INVENTORY EXPERT SERVICE REPORT

IES-ORDER NUMBER: 152304 REGISTRY NUMBER:[ 

# J

CA INDEX NAME

Please print the above CA Index Name on the appropriate page of your PMN.

CAS 

### If this box is checked, CAS has made correction(s) marked 

### in red to your IES order. Please make the same correction(s) to your PMN before submitting it to the EPA.

2540 Olentangy River Road P.O. Box 3343 Columbus, Ohio 43210-0334 USA 

# J

### ATTACHMENT HEADER SHEET

Attachment Number 004 Attachment Name Impurities Analysis on the New Chemical Substance 

### Associated PMN Section Number Pt.l, Sec.B, 3.
Does not contain CBI



### Report Number CMTP101105088166808TS-CMTPOl 

### Attachment#4 Pt. 1, Sec. B, 3 

### ImpuritiesAnalysis on theNew ChemicalSubstance

Impurity CASRN %(max.) Method of Comments Detection

### TS-CMTPOl

### ATTACHMENT HEADER SHEET

Attachment Number 005 Attachment Name Manufacturing and Processing Information on the New Chemical Substance at Industrial Sites Controlled by the Submitter- Manufacture 

### Associated PMN Section Number N/A
Does not contain CBI



### Report Number CMTP101105088166808

### TS-CMTP01 

### Attachment#5 Pt. 2, Sec. A, td 

### Manufacturing and Processing Information on the New Chemical Substance at IndustrialSites ControlledbytheSubmitter-Manufacture

# ~S-CMTP01



# L Attachment#5 

### ManufacturingandProcessing Information on the New Chemical Substance atIndustrial

### Sites Controlledbvthe Submitter- Manufacture

### ATTACHMENT HEADER SHEET

Attachment Number 006 Attachment Name Manufacturing and Processing Information on the New Chemical Substance at Industrial Sites Controlled by the Submitter- Processing 

### Associated PMN Section Number N/A
Does not contain CBI



### Report Number CMTP101105088166808

### TS-CMTPOl 

### Attachment#6 Pt. 2, Sec. 

### A, ld 

### Manufacturing and Processing Information on the New Chemical Substance at Industn'alSites ControUedbytheSubmitter- Processing

### ATTACHMENT HEADER SHEET

Attachment Number 007 Attachment Name Worker/Occupational Exposure Information on the New Chemical Substance at Industrial Sites Controlled by the Submitter- Processing 

### Associated PMN Section Number N/A
Does not contain CBI



### Report Number CMTP101105088166808

### TS-CMTP01 

### Attachment#7 Pt. 2, Sec. A, 2 

### Worker/Occupational Exposure Information on the New Chemical Substance at IndustrialSites ControUedbytheSubmitter- Processing

### ATTACHMENT HEADER SHEET

Attachment Number 008 Attachment Name Environmental Release and Disposal Information on the New Chemical Substance at Industrial Sites Controlled by the Submitter- Processing 

### Associated PMN Section Number N/A
Does not contain CBI



### Report Number CMTP101105088166808

### TS-CMTPOt 

### Attachment#8 Pt. 2, Sec. A, 3 EnvironmentalRelease andDisposalInformation on the New Chemical Substance atIndustrialSites ControUedbytheSubmitter- Processing

### ATTACHMENT HEADER SHEET

Attachment Number 009 Attachment Name Processing Information on the New Chemical Substance at Industrial Sites Controlled by Others 

### Associated PMN Section Number Pt.2, Sec.B, 1a.
Does not contain CBI



### Report Number CMTP101105088166808TS-CMTP01 

### Attachment#9 Pt. 

### 2, Sec. B, la 

### Processing Infonnation on the New Chemical Substance at Industrial Sites ControUedbyOthers

: <f\ _;TS-CMTP01 

### Attachment 9 Processing Information on the New Chemical Substance atIndustrial Sites Controlledbv Others

### ATTACHMENT HEADER SHEET

Attachment Number 010 Attachment Name Worker/Occupational Exposure and Environmental Release Information on the New Chemical Substance at Industrial Sites Controlled by Others 

### Associated PMN Section Number Pt.2, Sec.B, End
Does not contain CBI



### Report Number CMTP101105088166808

### TS-CMTPOl 

### Attachment#10 Pt. 2, Sec. B, End 

### Worker/Occupational Exposure and Environmental Release Information on the NewChemicalSubstance atIndustrialSites ControlledbyOthers

### ATTACHMENT HEADER SHEET

Attachment Number 011 Attachment Name Relevant Data on Analogous Chemical Substances 

### Associated PMN Section Number N/A
Does not contain CBI



### Report Number CMTP101105088166808

### TS-CMTPOl 

### Attachment#11 RelevantDataonAnalogous ChemicalSubstances

2

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### TS-CMTP01

4

6

6

### TS-CMTPOl

B

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11

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17
18 19
20

### ATTACHMENT HEADER SHEET

Attachment Number 012 Attachment Name Physical and Chemical Properties Data on the New Chemical Substance - Molecular Weight Data 

### Associated PMN Section Number Pt.l, Sec.B, 2a.
Does not contain CBI



### Report Number CMTP101105088166808TS-CMTPOt 

### Attachment#12 Physical and ChemicalProperties Dataonthe NewChemicalSubstance 

### Molecular WeightData

# 1-

### ATTACHMENT HEADER SHEET

Attachment Number013 Attachment Name Health Effects Data on the New Chemical Substance- Acute Oral Toxicity Test 

### Associated PMN Section Number N/A
Does not contain CBI



### Report Number CMTP101105088166808

### TS-CMTPOl 

### Attachment#13 Health Effects Dataon the NewChemicalSubstance 

### Acute Oral Toxicity Test

# L REPORT



## Acute Oral Toxicity Study 

## in Rats

Stutly Director: Test Facility: 

### Dr. C. Simon RCC Ltd Wolferstrasse 4 

# J

4414 

### FiillinsdorfI 

### Switzerland 

# [
Study Identification: RCC Study 

### Number B46462 Version: Final 

### Study Cmnplt-tion 

### Date: 05-NOV-2007 

### Page I of 18 

# J[ RCC Study**~tun!K'r** 1Ht•46l 

### Report Page 2


## CONTENTS C()N'IFN'i'S.,.,, ..,...........................................................................................................................2 TABLr: LIS·1·
..."
.............................................................................................................................2 

### SICiNAI'UHES
................................................................................................................................3 STATEMI.NI' OF ('0MPLIANCE
................................................................................................4 QUALITY ASSURANCE
..............................................................................................................

### 5 PREFACE
....................................................................................................................................... 6 (ienerallnfonnation
...............................................................................................................6 

### Responsibilities
......................................................................................................................6 Schedule
.................................................................................................................................6 Ciuichdincs
..............................................................................................................................

### 7 Ani1nal Welt(lrc
......................................................................................................................7 Archiving
...............................................................................................................................7 SUI\!Iiv1ARY
........................................................................................................................... 8 

### 2 l-·(JNl.'l.LJSION,.................**~~························**..**··~·~~···············~·····~·~········~~··~······~·······················** 9 

### 3 

### PURP()SF
............................................................................................................................ 10 

### 4 !VIi\ 

### I'ERIALS AND METHODS
......................................................................................... 11 4. I Test Systen1
............................................................................................................ II 

### 4.2 llusbanury
.............................................................................................................. II 4 .

### .1 Test lten1
................................................................................................................ 

### 12 

### 4.3.1 Test ltenl
.................................................................................................. 12 

### 4
..**~.2** 

### Vehicle
..................................................................................................... 12 

### 4.4 T>ose 

### Fonnulation
.................................................................................................. 13 4.5 Tn:at1nent
............................................................................................................... 

### 13 4.6 

### ()bservations
.......................................................................................................... 14 4.7 Pathology
............................................................................................................... 14 

### 4.7.1 Necropsy
.................................................................................................. 14 'l.X Statistical Analysis
................................................................................................ 

### 14 4.() [)ata 

### Con1pilation
................................................................................................... 14 

### 5 RESULTS
............................................................................................................................ 15 5.1 Mortality
................................................................................................................ 1

### S 5.2 

### Clinical Signs
......................................................................................................... IS 5.3 Body 

### Weights
........................................................................................................ 15 

### SA Macroscopic Findings
............................................................................................ 15 '> ') 

### Median Lethal Dose
............................................................................................... 15 6 INDIVIDUAl. FINDINGS
.................................................................................................. 16 7 

### CILP-CERTIFlCATE
........................................................................................................... 18 

## TABLE LIST Table I Mortality I 

### Clinical Signs
............................................................................................ 16 Table 2 Body Weights
.............................................................................................................. 16 

### Table 3 

### Macroscopic Findings
................................................................................................. 

### 17

# [ RCC Study Number 1346462 

# 1. Report 

### Page 3


## SIGNATURES Study 

### Director: Dr. C. Simon

Management: 

# (f'} Dr. H. Fankhauser

# ~ ·! ' 'i ' 

# .............**..~**

# ...\1-::,.'fr!.l, Date:
..\ lt

## ~ \.," 1 I
. . \ ) 1

. J v**lt~,,** V\.•**~.v-~** 

### . lL. .\...... 

# ·}

# [ RCC Study Number 846462 Report 

### Page4



### GOOD LAHORATORY PRACTICE 

# STATEMENT OFCOMPLIANCE RCC Study Number: B46462 Test 

### Item: 

# [ 

# J Study Director: 

### Dr. C. Simon Study Title: Acute Oral Toxicity Study in Rats

The stability of the test item dilutions under the test conditions is unknown. The formulation trials were performed before the study initiation date. Therefore, they are excluded from this statement. This study has been performed in compliance with the Swiss Ordinance relating to Good Laboratory Practice, adopted May 18th, 2005 [RS 813.112.1]. This Ordinance is based on the OECD Principles ofGood Laboratory Practice, as revised in 1997 and adopted November 26th, 1997 by decision ofthe OECD Council [C(97) 186/Final]. There were no circumstances that may have affectedthe quality or integrity ofthe data.
Study Director: Dr. C. Simon
'

# 1/

### [ RCC Study Number B46462 Report 

### Page 5 

# QUALITYASSURANCE RCC Ltd, W olferstrasse 4, 4414 FiillinsdorfI 

### Switzerland 

### STATEMENT RCC Study Number: B46462 Test 

### ltem: 

# [ 

# ;J Study Director: Dr. C. Simon Study Title: Acute Oral Toxicity Study in Rats The general facilities and activities are inspected periodically and the results are reported to the responsible person and the management. Study procedures, with exception ofthe formulation trials, were periodically audited. The study plan and this report were audited by the Quality Assurance. The dates are given below. 

### Dates ofReports to Dates andTypes ofQAInspections the StudyDirector and TestFacility

Management 30-AUG-2007 Study Plan 30-AUG-2007 22-AUG-2007 Process Based 22-AUG-2007 (Test System, Test Item, Raw Data, Dose Preparation, Treatment) 25-0CT-2007 Report 25-0CT-2007

This statement also confirms that this final report reflects the raw data.
Quality Assurance: 

### S. van Dongen**·~** '\ ' '



# ......:'
......**~**

# ..>:**.:·.~**

# ...:.**/~.:~~:~.~~. ~.~.~** 

# .... Dall':
. . _ , .. > 1 ·, . • • . • \,_,' '-·**~.** \ » •• 1._'-i.t

(:"-II ;J '.

# [ RCC Study Numb-:r B4Mb2


## PREFACE General Information Title: Sponsor:
Study 

### Monitor: Test Facility:


## Responsibilities Study Director: 

### Deputy Study Director: Tedmical Coordinator: He<id ofLead QA: 

## Schedule Experimental Starting Date: 

### Experimental Completion Date Delivery ofAnimals: 

### A

eelirnatization: Treatment: Observation: 

# [ 

# [ [ Report



### RCC Ltd Wolferstrasse 4 4414 FlillinsdorfI Switzerland



### Dr. C. Simon 

### G. Arcelin 

### F. Frickert I 

### M. Bernstein I. Wuthrich**04~SEP-2007** 

### 27-SEP-2007 04-SEP-2007 (females. 2000 mg/kg) 06-SEP-2007 (females. 2000 mg/kg) Page6 

# J 

# J

04-SEP-2007 to I0-SEP-2007 (females, 2000 mg/kg) 06-SEP-2007 to 12-SEP-2007 (females, 2000 mg/kg) ll-SEP-2007 (females, 2000 mglkg) 13-SEP-2007 (females, 2000 mg/kg) 04-SEP-2007 to 25-SEP-2007 (females, 2000 mg/kg) 06-SEP-2007 to 27-SEP-2007 (females, 2000 mg/kg)

# l RCC S1udy Numh.:r 046462 Report (>age 7


## (;uidelines The study procedures described in this report meet or exceed the requirements ofthe following guidelint:"s:
• Directive 2004/73/EC, 

### B. I tris "Acute Oral Toxicity-Acute Toxic Class Method", April 

### 29, 2004. • OFCD Guidelines for the Testing ofChemicals, Number423 "Acute Oral Toxicity**A~:utc** 

### Toxic Class Method", adopted 17th December 200 

### I. 

### Animal Welfare This study was perHmned in an AAALAC-approved laboratory in accordance with the Swiss Animal Protection Law under license no. 34. 

### Archiving RCC Ltd 

### (CII-4452 ltingcn I Switzerland) will retain the study plan, raw data, a sample oftest item(s) 

### and the linal repo11 ofthe present study for at least ten years. No data will be discarded without the Sponsor's written consent.**Pag~** 8


## I SUMMARY Two groups. each of 

### three female HanRcc:WlST (SPF) 

### rats, were treated 

### witl{ ]by 

### oral gavage administration at a dosage of 2000 

### mg/kg body weight. The tt'st item was diluted in vehicle (corn oil) at a concentration of0.2 g/mL and administered at a 

### dosing volume of I0 mL!kg. 

### The animals were examined daily during the acclimatization period and mortality, viability and dinical**~igns** 

### were**r~:cordcd.** 

### All 

### animals were examined for clinical signs within the 

### first 30 

### mi11nles 

### and approximately I, 2, 3 and Shours after treatment on day I and once daily during test days 2-1 ). 

### tvlortality/viability was recorded at approximately 30 

### minutes, I. 2. 

### 3 

### and 5 

### hours aftt·r administration on test day I (with the clinical signs) and twice daily during days 2-15. Body**wei~.hts Wt~n:**  

### recorded 

### on 

### day l (prior to administration) and on days 8 and 15. All animals were**necropsi~~d** 

### and examined macroscopically. AII animals survived until the end ofthe study period. No 

### clinical signs were observed during the course ofthe study. The body weight ofthe animals was within the range commonly recorded tor 

### this strain and age. No macroscopic findings were recorded at necropsy.Report**Pag~:** 9


## 2 CONCLUSION Tile median lethal 

### Jose o{ Jafter single oral administration 

### to female rats, obscrwd over 

### a period of 14 days is:
LDSO (temale rat): greaterthan 2000 mg/kg 

### body weight

··\ -, .j'',v**-~·** ;;_:Page 10


## 3 PlJRPOSE The purpose ofthis study was to assess the acute toxicity

### o~ ] when admin**istcrl~d** 

### hy a single oral gavage to rats. followed by an observation period of 14 days. This study provides inf(mnation 

### for both hazard assessment and hazard classification purposes.

# [ R('C Sl.udv Number B.to462 Report Page 11


# 4 MATERIALS AND METHODS 4.1 Test System Animals: Rationale:**Some~·**



### Number ofAnimals per Group: Total number ofAnimals: Age when treated: I

### denti**fi~.:ation:** 

### Randomization: 

### Aeel

### imatiz.ation:



### 4.2 Husbandry Room Number: Conditions:

Accommodation: Rat, HanRcc: WIST(SPF) Recognized by international guidelines as a recommended test system. RCC Ltd Laboratory Animal Services Wolferstrasse 4 4414 FullinsdorfI Switzerland 

### 3 females 6 

### females II 

### weeks Unique cage number and corresponding color-coded spots on the tail. The animals were marked at acclimatization start. Selected by hand at time ofdelivery. No computer generated randomization program. Under laboratory conditions, after health examination. Only animals without any visible signs ofillness were used for the study.

0I

### 05 I RCC Ltd, Fullinsdorf Standard Laboratory Conditions. Air-conditioned with 10-15 air changes per hour, and continuously monitored environment with ranges for room temperature 22 ± 

### 3 

# oc and for relative humidity between 30-70 % 

### (values above 70 % 

### during cleaning process possible), automatically controlled light cycle of 12 hours light and 12 hours dark, music during the daytime light period. In groups of three in Makrolon type-4 cages with wire mesh tops and standard softwood bedding ('Lignocel' Schill AG, CH-4132 Muttcnz/Switzerland).

# [ RCC Study Number fHMfi2



### Diet: Report Page 12



### Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 23/07 (Provimi Kliba AG, CH-4303 Kaiseraugst/Switzerland) ad libitum. Results ofanalyses for contaminants are archived at RCC Ltd. Water: Community tap water from Fiillinsdorf ad libitum.



## 4.3 Test Item 4.3.1 Tt•st Item 

### Results ofbacteriological, chemical and contaminant analyses are archived at RCC Ltd.

The following information was provided by the sponsor: Identification Synonym 

### Description Batch number Purity I 

### Formulation Stability oftest item 

### Expiry date Stability oftest 

### item dilution Storage coml

### itions Safetv 

### precautions

4.3.2 Vchicle 

# [ [ Solid; 

### light brown 

# ] 1 

### 00% active polymer Stable under storage conditions. 31-MAR-2008 

# J

Unknown in corn oil: is excluded fi·om the statement of compliance. At room temperature (range of20 :1 

### 5 

### °C), light protected. Routine hygienic procedures were used to ensure the health and 

### satcty ofthe personnel.

Corn oil was found to be a suitable vehicle. The vehicle was chosen after a non-GLP solubility trial which was performed before the study initiation date. This formulation trial is excluded from the GLP statement ofcompliance.[ RCC Sludy Numher B4M62 Page 13



### The f(JIIowing information was provided by RCC Ltd: ldenlilica1iou Dcscriplion Batch number Source Stability ofvehicle Storage conditions Safety precautions


## 4.4 Dose Formulation Corn oil Yellowish oily liquid 27572782 

### Carl Roth GmbH & 

### Co. D-76185 

### Karlsruhe I

### Germany Stable under storage conditions; expiration date: February 2008 At room temperature (range of20 ± 

### 5 °C}, 

### light protected. Routine hygienic procedures were used to ensure the health and safety ofthe personnel.



### l>osc levels are in terms ofthe test item as supplied 

### by the sponsor. The dose formulations were made shortly before each dosing. First the vehicle was heated up to I00-11

# ooc and the determined amount of the test item was added 

### while stirring on a 

### magnetic stirrer. Additionally, an Ultra-Turrax (Janke & 

### Kunkel, D7921 q Stauti.!n) was 

### used to get a 

### homogeneous suspension. Thereafter, the formulation cooled down 'vhik still 

### stirring on the magnetic stirrer and was applied at 

### a temperature of 20-30°C. The kmpcraturc 

### was 

### measured before each treatment and was found to 

### be 26.4 

### and 25.8°C. respt:·.:tivcly. 

### Homogcnt>ity of the test 

### item in the vehicle was maintained during administration using a magnetic stirrer. 

## 4.5 Treatment The animals received a single dose ofthe test item by oral gavage administration at 2000 mg/kg body weight alter 

### being fasted t{)r approximately 17 to 18 hours (access to water was permitted). Food \vas provided again approximately 3 hours after dosing. The dosing volume was I0 mL/kg body weight Rationale: Oral administration was considered to be an appropriate application method as it is a possible route 

### ofhuman exposure during manufacture, handling and use ofthe test item.[ RCC Sru.Jy ·I'Jwnher l:H6462



## 4.6 Observations Mortality I Viability



### Body Weights Clinical Signs



## 4.7 Pathology 4.7.1 Nc,:rotJsy Report Page 14

Daily during the acclimatization period, during the tirst 30 minutes and 

### at approximately 

### t, 

### 2, 3 and 

### 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 

### 2-15. On test days 

### I (priorto administration), 

### 8 and 

### 15. Daily during the acclimatization period, during the first 30 minutes and at approximately 

### 1, 2, 3 and 

### 5 hours after administration on test day 

### I. Once daily during days 

### 2-15. 

### All abnormalities were recorded.

All animals were killed at the end ofthe observation period by carbon dioxide asphyxiation and discarded attcr macroscopic examinations were performed. No organs ortissues were retained. 

## 4.8 Statistical Analysis No statistical analysis was used. 

## 4.9 l>a1a Compilation Body 

### \vcighls \vcre recorded on-line. Clinical sigus were recorded on data sheets. Mortality/viability 

### were recorded on data sheets. Macroscopic findings were compiled into the RCC Tox Computer System during recording. The RCC Tox Computer System (RCC-Tox-Lims) had been validated with respect to data collcdion. storage and rctrievability.RCC Studv Numhcr B4M62 

### Report Page 15 

# [ 5 RESULTS 5.1 Mortality No deaths occurred duringthe study. 

### 5.2 Clinical Signs No clinical signs were observed during the course ofthe study. 

## 5.3 Body 

### Weights The body weight ofthe animals was within the range commonly recorded tor this strain and age. 

### 5.4 Macroscopic Findings No macroscopic findings were recorded at necropsy. 

## 5.5 Median Lethal Dose The median lethal dose o{ to female rats, observed over a period of 14 days is: }fter 

### single oral administration
LDSO (1emale 

### rat): greaterthan 2000 

### mg/kgbody weight

# L RCC Study Numhcr B4M62


# 6 INDIVIDUAL FINDINGS Table 1 Mortality I 

### Clinical Signs**r)~~~~---~1\** mg/kg 111 N**~---------t** .-------- Test days

f!l- al Sex Si. I 2 3 4 5 6 7 

### 8 9 10 fL 0.5* I* 2* 

### 3* 5* 

### 2000 F 

### No clinical signs -.}**~ ~** **~** 

# " -.}**~ ~** **~** 

# ""**~ ~**  

# " -· ·f--··- c--------- 2 F No 

### clinical signs 

# " """"""""

# ""

# "

# "" F No 

### clinical signs 

# " 

# """

# ""

# """""

# " -.} 

# "

----- F No

### clinical signs 

# " """

# """"

# "**~** 

# "**~** 

# "" c--- 

### ----· F No 

### clinical signs ,j 

# "**-~** ,j 

# "

# "

# """**~** 

# "**~ ~**  

# "

·-·-- F No 

### clinical signs**~** 

# "

# " -.} 

# "**~** 

# "

# ""**~** 

# "

# " I -.} 

# " * Examinations wen: pcrtimned within the tirst 30 minutes and L 2. 3 and 5hours alter tr..:atment. 

### No climcal signs were evident in any animal during the acclimatization period.
Table 2 Body Weights mal**IDos;·--~**

### ~~g___ 

# ft<_l._**~---** ·-

-·-**--·~~·~** 

# -2-lli)o·---·],i

-- s 6



# --- ·-·---



# -- -· Sex 

### Dav 1 (

### tr~atment) 

# -- F 203.1



### -F 197.0



### -·--

# -- F 187.0 

## -· F 180.2 F 176.9**~--** F 180.0 

### Body weights are presented in grams. Day 8 Day 15 215.1 224.7 212.4 229.5 201.6 212.6 195.1 205.6 192.9 196.5 198.7 207.2 Page 16

11 12 13 14 15**~ ~**  -.}**~** 

# " "

# """" -.} -.} 

# ""

# " 

# ""

# """ 

# ""

# "**~** 

# " ,j**~ ~**  

# " ,j

# [ RCC**Stud~** Number 1346..162 Report Page 17



### Table 3 Macroscopic Findings**~·---** Dose Animal Sex Mode of Findings**~~g!~__g** 

### ____ No.**~ath** f.-----··- f.---- 

### 2000 I F 

## s No macroscopic findings f-· 

### 2 F 

## s No macroscopic findings .\ 

### r 

## s No macroscopic findings -·-·-· -·-·--·-**··~** ·- -----·-· 

## -- 2000**.~** F 

## s No macroscopic findings

---·------ --·-- ::; F 

## s No macroscopic findings 

## l__
______

# ---'~~--~~~~-=

# (. s No macroscopic lindings 

### S: scheduled necropsy

# [ RCC Study Numbt:r B4M62 Report


## 7 GLP-CERTIFICATE

l"he Sw1ss GLP Monitoring Authorities**sw~ss** Agency forthe Environment, Forests and Uodteapa 

# ;'
.**:=::~;!~cy**

# for SWISsmedlC TnorapoutcProduots 

# Statement of GLP Compliance

ll1s hereby confirmed that during the period of
the following Facilities of April 22. 25

- 29. 2005 May 09- 13, 2005 

### RCC ltd 4452 ltingen Switzerland
we1e mspected by the Federal Office of Public Health. the Swiss Agency for Therapeutic Products and the Sw1ss Agency for the Environment. Forests and Landscape with respect to the compliance with the Swiss legtslation on Good Laboratory Practice. Facilities -Test Facility: Toxicology
-Test Facility: Environmental Chemistry

& Pharmanalytics ·Archive Facilities Areas ofexpertise • TOX, ACC, OTH (Safety Pharmacology, Alternative Test Systems) ACC, ECT, ENF, EMN, PCT, RES, OTH (Animal Metabolism)

The tnspections were performed in agreement with the OECD Guidelines for National GLP Inspections and Audits. It was tound that the aforementioned test facilities were operating in compliance with the Swiss Ordinance relating to Good Laboratory Practice [RS 813.016.5) at the hme they were inspected.

Bern. November 2005 Federal Office of Public Health The Director

Prof. Th. Zeltner

·fOX=- Ton:olog:y, ACC"" APatyhcal and CluW·.at Chem1tUY: ECT=Env1ronmental tmuclty on aquatic and terrcstnal**orgamsrn~:** Et-.F" Behavtour mwattt(, toil .and 111. BioaccumvlatJcm: EMN =Studlfll on .affect& on mesoc.osms and nah.;ral eeosy!>tt!'tn$. PCl.:::. Physlcat.chemtcal testtng :RES" Residue stuclies; OTH; Other, to be spec;1fft!d Page 18

### ATTACHMENT HEADER SHEET

Attachment Number 014 Attachment Name Health Effects Data on the New Chemical Substance - Skin Irritation Test 

### Associated PMN Section Number N/A
Does not contain CBI



### Report Number CMTP101105088166808

### TS-CMTP01 

### Attachment#14 HealthEffects Dataonthe NewChemical Substance 

### SkinIrritation Test

## REPORT 

# J Primary Skin Irritation Study in Rabbits (4-Hour 

# Semi-OcclusiveApplication)

StudyDirector: Test Facility:



### Sponsor: 

# [ Dr. C. Simon Harlan LaboratoriesLtd. Wolferstrasse 4 

### 4414 FiillinsdorfI 

### Switzerland

StudyIdentification: Harlan Laboratories Study Cl5654



### Version: Final StudyCompletionDate: 06-Mar-2009 Page 1

### of32 

# harlanTM


# ....



# JHadan Laboratories StudyC25654 

# r J Report /


## --------·-**----~** 

## CONTENTS C()NTENTS
.......................................................................................................................... 2 SICiNATURES
...................................................................................................................... 4 STATEMENTOF COMPLIANCE
.................................................................. .................... 

### 5 QUALITY ASSURANCE GLP STATEMENT
................................................................... 6 PREFACE
............................................................................................................................. 7 General Infonnation
.... . ................. ........................................... . . ...... ............. ................. 7 Responsibilities
.............................................................................................................. 7 Schedule
......................................................................................................................... 8 Data 

### Requirements I

### Test Guidelines
............................................................................. 

### 8 Animal Welfare
.............................................................................................................. 8 Archtving
........................................................................................................................ 8 

### Classification Guidelines
................................................................................................ 9 l SUMMARY
..................................................................................................................... 10 2 PlJRPOSE
........................................................................................................................ 11 

### 3MATERIALS AND METHODS
..................................................................................... 12 3.1 Test System
............................................................................................................ 12 3.2 Allocation
............................................................................................................... 12 3.3 liusbandry
.............................................................................................................. 13 

### 3.4 TestItem
................................................................................................................. 13 3.5 Preparation ofDose Formulations
.......................................................................... 14 3.6Test 

### Iten1 Administration
....................................................................................... 14 

### 3.7 Rationale
................................................................................................................. 14 3.8 

### Observations
................................ ........ ................................................... . ............ ... 15 

### 3.9 Pathology
................................................................................................................ 15 

### 3.9. 1Necropsy
...................................................................................................... 15 3.10 Statistical Analysis
............................................................................................... 15 3.11 DataCotuplilation
................................................................................................ 15 3.12 Determination ofSkin Irritating Potential
............................................................ 16 

### 3.12.1 Observation and Scoring
............................................................................ 

### 16 4 RESULTS AND DISCUSSION
...................................................................................... 17 4.1 

### (Jbservations
........................................................................................................... 17 4.1.1 Viability 

### IMortality
..................................................................................... 17 4.1.2 

### Clinical Signs
............................................................................................... 17 4.1.3 BodyWeights
............................................................................................... 17 4.2 Pathology
................................................................................................................ 17 

### 4.2.1 Macroscopic Findings
.................................................................................. 17 4.3 Skin Reactions
......................................................................................................... 

### 17 4.3.1 Irritation
........................................................................................................ 17 4.3.2 Coloration
..................................................................................................... 18 4.3.3 Test ItemRemnants
...................................................................................... 18

09009e53805ec755 

### 2Harlan Laboratories Study C25654 Report 

# [ J

4.3.4 Corrosion
...................................................................................................... 18 5 CONCLUSION
................................................................................................................ 19 

### 6 SUMMARY TABLES
..................................................................................................... 20 Skin hTitation Scores
.................................................................................................... 

### 21 Skin Irritation scores- MeanValuesAfter24,48 and 72 Hours
................................ 21 Skin Irritation Scores Assessment according to ECGuidelines
... ................... ......... 21 7 lNDIVII>tJAL

### TABLES
.................................................................................................. 22 SkinR.eactions
... . . ......................... ................................................... ..... ........ . .............. . 23 Body

### Weights
.......... . ................................................................ .......... ................. . ...... . . 26 

### APPENDIX I- 

### COMMISSION REGULATION
............................................................... 28 Commission Regulation (EC)No440/2008, 30MAY 2008
...................................... 29 

### APPENDIX II

- COMMISSION DIRECTIVE
................................................................... 30 Commission Directive

### 2001159/EC, August

### 2001
....................................................... 31**09009e53805~c755** 

### 3Harlan Laboratories StudyC25654 

# [ J Report


## SIGNATURES Study Director: 

### Dr. C. Simon

Ma11agement: 

# ( f'ru) Dr. H. f'ankhauser

## ·?J~~l.vt. Date: 

# 0{).-**~ ~za?j** 

09009e5:18063517(i 

### 4Harlan Laboratories Study C25654 

### Report 

# r J
GOOD LABORATORY PRACTICE 

# STATEMENT OF COMPLIANCE Harlan Laboratories Study: TestItem: 

# [ StudyDirector: Study Title: 

### C25654 

# J Dr. C. Simon Primary Skin Irritation Study 

### in Rabbits 

### (4-Hour Semi-Occlusive

### Application)

The pH measurement of 

### the test item was performed before the 

### study initiation date. This procedure is, 

### therefore, excluded from this statement. This 

### study has been 

### performed in compliance with the Swiss Ordinance relating to Good Laboratory Practice, adopted 

### May 18th, 

### 2005 [SR 

### 813.112.1]. This Ordinance is based on the 

### OECD Principles ofGood Laboratory Practice, 

### as revised in 

### 1997 and adoptedNovember 

### 26th, 1997 

### by decision ofthe

### OECD Council 

### [C(97)186/Final].

Study Director:

09009e538063.St76 

### Dr. C. Simon 

# CJ(;---___**o~~~;c;;_;:·;:;;;:-:::(!(:0**

# !J

5Harlan Laboralmics Study C25654 

# L J Report



## ··---·---·--·-·--·----- 

## QUAl,ITYASSURANCE GLP 

## STATEMENT Harlan Laboratories Ltd., Zelgliweg 1, 4452 Itingen 

### I Switzerland Harlan Laboratories Study: TestItem: 

# [ Study Director: Studv Title: 

### C25654 

# J Dr. C. 

### Simon Primary Skin Irritation Study in Rabbits (4-Hour Semi-Occlusive Application) The general facilities and activities are inspected at least once ayear and the results are reported to the 

### responsible person and the management. Study 

### procedures, with 

### the exception ofthe 

### pH measurement 

### ofthe test item, were periodically inspected. The study plan and this report were audited by the 

### Quality Assurance. The dates are given below. Dates ofReports to Dates and Types ofQA

### Inspections the StudyDirector andTestFacility

Management 25-Nov-2008 Study 

### Plan 25-Nov-2008 17·-Nov-2008 Process Based 17-Nov-2008 (test system, test item, raw data, dose preparation, treatment) 17-Fcb-2009 Report 17-Feb-2009
This statementalso confirms that this final reportreflects the raw data.
Quality

### Assurance: N.

### Engelke

# 2:..~~11;

U9ll09c5380635t76 

### 6Harlan Laboratories Study C25654 

# [ l PREli'ACE General Information TestItem: 

### Study Title: 

### Sponsor:

Study 

### Monitor: Test Facility: QA:



## Responsibilities Study Director: Deputy Study Director: Laboratory 

### I Technical Coordinator: 

### QualityAssurance: HcadofQA:

09009e531!0635t76 Report


# [ J Primary Skin Irritation Study in Rabbits ( 4-Hour Semi-Occlusive Application)

HarlanLaboratories Ltd. WOlferstrasse 4 

### 4414 FilllinsdorfI

### Switzerland HarlanLaboratories Ltd. Quality Assurance 

### GLP Zelgliweg 1 

### 4452 Itingen I 

### Switzerland

Dr. C. Simon G. 

### Arcelin C. 

### Weng

T. Fink

7Harlan Laboratories Study C25654 Report 

# [ J

·------------------------------------- Schedule Experimental Starting Date: Tetmination: Delivery ofAnimals: Acclimatization:
Administration I

### Treatment: Experimental CompletionDate: 26-Nov-2008 05-Dec-2008 26-Nov-2008 26-Nov-2008 to 30-Nov-2008 (one female) 26-Nov-2008 to 01-Dec-2008 (one male andone female) 01-Dec-2008 (one female) 02-Dec-2008 (onemale and one female) 05-Dec-2008 

### Data Requirements 

### ITest Guidelines This study followed 

### the procedures indicated by the following internationally accepted guidelines and recommendations:
• OECD Guidelines for Testing of Chemicals, Section 4, number 404 "Acute Dermal Irritation 

### I Corrosion", adoptedApril24,2002.
• Commission Regulation (EC) No 440/2008 of30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Cmmcil on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), B.4 ACUTE TOXICITY: DERMAL IRRITATION/CORROSION (Official Journal No L 142,31/05/2008p. 0182-0190. 

### Animal Welfare This study was performed in an AAALAC-accredited laboratory in accordance with the Swiss Animal Protection Law underlicenseno. 

### 55. 

### Archiving Harlan Laboratories Ltd. (4452 Itingen/Switzerland), will retain the study plan, raw data, a sample ofthe test itemandthe final report ofthe present study for at least ten years. No 

### data will bediscarded without the Sponsor's writtenconsent.

09009eS380635f76 

### 8Harlan Laboratories Study C25654 Report 

# [ J**--·----·-···-~·--·----** 

# -------------------------- Classitlcation Guidelines Commission Directive 2001/59/EC adapting to technical progress for the 28th time Council Directive 67/548/EEC on the approximation of the laws, regulations and administrative provisions relating to the classification, packaging and labelling ofdangerous substances, August 

### 06, 20()1 (Official Journal ofthe European Communities Nr. L 225/1, August 21, 2001).

09009e.S:l8063**~fl6** 9Harlan Laboratories Study C25654 Report 

# [ J 1 SUMMARY The primary skin irritationpotential o{ lwas investigated according to OECD test guideline no. 404. The test item was applied by topical semi-occlusive application of0.5 mL 

### to the intact lett flank ofeach ofthree young adult New Zealand White rabbits. The duration of treatment was four hours. The scoring ofskin reactions was performed 1, 24, 48 and 72 hours after removal ofthe dressing. The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately. The mean erythema/eschar score ofthe three animals was 0.00, 0.33 and 0.00, respectively and the mean oedema score was 0.00 for each ofthe three animals. The application of[ ...,to the skin resulted in mild erythema in one animal. This effect was reversible and was no longerevident 48 hours after treatment. Thetest item caused no staining ofthe treated skin. No corrosive effects were noted on the treated skin ofany animal at any ofthe measuring intervals and no clinical signs were observed. Thus, the test item did not induce significantorirreversible damage to the skin. Based upon the referred classification criteria (Commission Directive 2001159/EC of August 200l

# )l Jisconsidered to be "notirritating" to rabbit skin.

09009e5380635n6 10

# [ Harlan Laboratories StudyC25654 Report --' -----------------------· 

# -------------------------- 2 PURPOSE The purpose ofthis pr_imary skin irritation study was to assess the possible irritation potential when a single dose of[ ·· ]was placed on the skin ofrabbits for approximately four hours. This study shouldprovide a rational basis for risk assessment

### in man as skin contact is one ofthe possible routes ofhuman exposure.

09009e538063Sf76 11Harlan Laboratories StudyC25654 [ I 

# J Report


## 3 MATERIALSAND METHODS 3.1 Test System Animals: Rationale: 

### Breeder:

Number ofAnimals per Test: Age (when treated): 

### Body Weight

### Range (when treated): Identification: Acclimatization:
Randomizatimt:


## 3.2 Allocation The animals 

### were distributed as follows: Sex: Male: Females:

09009cS380635f76 

### Young AdultNew Zealand White Rabbit, SPF Recognizedby international guidelines as a recommendedtest system. Harlan Laboratories B.V. Kreuzelweg 

### 53 NL-5961 

### NMHorst 

### IThe Netherlands Postbus 6174 

### 5960 AD Horst I

### TheNetherlands 3 

### (Animals ofboth sexes were used) 17 

### weeks (male) 16 weeks (females) 2364 

### g 

### (male) 2120- 2369 g

### (females) By unique cage number and corresponding ear number. Under laboratory conditions after health examination. Only animals without any visual signs 

### ofillness were used for the study. Selected by hand at time ofdelivery. No computer generatedrandomization program.



### AnimalNos.: 54 55- 56

:'**~** ! • (-;• .-:::)
,- ;,**~.,** 

### ..;_ -.,./ ' 

### 12Harlan**Laboratone~** Study C25654 

# [ J
3.3 Husbandry Room Number: Conditions:

Accommodation:



### Diet:

Water:



## 3.4 Test Item Report



### 0501 I

### Harlan Laboratories Ltd., Filllinsdorf Standard Laboratory Conditions. Air-conditioned with 

### 10-15 air changes per hour, and continuously monitored environment with ranges for room temperature 17-23 

# oc and for relative humidity between 30-70 %, automatically controlled light cycle of 12 hours light and 12 hours dark, music during the daytime lightperiod. Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks (Harlan Laboratories Ltd, Fullinsdort) and haysticks 

### 4642 (batch no. 

### 69/08, Provimi Kliba AG) wereprovidedfor gnawing. Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitwn (batch no. 

### 55/08) provided by Provimi K.liba AG, 4303 Kaiseraugst I 

### Switzerland. Results ofanalysis for contaminants are archived atHarlan Laboratories Ltd. Community tap water from Filllinsdorf ad libitum. Results ofbacteriological, chemical and contaminant analyses are archived atHarlan Laboratories Ltd.

The 

### following infonnation was providedby the sponsor: Identification: Description: 

### Batch Number: Purity: Stability

### ofTestItem: Expiry

### Date: Storage Conditions: Safety Precautions:

09009e538063flb4 

# [ J 100% Stableunder storage conditions. 28-0ct-2009 Atroom temperature (range of20 ± 

### 5 °C}, 

### light protected. Additionally to routine hygienicprocedures, a helmetmask was used to ensurethehealth and safety ofthepersonnel.



### 13Harlan**Laboratodc~** Study C25654 Report 

# [ J

r L -**----~-** 

# ·----------------------------- 3.5 Preparation ofDose Formulations jwas used as deliveredby the Sponsor. The pH ofthe test item was measured beforethe study initiation date and was found to be 4. According to Commission Regulation (EC) No 440/2008, 

### 8.4. and OECD Guidelines 404, a test item is not required to be tested ifthe pH-value 

### is less than 2 or greater than ll.S, owing to its predictable corrosive properties. 

### 3.6 Test Item Administration The test item was administered at 0.5 mL/animal, the dose specified in the test guidelines for a liquid test item. Four days before treatment, the left flank was clipped with an electric clipper, exposing an area of approximately 100 cm2 

### (10 em x 10 em). The skin of the animals was examined one day beforetreatment, and regrownfur ofall animals was clipped again. Animals with overt signs ofskin injury or marked irritation which may have interfered with the interpretation oftheresults were notused in the test. Onthe day oftreatment, 0.5 

### mLot{_ }asplaced on a surgical gauze patch (ca. 2.5 em x 2.5 

### em) held in contactwith the skinbymeans ofan adhesive hypoallergenic aerated semiocclusive dressing and a restrainerbandage wrapped around the abdomen. The duration oftreatment was 4 hours. Then the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site so that any reactions (erythema) were clearlyvisible at that time. As 

### it was suspected that the test item mightproduce irritancy, a single animal (one female) was treated first. As neither a corrosive effect nor a severe irritant effect was observed after the 4- hour exposure, the test was completedusing the two remaining animals for an exposure period of fourhours. 

## 3.7 Rationale The application form and dose were used to detect a possible irritating potential ofthe test item applied.

09009c53SO!'d5t76 

### 14Harlan Laboratories Study C25654 

# l J
3.8 Observations Viability 

### IMortality: Clinical Signs (systemic): Body

### Weights: Skin Reactions: 

## 3.9 

## Pntbology 3.9.1 Necropsy Report



### Daily from acclimatization ofthe animals to the termination oftest. Dailyfrom acclimatization ofthe animals to the tennination oftest. 

### At start ofacclimatization, onthe day ofapplication and

### at tennination ofobservation. See Section 

### 3.12

No necropsy was perfonned on the animals sacrificed attermination ofobservation. All rabbtts were sacrificed by an intravenous injection ofpentobarbitone into the ear vein at a dose ofat least 1mL!kgbodyweightand discarded. 

## 3.10 StatisticalAnalysis No statistical 

### analysis was perfonned. 

## 3.11 Uata Complilation Viability/mortality, clinical 

### signs andskinreactions wererecorded on data sheets and transcribed 

### for compilation andanalysis. Body weights wererecorded on-line. The RCC Tox Computer System (RCC-Tox-Lims) had been validated with respect to data collection, storage and retrievability.**09009e5~81l635!76** 

### 15

# [ Harlan Laboratories Study C25654 Report 

# J
3.12 Determination ofSkin Irritating Potential 3.12.1 Observation 

### and Scoring The skin reaction was assessed according to the numerical scoring system listed in the Commission Regulation (EC) No 440/2008, B.4, at approximately 1, 24, 48 and 72 hours after exposure (removal ofthe uressing, gauzepatch andtest item). (See Appendix 

### 1onp. 

### 28 ) Data were summarized in tabular form, showing for each individual animal the irritation scores tor erythema and oedema

### at all measurement intervals. Any lesions were described, including the degree and nature of irritation, corrosion or any other toxic effects observed, and their reversibility. The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patchremoval) for each animal for erythema/eschar grades and for oedema grades, separately. 

### An animal is positive when the mean score is 2 or greater. The test is positive for irritation when at least 2 animals arepositive for the same endpoint (erythema/escharoroedema). (See Appendix IJ 

### on p. 30 )**090fl9e~3806J5fl6** 16Harlan Laboratories Study C25654 

# [ J Report**·------~----------------------------** 

## 4 RESULTS AND DISCUSSION 4.1 Observations 4.1.1 Viability I 

### Mortality No 

### mortality occurred.


### 4.1.2 Clinical Signs No clinical signs were observed during the course ofthe study.
4.1.3 BodyWeights 

### (See Individual Tables on p. 26 ) One female lost slight body weight (1.3%) during the acclimatization period but recovered until the end ofthe study. Otherwise, the body weights ofall rabbits were considered to be within the nonnal range ofvariability. 

## 4.2 Pathology 4.2.1 Macroscopic Findings No necropsy was performed at the end ofthe study. 

## 4.3 Skin Reactions 4.3.1 Irritation (See SummaryTables on p. 21 and IndividualTables on p 23 ) The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately. The mean ctythemaleschar score ofthe three animals was 0.00, 0.33 and 0.00, respectively and the mean oedema score was 0.00 for each ofthe three animals. A very slight erythema was observed in the first treated female 1 hour after test item exposure and persisted up to the 24-hour reading. Otherwise, no local

### signs were observed in any animal at any observation. No abnom1al findings were observed on the treated skin ofone animal48 hours aftertreatment.

09009e5380635f76 

### 17Harla•1 Laboratories Study C25654 

# [ J Report



### 4.3.2 Coloration No staining producedbythe test item ofthe treated skinwas observed. 

### 4.3.3 Test Item Remnants Test item rcnmants were evident on the application site of one female l hour after test item 

### exposure. 

### 4.3.4 Corrosion Neither alterations ofthe treated skin wereobservednorwere corrosive effects evident on the skin.

09009e5JROf..l5f76 18Ha1lan**Lab01atoric~** Study C25654 

# [ J Report


## 5 CONCLUSION Based upon the referred classification criteria (Conunission Directive 2001/59/EC of August 200l

### )~ 1

### is consideredto be "not irritating" to rabbit skin.**09009e51806J~f76** 

### 19Harlan Laboratories Study C25654 Report 

# [ J

6 SUMMARYTABLES**{)900~Jd3&(!(.35f76** 

### 20

.n. j
.**·~** \..,. .,.'}' ....**~**Hatlan Laboratc•ries Study C25654 Report 

# [_ J
Skin Irritation Scores Animal Evaluation Number

### Su Interval* 

### Erythema Oedema -------" "" 

### 54 M 

### 0 0 

### 55 F I 

### l hour 1 

### 0 I I 

### 56

## --~~t-- 0 

### 0

·- .. 

### 54 ,MI 

### 0 

### 0 I
. 

### 55 

### i F 

# I 24 hours l 

### 0 I 

### 56 i 

### F 

### 0 0 I**------·~~-** 

# "rl'vr ·---- . """"'t-· 

### 54 0 0 

### 55 1 

### F 41i 

### hours 0 

### 0 

# S6 i 

### F 

# i 0 0



# - s-4 

# ···1M1 --

--- c···"· 

### 0 

### 0 

# ss 

# If 

## ! 72 

### hours 0 

### 0 56 F 

# l 0 

### 0 

## *Exammatwm; wereperformed atthe specifiedtimes afterremoval ofthe dressing. 

## Skin Irritation scores- Mean Values After 24, 48 and 72 Hours Animal! !

# ~-tl_l!l_l_J_~lJl;Se._~LE~)'!!t~l~~-**_Q~_em** __

a_-'-N-1 

### 54 M i 

### 0.00 3 0.00 3 I 

### 55 

# If: II 

### 0.33 3 0.00 3 56 [< 

### 0.00 3 0.00 3 N=numberofavailable data

### points. 

### Skin Irritation Scores 

## Assessment according to EC Guidelines

Evaluated 

# I

,___**it!t<~rvals** 

## !--·-**Et")t~~~~~_a** 

### __ -+-_O_e
__ d_e_m_a_-1 24 hon.-s 1 

### 48 hours 

## I Not Irritating NotIrritating

# _]!.hou~l__--------~--------~

'·:.**~--,**

<;,..,! 'v -'-. -· 

### 21Harlan Laboratories Study C25654 Report 

# [ J

---**-~·-----**



## 7 

# INDIVIDUALTABLES**09009c53~06J5f76** 

### 22Harlan Labor**atone~** Study C25654 Report 

# [ 

# J**~--"'-~---"-------~--** 

## Skin Reactions Animal No. 54, Male After I hour: 

### Erythema: NO ABNORMAL FINDINGS NOTED Oedema: NO ABNORMAL FINDINGS NOTED Scaling: NO ABNORMALFINDINGS NOTED Staining: NO STAINING PRESENT Aftc:r 24 hours: Erythema: NO ABNORMAL FINDINGS NOTED Oedema: NO ABNORMAL FINDINGS NOTED Scaling: NO ABNORMAL FINDINGS NOTED Staining: NO STAININGPRESENT After48 hours: Erythema: NO ABNORMAL FINDINGS NOTED Oedema: NO ABNORMAL FINDINGS NOTED Scaling: NO ABNORMALFINDINGS NOTED Staining: NO STAINING PRESENT
After72 hours: 

### Erythema: NO ABNORMAL FINDINGS NOTED Oedema: NO ABNORMALFINDINGS NOTED Scaling: NO ABNORMAL FINDINGS NOTED Staining: NO STAINING PRESENT**09009e538063~f76** 

### 23Harlan Laboratories Study C25654 Report 

# r "1 L

. .J

..**--.---~--** -·-**----·----~--~-""~-----** 

### Animal No. 55, Female 

### After 1hour: Erythema: Very slight erythema Oedema: NO ABNORMAL FINDINGS NOTED Scaling: NO ABNORMAL FINDINGS NOTED Staining: NO STAINING PRESENT Remnants: 

### Test itemremnants evidenton the application site
After 24 hours: 

### Erythema: Very slight erythema Oedema: NO ABNORMALFINDINGS NOTED Scaling: NO ABNORMAL FINDINGS NOTED Staining: NO STAINING PRESENT After48 hours: Erythema: NO ABNORMALFINDINGS NOTED Oedema: NO ABNORMAL FINDINGS NOTED Scaling: NO ABNORMAL FINDINGS NOTED Staining: NO STAINING PRESENT
After 72 hours: 

### Erythema: NO ABNORMAL FINDINGS NOTED Oedema: NO ABNORMAL FINDINGS NOTED Scaling: NO ABNORMAL FINDINGS NOTED Staining: NO STAINING PRESENT**09!10'1eS3806~5fl6** 24

,,**?~** ,, \ ; . f".j,

· t -;.j · .........t,**r-~~**Harlan Laboratones Study C25654 Report 

# [ 

# J**~---- ---~-~~-**  --------"----·· 

### Animal No. 56, Female Atkr l hour: 

### Erythema: NO ABNORMAL FINDINGS NOTED Oedema: NO ABNORMAL FINDINGS NOTED Scaling: NO ABNORMAL FINDINGS NOTED Staining: NO STAINING PRESENT
After 24 hours: 

### Erythema: NO ABNORMAL FINDINGS NOTED Oedema: NO ABNORMAL FINDINGS NOTED Scaling: NO ABNORMAL FINDINGS NOTED Staining: NO STAINING PRESENT After 48 hours: 

### Erythema: NO ABNORMAL FINDINGS NOTED Oedema: NO ABNORMAL FINDINGS NOTED Scaling: NO ABNORMAL FINDINGS NOTED Staining: NO STAINING PRESENT
After 72 hours: 

### Erythema: NO ABNORMAL FINDINGS NOTED Oedema: NO ABNORMAL FINDINGS NOTED Scaling: NO ABNORMAL FINDINGS NOTED Staining: NO STAINING PRESENT**09009eS3~06J5f76** 

### 25IIallan Laboralorieh Study C256S4 Report 

# [ 

# ]

--··------·---------------------

BODY WEIGHTS {GRAM) MALES GROUP 1 (GROUPl)**DAY(:~** WFB!<S ANlMI\.L l 1 2 1 5 1


### ------·-··

## -------·····-·---------------···--------------------- 2:1:15 2364 2458

09009e'iJS05c9ef.9 

### 26HarLm I. aboratorics Study C25654 Report 

# [ 1

···-··---·-------------------------------------- BODY WEIGHTS (GRAM) FEMAI,ES GROUP l (GROUPl)



# -----------------------------------------
LJAYS WEEKS ANIMAl, IVTL11>11\T.l ?.ATln-' TREATMENT 1 1 2 1 s 1

------·-----·------------------------ 

# ---------------------- 214? 2170

O'l009e5330:\e9e69 2120 2369 2242 2427



### 27Harlan Laboratories Study C25654 

# [ l Report



## APPENDIX l- COMMISSIONREGULATION

090H9c53N06J.Sf!8 

### 28Harlan Laborat\,ries Study C25654 

# [ J Report



### Commission Regulation (EC) No 440/ 2008, 30 MAY 2008 GradingofSkin Reactions ERYTliEMA AND ESCHAR FORMATION No erythema Very slight erythema (barely 

### perceptible) Well-def:ined erythema 

### Modcrak to severe erythema Severe erythema (beefredness) or eschar formation**(i1~jurics** 

### in depth preventing erythema) reading 

### OEDEMA FORMATION No oedema Very slight oedema(barely perceptible) Slightoedema (edges ofarea well-definedbydefiniteraising) Moderate oedema (edges raised approximately 1mm) 

### Sl:lvere oedema (raised more than l rnm and extendingbeyondthe area ofexposure)

09009e5J80635fl8 

### 0 I 

### 2 3 

### 4



### 0 1 2 

### 3 

### 4



### 29Harlan Laboraturies Study C25654 

# [ J Report



## APPENDIX II- COMMISSION DIRECTIVE

30

# [ Harlan Labmatoncs Study C25654 

# J
. Report



### Commission Oirective 

### 2001159/EC, August 2001 Inflammation ofthe skin The following riskphrase shall be assigned in accordance with the criteriagiven: "R38 

### Irritating to skin" Substances and 

### preparations which cause significant inflammation ofthe skin which persists for at least 24 

### hours after 

### an exposure period ofup to four hours determined on the rabbit according 

### to the 

### cutaneous irritation testmethod cited in Annex V. 

### lnflanmmtion of

### the skin is significant if: (a) the mean 

### value of the 

### scores for either erythema and eschar formation or oedema formation, calculated over all the animals tested, is 2or more, or (b) 

### in the case where 

### the Annex V 

### test has been completed using three animals, either erythema and eschar 

### formation or oedema formation equivalentto a mean value of2 or more calculated for each animal separately has been observedin two ormore animals. Jn both cases all scores at each ofthe reading times (24, 48 and 72 hours) for an effect should be used in calculatingthe respective mean values. Inflammation ofthe skin is also significant ifit persists in at least two animals at the end of the observation time. Particular effects e.g. hyperplasia, scaling, discoloration, fissures, scabs and alopecia 

### shouldbe taken into account. Substances and 

### preparations which cause significant inflammation of the 

### skin, based on practical observation inhumans on immediate, prolonged orrepeated contact. Organic peroxides, except where evidence to the contrary is available. Cm·rosivc The substances or

### preparation shallbe classified as corrosive and assigned the symbol 'C' 

### and lhe indication of

### danger 'corrosive' in accordance with the following criteria: •a substance or a 

### preparation is considered to be corrosive if, when it 

### is applied to healthy intact animal 

### skin, it 

### produces full 

### thickness destruction ofskin tissue on at least one animal during the test 

### for skin irritation cited inAnnex X 

### orduring an

### equivalentmethod, •classification can be based on the results of a validated in vitro test, such as that cited in AJmex V (B.40. Skin corrosion: rat skin transcutaneous declricalresistance assay and human skin model 

### assay), •a substance or a preparation should also be considered corrosive ifthe result canbe predicted, for example fwm strongly acid or alkaline reactions indicated by a pH of2 or less or 11,5 or greater. However, where extreme pH is the basis for classification, acid/alkali reserve may also be taken into consideration. Ifconsideration ofalkali/acid reserve suggests the substance or preparation may not be corrosive then further testing should be carried out to confirm this, preferably by usc ofan appropriate validated in vitro test. Consideration ofacid/alkali reserve should not be used alone to exonerate substances or preparations from classification as corrosive.

09009e53805r•;71a 

### 31Harlan Laboratories Study C25654 

# [ J Report



### Risk phrases shall be assigned in accordance with the following criteria: R35 Causes severe

### burns •if, when 

### applied to healthy intact animal skin, full 

### thickness destruction of

### skin tissue occurs as a result ofup to three minutes exposure, or 

### ifthis result canbe predicted. R34 Causes burns •if. when appii;:d to healthy 

### intact animal skin, full 

### thickness destruction of

### skin tissue occurs as a result ofup 

### to four hours exposure, or

### ifthis result canbe predicted. •Organic hydroperoxides, exceptwhere evidence to the contrary is available. Notes: Where. classificat.ion is based on results of 

### a validated 

### in vitro test R35 or R34 should be applied according to the capacity ofthe testmethod to discriminatebetween these. Where classification is based upon consideration ofextremepH alone, R35 shouldbe applied.

0900?r5:1805ec7!a 

### 32

### ATTACHMENT HEADER SHEET

Attachment Number 015 Attachment Name Health Effects Data on the New Chemical Substance- Eye Irritation Test 

### Associated PMN Section Number N/A
Does not contain CBI



### Report Number CMTP1 011 05088166808

### TS-CMTPOl 

### Attachment#15 HealthEffects Dataonthe NewChemicalSubstance 

### EyeIrritation Test

# L REPORT 

# J Primary EyeIrritation Study in Rabbits

StudyDirector: TestFacility:

Sponsor: 

# [ Dr. C. Simon Harlan.Laboratories Ltd. 

### Wolferstrasse4 4414 Fullinsdorf

### ISwitzerland

Studyldentificatlon: 

### HarlanLaboratories Study C2566S

Version: 

### Final Study Completion Date: 11-Mar-2009 Page 1of31 

# J

# [ Harlan Laboratories Study C25665 Report
..J


# CONTENTS CONTENTS
.......................................................................................................................... 2 SIGNATURES
...................................................................................................................... 4 

### STATEMENT OF COMPLIANCE
...................................................................................... 5 

### QUALITY ASSURANCE GLP STATEMENT
................................................................... 6 

### PREFACE
............................................................................................................................. 7 

### General Information
....................................................................................................... 7 

### Responsibilities
............................. ..................................................... .......... .................. 

### 7 Schedule
......................................................................................................................... 

### 7 Data Requirements ITest Guidelines
............................................................................. 8 

### Animal Welfare
.............................................................................................................. 8 

### Archiving
. .... . . ... ... . . . . . .......... .... . . . . . . ................... . . . . . . . . . . . . . . . ..... . .... . . . . .... . . . . . . . . . . . . . . . . . . . . . . . . . . .... 8 

### Classification Guidelines
............................................... ........................... . . . ................... 8 1 

### SUMMARY
....................................................................................................................... 9 2

### PURPOSE
.. ...................... ... ............................................................ ... .. .. .......................... 10 

### 3MATERIALS AND METHODS
..................................................................................... 

### 11 3.1 Test System
............................................................................................................ 11 3.2 Allocation
..... ..... .. .............................................................................................. ..... 11 3.3 

### liusbandry
.............................................................................................................. 12 3.4

### Test Item
................................................................................................................. 12 3.4.1 

### Test Item
....................................................................................................... 12 

### 3.5 Preparation ofDoseFormulations
.......................................................................... 13 3.6 TestItem Administration
....................................................................................... 13 3.7 Rationale
................................................................................................................. 13 3.8 

### Observations
........................................................................................................... 14 

### 3.9 Pathology
................................................................................................................ 

### 14 3.9.1 Necropsy
...................................................................................................... 14 3.10 

### Statistical Analysis
.................................................. . . . . . ... ...................... ....... .... .. .. 14 3.11 Data

### Compilation
................................................................................................. 14 3.12 Determination ofEye Irritating Potential
............................................................. 15 3.12.1 Observation and Scoring
............................................................................ 15 4 RESULTS AND DISCUSSION
...................................................................................... 16 4.1 Observations
........................................................................................................... 16 

### 4.1.1 Viability 

### IMortality
..................................................................................... 16 

### 4.1.2 Clinical Signs
............................................................................................... 16 4.1.3 BodyWeights
............................................................................................... 16 4.2 Pathology
................................................................................................................ 16 4.2.1 Macroscopic Findings
.................................................................................. 16 4.3 Eye Reactions
......................................................................................................... 16 4.3.1 Irritation
........................................................................................................ 16 4.3.2 Coloration
..................................................................................................... 17

09009e538060dc33 

### 2Harlan Laboratories StudyC25665 Report 

# [ l

4.3.3 Corrosion
...................................................................................................... 17 5CONCLUSION
................................................................................................................ 18 6 SUMMARY TABLES
..................................................................................................... 19 Eye Irritation Scores
..................................................................................................... 20 Eye Irritation Scores-Mean Values after24, 48 and 72 Hours
.................................. 20 Eye Irritation Scores

- AssessmentAccording to 

### EC Guidelines
............................... 20 7INDIVIDUALTABLES
.................................................................................................. 21 Eye Reactions
............................................................................................................... 21 BodyWeight
................................................................................................................ 25 APPENDIX I- COMMISSION REGULATION
............................................................... 27 Commission Regulation (EC) No 440/2008, 30May 2008
........................................ 28 APPENDIX II

- COMMISSION DIRECTIVE
................................................................... 30 Commission Directive 200l/59/EC,August2001
....................................................... 31

09009eS38060dc33 

### 3Harlan Laboratories Study C25665 

# [ J SIGNATURES Study Director:

Management:

09009e$38060dd4b Report



### Dr. C. Simon 

# c_·J·,er----__**n~i~;d(·:.:··Jrw·~·:zao**

# 5 

# (ff11) Dr. H. Fankhauser**/>.~·tvL** 

# ..........**~**

## ....**~····················** 

### Date:**//~-** 

## h4'1(,...**~00!1**



### 4Harlan Laboratories StudyC25665 

# [ l Report



### GOOD LABORATORY PRACTICE 

## STATEMENT OFCOMPLIANCE Harlan Laboratories Study: 

### C25665 TestItem: 

# J Study Director: Dr. C. Simon Study Title: PrimaryEye Irritation StudyinRabbits

This study has been performed in compliance with the Swiss Ordinance relating to Good Laboratory Practice, adopted May 18th, 2005 [SR 813.112.1]. This Ordinance is based on the OECD Principles ofGood Laboratory Practice, as revised in 1997 and adoptedNovember 26th, 1997 by decision ofthe OECD Council [C(97)186/Final].

Study Director:**09009e53806450f~;** 

### Dr. C. Simon 

# C.}t\.t--**D~t~;;;jt::···'ffc:r··:::·zws**

5Harlan Laboratories StudyC25665 Report 

# [ J QUALITYASSURANCE GLPSTATEMENT Harlan Laboratories Ltd., Zelgliweg 1, 4452 Itingen I

### Switzerland Harlan Laboratories Study: C25665 Test Item: 

# J Study Director: Dr. C. 

### Simon Study Title: Primary Eye Irritation Studyin Rabbits The general facilities and activities 

### are inspected at leastonce 

### ayear and the results are reported to the responsible person andthe management. Study procedures were periodically inspected. The study plan and this report were auditedby the Quality 

### Assurance. The dates are givenbelow. Dates ofReports to 

### Dates and Types ofQA

### Inspections the

### StudyDirector andTestFacility

Management 04-Dec-2008 Study Plan 04-Dec-2008 26-Nov-2008 Process based 26-Nov-2008 (testsystem, testitem, raw data, dose preparation, treatment) 05-Mar-2009 Report 05-Mar-2009
This statement also 

### confirms thatthis final reportreflects the

### raw data.
Quality

### Assurance:

09009e538064SOfc 

### 6Harlan Laboratories Study C25665 

# [ l PREFACE GeneralInformation Test Item: StudyTitle: Sponsor: Study Monitor: Test Facility: QA:



### Responsibilities Study Director: Deputy Study Director: Laboratory I

### Technical Coordinator: QualityAssurance: 

### HeadofQA: 

### Schedule Experimental Starting Date: Termination: Deliveryof

### Animals: Acclimatization:
Administration ITreatment: Experimental CompletionDate:

09009eS38060dd4b 

# [

# l Report



### PrimaryEye Irritation StudyinRabbits



### Harlan Laboratories Ltd. 

# J
Wolferstrasse 4 4414 FUllinsdorf

### ISwitzerland 

### HarlanLaboratories Ltd. QualityAssurance 

### GLP Zelgliweg 1 4452 Itingen I 

### Switzerland

Dr. C. Simon G. 

### Arcelin C. 

### Weng

T. Fink

10-Dec-2008 22-Dec-2008 03-Dec-2008 10-Dec-2008 to 14-Dec-2008(onefemale) 10-Dec-2008

### to 15-Dec-2008 (one male and one female) 15-Dec-2008 (one female) 16-Dec-2008 (one male and one female) 22-Dec-2008
7

# [ Harlan Laboratories Study

### C2S665 Report 

# J
DataRequirements I

## TestGuidelines This study followed the procedures indicated by the following internationally accepted guidelines and recommendations:



### • OECD Guidelines for Testing of Chemicals, Section 4, number 405 "Acute Eye Irritation I 

### Corrosion", adopted April 24, 2002.

• COMMISSION REGULATION (EC) No 440/2008 of 30 May 2008, B.S. "Acute ToxicityEye Initation!Corrosion". 

## Animal Welfare TI1is study was perfonned in an AAALAC-accredited laboratory in accordance with the Swiss Animal ProtectionLawunder license no. 54. 

## Archiving Harlan Laboratories Ltd. (4452 Itingen!Switzerland), will retain the study plan, raw 

### data, a sample ofthe test item and the final report ofthepresent study for atleastten years. No 

### data will be discardedwithoutthe Sponsor's written consent. 

## Classification Guidelines Commission Directive 2001159/EC adapting 

### to technical progress for the 28th time Council Directive 67/548/EEC on the approximation of the laws, regulations and administrative provisions relatingto the classification, packaging and labelling ofdangerous substances, August 06,2001 (OfficialJournal ofthe European CommunitiesNr. L 225/1, August 21, 2001).

09009e5380624ce4 

### 8

# [ Harlan Laboratories Study C2566S Report 

# J 1 SUMMARY The primary eye irritationpotential o1 !]was investigated according to OECD test guideline no. 405. 

### The test itemwas appliedby instillation ofO.l 

### mL into the left eye ofeach of three young adult 

### New Zealand White rabbits. 

### Scoring of irritation effects was performed approximately 1, 24, 48

### and 72

### hours aftertestitem instillation. The mean score was calculated across 3 scoring times (24, 48 and 72 hours afterinstillation) for**ea<.~h** 

### animal for corneal opacity, iris, redness and chemosis ofthe conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for the conjunctivae were 0.00 for reddening and for chemosis for each ofthe

### three animals. The instillation

# o{ =into the 

### eye resulted 

### in mild, early-onset and transient ocular changes, such as reddening 

### ofthe conjunctivae and sclerae. These effects were reversible and were no longer evident 24 hours after treatment. No abnormal findings were observed in the cornea or 

### iris ofany animal at 

### any ofthe examinations. No corrosion was observed at

### any ofthe 

### measuring intervals. No staining ofthe treated eyes by the test item was noted and no clinical signs were observed. Thus, the test item didnotinduce significantorirreversible damageto the rabbiteye. Basedupon the referred classification criteria (Commission Directive 2001159/EC ofAugust06, 2001

# >[**~s** consideredto be "notirritating"to the rabbiteye.

0900!le5380624ce4 

### 9

J\ '**.~** . ;•**~. ~~** 

J
....Y "-' -·

# [ Harlan Laboratories StudyC25665 

### Report


## 2 PURPOSE The purpose of this primary eye irritation study was to assess the possible irritation potential when a single dose

# ot ··Jvasplacedinthe conjunctival sac ofrabbiteyes. This study should provide a rational basis for risk assessment in man as ocular contact is one of 

### thepossible routes ofhuman

### exposure.

09009eS:J8000dd4b 

### 10Harlan Laboratories Study C25665 Report [ I 

# J 3 MATERIALSANDMETHODS 3.1 TestSystem Animals: Rationale: Breeder:

NumberofAnimals perTest: Age (when treated): Body Weight Range (when treated): Identification: 

### Acclimatization:
Randomization:


## 3.2 Allocation Young AdultNew ZealandWhite Rabbit, SPF Recognizedby international guidelines as a recommendedtest system. Harlan Laboratories 

### B.V. Kreuzelweg 53 5961 NM Horst I

### The Netherlands Postbus 6174 5960 AD HorstITheNetherlands 

### 3 (Animals ofboth sexes were used) 18 

### weeks (male) 12 

### weeks (females) 2700 g (male) 2570-2797 g (females) By unique cage number and corresponding ear number. Under laboratory conditions after health examination. Only animals without any 

### visual signs of

### illness wereused forthe study. Selectedby hand attime ofdelivery. No computer generatedrandomization program.

The animals were distributed as follows: Sex: 

### AnimalNos.: Male: 57 Females: 58-59

09009e53R060dd4b 11Harlan Laboratories Study C25665 

# l J
3.3 Husbandry RoomNumber: Conditions:

Accommodation:

Diet:

Water:



## 3.4 TestItem 3.4.1 TestItem Report



### 0501 

### IHarlan LaboratoriesLtd., Ftillinsdorf Standard Laboratory Conditions. Air-conditioned with 

### 10-15 air changes per hour, and continuously monitored environment with ranges for room temperature 

### 17-23 °C and for relative humidity between 30-70 % 

### (values above 70 % 

### during cleaning process possible), automatically controlled light cycle of 12 hours light and 12 hours dark, music during the daytime lightperiod. Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks (Harlan Laboratories Ltd, Ftillinsdort) and haysticks 

### 4642 (batch no. 

### 69/08, Provimi Kliba AG) wereprovided for gnawing. Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 55/08) 

### provided by Provimi Kliba AG, 4303 Kaiseraugst 

### I Switzerland. Results ofanalysis for contaminants are archived atHarlanLaboratories Ltd. Community tap water from Ftillinsdorf 

### ad libitum. Results ofbacteriological, chemical and contaminant analyses arearchivedatHarlanLaboratories Ltd.



### The following information

### was providedbythe sponsor: Identification: Description: BatchNumber: Purity: StabilityofTestItem: ExpiryDate: Storage Conditions: Safety Precautions:

09009c53806450fe 

# [ J
100% Stableunderstorage conditions. 28-0ct-2009 

### Atroomtemperature (range of20± 

### 5 °C), light protected. Additionallyto 

### routine hygienicprocedures, a helmetmaskwasusedto ensure the health

### and safety of

### thepersonnel. 12Harlan Laboratories Study C2S665 Report 

# C I

[ 3.5 Preparation ofDose Formulations ]was measured with a syringe and applied undiluted as it was delivered by the Sponsor. The pH ofthe test itemwas measured for Harlan Laboratories Study No. C25654 and was found to be4. 

### According to Commision Regulation 

### (EC) No 440/2008, 

### B.5. and 

### OECD Guidelines 405, a test item needs not to be 

### tested if 

### the pH-value is less than 2 

### or greater than 11.5, 

### owing to its predictable corrosiveproperties. 

## 3.6 Test Item Administration The test item was applied at 0.1 mL/animal, the dose specified in the test guidelines for a liquid testitem. The eyes ofthe animals wereexamined one dayprior to testitemadministration. Animals with overt signs of ocular injury or irritation which may have interfered with the interpretation of

### the: results were notused in the test. On the day oftreatment, 0.1 

### mL o{ was placedin the cof\iunctival sac ofthe left eye ofeach 

### animal after gently pulling the lower ltd away from the eyeball. The 

### lids were then gently held together for about one second to prevent loss oftest item. The right eye 

### remained untreated and served as the reference control. The treated eyes were notrinsed afterinstillation. As it was suspected that the test item mightproduce irritancy, a single animal (one female) was treated first. As neither a corrosive effect nor a severe irritant effect was observed after the land 24-hourexaminations, the testwas completedusingthe two remaining animals. 

## 3.7 Rationale The application form and dose were used 

### to detect a possible irritating potential ofthe test item applied.



### 13Harlan Laboratories StudyC25665 

# l 

# J
3.8 Observations Viability I

### Mortality: Clinical Signs (systemic): Body Weights: 

### Eye Reactions: 

## 3.9 Pathology 3.9.1 

### Necropsy Report



### Daily from acclimatization ofthe animals to the termination of

### test. Daily from acclimatization ofthe animals to the termination oftest. At startofacclimatization, onthe day ofapplication and attermination ofobservation. See Section 3.12

No necropsy was performedon the animals sacrificed at termination ofobservation. The animals were killed by intravenous injection ofpentobarbitone into the ear vein at a dose of at least l 

### mL/kg bodyweight (162 

### mg sodiumpentobarbitone/kgbody weight) anddiscarded. 

## 3.10 Statistical Analysis No statistical analysis was performed. 

## 3.11 Data Compilation Viability/mortality and local I 

### clinical signs were recorded on 

### data sheets and transcribed for compilation and analysis. Body weights were recorded on-line. The RCC Tox Computer System (RCC-Tox-Lims) had been validated with respect to data collection, storage andretrievability.

09009e538060dd4b 14Harlan Laboratories StudyC25665 

# [ ] Report


## 3.12 Determination ofEye 

## IrritatingPotential 3.12.1 Observation and Scoring (See Appendix I 

### on p. 27 ) The eye reactions were assessed according to 

### the numerical scoring system listed in the Commision Regulation (EC) No 440/2008, B.5, 

### at approximately l, 

### 24, 48, 72 

### hours after administration. Scleral reddening and ocular discharge were also assessed. Eye examinations were made with a VartaCliptrix diagnostic-lamp (Roth AG, CH-4153 Reinach/Switzerland). Data were summarized in tabular form, showing for each individual animal the irritation scores 

### for the designated observation time, 

### a description of the degree and nature of irritation, the presence 

### of serious lesions and non-ocular effects. The scores of each animal at the following reading times (24, 48, 72 hours) were used in calculating the respective mean values (with the exception ofthe sclerae) for each

### type oflesion. (See Appendix 

### II on p. 

### 30 ) The classification system was based upon the referred classification criteria Commission Directive 2001/59/EC ofAugust 06, 2001.

09009e5380624ce4 

### 15Harlan Laboratories Study C25665 

# [ l Report


## 4 RESULTSAND DISCUSSION 4.1 Observations 4.1.1 Viability I 

### Mortality No mortality occurred.
4.1.2 Clinical Signs Noclinical signs were observed during the course ofthe study. 

### 4.1.3 Body Weights 

### {See Individual Tables on p. 25 ) Thebodyweights ofall rabbits were consideredto 

### be withinthe normalrange ofvariability. 

## 4.2 

## Pathology 4.2.1 MacroscopicFindings Nonecropsywasperformed at the endofthe study. 

## 4.3 Eye Reactions 4.3.1 Irritation (See Summary Tables on p. 19 andIndividualTalbes onp. 21 ) The mean score was calculated across 3 scoring times (24, 48 and72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis ofthe conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for the conjunctivae were 0.00 for reddening and for chemosis for each ofthe three animals. No abnormal· findings were observed in the cornea or iris of any animal at any of the measurementintervals. A slight reddening of the conjunctivae was noted in the three animals 1 hour after test item instillation. Additionally, one female was observed with a slight reddening ofthe sclerae at the same observation time point.
09009eS38060dd4b 

### 16Harlan Laboratories StudyC25665 Report 

# L l

No abnom1al findings were observedin the treated eye ofany animal 24hours aftertreatment. 4.3.2 

### Coloration No staining produced by 

### the test item ofthe treated eye was observed. 4.3.3 Corrosion 

### No corrosion ofthe corneawas observed at any ofthe reading times.

09009eS38060dd1b 

### 17

i \ 1 '
..**~**....**~~** :, t ,)·l·**.J~.**_[Harlan Laboratories Study C25665 Report 

# [ J 5 CONCLUSION Based upon the referred classification (Commission Directive 2001/59/EC ofAugust 

### 06, 2001), [ }is consideredto be "notirritating"

### totherabbiteye.

09Q09e538060dd4b 

### 18Harlan Laboratories StudyC25665 Report 

# [ J


# _______..

## ____ . 

# ___________________________

6 SUMMARYTABLES

09009e53ROfiOdd4b 

### 19Harlan Laboratories Study C25665 

# [ J Report 

## Eye Irritation Scores

Animal Evaluation Corneal Area

### or Conjunctivae**-~ t-~U_!II_!!~t:_**  Set Interval* Opacity CornealOpacity Iris Redness Chemosis Sclera**~M- ··-----·~~---·-**  -·-

57 0 0 0 I 0 0 5!1 F 1 hour 0 0 0 I 0 0**f--~_5~-** F 0 0 0 I 0 1**··-·-~ -~---**  ·-**--~~.--** 

## --- 57 M 0 0 0 0 0 0 58 F 24 hours 0 0 0 0 0 0 59 

### !
__. 0 0 0 0 0 0

- 57 M 0 0 0 0 0 0 58

# -fJ-':''""~ 0 0 0 0 0 0 59 0 0 0 0 0 0 

# -· 57 0 0 0 0 0 0 58 F \ 72 hours 0 0 0 0 0 0 59 l F 0 0 0 0 

# l 0 I 0 • Exammat1ons were performed atthe specified times aftermstlllatJon ofthetest 1tem. 

## Eye Irritation Scores- MeanValues after24,48 and 72 Hours

Animal 

### ! Corneal Conjunctivae Number Sex Opacity 

### N Iris N 

### Redness N Chemosis 

### N**---~------- -·-~**  !--·------·· 

### 57 M 0.00 

### 3 

### 0.00 3 0.00 3 0.00 

### 3 58 F 0.00 3 

### 0.00 3 

### 0.00 3 

### 0.00 3 59 F 0.00 3 

### 0.00 3 0.00 3 

### 0.00 3 N ""' numberofavailable datapoints. 

## Eye Irritation Scores

- AssessmentAccordingto EC Guidelines Evaluated Corneal Conjunctivae

intervals Opacity Iris Redness Chemosis 24 hours 48 hours NotIrritating NotIrritating NotIrritating NotIrritating 72 hours

09009e53S060dd4b 20Harlan Laboratories StudyC25665 

### Report 

# [ l

7 INDIVIDUALTABLES

09009e538060dd4b 

### 21

# [ Harlan Laboratories Study C25665 Report 

# l ..J 

## EyeReactions Animal No. 57, Male 

### After l hour Cornea: NO ABNORMAL FINDINGS NOTED 

### Iris: NO ABNORMALFINDINGS NOTED Conjunctivae: Slightreddening Discharge: NO ABNORMAL FINDINGS NOTED Sclerae: NO ABNORMALFINDINGS NOTED Test item: NO REMNANTS EVIDENT After 24 hours: Cornea: NO ABNORMALFINDINGS NOTED Iris: NO ABNORMAL FINDINGS NOTED Conjunctivae: NO ABNORMAL FINDINGS NOTED Discharge: NO ABNORMAL FINDINGS NOTED Sclerae: NO ABNORMAL FINDINGS NOTED Test item: NO REMNANTS EVIDENT After48 hours: Cornea: NO ABNORMAL FINDINGS NOTED Iris: NO ABNORMALFINDINGS NOTED Conjunctivae: NO ABNORMALFINDINGS NOTED Discharge: NO ABNORMAL FINDINGS NOTED Sclerae: NO ABNORMAL FINDINGS NOTED Test item: NO REMNANTS EVIDENT After 72 hours: Cornea: NO ABNORMALFINDINGS NOTED Iris: NO ABNORMALFINDINGS NOTED Conjunctivae: NO ABNORMALFINDINGS NOTED Discharge: NO ABNORMAL FINDINGS NOTED Sclerae: NO ABNORMALFINDINGS NOTED Testitem: NO REMNANTS EVIDENT

Observations included: cornea, conjunctivae (including nictitating membrane), sclera and iris. The presence (or absence, as appropriate} ofopacity, vascularization, reddening, oedema, discharge, staining and test item remnants were assessed. 09009e538060d,l4b 

### 22

# [ Harlan Laboratories StudyC25665 Report 

# J

----· Animal No. 58, l<'emale 

### After 1hour: Cornea: NO ABNORMAL FINDINGS NOTED Iris: 

### NO ABNORMAL FINDINGS NOTED Conjunctivae: Slightreddening Discharge: NO ABNORMAL FINDINGS NOTED Sclerae: NO ABNORMAL FINDINGS NOTED Testitem: NO REMNANTS EVIDENT After 24 hours: Cornea: NO ABNORMAL FINDINGS NOTED Iris: NO ABNORMAL FINDINGS NOTED Conjunctivae: NO ABNORMAL FINDINGS NOTED Discharge: NO ABNORMAL FINDINGS NOTED Sclerae: NO ABNORMAL FINDINGS NOTED Testitem: NO REMNANTS EVIDENT After48 hours: Cornea: NO ABNORMAL FINDINGS NOTED Iris: NO ABNORMAL FINDINGS NOTED Conjunctivae: NO ABNORMAL FINDINGS NOTED Discharge: NO ABNORMALFINDINGS NOTED Sclerae: NO ABNORMAL FINDINGS NOTED Testitem: NO REMNANTS EVIDENT After 72 hours: Comea: NO ABNORMAL FINDINGS NOTED Iris: 

### NO ABNORMAL FINDINGS NOTED Conjunctivae: NO ABNORMAL FINDINGS NOTED Discharge: NO ABNORMAL FINDINGS NOTED Sclerae: NO ABNORMAL FINDINGS NOTED Testitem: NO REMNANTS EVIDENT

Observations included: cornea, conjunctivae (including nictitating membrane), sclera and iris. The presence (or absence, as appropriate) ofopacity, vascularization, reddening, oedema, discharge, staining andtest item remnants were assessed. 09009e.538ll60dd4b 

### 23

# [ Harlan Laboratories StudyC25665 Report 

# l

--

....**~------·-·--** 

### Animal No. 59, Female After t 

### hour: Cornea: NO ABNORMALFINDINGS NOTED Iris: NO ABNORMAL FINDINGS NOTED Conjunctivae: Slightreddening Discharge: NO ABNORMAL FINDINGS NOTED Sclerae: Slightreddening Test item: NO REMNANTS EVIDENT After 24 hours: 

### Cornea: NO ABNORMAL FINDINGS NOTED Iris: NO ABNORMAL FINDINGS NOTED Conjunctivae: NO ABNORMALFINDINGS NOTED Discharge: NO ABNORMAL FINDINGS NOTED Sclerae: NO ABNORMAL FINDINGS NOTED Testitem: NO REMNANTS EVIDENT After 48 hours: Cornea: NO ABNORMALFINDINGS NOTED Iris: NO ABNORMAL FINDINGS NOTED Conjunctivae: NO ABNORMAL FINDINGS NOTED Discharge: NO ABNORMALFINDINGS NOTED Sclerae: NO ABNORMALFINDINGS NOTED Testitem: NO REMNANTS EVIDENT After 72 hours: Cornea: NO ABNORMALFINDINGS NOTED Iris: NO ABNORMAL FINDINGS NOTED Conjunctivae: NO ABNORMAL FINDINGS NOTED Discharge: NO ABNORMAL FINDINGS NOTED Sclerae: NO ABNORMAL FINDINGS NOTED Testitem: NO REMNANTS EVIDENT

Observations included: cornea, conjunctivae (including nictitating membrane), sclera and iris. The presence (or absence, as appropriate) ofopacity, vascularization, reddening, oedema. discharge, staining andtest itemremnants were ussessed. 09009e538060dd4b 

### 24Hatlan Lahorntories Study C25665 

# l J

BODY WEIGHTS (GRAM) MALES GROUP 1 (GROUPl)

IICCl·lMA'I'IZ11.T!ON DAYS 1 WEI':KS l ANIMAI.**---~--·-"**

57 2620

(19009e538060d9cc TREATMENT 1 l Report

2 8 1 2

2700 2716



### 25

# [ Harlan Laboratories Study C25665 

# J

BODY WEIGHTS (GRAM) FEMALES GROUP 1 (GROUPl)
1\CCT,UIAT!ZATION TREATMENT DAYS 1 l WEEKS 1 l ANIMAL**·---~·· ---~·-~-·---** 

SB 2745 2797 59 249l

0900'Jc538060d9C<: Report

2 a l 2
3041 2570 2655



### 26Harlan Laboratories Study C2566S 

# [ J Report



## APPENDIX I- COMMISSION REGULATION

27

09009eS380624cfllHarlan Laboratories StudyC25665 

# [ l Report



# --------------------------------- Commission Regulation (EC) No 440/2008, 30 May 2008 GradingofOcularLesions CORNEA 

### Opacity: degree ofdensity (areamost dense taken torreading) No ulceration oropacity 0 Scatteredor diffuse areas 

### ofopacity (otherthan slight

### dulling ofnormal luster), details ofiris clearly visible 1 Easily discernible translucentarea, details ofiris slightly obscured 2 Nacreous area, no details ofiris visible, size ofpupil barelydiscernible 

### 3 Opaque cornea, iris uot discernible through the opacity 4 Area of

### cornea involved Zero 0 

### One quarter (or less) butnot zero 1 

### Greater than one quarter, butless than half 2 Greater than half, but less than three quarters 3 

### Greaterthan three quarters, up to

### whole area 4 

### IRIS 

### Normal Markedly deepened mgae, congestion, swelling, moderate circumcorneal hyperemia, orinjection, any ofthese orcombinationofany thereof, iris stillreacting to light (sluggish reaction is positive) No reaction to light, hemorrhage, gross destruction (any or all of

### these) CONJUNCTIVAE 

### Redness (refers 1o 

### most severe reading ofpalpebral andbulbarconjunctivae when compared with control eye) Blood vessels normal Someblood vessels definitelyhyperemic (injected) Diffuse, crimson color, individual vessels noteasily discernible Diffuse beefy

### red Chemosis: 

### lids and/or nictitatingmembranes No swelling Any 

### swelling abovenormal (includingnictitating membranes) Obvious swelling with partial eversion oflids Swelling with lids abouthalf-closed Swelling with lids more than half-closed**09009e5380624~ffi** 

### 0 

### 1 

### 2

0 1 

### 2 

### 3 0 

### 1 

### 2 

### 3 4



### 28

# [ Harlan Laboratories Study C25665 

## l .J Report



### ADDITIONALLY, OCULAR DISCHARGE, REDDENING OF THE SCLERAE AND STAINING 

### 01<, CONJUNCTIVAE, SCLERAE AND CORNEA 

### BY THE TEST ITEM WILL BE ASSESSED ACCORDINGTOTHEFOLLOWINGSCHEME: OCULARDISCHARGE No discharge 0 Slight: Any 

### amount different to nonnal (does notinclude small amount observed in inner

### canthus ofnormal animal) 1 

### Moderate: 

### Discharge with moistening

### ofthe lids and 

### hair

### justadjacentto 

### the lids 

### 2 Marked: 

### Discharge with moistening

### ofthe lids andhairs, and 

### aconsiderable 

### area around the eye (rUJming) 3 

### SCLERAE Redness Nonnal: Blood

### vessels nonnal 

### 0 Slightreddening: Some blood

### vessels definitelyhyperemic (injected) l 

### Moderatereddening: Diffuse, crimson color, individualvessels noteasily discernible 2 Marked reddening: Diffuse beefy

### red 3 ST 

### AfNJNG OFCONJUNCTIVAE,SCLERAEAND CORNEA

### BY THETEST ITEM Notobserved 0 

### Slight staining 1 Marked staining 2

09009e538060c:ut4 29HarlanLaboratories StudyC25665 

# [ J Report



## APPENDIX II- COMMISSION DIRECTIVE

09009e538060dc2e 

### 30Harlan Laboratories Study C25665 

# [ J Report



## Commission Directive

## 2001/59/EC, August 2001 The following risk phrases shall also be assigned in accordance with the criteria given: "R36**~** 

### Irritatingto eyes" 

### Substances and preparations which, when applied to the eye of the animal, cause significant ocular lesions which occurred within 72 

### hours after exposure and which persist for at least 24 hours. Ocular lesions are significant ifthe mean scores ofthe eye irritation test cited in Annex V 

### have any ofthe following values: • •comeaopacity equal to orgreaterthan 2 

### butless than 3; 

### • •iris lesion equal to orgreaterthan 

### 1butnot greaterthan 

### 1,5; • •redness of

### the conjunctivae equal to or greaterthan 2,5; 

### • •oedema of

### the conjunctivae (chemosis) equal to 

### orgreaterthan 2; 

### or, in the 

### case where the Annex V 

### test has been completed using three animals ifthe lesions, on two or more animals, are equivalent to any ofthe above values except that for iris lesion the value should be equal to or greater than l butless than 2 and for redness ofthe conjunctivae the value shouldbe equal to or greaterthan 2,5. In both cases all scores ateach ofthe reading times (24, 48 and 72 hours) for an effect shouldbe used 

### in calculating 

### the respective mean values. Substances or preparations which cause significantocular lesions, based on practical experience in humans. Organic peroxides except where evidenceto the contrary is available. "R41**~Risk** 

### ofserious

### damagetoeyes" Substances and preparations which, when applied to the eye ofthe animal, cause severe ocular lesions which occurwithin 72 hours afterexposure andwhichpersistfor atleast 24 hours. Ocular lesions are severe if

### the mean scores ofthe eye irritation test in Annex V have any ofthe following values: • •comea opacity equal to orgreaterthan 3; • •iris lesion greaterthan 1,5. The same shall be the case where the test has been completedusing three animals ifthe lesion, on two ormore animals, have any ofthevalues: • •cornea opacity equal to orgreaterthan 3; • •iris lesion equal to 2. In both cases all scores ateach ofthe reading times (24, 48 and 72 hours) for an effect shouldbe used in

### calculating the respectivemean values. Ocular lesions are also severewhenthey are stillpresentatthe end ofthe observation time. Ocular lesions are also severe if

### the substance or preparation causes irreversible colouration of the eyes. Substances and preparations which cause severe ocular lesions, based on practical experience in humans.

09009e538061.ldc2e 

### 31

### ATTACHMENT HEADER SHEET

Attachment Number 016 Attachment Name Health Effects Data on the New Chemical Substance- Ames (Bacterial Reverse Mutation) Test 

### Associated PMN Section Number N/A
Does not contain CBI



### Report Number CMTP101105088166808

### TS-CMTP01 

### Attachment#16 HealthEffects DataontheNewChemicalSubstance 

### Ames(BacterialReverseMutation) Test

# r

RCC - CCR STUDY NUMBER 1111700
SALMONELLA TYPHIMURIUM AND ESCHERICHIA COLI REVERSE MUTATION ASSAY WITH

FINAL REPORT
STUDY COMPLETION DATE: September20, 2007 

# ]

# [ RCC-CCR Study Number 

### 1111700

# J Final Report Page 2 of 33



# 1 COPY OF GLP CERTIFICATE



# • HESSEN 

# •

st4ecdr.doc GuteLaborpra:ds/GoodLaboratoryPraetiee GLP-BeseheiDigaag/StatemeatofGLPCompliance (gemllllliCcordingto § 19bAbs. 1Chtmlk.alierogesctz)

Einc GLP-IMpektion rur Oberwac:hungderEinhaltung dcr(lLP-Grundsltze gemllll Chetnikaliengesetzbzw. Rlchtlinic**~8/320/EG** wurde durchgefllhrt in 1:1!:1 Prllfeinrichtung/Testfacility Assessment ofconfonnity with GLP accordillg to Chemikalifngesetz and Directive 881320/EEC at: 0 Prllf•tandottrresl site RCC-C)totestCelRoMa.UG•ltH RCC- CytolcstCell Resean:b GmbH In denLcppsleinswiesen 19 64380Rossdorf (Unverwecbselbm Beuichrumgund Adrcstc!Untquivocal name and**~~dress)** Prt
.....aado Kaleprieii/ANUofEspenlle (gemlll/according chemVwV-GLPNr. 5.3/0I!COguidance) :a Prf1funatn t.lll llt'.aU.nununc**d~r** tox.ikolopac.:hen K-igf!n!\{, htdteo 3 ,..,.nruogt-n wr Bll"'ahmmunc drr**erbl'l'tverin~** df!!rndrn Eigf'nschaften (in vitro und in vivo) 6**PrClfung~n** aur BeMimmuna von Rllc:kaU\nden • Ann.lytt!lcht: PrUfuncen an biolostar.hc:n Materialien t VttUI'J!Iichc-rhdtopnlfungen ;;a Toxi<.tty atudit-a 3 Mutagenicity studies 6 Relliduea a Analytical**studJ~s** on biologital materials 9 Viruavalidation atudies 02.09.%GI6 Datum derlnspcktion/Datc oflnsptttlon

(Tag Mona!Jahrlday month year)
Die gc....,nlc l'rUfeinri,btungbellndel si<:h im nationalen GLP-Obc:rwachungsverfabnm und wird regelmllllig aufEinhalt1111gder GLP-Grundsll7.e l!berwacbt. Auf.Jer Gmrnllage de• lnspektioosbericllte& wirdhiermit ooMJ!tigt, da-.in dieser Prtlfeinrichturog dieoben genannlen Prtlfungen unter F.inhaltungder GLP- Grund-**~611.<** durchgcfUhrt werden lconnen, The above mentioned lett facility is included inthe national GLPCompliance Programme and is inspected on aregularbasis. Based on lhe inspectionreport it can be confllTiled, thatthistest facility is able to**cond~~<:t** the aforementioned studiesin compliance with the PrinciplesofGLP. lmAul\rag

Th.

### z· c:rmann, Referent, Wiesbaden,den 19.Jallur%0117 (Name und FunlttiollderYe1'11ltwortlicbcn Person/ Name and function ofresponsiblepmon)

Hen. Mlollter1a• mrU•wclt,lllldklleaRaam alld Verbraudoenchlz,**Mai~~ZB**Stnle841 065119Wlcsbadea (Name und Adresse der OLP-ObetwaebungsbehOrde/Name and addressoftheGLP Monitoring Authority

# [ RCC-CCR Study Number 1111700


## 2 CONTENTS 1 COPY OF GLP CERTIFICATE 2 CONTENTS 3 PREFACE 

### 3.1 General 3.2 Responsibilities 3.3 Schedule 34 Project Staff Signatures 3.5 Good Laboratory Practice 3 6 Guidelines 3.7 Archiving 3.8 Deviations to Study Plan 

### 4 STATEMENT OF COMPLIANCE Final Report

5 STATEMENT OF QUALITY ASSURANCE UNIT 6 SUMMARY OF RESULTS 6.1 Conclusion 7 OBJECTIVE 7. 

### 1 A1ms of the Study 7.2 Reasons for the Study 8 MATERIALS AND METHODS 8.1 Controls 8.2 Test System 8.3 Mammalian Microsomal Fraction S9 Mix 84 Pre-Experiment for Toxicity 8.5 Dose Selection 8.6 Experimental Performance 8 7 Data Recording 8.8 Acceptability of the Assay 8.9 Evaluation of Results 8.10 Biometry 9 DISCUSSION OF RESULTS 10 REFERENCES 

### 11 DISTRIBUTION OF THE REPORT 12 SUMMARY OF RESULTS 12.1 Summary of Results Pre-Experiment and Experiment I 

### 12.2 Summary of Results Pre-Experiment and Experiment I a 12.3 Summary of Results Experiment II ·13 HISTORICAL CONTROL DATA 

### 14 ANNEX 

### 1: TABLES OF RESULTS (9 PAGES)

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### 2 3 4 4 

### 4 4 

### 5 5 

### 5 6 6 7 

### 8 9 9 

### 10 10 10 

### 11 12 

### 13 14 

### 15 16 16 16 

### 17 17 17 18 

### 19 19 

### 20 20 21 

### 22 23 24[ RCC-CCR Study Number 

### 1111700 

# ·J Final Report 

### Page 4of 33 

# 3 PREFACE 3.1 General Title Salmonella typhimurium and Escherichia coli Reverse Mutation Assay with 

# [ J Sponsor
Study Monitor

Test Facs!ity·

Contracting Institute: Reference Number: 

## 3.2 Responsibilities Study Dtrector. Deputy Study Director: Management Head of Quality Assurance Unit: 

## 3.3 Schedule Date ofthe Study Plan Experimental Starting Date: Experimental Completion Date: Date of Draft Report: Date of Final Report:



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# [ ]

# ]

RCC Cytotest Cell Research GmbH (RCC-CCR) In den Leppsteinswiesen 19 D-64380 Rossdorf 

### RCC Ltd CH-4452 ltingen 846473

Dipl. Bioi. Andrea Sokolowski Dr. Hans-EricWollny Dr. Wolfgang VOikner Frauke Hermann

July July August September September 

### 02,2007 10.2007 07,2007 05,2007 20,2007

# [**r~CC·CCH** Study Niimber 1111700 Final Repo1i Page !:i of 33


## 3.4 Project Staff Signatures
Stttdy Director Dip!. Bioi. Andrea Sokolowski

Date: September 20, 2007 Manaqement Dr. Wolfgang Volkner

Date: September 20. 2007 

## 3.5 Good Laboratory Practice The study was performed in compliance with: "Chenukaliengesetz" (Chemicals Act) of the Federal Republic of Germany, "Anhang 1" (Annex 

### 1) dated July 

### 25, 1994 ("BGBI. 

### I 1994", pp. 1703), last revision dated ,June 27, 2002 "OCCD Principles of Good Laboratory Practice", as revised in 1997 fC(97)186/Final] 

## 3.6 Guidelines**Tlli~, ~,tudy**  followed the procedures indicated by the following internationally accepted guidelines and recommendations: "Ninth Addendum to OECD Guidelines for Testing of Chemicals", Section 4, No. 47·1: "Bncteri::ll Heverse Mutation Test", adopted July 21, 1997 "Commission Directive 2000/32/EC, L1362000, Annex 4D", dated May 19, 2000 Japanese GuidelinP.s: "Kanpoan No. 

### 281

-- Environment Protection Agency" "Eisei No. 127 --Ministry of Health & Welfare" "Ht;lisei mJ/10/31 Kikyoku No.2-- Ministry of International Trade & Industry"

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# [ RCC-CCR Study Number 

### 1111700 

# ; J Final Report Page 6 of33


## 3.7 Archiving RCC Cytotest Cell Research GmbH will archive the following data for 15 years: Raw data, study plan, final report, and a sample ofthe test item. No data will be discarded without the sponsor's consent. 

## 3.8 Deviations to Study Plan There were no deviations to the study plan.

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## 4 STATEMENT OF COMPLIANCE Study Nurnher: 

### 1111700 Test !tern 

# [ 

# J Sllrdy Di1 ector: Dip!. Bioi. Andrea Sokolowski 

### Title: Salmonella typhimurium and Escherichia coli [ Reverse Mutation Assay with 

# J

This study petiormed in the test facility of RCC Cytotest Cell Research GmbH was conducted 

### in cornphance with Good Laboratory Practice Regulations: "Chemikalien9esetz" (Chemicals Act) of the Federal Republic of Gem1any. "Anhang 

### 1" (Annex l) dated July 

### 25, 1994 ("BGBL I 1994", pp. 

### 1703), last 

### revrsion dated June 27, 2002. "OECD Princ:ipk;s of Good Laboratory Practice'', as revised in 1997 [C(97)186/Finall There were no circumstances that may have affected the quality or integrity ofthe study. 

### f.>t11dy 

### Direc;tor 

### RCC-CCR Dip!. 

### Bioi. Andrea Sokolowski

.•**~:~-!** .......**~···············~··········** ·•·•••••·····•• Date: September 20, 2007

## l**RCC-CCI~** Study Number L 111 1'/00 

### 5 

# J Final Report Page 8 of 33

''**-----------~---------------** 

## 5 STATEMENT OF QUALITY ASSURANCE UNIT Gtudy 

### Numlwt:

SttHiV Du edor Tttle 

### 1111700 

# [ 

# J Dipl. BioL Andrea Sokolowski Salmonella typhimurium and 

### Escherichia coli Reverse Mutation 

### Assay with 

# [ J
The**g(mer;:~l** facilities and activities of RCC Cytotest Cell Research GmbH are inspected penodically and the results are reported to the responsible person and the management 

### Study procedures were inspected periodically. 

### The study plan and 

### this report 

### were audited 

### by**!IH~** Cluality Assurance Unit. The dates are given below. r-·

PhaS£':S 

### attd DRtes of OAU Inspections/Audits Dates of Reports to 

### the 

### Study Director and 

### to Management



## -----**·~---·--** ---·-·-·------··--- Study Pl<ln**(Drt.~ft):** 

### June 

### 29,2007 June 29, 2007**1~-- ------~-~-**  Study Plan: 

### ,July 

### 03,2007 .. ···-· ·----- 

## - ------ ··---- Process**ln~.;pr:ctinrr** July 

### 19, 2007 July 19, 2007 August 

### 01, 2007 August 

### 01, 2007

--- '"'' '. ·-·-·-" . -**-·---·-·~** . 

### Draft 

### Repo1i September 

### 13,2007 September 

### 13. 2007
Thts ::;t;1ten1ent is to contirm that the present final repoti reflects the raw data.
Head of Quality Assurance Unit Frauke Hermann



### l 

# '·''·- ". _;; 

# .., . 

### . . .

### .

## . . . .

### .' 

### .' '**'~**

### .... ' ..... '

### .. ' . ' .... ' .... Date: September 20. 2007

# [ RCC-CCR Study Number 1111700 

# >] Final Report Page 9 of33


# 6 SUMMARY OF RESULTS This study was performed to investigate the potential o( ]to induce gene mutations in the plate incorporation test (experiment I) and the pre-incubation test (experiment II) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, and TA 100, and the Escherichia coli strain WP2 uvrA. The assay was performed in two independent experiments both with and without liver microsomal activation. The plate incorporation test with strain TA 1537 with metabolic activation was repeated, since questionable results were obtained (reported as experiment Ia). In the pre-incubation test data were not evaluate for strain TA 1537, due to contamination ofthe strain. This part was repeated and is reported as part ofexperiment II. Each concentration, including the controls, was tested in triplicate. The test item was tested at the following concentrations: Pre-Experiment/Experiment I and Ia: 3; 10; 33; 100; 333; 1000; 2500; and 5000 IJg/plate Experiment II: 33; 100; 333; 1000; 2500; and 5000 IJg/plate The plates incubated with the test item showed normal background growth up to 5000 IJg/plate with and without metabolic activation in both independent experiments. No toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), occurred in all experiments with and without metabolic activation. No substantial increase in revert?Dt colony numbers of any of the five tester strains was observed following treatment wit'L_ ]at any dose level, neither in the presence norabsence of metabolic activation (S9 mix). Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies. 

## 6.1 Conclusion In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes orframeshifts in the genome ofthe strains used. Therefore.[ ;Js considered to be non-mutagenic in this Salmonella typhimurium and Escherichia coli reverse mutation assay.



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# 7 OBJECTIVE 7.1 Aims ofthe Study The experiments were performed to assess the potential of the test item to induce gene mutations by means of two independent Salmonella typhimurium and Escherichia coli reverse mutation assays. Experiment I was performed as a plate incorporation assay. Since a negative result was obtained in this experiment, experiment II was performed as a pre-incubation assay. 

## 7.2 Reasons for the Study The most widely used assays for detecting gene mutations are those using bacteria (3). They are relatively simple and rapid to perform, and give reliable data on the ability of an agent to interactwith DNA and produce mutations. Reverse mutation assays determine the frequency with which an agent reverses or suppresses the effect of the forward mutation. The genetic target presented to an agent is therefore small, specific and selective. Several bacterial strains, or a single strain with multiple markers are necessary to overcome the effects of mutagen specificity. The reversion of bacteria from growth-dependence on a particular amino acid to growth in the absence of that amino acid (reversion from auxotrophy to prototrophy) is the most widely used marker. The Salmonella typhimurium histidine (his) and the 

### E. coli tryptophan (trp) reversion system measures his·**~** 

### his• and trp-**~** 

### trp+ reversions, respectively. The S. typhimurium and Escherichia coli strains are constructed to differentiate between base pair (TA 1535, TA 100. and WP2 uvrA) and frameshift (TA 1537, TA 98) mutations. According to the direct plate incorporation or the pre-incubation method the bacteria are exposed to the test item with and without metabolic activation and plated on selective medium. After a suitable period of incubation, revertant colonies are counted. To establish a dose response effect at least 6 dose levels with adequately spaced intervals were tested. The maximum dose level was 5000 j.Jg/plate. To validate the test, reference mutagens were tested in parallel to the test item.

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### 1111700 

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### 11 of33 

## 8 MATERIALS AND METHODS
Internal RCC-CCR Test Item Number: S 768911 

### The test item and the information concerning the test item were provided by the sponsor. Identity: Batch No.: Aggregate state at room temperature: Colour: Purity: Stability in solvent: Storage: Expiration Date: 

# [ [ 

# J
Solid Light brown 100 

### % active polymer Not indicated by the sponsor At room temperature, light protected March 31 , 2008 

# J

On the day of the experiment, the test iten( Jwas suspended in THF (MERCK, D-64293 Darmstadt; purity > 99 %). The solvent was cnosen because of its solubility properties and its relative non-toxicity to the bacteria (5}. Precipitation of the test item occurred in the test tubes (all experiments) and in the overlay agar on the incubated agar plates in experiment II from 2500 J.Jg/plate up to 5000 J,Jg/plate with and without metabolic activation. The undissolved particles had no influence on the data recording.

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# [ RCC-CCR Study Number 11117()0 Final Report Page 12 of 33 

# ;J
8.1 Controls 8.1.1 Negative Controls Concurrent untreated and solvent controls were performed. 

### 8.1.2 Positive Control Substances Without metabolic activation Strains: Name: Supplier: Catalogue No.: Purity: Dissolved in: Concentration: Strains Name: Supplier: Catalogue No.: Purity: Dissolved in: Concentration: Strain: Name: Supplier: Catalogue No.: Purity: Dissolved in: Concentration: 

### With metabolic activation Strains: Name: Supplier: Catalogue No.: Purity: Dissolved in: Concentration: TA 1535, TA 100 sodium azide, NaN3 SERVA, D-69042 Heidelberg 30175 at least 99% water deionised 10 IJQ/plate TA 1537, TA 98 4-nitro-o-phenylene-diamine, 4-NOPD SIGMA, D-82041 Deisenhofen N 9504 > 99.9% DMSO (MERCK, D-64293 Darmstadt; purity> 99 %) 10 IJQ/plate in T A 98, 50 IJQ/plate in TA 1537 WP2 uvrA methyl methane sulfonate, MMS Merck-Schuchardt. D-85662 Hohenbrunn 820775 > 99.0% waterdeionised 3 IJL/plate

TA 1535, TA 1537, TA 98, TA 100, WP2 uvrA 2-aminoanthracene, 2-AA SIGMA, D-82041 Deisenhofen 

### A 1381 97.5% DMSO (MERCK, D-64293 Darmstadt; purity> 99 %) 2.51Jg/plate (TA 1535, TA 1537, TA 98, TA 100), 10 tJg/plate (WP2 uvrA) The stability ofthe positive control substances in solution is unknown but a mutagenic response in the expected range will be sufficient evidence ofbiological stability.

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## 8.2 Test System 8.2.1 Characterisation ofthe Salmonella typhimurium Strains and E. coli Strain The histidine dependent strains are derived from S. typhimurium strain 

### L T2 through a mutation in the histidine locus. Additionally due to the "deep rough" (rfa-minus) mutation they possess a faulty lipopolysaccharide envelope which enables substances to penetrate the cell wall more easily. A further mutation causes a reduction in the activity ofan excision repair system. The latter alteration includes mutational processes in the nitrate reductase and biotin genes produced in a UV-sensitive area of the gene named "uvrB-minus". In the strains TA 98 and T A 100 the R-factor plasmid pKM 101 carries the ampicillin resistance marker (6). Strain WP2 (4) and its derivatives all carry the same defect in one of the genes for tryptophan biosynthesis. Tryptophan-independent (Trp+) mutants (revertants) can arise either by a base change at the site ofthe original alteration or by a base change elsewhere in the chromosome so that the original defect is suppressed. This second possibility can occur in several different ways so that the system seems capable of detecting all types of mutagen which substitute one base for another. Additionally, the uvrA derivative is deficient in the DNA repair process (excision repair damage). Such a repair-deficient strain may be more readily mutated by agents. When summarised the mutations ofthe T A strains and the 

### E. coli strain, used in this study can be described as follows:

.-----·--·

--·---- Salmonella typhimurium Strains Genotype Type of mutations indicated 

### TA 1537 his C 3076; rta·; uvrs·: frame shift mutations

-- 

### ITA98 his D 3052; rta·; uvrB-;R-factor 

### " " 

### TA 1535 his G 46; rta·; uvrs·:**base~air** 

### substitutions TA 100 his G 46; rta·; uvrB-;R-factor 

### " II Escherichia coli**~--·-~·-·-·~-~--~~·~-** 

### WP2 uvrA trp·; uvrA·: base-pair substitutions and others Regular checking ofthe properties of the strains regarding the membrane permeability and ampicillin resistance as well as spontaneous mutation rates is performed in RCC Cytotest Cell Research according to B. Ames et al. (1) and D. Maron and 

### B. Ames (6). In this way it was ensured thatthe experimental conditions set down by Ames were fulfilled. The bacterial strains TA 1535, TA 1537, TA 98, TA 100, and WP2 uvrA were obtained from Trinova Biochem GmbH (35394 GieBen, Germany).

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### Number 1111700 

# ·] Final Report Page 14 of33



### 8.2.2 Storage The strain cultures were stored as stock cultures in ampoules with nutrient broth + 

### 5% DMSO (MERCK, D-64293 Darmstadt) in liquid nitrogen. 

### 8.2.3 Precultures From the thawed ampoules of the strains 0.5 ml suspension was transferred into 250 ml Erlenmeyer flasks containing 20 ml nutrient medium. A solution of 20 !JL ampicillin (25**~Jg/ml)** 

### was added to the strains TA 98 and TA 100. This nutrient medium contains per litre: 8 g Merck Nutrient Broth (MERCK, D-64293 Darmstadt) 5 g NaCI (MERCK, D-64293 Darmstadt) The bacterial cultures were incubated in a shaking water bath for 4 hours at 3r C. 

### 8.2.4 Selective Agar The plates with the selective agar were obtained from 

### E. Merck, D-64293 Darmstadt 

### 8.2.5 Overlay Agar The overlay agar contains per litre: for Salmonella strains:

6.0 g MERCK Agar Agar• 6.0 g NaCI* 10.5mg l.-HistidinexHCixH20* 12.2 mg Biotin* * 

### (MERCK, D-64293 Darmstadt) for Escherichia coli: 6.0 g MERCK AgarAgar* 6.0 g NaCI* 2.5 mg Tryptophan*

Sterilisations were performed at 121° C in an autoclave. 

## 8.3 Mammalian Microsomal Fraction 59 Mix The bacteria used in this assay do not possess the enzyme systems which, in mammals, are known to convert promutagens into active DNA damaging metabolites. In order to overcome this major drawback an exogenous metabolic system is added in form of mammalian microsome enzyme activation mixture.

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### 8.3.1 S9 (Preparation 

## by R C C - 

### C C R)**Phenobarbitai/~-Naphthoflavone** 

### induced rat liver 59 is used as the metabolic activation system. The 59 is prepared from 8 - 12 weeks old male Wistar Hanlbm rats, weight approx. 220 - 320 g induced by applications of 80 mg/kg b.w. Phenobarbital i.p. (Desitin; D-22335 Hamburg) and B-Naphthoflavone p.o. (Aldrich, D-89555 Steinheim) each on three consecutive days. The livers are prepared 24 hours after the last treatment. The 59 fractions are produced by dilution of the liver homogenate with a KCI solution (1+3) followed by centrifugation at 9000 g. Aliquots of the supernatant are frozen and stored in ampoules at -80° C. Small numbers of the ampoules can be kept at -20°C for up to one week. Each batch of 59 mix is routinely tested with 2-aminoanthracene as welt as benzo(a)pyrene. The protein concentration in the 59 preparation was 27.6 mg/ml (lot no. R 210607) in all experiments. 

### 8.3.2 S9 Mix Before the experiment an appropriate quantity of 59 supernatant was thawed and mixed with 59 co-factor solution. The amount of 59 supernatant was 10% v/v in the 59 mix. Cofactors are added to the 59 mix to reach the following concentrations in the 59 mix: 8 mM MgCI2 33 mM KCI

5 mM Glucose-6-phosphate 5 mM NADP in 100 mM sodium-ortho-phosphate-buffer, pH 7.4. During the experiment the 59 mix was stored in an ice bath. The 59 mix preparation was performed according to Ames et al.(1 ). 

## 8.4 Pre-Experiment for Toxicity To evaluate the toxicity of the test item a pre-experiment was performed with strains TA 1535, TA 1537, TA 98, TA 100, and WP2 uvrA Eight concentrations were tested for toxicity and mutation induction with three plates each. The experimental conditions in this pre-experiment were the same as described below for the experiment I (plate incorporation test). Toxicity ofthe test item results in a reduction in the number ofspontaneous revertants or a clearing ofthe bacterial background lawn. The pre-experiment is reported as experiment I, ifthe following criteria are met: Evaluable plates (>0 colonies) at five concentrations or more in all strains used.

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# [ RCC-CCR Studv 

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## 8.5 Dose Selection In the pre-experiment the concentration range ofthe test item was 

### 3- 5000 1-1gfplate. The pre-experiment is reported as experiment 

### I. Since no toxic effects were observed 5000 j.lgfplate were chosen as maximal concentration. Tile concentration range included two logarithmic decades. The following concentrations were tested in experiment II: 33; 100; 

### 333; 1000; 2500; and 5000 J,Jg/plate 

## 8.6 Experimental Performance For each strain and dose level including the controls, three plates were used. The following materials were mixed in a test tube and poured onto the selective agar plates: 251JL Test solution at each dose level, solvent control 100 IJL Reference mutagen solution (positive control}, 500 !JL S9 mix (for testwith metabolic activation) or S9 mix substitution buffer (for test without metabolic activation), 100 IJL Bacteria suspension 

### (cf. test system, pre-culture ofthe strains), 2000 IJL Overlay agar In the pre-incubation assay 25 IJL test solution, solvent control, 100 J.il reference mutagen solution (positive control), 500 JJL S9 mix 

### I S9 mix substitution buffer and 100 IJL bacterial suspension were mixed in a test tube and shaken at 37° C for 60 minutes. After preincubation 2.0 ml overlay agar (45° 

### C) was added to each tube. The mixture was poured on selective agar plates. After solidification the plates were incubated upside down for at least 48 hours at 

### 3r C in the dark (2). 

## 8.7 Data Recording The colonies were counted using the Petri Viewer Mk2 (Perceptive Instruments Ltd, Suffolk CB 7BN, UK) with the software program Ames Study Manager. The counter was connected to an IBM AT compatible PC with printer to print out the individual values and the means from the plates for each concentration together with standard deviations and enhancement factors as compared to the spontaneous reversion rates (see tables of results) Due to precipitation of the test item and air bubbles some plates were counted manually.**~h.h,** 

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## 8.8 Acceptability ofthe Assay The Salmonella typhimurium and Escherichia coli reverse mutation assay is considered acceptable if it meets the following criteria: regular background growth in the negative and solvent control the spontaneous reversion rates in the negative and solvent control are in the range of our historical data the positive control substances should produce a significant increase in mutant colony frequencies 

## 8.9 Evaluation of Results A test item is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice {strains TA 98, TA 100, and WP2 uvrA) or thrice (strains TA 1535 and TA 1537) the colony count ofthe corresponding solvent control is observed (3}. A dose dependent increase is considered biologically relevant 

### ifthe threshold is exceeded at more than one concentration (2). An increase exceeding the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment. A dose dependent increase in the number of revertant colonies below the threshold is regarded as an indication of a mutagenic potential if reproduced in an independent second experiment. However, whenever the colony counts remain within the historical range of negative and solvent controls such an increase is not considered biologically relevant. 

## 8.10 Biometry According to the OECD guideline 471, a statistical analysis ofthe data is not mandatory.

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# J Final Report Page 18 of33 

# 9 DISCUSSION OF RESULTS The test item[ )was assessed for its potential to induce gene mutations in the plate incorporation test \experiment I) and the pre-incubation test (experiment II) using Salmonella typhimurium strains TA 1535, TA 1537, TA 98, and TA 100, 

### and the Escherichia coli strain WP2 uvrA The assay was performed in two independent experiments both with and without liver microsomal activation. The plate incorporation test with strain T A 1537 

### with metabolic activation was repeated, since questionable results were obtained (reported as experiment Ia). In the pre-incubation test data were not evaluate for strain TA 1537, due to contamination ofthe strain. This part was repeated and is reported as part ofexperiment II. Each concentration, including the controls, was tested in triplicate. The test item was tested at the following concentrations: Pre-Experiment/Experiment I and Ia; 3; 10; 33; 100; 333; 1000; 2500; 

### and 5000 

### !Jglplate Experiment II: 33; 100; 333; 1000; 2500; 

### and 5000 

### J.Jg/plate The plates incubated with the test item showed normal background growth up to 5000 

### J.Jg/plate with and without metabolic activation in both independent experiments. No toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), 

### occurred in all experiments with and without metabolic activation. No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with[ ]at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). In Experiment I with metabolic activation strain TA 1537 

### showed a minor increase. The number of colonies exceeded the threshold of thrice the number of the corresponding solvent control at concentration 1000 

### IJQ/plate. To clarify the results a confirmatory experiment was performed under identical conditions. In the confirmatory experiment no increase was observed. Therefore this minor increase is judged ad biologically irrelevant. Appropriate reference mutagens were used as positive controls. They showed a distinct increase of induced revertant colonies. In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes orframeshifts in the genome ofthe strains used.

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### 1111700

# J Final Report Page 19 of33


## 10 REFERENCES 1. Ames, B.N, J. 

### McCann, and 

### E. Yamasaki (1977) Methods for detecting carcinogens and mutagens with the Salmonella/mammalian microsome mutagenicity test In: B J 

### Kilbey et al. (Eds.)"Handbook of Mutagenicity Test Procedures" Elsevier, Amsterdam, 1-17 2. de Serres F.J. and M.D. Shelby (1979) Recommendations on data production and analysis using the Salmonella/microsome mutagenicity assay Mutation Res. 64, 159-165 3 Hollstein,M., 

### J. McCann, 

### FA Angelosanto and W.W. Nichols (1979) Short-term tests for carcinogens and mutagens Mutation Res. 65, 133-226 4. Green, M.H.L and WJ. Muriel (1976) Mutagen Testing Using TRP+ Reversion in Escherichia Coli Mutation. Res. 38, 3- 32 5. Maron D.M., J. Katzenellenbogen and B.N. Ames, (1981) Compatibility of organic solvents with the Salmonella/Microsome Test Mutation Res. 88, 343-350 6. Maron D.M., Ames, B.N. (1983} Revised methods for the Salmonella mutagenicity test Mutation Res. 113, 173-215



## 11 DISTRIBUTION OF THE REPORT Sponsor Study Director

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### 3 x 

### (2x copy, 1x electronic copy (pdf)) 1 x 

### (original)

# [**RCC~CCR** Studv Number 1111700 Final Report Page 20 of 33 

# J 12 SUMMARY OF RESULTS 12.1 Summary of Results Pre-Experiment and Experiment 

## I Study Name 1111700 Study Code: RCC-CCR 1111700 Experiment: 1111700 W Plate Date Plated: 10/07/2007 Assay Conditions Date Counted: 1310712007 Metabolic Test Dose Level Revertant Colony Counts (Mean ±SD) ACIIVUII.Q!! Group {b!glplate} TA 1535 TA 1537 TA98 TA 100 WP2 uvrA Without THF 20±4 12 ± 5 26 ±6 162 ± 16 49 ± 4 Activation Untreated 19 ± 3 10 ±6 34±17 153 ± 10 45 ± 8 

# [ J 3119 16 ± 1 10 ± 4 34±7 143 ± 10 40±6 10 1J9 17 ± 7 8±3 28 ± 12 171 ±9 47 ± 11 33119 17 ± 9 11 ± 2 30 ± 13 153 ± 7 56 ±4 100 119 22 ±8 12 ± 1 33±8 155 ± 9 57 ±7 333 1J9 17 ± 3 10 ± 2 28 ± 2 151 ± 10 46 ±4 1000 119 20±2 14 ±2 29 ± 2 159 ± 30 52 ±8 2500119 20 ±3 10 ±4 36 ±3 160 ± 10 51±21 5000119 15 ± 2 9±3 36±4 145 ± 19 43 ± 2 NaN3 10119 2019±55 2218 ± 80 4--NOPD 10119 464 

### ±17 4-NOPD 50119 108 ± 16 MMS 3.0 11L 1197 ±72 With THF 20 ± 3 11 ±4 41 :I: 10 188 :I: 12 76 ± 15 Activation Untreated 22 ±3 16 :t 3 28 ±3 170 :t 15 59±3 1
.. 

# J 3 1J9 18

### ±7 14 ± 1 36 

### ±7 164 ± 13 62 ±4 

### i r 10 119 17:1:5 10:1:2 40±9 152 ± 25 65 ± 11 

# L 33119 19 ±4 15 ± 3 47:1:2 163 ± 10 67 ±7 100 119 17 ± 5 14 ± 3 40 ± 5 174 ± 11 67 ±6 333119 20±4 16 ±2 43 ±7 146 ± 10 61 ± 8 1000 ll9 26 ± 10 37 ± 6 46:1:10 171 :1:5 77 ±6 2500119 28 :t 8 20±7 51±9 175±15 69 ± 10 5000 1J9 23±2 23 ± 1 43 ± 2 174 ± 13 62 ± 12 2-AA 2.5 1J9 289±8 252 :t 29 1467± 2128 ± 138 196 2-AA 10.0 119 296 ± 18**~~~Posit~lv~e~C~o~nt~rocl~s~---------------------------------------------------** NaN3 2-AA 4-NOPD MMS sodium azide 2-aminoanthracene 4-nitro-o-phenylene-diamine methyl methane sulfonate

s4c2dr.doc

# L RCC-CCR Study Number 

### 1111700

# J Final Report Page 21 of 33


## 12.2Summary of Results Pre-Experiment and Experiment I a Study Name: '1111700 Experiment. 1111700Wa Plate Assay Conditions: Metabolic Test Activation Qroup
With THF 

### Actl'"l'on [ Uot.eatod

2-AA Dose Level lug/plate) 

# J 31Jg 10 1-19 331Jg 100 IJ9 3331JQ 1000 1-19 2500 IJ9 50001-19 2.51JQ Study Code: RCC-CCR 1111700 Date Plated: 17/07/2007 Date Counted: 20!07/2007 Revertant Colony Counts (Mean ±SO) TA 1537 15±4 16 ±4 19 ±4 14 ±6 15 ±2 18 ± 1 16 ±6 16 ±3 16 ± 1 p 18 ± 2p 363 ± 15**~X~~ositiveCon~t~ro~ls~--------------------~K~ey~ro~P~Ia~t~e~P~os~~~·x~C~o~d~e~s**______________ __ 4-NOPD 4-nitro-o-phenylene-diamine p Precipitate 2-AA 2-aminoanthracene

s4c2dr.docRCC-CCR Study Number 1111700 Final Report Page 22 of33 

# [ 5 

# J
12.3Summary of Results Experiment II Study Name: 1111700 Study Code: RCC-CCR 1111700 Expenment 1111700 HV2 Pre Date Plated 26/07/2007 I 02/0812007* Assay Condttions: Date Counted: 01108/2007 f0710812007* Metabolic Test Dose level Revertant Colony Counts (Mean ±SD) 8.c;tiy9tion 

### QlQY.Q (yg/(21ate} TA 1535 TA 1537 TA98 TA 100 WP2 uvrA Without THF 15 ± 5 9±1 39 ±4 140 ± 18 35±6 Activation Untreated 13 ±4 10 ±3 41 ± 7 132 ± 15 58± 13 

# [ 

# l331J9 12 ± 3 11 ± 3 27:1:3 135 ± 9 39 ± 5 100 IJ9 14 ±2 8±4 37 ± 8 125 ± 10 45 ± 13 333 IJ9 15 ±4 9±3 29±6 130 ± 13 41 ± 9 1000 IJ9 16 ±4 13 ± 2 34 ±4 129 ±0 44 ±4 2500 IJ9 13 ± 3p 9 ± 3p 28 ±4p 79 ± 9P 42 ± 3p 5000 IJ9 14 ± 9P 10 ± 2p 32 ±

### sP 106±2p 41 ±

### aP NaN3 10 IJ9 1919± 67 1845 ± 48 4-NOPD 10 IJ9 741 ±67 4-NOPD 50 IJ9 145 ± 16 MMS 3.0 !JL 344 ± 13 With THF 18 ::1: 1 18 ±4 47 ±8 142 ± 8 62 ± 10 Activation Untreated 20±7 12 ±2 49±8 159 ± 11 62±9 

# [ J 331Jg 19:1:4 18 ±4 44:1:10 150

### ±20 63 ± 12 100 IJ9 18 ±2 15:1: 1 42:1:11 136 ± 16 59:1:3 333j.lg 20 ± 1 16 ±2 42 ±6 145:1: 13 55:1:1 1000 IJ9 16:1:3 15±2UM 34:1:3 143 ± 4 61 ± 3 2500 IJ9 18:1: 3p 12 ±4 PM 45 ±5p 117±14p 53:1:5 PM M 5000 IJ9 18 ± 2 p 9 ± 1PM 41 ±2p 109±14p 54 :1:6PM M 2-AA 2.5 IJ9 233:1:3 159:1:8 1277:1:85 1939 ± 144 2-AA 10.0 IJ9 415 ± 22**~eyjo_F:()~'!}~~gon~!<!l?** Ke:f to Plate Postfix Codes NaN3 sodium azide p Precipitate 2-AA 2-aminoanthracene M Manual count 4-NOPO 4-nttro-o-phenylene-diamine 

### u Air bubbles MMS methyl methane sulfonate

s4c2dr.doc[ RCC-CCR Study Number 1111700

# J Final Report Page 23 of33 

# 13 HISTORICAL CONTROL DATA These data represent the laboratory's historical control data from May 2005 until June 2006 representing approx. 200 experiments (WP2 uvrA the historical data are based on approx. 100 experiments). Strain without S9 mix with S9 mix Mean 

## so Min Max Mean 

## so Min Solvent control 20.8 4.7 9 35 24.7 5.9 7 TA 1535 Negative control 20.4 4.4 11 31 24.2 5.5 10 Max 43 38 Positive control 1422.0 464.7 781 4900 332.0 95.3 107 695 Solvent control 11.2 3.7 5 28 16.2 5.0 6 36 TA1537 Negative control 11.6 4.0 4 28 17.1 5.4 7 34 Positive control 99.8 32.5 53 425 276.8 132.6 59 746 Solvent control 28.1 6.1 15 49 37.9 7.4 20 57 TA98 Negative control 30.2 6.6 16 60 39.0 7.5 18 64 Positive control 439.0 155.2 176 1818 1839.4 898.6 407 4891 Solvent control 130.7 20.8 87 197 147.0 25.5 84 255 TA 100 Negative control 138.2 21.6 86 216 150 1 24.2 96 214 Positive control 2083.1 281.3 616 2872 2372.9 958.4 417 5230 Solvent control 55.8 7.2 31 74 63.9 9.1 34 84 WP2uvrA Negative control 55.9 8.6 36 76 65.6 10.4 33 91 Positive control 991 0 522.9 249 1810 3194 84.8 211 930 Mean = mean value of revertants/plate SD 

# = standard deviation Min 

### =minimal value/Max

# = maximal value



### s4c2dr.docRCC-CCR Study 

### Number 1111700 

# [ J Final Report Page 24 of 33


## 14ANNEX 

# 1: TABLES OF RESULTS (9 PAGES) Pre-Experiment and Experiment 1: 

### 1111700 W Plate Incorporation (4 pages) Pre-Experiment and Experiment I a: 

### 1111700 W Plate Incorporation (

### 1 page) Experiment II 

### 1111700 HV2 Pre-Incubation (4 pages)

s4c2dr.doc[ RCC-CCR Study Number 

### 1111700

# J Final Report Page 25 of 33



## ---------------------------------------------- Study Name 1111700 Study Code RCC-CCR 1111700 Expemnent 1111 700 W Plate Date Plated: 10/07/2007 Assay Condit•ons. Date Counted: 13107/2007 Without metabolic activation



### -----'stiain
_______ 

### COmpound Dose level- Mean Standard Individual revertant per plate revertants Deviation counts TA 1535 

# [ 3**~g** 16.3 0.6 0.8 16, 17, 16 10 IJ9 17.0 7.2 0.9 19,9, 23 331J9 16.7 9.1 0.8 27, 13, 10 100j.Jg 22.0 8.2 1 1 13, 29, 24 3331J9 17.0 35 0.9 19, 19, 13 1000 IJQ 20.3 1.5 1.0 19,22.20 2500 IJ9 19.7 2.5 1.0 20, 17, 22 5000 tJQ 14.7 2.3 07 16, 12. 16 THF 19.7 4.0 19,16,24 Untreated Control 18.7 2.5 16**---·~~··-** -·----···------- TA 1537 [ 

# l 3JJg 10.0 40 0.8 6, 14. 10 101.19 8.0 3.5 0.7 6, 12,6 33!-19 113 2.1 0.9 12, 13. 9 100 1-19 12.3 0.6 1.0 12, 13. 12 3331J9 10.3 1.5 09 10, 9, 12 10001-19 13.7 1.5 1.1 14, 15, 12 25001-19 10.0 3.6 08 7, 14, 9 50001-19 9.0 2.6 0.8 12, 7, 8 THF 12.0 50 12, 7. 17 Untreated Control 9.7 5.8 13, 3, 13**---~------·---**

------ TA 98 

# J 31J9 34.3 6.7 13 31,30,42 10 1-19 28.3 119 1.1 23,42.20 331Jg 29 7 12.9 1.1 44,26,19 100119 33.3 8.1 1.3 24,38,38 3331JQ 28.0 1.7 1.1 26,29,29 1000 IJQ 28 7 2.1 1.1 28,31,27 2500 IJQ 36.3 32 14 34, 35,40 5000 IJ9 363 4.0 1.4 41, 34, 34 THF 26.0 6.2 21,33,24 Untreated Control 34.0 16.7 19, 52, 31



# ---------- TA 100 

# ] 3 f-19 143.0 101 0.9 152, 145, 132 10 IJ9 171.0 9.2 1.1 161, 173, 179 331JQ 153.3 7.4 0.9 156, 159, 145 1001-JQ 154.7 9 1 1.0 165, 151, 148 3331J9 151.3 10.1 09 150, 142, 162 1000 IJQ 158.7 297 1.0 126, 184, 166 2500 IJQ 159.7 10.1 1.0 148, 166, 165 5000 fJQ 144.7 186 0.9 125, 162, 147 THF 162.0 157 155, 180, 151 Untreated Control 153.0 9.5 154. 143, 162

---**·----~~-~~-**

s4c2dr.doc[ RCC-CCR Study Number 

### 1111700 Final Report ..J Page 26 of 33**--·-·---~--------·------------------------------** Study Name: 1111 700 Experiment 1111700 W Plate Assay Conditions 

### ----strai;,-----·- Compound

WP2uvrA

THF Untreated Control TA 1535 NaN3 TA 1537 4-NOPD

TA 98 4-NOPD TA 100 NaNJ WP2 uvrA MMS 

# ]Dose level per plate

31.19 101.19 331JQ 1001J9 3331.19 10001.19 25001.19 5000 IJ9

10 IJ9 501JQ 10 IJ9 101-19 3.01JL

4-NOPD 4-nitro-o-phenylene-diamine MMS methyl methane sulfonate



### s4c2dr.doc Study Code RCC-CCR 1111700 Date Plated 10/0712007 Date Counted 1310712007 Without metabolic activation

Mean Standard Ratio Individual revertant revertants Deviation treated I colony counts per plate solvent

40.0 5.6 0.8 41, 34,45 46 7 11.0 1.0 34,52,54 55.7 4.2 1.1 51, 59, 57 57.3 6.5 1.2 64, 51,57 45.7 3.8 0.9 44,50,43 51 7 8.4 1.1 42, 57, 56 51.0 207 10 74,34,45 42.7 1.5 0.9 44,43,41 48.7 42 50,52,44 450 8.2 38, 43,54 2019.0 54.6 102.7 1988,2082,1987

108.0 15.9 9.0 120,114,90 464.3 16 7 179 474,445,474 2217.7 79.9 13.7 2183, 2309, 2161 1197.3 71.8 24.6 1224, 1116, 1252

# [ RCC-CCR Study Number 

### 1111700

# J Final Report Page 27 of 33

Study Name 1111 700 Study Code: RCC-CCR 1111700 Experiment 1111700W Plate Date Plated 10107/2007 Assay Conditions Date Counted. 13/07/2007 With metabolic activation --Strain**--··~--COmpound** Dose level Mean Standard Ratio Individual revertant per plate revertants Deviation treated I colony counts TA 1535 31J9 18.0 7.2 0.9 16,12,26 10119 17.0 4.6 0.9 16,22,13 331Jg 18.7 4.0 0.9 21, 21, 14 100 ll9 17.3 5.1 0.9 16,23,13 333119 19 7 4.0 1.0 22,15,22 1000119 260 9.5 1.3 20,37,21 2500119 28.0 8.2 1.4 35, 30, 19 5000119 22.7 1.5 1.1 23,21,24 THF 20.0 3.0 17,20,23 Untreated Control 22.0 2.6 19,24, 23**--~-----·-·~----** TA 1537 [ 3119 14 0 10 1.3 14, 13, 15 10119 10.0 1.7 0.9 9,9, 12 331J9 15.3 2.9 1.4 17, 17, 12 100119 14.0 3.5 13 10, 16, 16 3331Jg 15.7 1.5 1.4 17, 16, 14 1000 119 37.3 59 3.4 44,33,35 2500119 20.3 65 18 27, 14, 20 5000 IJ9 22.7 1.2 2.1 22,22,24 THF 110 3.6 8, 10, 15 Untreated Control 16.0 3.0 19. 16
TAM [ 3119 35.7 7.1 0.9 28,42,37 10 IJ9 39.7 8.5 1.0 31.48,40 331Jg 47.0 1.7 1.2 48,48,45 1001Jg 39.7 4.5 1.0 40,35,44 3331J9 42.7 6.5 1.0 43,36,49 1000 1!9 46.3 9.7 1.1 44,38, 57 2500 ll9 510 8.9 1.3 58, 41, 54 5000 IJ9 43.3 2.1 1.1 44,45,41 THF 40.7 10.0 51,31,40 Untreated Control 28.0 3.5 30, 30 TA 100 

# 1 3119 163.7 13.3 0.9 157, 155, 179 10 IJ9 152.3 24.7 08 125, 173, 159 33119 163.0 10.4 09 168, 170. 151 100119 173.7 11.4 0.9 177, 161' 183 333119 146.3 10.0 0.8 156, 136, 147 1000 IJ9 171.0 4.6 0.9 172, 166, 175 2500 l.l9 174.7 153 0.9 169. 163, 192 5000 IJ9 1737 13.1 09 159, 184, 178 THF 188.0 12.2 182,180,202 Untreated 170.0 15.0 170

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# [ 1111700

# _] Final Report Page 28 of33



## ---------------------------------------------------
Study Name_ 1111700 Experiment 1111700W Plate Assay Condttions_ Study Code: RCC-CCR 1111700 Date Plated: 10/07/2007 Date Counted: 13/07/2007 With metabolic activation

- ---COmpound ----Doselevel Mean per plate revertants



## --------- WP2•"A [ 

# l 3119 62_0 10119 65.0 33119 66 7 100119 67.3 3331J9 61.0 1000119 77.3 2500 IJ9 69.0 5000 I.JQ 61.7 THF 75.7 Untreated Control 59.0**-------··~·--.-** TA 1535 2-AA 2.51.JQ 2887 TA 1537 2-AA 2.51.JQ 252.0

TA98 2·AA 2.5119 1467.0 TA 100 2-AA 2.51J9 2127.7 WP2 uvrA 2-AA 10.0 IJQ 295.7 Key to Posillve Controls 2-M 2-aminoanthracene



### s4c2dr.doc Standard Deviation

3_6 10.5 67 5.5 7.9 6.1 9.5 12.3 14.8 3.5 8.1 28.6 137.8 1963 18.1

....... ,. Ratio Individual revertant treated I colony counts



## o_a 65,58,63 0.9 55,64, 76 09 65,74,61 09 61, 70, 71 08 55,58, 70 1.0 84, 76, 72 0.9 70,59, 78 08 72,48,65 92,63, 72 63, 57, 57 14.4 285,298,283 22.9 249,282,225 36.1 1562, 1530, 1309 11.3 1925,2317.2141 3.9 279. 293, 315[ RCC-CCR Study Number 

### 1111700

# J Final Report Page 29 of 33**-·----····----~~----------------------------------------** Study Name 1111700 Experiment: 1111700Wa Plate Assay Condi!lons

TA 1537

THF Untreated Control TA 1537 2·AA Key to Positive Controls 2-AA 2-aminoanlhracene

s4c2drdoc 

# J 31JQ 10**~IQ** 331Jg 1001-19 3331-Jg 1000 1-19 25001-19 5000 IJ9
2.51)9 With metabolic activation

19.3 14.3 14.7 17.7 163 16.0 16.0 17.7 15.3 15.7 363.3 Standard Deviation
3.5 5.7 2.3 12 6.0 2.6 1.0 2.1 4.2 4.0 15.1 1.3 09 1.0 1.2 1.1 1.0 1.0 1.2

23.7 Study Code RCC-CCR 1111700 Date Plated: 26/071200·1 I Date Counted: 01/08/2007 Individual revertant counts 19, 16, 23 8, 16, 19 16. 16. 12 19, 17, 17 10, 17, 22 19, 14. 15 15P,16P.17P 16P, 20 P, 17 P 14,12,20 12,20 374,346,370 Key to Plate Postfix Codes p Preapitate

# [ RCC·CCR Study Number 

### 1111700 

# J Final Report 

### Page 30 of 33

Study Name 1111 700 Study Code RCC-CCR 1111700 Expenment 1111 700 HV2 Pre Date Plated 26/07/2007 I 0210612007* Assay Comlltions Date Counted: 01/08/2007 I0710812007* Without metabolic activation

----· -------**~-~-.** 

### coifiiXlillid Stra•n Dose level Standard Ratio Individual revertant Deviation treated I counts**-----~~·...,_.._.._....,_~-----~------** TA 1535 

## f 

# J 331.19 12.0 2.6 08 14, 9. 13 

# L 1001-19 14.0 1.7 0.9 13, 13. 16 333119 14.7 4.0 1 0 17, 17, 10 1000 119 16.3 4.0 1.1 20, 12, 17 2500119 13.3 2.9 0.9 10P,15P,15P 5000119 140 8.5 0.9 23 P, 6 P. 13 P THF 15.0 4.6 19, 10. 16 Untreated Control 13.3 3.5 17



### -------- 

# TA1537l 1 331J9 11.3 3.1 1.2 14, 12, 8 100 IJQ 8.3 40 0.9 6,6. 13 3331.19 9.3 2.5 1.0 12, 9, 7 10001-19 12.7 2.3 1.4 14, 14. 10 2500119 8.7 31 0.9 8 P, 12 P, 6 P 5000 IJ9 10.3 1.5 1.1 12 P, 10 P, 9 P THF 9.3 12 10, 8, 10 Untreated Control 103 25 10,13,8**---------·---~**
·--------
..**~-·· ~--~-** **----~----** TA 98 

# ] 33 IJQ 270 3.5 0.7 23.29,29 100 1.19 37.3 8.3 1.0 44,40, 28 3331)g 28.7 5.9 0.7 31,22,33 10001.19 33.7 4.2 0.9 37.35,29 2500 IJQ 27.7 3.5 0.7 31 P. 28 P, 24 P 5000119 32.0 5.2 08 29P, 29 P, 38 P THF 387 3.8 37,43, 36 Untreated Control 40.7 7.0 48,40,34 

# __ ,

## _______**·-----~-----~---------~·-·** 

## TAiOor-

---- 331J9 134.7 8.7 1.0 142, 137, 125 100 IJ9 1250 96 0.9 129, 132, 114 3331.19 130.3 12.7 0.9 128, 144, 119 10001.19 129.0 00 0.9 129, 129. 129 25001.19 78 7 9.1 0.6 87 P, 80 P, 69 P 5000 IJQ 105.7 2.1 0.8 105 P, 108 P, 104 P THF 139.7 17.9 149,119,151 Untreated Control 131.7 15.3 140, 141' 114 Key to Plate Postfix Codes

s4c2dr.doc[ RCC-CCR Study Number

Study Name 1111700 Expenment. 1111700 HV2 Pre Assay Cond•tions 

### 1111700

# J Final Report

Without metabolic activation Page 31 of 33

Study Code RCC-CCR 1111700 Date Plated 2610712007 I 02108/2007* Date Counted 01/08/2007 I 07/08/2007• 

### ---Sirain-- --------Compound -·- Dose level Mean Standard Ratio Individual revertant per plate revertants Deviation treated I colony counts
WP2 uvrA 33f.l9 38.7 5.1 1 1**33,40,4~** 100 f.l9 45.3 12.5 1.3 54, 51, 31 3331Jg 41.0 8.7 1.2 51,36,36 1000 ll9 43.7 38 1.2 42,41,48 2500 il9 42.0 2.6 1.2 45 P, 40 P, 41 P 50001-19 410 7.5 1.2 34 P. 40 P, 49 P THF 35.0 6.2 37,28,40 Untreated Control 58.0 13.0 50, 73, 51**-----·__.~.------** TA 1535 NaN3 101-19 1918.7 67.2 127.9 1996, 1886, 1874 TA 1537* 4-NOPD 501Jg 145.3 16.2 156 137, 164, 135

TA96 4-NOPD 10 ll9 740.7 673 19.2 709,695,818 TA 100 NaN3 101Jg 1844.7 47.6 13.2 1802. 1836, 1896 WP2 uvrA MMS 301JL 344.0 13.5 9.8 355,329,348 Key to Positrve Co•1lrols Key to Plate Postfix Codes**--------~··----·-** NaN3 sodrum azide p Prectprtate 4-NOPD 4-n1tro-o-phenylene-diam1ne MMS methyl methane sulfonate

s4c2dr.doc

# [ RCC-CCR Study Number 

### 1111700

# J Final Report Page 32 of 33

Study Name 1111700 Study Code: RCC-CCR 1111700 Experiment 1111700 HV2 Pre Date Plated 26/07/2007 I 0210812007* Assay Conditions Date Counted 01/08/2007 I 0710812007* With metabolic activation**------strair\~---** compound Dose level Mean Standard Ratio Individual revertant per plate revertants Deviation treated I colony counts
TA 1535 33119 19.0 4.4 1.1 21,22,14 100119 18.3 2.1 1.0 19,16,20 333 pg 19.7 06 1.1 19,20, 20 1000119 16.3 3.1 0.9 19. 17, 13 2500119 17.7 29 1.0 21 P, 16 P, 16 P 5000 IJ9 17 7 2.1 1.0 17P,16P.20P THF 17.7 1.2 17, 19, 17 Untreated Control 203 7.4 23 TA 1537* 33119 18.0 4.4 1.0 23, 15. 16 100119 15.3 1.2 0.8 16, 14, 16 333119 16.0 1.7 0.9 17, 14, 17 10001-19 15.3 1.5 0.8 15, 17 U M. 14 2500119 12.3 3.5 0.7 16 PM, 12 PM, 9 PM 5000119 9.3 0.6 0.5 10 P M, 9 P M. 9 P M THF 18.3 3.8 21, 20, 14 Untreated Control 12.0 2.0 12, 10. 14



## ------ TA98 331Jg 43.7 9.8 0.9 38,38, 55 100 IJ9 42.0 11 4 0.9 29,47, 50 3331Jg 42.3 5.7 09 36,44, 47 1000 IJ9 34.0 2.6 0.7 31, 35, 36 2500 IJ9 44.7 4.5 1.0 49 P, 45 P, 40 P 5000119 41.3 2.3 09 44 P, 40 P, 40 P THF 46.7 8.1 42,42, 56 Untreated Control 48.7 7.6 40,54 TA 100 [ 331-19 150.3 19.7 1.1 158, 128. 165 1001-19 136.3 16.3 1.0 122, 133, 154 333 IJ9 144.7 12.5 1.0 132, 157. 145 10001-19 1433 35 1.0 143, 147. 140 2500 IJ9 117.0 14.4 0.8 121 PM, 101 PM, 129 PM 5000 f-19 109.0 14.2 0.8 125 PM, 98 P M, 104 PM THF 142.0 7.5 141, 135, 150 Untreated Control 159.3 10.5 170 Key to Plate Postfix Codes M Manual count u Air bubbles

s4c2dr.doc[ RCC-CCR Study Number

Study Name 1111700 Expenment 1111700 HV2 Pre Assay Cond1\lons·

THF 

# J

Untreated Control TA 1535 2-AA TA 1537* 2-AA

TA98 2-AA TA 100 2-AA WP2 uvrA 2·AA Key to Positive Controls 2-AA 2-aminoanthracene

s4c2dr.doc 

### 1111700

# J Final Report

With metabolic activation Page 33 of33

Study Code: RCC-CCR 1111700 Date Plated 26/07/2007 I 02/08/2007* Date Counted: 01/08/2007 I 0710812007* Dose level Mean Standard Ratio Individual revertant per plate revertants Deviation treated I colony counts

331J9 63.3 11.5 1.0 70, 70, 50 100 IJ9 59.3 3.2 10 63,57,58 3331Jg 55.0 1.0 0.9 55,54,56 1000 IJ9 61.0 3.5 1.0 65,59, 59 2500 IJ9 53.0 5.3 0.9 49 PM, 51 PM, 59 PM 5000 IJ9 53.7 5.5 0.9 54 PM, 48 PM, 59 PM 61.7 9.5 62, 71, 52 61.7 9.3 54 2.51J9 2327 2.5 13.2 235, 230, 233 251)9 159.0 8.2 8.7 152, 157, 168 2.51Jg 1277.3 85.3 27.4 1291, 1355, 1186 2.51Jg 19393 144.0 13.7 2055, 1778, 1985 10.0pg 415.3 22 1 6.7 418,392,436 Key to Plate Postfix Codes p Precipitate M Manual count