February 13, 2009 

TSCA Document Control Officer (7407M) 

Office of Pollution Prevention and Toxics (New Chemicals Program) 

U.S. Environmental Protection Agency 

EPA East Building, Room 4133 

1201 Constitution Ave. 

Washington, DC 20004-3302 

Subject: Premanufacture Notification Submission for TS-M034A0 

Dear Sir/Madam: 

-iP. 1 

Enclosed please find a new copy of sanitized pages 14-247 of the Premanufacture Notice TSM034A0 . The new copy of these pages is being sent at the request of Bernice Mudd with EPA 

and better hide the confidential information . 

Sincerely, 
p. 2 

9 0 C 

U.S . ENVIRONMENTAL PROTECTION AGENCY 

r i,

nter Me total number of-pAges 

n the Premanufacture Notice 

Form Approved . O.M.B . No. 2070-0012 . Approval Expires 10-31-96 . 

t P as _ r 

a ' 

GENERAL INSTRUCTIONS TS - 

. 

You must provide all information requested in this form to the extent that it is known to or reasonably ascertaiitable by you. Make reasonable estimates if you do 

not have actual data. 

Before you complete this form, you should read the "Instructions Manual for Premanufacture Notification" (the Instructions Manual is available from the Toxic 

Substances Control Act (TSCA) Information Service by calling 202-554-1404, or faxing 202-554-5603). 

Pittsburgh, PA 15251-6399, Attn. TSCA User fee. 

If a user fee has been remitted for this notice (40 CFR 700.45), indicate in the boxes above the TS-user fee identification number you have generated. Remember, you 

user fee ID number must also appear on your corresponding fee remittance, which is sent to EPA, HQ Accounting Operations Branch (PM-226), P.O . 360399M, 

Part I-GENERAL INFORMATION 

You must provide the currently correct Chemical Abstracts (CA) Name of the new 

chemical substance, even if you claim the identity as confidential. You may authorize 

another person to submit chemical identity information for you, but your 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 . 

You must submit an original and two copies of this notice including all test data. If you 

claimed any information as confidential, a single sanitized copy must also be submitted. 

Part II-HUMAN EXPOSURE AND ENVIRONMENTAL RELEASE 

If there are several manufacture, processing, or use operations to be described in Part II, 

sections A and B of this notice, reproduce the sections as needed. 

Part III-LIST OF ATTACHMENTS 

Attach additional sheets if there is not enough space to answer a question fully . Label each 

continuation sheet with the corresponding section heading. In Part III, 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 of this form, space has been provided for you to described 

pollution prevention and recycling information you may have regarding the new substance. 

So-called "binding" boxes are included throughout this form for you to indicate your 

willingness to be bound to certain statements you make in this section, such as use, 

production volume, protective equipment . . This option is intended to reduce delays that 

routinely accompany the development of consent orders or Significant New Use Rules. 

Except in the case of exemption applications (such as TMEA, LVE, LOREX) where 

certain information provided in such notification is binding on the submitter when the 

Agency approves the exemption application, checking a binding box in this notice does not 

by itself prohibit the submitter from later deviating from the information (except chemical 

identity) reported in the form. 

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 of the notice. (including attachments) . For additional instructions on 

claiming information as confidential, read the Instructions Manual . 

® Mark (x) if any information in this notice is claimed as confidential. 

EPA FORM 7710-25 (Rev. 5-95) 

TEST DATA AND 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, if these data are related to the health and environmental effects on the 

manufacture, processing, distribution in commerce, use, or disposal of the 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 or on an analog . Also, the 

chemical composition of the tested material should be characterized . Following 

are examples of test data and other data. Data should be submitted according to 

the requirements of §720 .50 of the Premanufacture Notification Rule (40 CFR 

Part 720). 

Test Data (Check Below any included in this notice) 

" Environmental fate data El Yes " Other data ® Yes 

" Health effects data ® Yes Risk assessments 

" Environmental effects data 0 Yes 

" Physical/Chemical Properties* ® Yes 

M 0 L3] 

* A physical and chemical properties worksheet is located on the last page of this form . TYPE OF NOTICE (Check Only One) 

® PMN (Premanufacture Notice) 

INTERMEDIATE PMN (submitted in sequence with final product PUN) 

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 

on 

IS THIS A CONSOLIDATED PMN? 1:1 Yes 

# of chemicals _ 

(Prenotice Communication 

9 

Structure/activity relationships 

Test data not in the possession 

or control of the submitter 

LOREX Modification 

age 3) 

Replaces previous editions of EPA Form 77T0-25 . 
p. 3 

Public reporting burden for this collection of information is estimated to average 110 hours per response, including time for reviewing 

instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing the collection of 

information. Send comments regarding the burden estimate or any other aspect of this collection of information, including suggestions for 

reducing this burden, to Chief, lnformation Policy Branch, PM-223, U.S . Environmental Protection Agency, 401 M. St., S.W., Washington, D.C . 

20460; and to the Office of Management and Budget, Paperwork Reduction Act (2070-0012), Washington, D.C . 20503. 

CERTIFICATION 

I certify that to the best of my knowledge and belief: 

1. The company named in Part I, section A, subsection 1 a of this notice form intends to manufacture or import 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 of the date of submission . 

3. 1 am submitting with this notice all test data in my possession or control and a description of all other data known to or 

reasonably ascertainable by me as required by §720.50 of the Premanufacture Notification Rule. 

Additional Certification Statements: 

If you are submitting a PMN, Intermediate PMN, Consolidated PMN, or SNUN, check the following user fee certification 

statement that applies : 

® The Company named in Part I, Section A has remitted the fee of $2500 specified in 40 CFR 700.45(b), or 

0 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 

0 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) . 

If you 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 : 

0 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 of 40 CFR 723 .50. 

F-1 The manufacturer is familiar with the terms of this section and will comply with those terms; and 

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

F~ If this 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 1 year of the date of the expiration of the 30 day 

review period. 

The accuracy of the statements you make in this notice should reflect your best prediction of the anticipated facts regarding the chemical substance 

described herein . An knowin g and willful misinterp retation is subject to criminal penal ty pursuant to 18 USC 1001. Confidential 

Signature and title of Authorized Official (Original Signature Required) Date 

X 

Signature of agent -(if applicable) Date 

EPA FORM 7710-25 (Rev . 5-95) Page 2 
Part I -- GENERAL INFORMATION 

Section A -- SUBMITTER IDENTIFICATION 

la. Person Name of authorized official Position 

Submitting 

X Notice (in U.S.) 

Company 

Mailing address (number and street) 

City, State, ZIP Code 

b. Agent (if Name of authorized official Position 

applicable) 

Company 

Mailing address (number and street) 

City, State, ZIP Code Telephone ~ 

' 

Area Code ~ Number 

c . If you are submitting this notice as part of a joint submission, mark (X) this box. ' 

0 

Joint Submitter (if Name of authorized official Position 

applicable) 

Company 

Mailing address (number and street) 

City, State, ZIP Code Telephone ~ Area Code ~ Number 

2 . Technical Name of authorized official Position 

Contact (in 

X U.S .) 

Company 

Mailing address (number and street) 

City, State, ZIP Code Telephone ~ Area Code ~ Number 

3 . If you have had a prenotice communication (PC) concerning this notice and 

EPA assigned a PC Number to the notice, enter the number. 

10 Mark (X) -01 ® 

if none 

4 . If you previously submitted an exemption application for the chemical 

substance covered by this notice, enter the exemption number assigned by Mark (X) ~-10 ® 

EPA. If you previously submitted a PMN for this substance enter the PMN if none 

5 . 

number 

If you have 

assigned 

submitted 

by EPA 

a notice 

(i.e . withdrawn 

of Bona fide 

or 

intent 

incomplete)

to 

. - 

manufacture or import 

for the chemical substance covered by this notice, enter the notice number Mark (X) --01 ® assigned by EPA. ----~ if none 

6. Type of Notice - Mark (X) 

Mark () the "Confidential" box next to any subsection you claim as confidential 

® 

0 

Import 

Only 

Binding Option 

Mark (X) I Mark (X) 

1 . U Manufacture ~ 2. 

Only 

Binding Option 

~ 3 . F-1 Both 

Confidential 

EPA FORM 7710-25 (Rev. 5-95) Replaces previous editions of EPA Form 7710-25. 
Part I -- GENERAL INFORMATION -- Continued 

Section B -- CHEMICAL IDENTITY INFORMATION: You must provide a currently correct Chemical Abstracts (CA) name of the substance based on 

the ninth Collective Index (9C of CA nomenclature rules and conventions . 

Mark (X the "Confidential" box next to any item you claim as confidential 

Complete either item 1 (Class 1 or 2 substances) or 2 (Polymers) as appropriate . Complete all other items . 

If another person will submit chemical identity information for you (for either Item 1 or 2), mark (X) the box at the right. ConfiIdenti the name, company, and address of that person in a continuation sheet . 10 El dential 

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

a. Class of substance - Mark (X) 1 [ :] Class 1 or 2 [:] Class 2 X 

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

For Class 1 substances a CA Index Name must be provided . For Class 2 substances either a CA Index Name or CA Preferred Name must X 

be provided, which ever is appropriate based on CA 9CI nomenclature rules and conventions). 

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 of the Identification ® Method 2 (Other Source) 

]I 

report obtained from the CAS Inventory Expert Services must be 

submitted as an attachment to this notice) 

d. Molecular formula and CAS Registry Number (if a number already exists for the substance) 

AS# 

F X 

e . For a class 1 substance, provide a complete and correct chemical structure diagram . For a class 2 substance - (1) List the immediate 

precursor substances with their respective CAS Registry Numbers . (2) Describe the nature of the reaction or process . (3) Indicate the X 

range of composition and the typical composition (where appropriate) . (4) Provide a correct representative or partial chemical structure 

diagram, as complete as can be known, if one can be reasonably ascertained . 

® Mark (X) this box if you attach a continuation s heet. Attachment 2. 

EPA FORM 7710-25 (Rev . 5-95) Page 4 
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 of the lowest molecular weight composition of the polymer you intend to manufacture . 

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 of that composition . 

Describe the methods of measurement or the basis for your estimates : GPC ~ Other []: (Specify) 

i) lowest number average molecular weight : 

ii) maximum weight % below 500 molecular weight: 

iii) maximum weight % below 1000 molecular weight: 

Mark (X) this box if you attach a continuation sheet. 

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) of each monomer or other reactant used in the manufacture of 

the polymer . 

(2) - Mark (X) this column if entry in column (1) is confidential. 

(3) - Indicate the typical weight percent of each monomer or other reactant in the polymer. 

(4) - Mark (X) the identity column if you 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 if entries in columns (3) and (4) are confidential . 

(6) - Indicate the maximum weight percent of each monomer or other reactant that may be present as a residual in the polymer as manufactured for 

commercial purposes . 

(7) - Mark this column if entry in column (6) is confidential. 

Monomer or other reactant and CAS Registry Number Confr- Typical Identity Confi- Maximum Confidential composition Mark (X) dential residual dential 

(1) (2) (3) (4) (5) (6) 7 

% % 

I ~ Mark (X) this box if you attach a continuation sheet. 

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 of the identification report F Method 2 (other source) 

obtained from CAS Inventory Expert Service must be submitted as 

as attachment to this notice) 

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

e. Provide a correct representative or partial chemical structure diagram, as complete as can be known, if one can be reasonably ascertained . 

THIS PAGE IS LEFT BLANK INTENTIONALLY. 

E]Mark (X) this box if you attach a continuation sheet . 

EPA FORM 7710-25 (Rev . 5-95) Page 5 
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 

the CAS Registry Number if available . If there are unidentified impurities, enter "unidentified ." 

(b) - Estimate the maximum weight % of each impurity . If there are unidentified impurities, estimate their total weight %. 

. Provide 

Impurity and CAS Registry Number 

(a) 

Maximum 

percent 

(b) 

Confidential 

% 

% 

% 

% 

% 

% 

% 

Mark (X) this box if you attach a continuation sheet. 

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

Mark (X) this box if you attach a continuation sheet. 

X 

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

Mark (X) this box if you attach a continuation sheet. 

- 

X 

6. Generic chemical name - If you claim chemical identify as confidential, you must provide a generic name for your substance that reveals 

the specific chemical identity of the 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. 

Carbon nanomaterial 

Mark (X) this box if you attach a continuation sheet . 

7 . Byproducts - Describe any byproducts resulting from the manufacture, processing, use, or disposal of the new chemical substance . Provide the CAS Registry 

Number if available. 

Byproduct 

(1) 

CAS Registry Number 

(2) 

Confidential 

None known 

Mark (X) this box if you attach a continuation sheet . 

EPA FORM 7710-25 (Rev. 5-95) Page 6 
Part I -- GENERAL INFORMATION -- Continued 

Section C -- PRODUCTION, IMPORT, AND USE INFORMATION: 

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, if you 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. If granted, you are bound to this volume 

Maximum first 12-month production (kg/yr) Maximum 12-month production (kg/yr) Confi- Binding 

(100% new chemical substance basis) (100% new chemical substance basis) dential Option 

Mark (x) 

X 

2. Use Information -- You must make separate confidentiality claims for the description of the category of use, the percent of production volume 

devoted to each category, the formulation of the new substance, and other use information . Mark (X) the "Confidential" Box next to any item 

you claim as confidential . 

a. (1) -- Describe each intended category of use of the 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 of total production for the first three years devoted to each category of use. 

(5) -- Mark (X) this column if entry in column (4) is confidential business information (CBI). 

(6) -- Estimate the percent of the new substance as formulated in mixtures, suspensions, emulsions, solutions, or gels as manufactured for 

commercial purposes at sites under your control associated with each category of use. 

(7) -- Mark (X) this column if entry in column (6) is confidential business information (CBI). 

(8) -- Indicate % of product volume expected for the listed "use" sectors . Mark more than one box if appropriate. Mark (X) to indicate 

your willingness to have the use type provided in (8) binding . 

(9) -- Mark (X) this column if entry(ies) in column (8) is (are) confidential business information (CBI). 

Category of use (1) CBI Binding Produc- CBI % in CBI % of substance expected per use CBI 

Option 

tion % Form- (8) 

(by function and application i.e . a dispersive dye Mark (X) ulation Site- Con-* Indus- Com- Binding 

for finishing polyester fibers (2) (3) (4) (5) (6) (7) limited sumer trial mercial Option (9) 

X % X % X X 

X % X % X X 

X % X % X X 

X % X % X X 

o, o, 

% 

* If you have identified a "consumer" use, please provide on a continuation sheet a detailed description of the use(s) of this chemical substance in consumer products . 

In addition include estimates of the concentration of the 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 if you attach a continuation sheet. 

b. Generic If you claim any category of use description in subsection 2a as confidential, enter a generic description of that category . Read the 

use Instructions Manual for examples of generic use descriptions . 

description Component of paints, coatings, and industrial composites 

~ Mark (X) this box if you attach a continuation sheet. 

3. Hazard Information -- Include in the notice a copy of reasonable facsimile of any hazard warning statement, label, material safety data sheet, or other Binding 

information which will be provided p to any y person p who is reasonably y likely y to be exposed to this substance re g ardin g protective equipment or practices Option 

Muk Ixl for the safe handing, transport, use, or disposal of the new substance . List in part III hazard information you include . 

® Mark (X) this box if you attach hazard information. Attachment 3 . 

EPA FORM 7710-25 (Rev . 5-95) Page 7 
Part II-- HUMAN EXPOSURE AND ENVIRONMENTAL RELEASE 

Section A -- INDUSTRIAL SITES CONTROLLED BY THE SUBMITTER Mark (X) the "Confidential" box next to any item you 

claim as confidential 

Complete section A for each type of manufacture, 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 if 

there are further industrial processing or use operations after import. You must describe these operations . See instructions manual 

1 . Operation description Confia. Identity -- Enter the identi ty of the site at which the operation will occur. dential 

Name 

Site address (number and street) 

City, County, State, ZIP code 

If the same operation will occur at more than one site, enter the number of sites. Identify the 

additional sites on a continuation sheet, and if any of the sites have significantly different 

production rates or operations, include all the information requested in this section for those ; 

sites as attachments . ' 

Mark (X) this box if you attach a continuation sheet. 

b. Type -- 

Mark (X) ~ Manufacturing ~ Processing ~ Use 

c. Amount and Duration -- Complete I or 2 as appropriate 

Maximum kg/batch (i ooi new chemicat ~ Hours/batch ~ Batches/year 

substance) i i 

1 . Batch ' 

Maximum kg/batch (tooi new chemical , Hours/batch , Batches/year 

substance) ~ 

i 

2 . Continuous 

d . Process description 1:1 Mark (X) to indicate your willingness to have your process description binding . 

(1) Diagram the major unit operation steps and chemical conversions . Include interim storage and transport containers (specify- e .g . 5 gallon 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 of all starting materials and 

feedstocks (including reactants, solvents, catalysts, etc.), and of all products, recycle streams, and wastes . Include cleaning chemicals (note frequency if not 

used daily or per batch .) . 

(3) Identify by number the points of release, including small or intermittent releases, to the environment of the new chemical substance. 

® Mark (X) this box if you attach a continuation sheet. Attachment 4. 

EPA FORM 7710-25 (Rev. 5-95) Page 8 
p. 10 

Part II-- HUMAN EXPOSURE AND ENVIRONMENTAL RELEASE -- Continued 

Section A -- INDUSTRIAL SITES CONTROLLED BY THE SUBMITTER - Continued 

2. Occupational Exposure -- You must make separate confidentiality claims for the description of worker activity, physical form of the new chemical substance, 

number of works exposed, and duration of activity. Mark (X) the "Confidential" box next to 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 if entry in column (1) is confidential business information (CB1) . 

(3) --Describe any protective equipment and engineering controls used to protect workers . 

(4) and (6) -- Indicate your willingness to have the information provided in column (3) or (5) binding . 

(5) -- Indicate the physical form(s) of the new chemical substance (e .g ., solid : crystal, granule, powder, or dust) and % new chemical substance (if part of a 

mixture) at the time of exposure . 

(7) --Mark (X) this column if entry in column (5) is confidential business information (CBI) . 

(8) --Estimate the maximum number of workers involved in each activity for all sites combined. 

(9) --Mark (X) this column if entry in column (8) is confidential business information (CBI) . 

(10) and (11) -- Estimate the maximum duration of the activity for any worker in hours per day and days per year. 

(12) -- Mark (X) this column if entries in columns (10) and (11) are confidential business information (CBl) . 

Worker activity CBl Protective Equipment/ Binding Physical forms(s) Binding CBI # of CBl Maximum duration ~ CBI 

(i .e ., bag dumping, filling drums) 

(1) (z) 

Engineering Controls 

(3) 

Option 

Mark (x) 

(4) 

and % new 

substance 

(5) 

Option 

Mark (x) 

(6) (7) 

Workers 

Expose 

d (9) 

Hrs/day 

(10) 

Days/yr 

(11) (12) 

(8) 

THIS PAGE IS 

LEFT BLANK 

INTENTIONALLY. 

® Mark (X) this box if you attach a continuation sheet. See attachment7 . 

3. Environmental Release and Disposal -- You must make separate confidentiality claims for the release number and the amount of the 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 l d(3) . 

(2) -- Estimate the amount of the new substance released (a) directly to the environment or (b) into control technology (in kg/day or kg/batch) . 

(3) --Mark (X) this column if entries 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, POTW, or other (specify)) to 

which the new substance will be released from that release point. 

(5) -- a . Describe control technology, if any, and control efficiency that will be used to limit the release of the 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 whether the waste is subject to 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 if entries 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 of the POTW (Publicly Owned Treatment Works) . Mark if the POTW name or NPDES # is confidential business information (CBI) . 

Release Amount of new substance CBI Media of Control technology and efficiency (you may wish to optionally attach efficiency data) CBI 

Number released release 

(1) (2a) (2b) 

(3 

e.g. stack air 

(4) 

~ Binding 

(5a) 1 Mark (x) (Sb) (6) 

. ' 

(7) Mark (X) the 

0 POTW provide name(s) below: 

CBI ~ Navigable FlOther - Specify provide NPDES # CBI 

destination(s) of waterway 

releases to water. 

10 Mark (X) this box if you attach a continuation sheet. 

EPA FORM 7710-25 (Rev . 5-95) Page 9 
p. 11 

Part II-- HUMAN EXPOSURE AND ENVIRONMENTAL RELEASE -- Continued 

Section B -- INDUSTRIAL SITES CONTROLLED BY OTHERS 

Complete section B for typical 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 B for each type ofprocessing, or use operation involving the new chemical substance. If the same operation is performed at more than one site describe the 

typ ical operation common to these sites . Identify additional sites on a continuation sheet. 

1 . Operation Description -- To claim information in this section as confidential, circle or bracket 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. 5 gallon pails, 55 gallon 

drums, rail cars, tank trucks, etc) . On the diagram, identify by letter and briefly describe each worker activity. (2) -- 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 if not used daily or per batch) . (3) -- Identify by number the points of release, 

including small or intermittent releases, to the environment of the new chemical substance . (4) Please enter the # of sites (remember to identify the locations of 

these sites on a continuation sheet) : 

# of sites 

® Mark (X) this box if you attach a continuation sheet. Attachments 6 and 7 

2. Worker Exposure/Environmental Release 

(1) --From the diagram above, provide the letter for each worker activity . Complete 2-8 for each worker activity described . 

(2) --Estimate the number of workers exposed for all sites combined . 

(4) -- Estimate the typical duration of exposure per worker in (a) hours per day and (b) days per year. 

(6) --Describe physical form of exposure and % new chemical substance (if in mixture), and any protective equipment and engineering controls, if any, used to 

protect workers . 

(7) -- Estimate the percent of the new substance as formulated when packaged or used as a final product. 

(9) -- From the process diagram above, enter the number of each release point . Complete 9-13 for each release point identified . 

(10) -- Estimate the amount of the new substance 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, if any, that will be used to limit the release of the 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 of the proceeding entries are confidential business information (CB l) . 

Letter # of CBI Duration CBI Protective Equip . / % in CBI Release Amount of CBI Media of Release CBI 

of Workers of Engineering Controls/ Form- Number New & Control 

Act- Exposed Exposure Physical Form and % ulation Substance Technology 

ivity 

new substance Released 

(1) (2) (3) (4a) i (4b) (5) (6 (7) (8) (9) (l 0a) , (lOb) (I1) (12) (13) 

i 

i 

i 

i 

i 

(14) -- Byproducts : - (15) 

None known 

® Mark (X) this box if you attach a continuation sheet. See attachment 8 . 

EPA FORM 7710-25 (Rev . 5-95) Page 10 
p. 12 

OPTIONAL POLLUTION PREVENTION INFORMATION 

To claim information in this section as confidential circle or bracket the specific information that you 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 

of the PMhT substance. Please include new information pertinent to pollution prevention, including source 

reduction, recycling activities and safer processes or products available due to the new chemical substance. 

Source reduction includes the reduction in the amount or toxicity of chemical wastes by technological 

modification, process and procedure modification, product reformulation, raw materials substitution, and/or 

inventory control. Recycling refers to the reclamation of useful chemical components from wastes that would 

otherwise be treated or released as air emissions or water discharges, or land disposal . Descriptions of pollution 

prevention, source reduction and recycling should emphasize potential risk reduction subsequent to compliance 

with existing regulatory requirements and can be either quantitative or qualitative. 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 environmental media or non-environmental areas (e.g., occupational or consumer exposure). In 

addition, information on the relative cost or performance characteristics of the P1V1N substance to potential 

alternatives may be provided. 

All information provided in this section will be taken into consideration during the review of this 

substance. See Instructions Manual and Pollution Prevention Guidance manual for guidance and 

examples. 

Describe the expected net benefits, such as (1) an overall reduction in risk to human health or the environment; (2) a reduction in the 

volume manufactured; (3) a reduction in the generation of waste materials through recycling, source reduction or other means; (4) a 

reduction in potential toxicity or human exposure and/or environmental release; (5) an increase in product performance, a decrease in 

the cost of production and/or improved operation efficiency of the new chemical substance in comparison to existing chemical 

substances used in similar application; or (6) the extent to which the new chemical substance may be a substitute for an existing 

substance that poses a greater overall risk to human health or the environment. 

0 Mark (X) this box if you attach a continuation sheet. 

EPA FORM 7710-25 (Rev . 5-95) Page 11 
Part III -- LIST OF ATTACHMENTS 

Attach continuation sheets for sections of the form and 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 of the form to 

which it relates, if appropriate . Number consecutively the pages of the attachments. In the column below, enter the inclusive page 

numbers of each attachment . 

Mark (X) the "Confidential" box next to any attachment name you claim as confidential . Read the Instructions Manual for guidance on 

how to claim any information in an attachment as confidential . You must include with the sanitized copy of the notice form a sanitized 

version of an attachment in which you claim information as confidential. 

Attachment name Attachment 

page number(s) 

Confidential 

1 . Material Safety Data Sheets for the PMN Substance 14-43 

2 . Continuation Sheet for Part I, Section B, 1 .e ., Chemical Identity Information 44-46 

3 . Toxicity Review and Hazard Assessment of the PMN Substance 47-53 

X 

X 

X 

X 

X 

X 

X 

X 

X 

X 

X 

X 

~ Mark (X) this box if you attach a continuation sheet. Enter the attachment name and number. 

EPA FORM 7710-25 (Rev. 5-95) Page 12 
p. 14 

PHYSICAL AND CHEMICAL PROPERTIES WORKSHEET 

To assist EPA's review of physical and chemical properties data, please complete the following worksheet for data you provide and 

include it in the notice . Identify the property measured, the page of the notice on which the property appears, the value of the 

property, the units in which the property is measured (as necessary), and whether or not the property is claimed as confidential . The 

physical state of the 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 review and ensure that 

confidential information is properly protected . You should submit this worksheet as a supplement to your submission of test data. 

This worksheet is not a substitute for submission of test data . 

Property Mark (x) if Page Value Measured o coniiprovided number Estimate dential 

Mark N 

(a) (b) (e) (M or E) (d) 

X M 

Physical state of neat substance X_ (s) - (1) - (g) 

Vapor pressure 

Temperature °C Torr 

Density/relative density g/cm3 

Solubility Insoluble in water. Not soluble in E 

@ Temperature oC solvents . The PMN substance does form 

stable suspensions in dimethylformamide 

Solvent (after high power sonication) but in very 

limited quantity (about 1-10 mg/L). 

Solubility in water @ Temperature °C g/L 

Meltin g temperature oc 

Boiling / sublimation temperature@ torr pressure oC 

Spectra 

Specific Gravity 2.0 /cm3 E 

Particle size distribution X 99.15% particles are > 50 ~tm. M X 

99.15% are < 1 mm. No particles 

below 3.6 pm. 0.10% are < 10 jim. 

0 .13% are < 12 pm. 0.17% are < 15 

~m. 

X 1 .200 Gp E 

Young's Modulus 

X 100-250 m2/g M X 

Surface area 

X 5-15 M X 

Mean Number of Walls 

X - 15 M X 

Outer Mean Diameter 

X - 15 M X 

Outer Diameter Distribution 

X - 10 M X 

Inner Mean Diameter 

X - 10 M X 

Inner Diameter Distribution 

X 0 .1-0 .4 g/cm3 M X 

Bulk Density 

Agglomeration/Aggregation State X Agglomerate rope-like structures M X 

typically 300-400 m in diameter. 

Tensile Strength X 150 GP E 

Molecular Weight X statistical analysis of a sample of 50 M X 

kema carbon nanotubes indicate that 

90% diameters span from 7 to 15 Dm 

and that 90% of number of walls spans 

from 5 to 11 walls. Therefore for this 

sample, molecular weights span from 

3 .94x106 g/mol to 1 .82 x 109 /mol. 

ATTACHMENT 

PAGE 13 
p. 15 

Material Safety Data Sheet 

11 . PRODUCT AND COMPANY IDENTIFICATION 

_.~;~_ 

Customer Service Telephone Number: ~ - _ 

Emergency Info rmation 

Transportation : CHEMTREC: (800) 424-9300 

(24 hrs ., 7 days a week) 

Medical: Rocky Mountain Poison Center: (303) 623-5716 

(24 hrs., 7 days a week) 

Product Information 

Product name: 

Synonyms: 

Molecular formula: 

Chemical family : 

Product use: 

2. HAZARDS IDENTIFICATION 

Emergency Overview 

Color: black 

Physical state: solid 

Form: powder 

Odor: none 

coa~ings, composities, and thermoplastic polymers. 

WARNINGI 

MAY FORM COMBUSTIBLE DUST AIR MIXTURES. 

MAY CAUSE EYE, SKIN AND RESPIRATORY TRACT IRRITATION . 

Potential Health Effects 

Primary routes of exposure : 

Inhalation and skin contact. 

Signs and symptoms of acute exposure : 

Dust: May cause eye irritation . May cause irritation of respiratory tract. Prolonged or repeated contact may dry skin and cause irritation . 
p. 16 

r3. COMPOSITION/INFORMATION ON INGREDIENTS 

Chemical Name CAS-No. Wt/Wt OSHA 

Hazardous 

The substance(s) marked with a "Y" in the Hazard column above, are those identified as hazardous chemicals under 

the criteria of the OSHA Hazard Communication Standard (29 CFR 1910.1200) . 

This material is classified as hazardous under Federal OSHA regulation . 

4. FI RST AID MEASURES 

Inhalation : 

If inhaled, remove victim to fresh air. If not breathing, give artificial respiration . If breathing is difficult, give oxygen. 

Get medical attention. 

Skin: 

In case of contact, immediately flush skin with plenty of water. Remove contaminated clothing and shoes. Get 

medical attention if irritation persists. Wash clothing before reuse. Thoroughly clean shoes before reuse. 

Eyes: 

Immediately flush eye(s) with plenty of water. Get medical attention if irritation persists . 

Ingestion: 

If swallowed, DO NOT induce vomiting . Get medical attention. Never give anything by mouth to an unconscious 

person . 

5. FIRE-FIGHTING MEASURES 

Flash point: Not determined 

Auto-ignition temperature : > 752 °F (> 400 °C) (Method : Standard NF EN 50281-2-1) 

Lower flammable limit (LFL): Not determined 

Upper flammable limit (UFL): Not determined 

Extinguishing media (suitable) : 

Water spray, Carbon dioxide (C02), Dry powder 

Protective equipment: 

Fire fighters and others who may be exposed to products of combustion should wear full fire fighting turn out gear 

(full Bunker Gear) and self-contained breathing apparatus (pressure demand / NIOSH approved or equivalent). 

Fire fighting equipment should be thoroughly decontaminated after use. 
p. 17 

Further fireflghting advice: 

Do not use a solid stream of water. 

A solid stream of water can cause a dust explosion. 

Fire and explosion hazards: 

Dust clouds generated during handling and/or storage can form explosive mixtures with air. Dust explosion 

characteristics vary with the particle size, particle shape, moisture content, contaminants, and other variables. 

Note : Check that all equipment is properly grounded and installed to satisfy electrical classification requirements. As 

with any dry material, pouring this material or allowing it to free-fall or to be conveyed through chutes or pipes can 

accumulate and generate electrostatic sparks, potentially causing ignition of the material itself, or of any flammable 

materials which may come into contact with the material or its container. 

6. ACCIDENTAL RELEASE MEASURES 

In case of spill or leak: 

Sweep up and shovel into suitable containers for disposal . Use only non-sparking tools. Wet down dust with water 

spray jet. Spills should be contained and placed in suitable containers for disposal . Consult a regulatory specialist to 

determine appropriate state or local reporting requirements, for assistance in waste characterization and/or 

hazardous waste disposal and other requirements listed in pertinent environmental permits. 

7. HANDLING AND STORAGE 

Handling 

General information on handling: 

Keep away from heat, sparks and flames. 

Avoid contact with the skin, eyes and clothing . 

Avoid breathing dust . 

Keep container closed . 

Avoid creating dust in handling, transfer or clean up. 

Prevent dust accumulation . 

Check that all equipment is properly grounded and installed to satisfy electrical classification requirements . 

Use only with adequate ventilation . 

Wash thoroughly after handling . 

Container hazardous when empty. 

Emptied container retains product residue. 

Follow label warnings even after container is emptied. 

RESIDUAL DUSTS MAY EXPLODE ON IGNITION . 

DO NOT CUT, DRILL, GRIND, OR WELD ON OR NEAR THIS CONTAINER. 

Improper disposal or reuse of this container may be dangerous and/or illegal . 

Storage 

General information on storage conditions : 

Store in cool, dry, well ventilated area away from sources of ignition such as flame, sparks and static electricity . 

Ensure that all storage and handling equipment is properly grounded and installed to satisfy electrical classification 

requirements. Static electricity may accumulate when transferring material . All storage containers, including drums, 

cylinders and IBCs, must be bonded and grounded during filling and emptying operations . 

Version 1.0 Issued on: 0712512008 Page: 3 1 10 
p. 18 

Material Safety Data Sheet 

Storage incompatibility-General: 

Store separate from oxidizers. 

Storage incompatibility-Segregation (specific) : 

Strong oxidizing agents 

8. EXPOSURE CONTROLS/PERSONAL PROTECTION 

Airborne Exposure Guidelines : 
p. 19 

Material Safety Data Sheet 

Only those components with exposure limits are printed in this section . Limits with skin contact designation above have skin contact effect. Air sampling alone is insufficient to accurately quantitate exposure . Measures to prevent significant cutaneous absorption 

may be required. Limits with a sensitizer designation above mean that exposure to this material may cause allergic reactions . 

Engineering controls : 

Investigate engineering techniques to reduce exposures below airborne exposure limits . Provide ventilation if 

necessary to control exposure levels below airborne exposure limits (see above) . If practical, use local 

mechanical exhaust ventilation at sources of air contamination such as open process equipment. Consult ACGIH ventilation manual or NFPA Standard 91 for design of exhaust systems. 

Respiratory protection : 

Avoid breathing dust. Where airborne exposure is likely or airborne exposure limits are exceeded (if applicable, 

see above), use NIOSH approved respiratory protection equipment appropriate to the material and/or its 

components (full facepiece recommended) . Consult respirator manufacturer to determine appropriate type 

equipment for a given application. Observe respirator use limitations specified by NIOSH or the manufacturer . 

For emergency and other conditions where there may be a potential for significant exposure or where exposure limit may be significantly exceeded, use an approved full face positive-pressure, self-contained breathing apparatus or positive-pressure airline with auxiliary self-contained air supply . Respiratory protection programs 

must comply with 29 CFR § 1910 .134 . 

Skin protection : 

Wear appropriate chemical resistant protective clothing and chemical resistant gloves to prevent skin contact. 

Consult glove manufacturer to determine appropriate type glove material for given application. Rinse 

immediately if skin is contaminated . Wash contaminated clothing and clean protective equipment before reuse. Wash thoroughly after handling . 

Eye protection: 

Where eye contact may be likely, wear chemical goggles and have eye flushing equipment available. 

9. PHYSICAL AND CHEMICAL PROPERTIES 

Color: black 

Physical state: solid 

Form ; powder 

Odor: none 

pH: no data available 

Density: no data available 

Specific Gravity (Relative no data available 

density): 

Bulk density: 100 - 400 kg/m3 

Vapor pressure: no data available 

Vapor density: no data available 
p. 20 

Boiling point/boiling no data available 

range: 

Freezing point: no data available 

Melting point/range: no data available 

Solubility in water: insoluble 

10. STABILITY AND REACTIVITY 

Stability: 

This material is chemically stable under normal and anticipated storage, handling and processing conditions . 

Materials to avoid: 

Strong oxidizing agents 

Conditions I hazards to avoid : 

To avoid thermal decomposition, do not overheat . Hazardous decomposition products: 

Thermal decomposition giving toxic products : 

Carbon oxides 

11 . TOXICOLOGICAL INFORMATION 

Acute toxicity 

4 

__ Acute toxicity 

Oral : 

Slightly toxic. (rat) LD50 = 1,600 mg/kg. 

Oral : 

Practically nontoxic . (rat) LD50 > 5,000 mg/kg. 

Skin Irritation : 

Non-irritating . (rabbit) 

Eye Irritation : 

Non-irritating . (rabbit) 

Dermal : 

No more than slightly toxic. (rabbit) LD50 > 1,000 mg/kg. 

Carcinocenicitv 
p. 21 

Material Safety Data Sheet ° 

Chronic inhalation, intratracheal injection administration to rat, mouse, hamster, guinea pig / No 

in tumor incidence was reported . 

Carcinoaenicitv 

Repeated inhalation, intratracheal injection administration to rat / affected organ(s) : lung / signs: fibrosis 

Human experience 

Inhalation : 

Respiratory tract: Epidemiology studies of workers indicate pulmonary function is not affected ; no significant 

increases in cancer. 

Oral : 

No more than slightly toxic. (rat) LD50 > 2,000 mg/kg. 

Dermal: 

No more than slightly toxic. (rat) LD50 > 2,000 mg/kg. 

Skin Irritation : 

Slightly irritating . (rabbit) 

Eye Irritation : 

Moderately irritating. (rabbit) 

Other routes : 

Skin Sensitization : 

Not a skin sensitizer. (mouse) 

Genotoxicity 

Assessment in Vitro: 

No genetic changes were observed in laboratory tests using: bacteria, human cells 

12. ECOLOGICAL INFORMATION 

Chemical Fate and_Pathway 

No data are available. 

Ecotoxicolo9v 

Aquatic toxicity data : 

Version 1 .0 Issued on: 07125/2008 Page: 7 l 10 
p. 22 

Material Safety Data Sheet 

Practically nontoxic. Leuciscus idus (Golden orfe) 48 h LCO = 1,000 mg/I 

Microorganisms: 

Practically nontoxic. Bacteria ECO = 5,000 mg1I 

13. DISPOSAL CONSIDERATIONS 

Waste disposal : 

Where possible recycling is preferred to disposal or incineration . If recycling is not an option, incinerate or dispose of in accordance with federal, state, and local regulations. Consult a regulatory specialist to determine appropriate state or local reporting requirements, for assistance in waste characterization and/or hazardous waste disposal and other requirements listed in pertinent environmental permits. Note: Chemical additions to, processing of, or otherwise altering this material may make this waste management information incomplete, inaccurate, or otherwise inappropriate. Furthermore, state and local waste disposal requirements may be more restrictive or otherwise different from federal laws and regulations. 

14. TRANSPORT INFORMATION 

US Department of Transportation (DOT): not regulated 

International Maritime Dangerous Goods Code (IMDG) : not regulated 

15. REGULATORY INFORMATION 

Chemical Inventorv Status 

EU. EINECS EINECS Conforms to 

US. Toxic Substances Control Act TSCA The components of this product are all on 

the TSCA Inventory. 

Australia. Industrial Chemical (Notification and AICS Conforms to 

Assessment) Act 

Canada. Canadian Environmental Protection Act DSL All components of this product are on the (CEPA) . Domestic Substances List (DSL). (Can. Gaz. Canadian DSL list . Part 11, Vol. 133) 

Japan. Kashin-Hou Law List ENCS (JP) Conforms to 

Korea. Toxic Chemical Control Law (TCCL) List KECI (KR) Conforms to 

Philippines. The Toxic Substances and Hazardous PICCS (PH) Conforms to and Nuclear Waste Control Act 

China. Inventory of Existing Chemical Substances INV (CN) Conforms to 

New Zealand. Inventory of Chemicals (NZIoC), as NZIOC Conforms to published by ERMA New Zealand 

Version 1 .0 Issued on: 07I25I2008 Page: 8110 
p. 23 

United States - Federal Reaulations 

SARA Title III - Section 302 Extremely Hazardous Chemicals: 

SARA 302: No chemicals in this material are subject to the reporting requirements of SARA Title III, Section 

302. 

SARA Title III - Section 311/312 Hazard Categories : 

Acute Health Hazard 

SARA Title III -Section 313 Toxic Chemicals: 

Chemical Name CAS-No. De minimis Reportable threshold : 

Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) - Reportable 

Quantity (RQ): 

The components in this product are either not CERCLA regulated, regulated but present in negligible concentrations, 

or regulated with no assigned reportable quantity. 

OSHA Regulated Carcinogens (NTP, IARC, OSHA Listed): 

NTP: 

No component of this product present at levels greater than or equal to 0.1 % is identified as a known or 

anticipated carcinogen by NTP. 

IARC: 

No component of this product present at levels greater than or equal to 0.1 % is identified as probable, 

possible or confirmed human carcinogen by IARC. 

OSHA: 

No component of this product present at levels greater than or equal to 0.1 % is identified as a carcinogen 

or potential carcinogen by OSHA. 

United States - State Reaulations 

Massachusetts Right to Know 

Chemical Name 

Product code: HES1000 Version 1 .0 Issued on: 0712512008 Page: 9 110 
Material Safety Data Sheet 

New Jersey Right to Know 

Pennsylvania Right to Know 

Pennsylvania Right to Know - Environmentally Hazardous Substance(s) 

ifornia Prop. 65 

This product does not contain any chemicals known to the State of California to cause cancer, birth 

defects, or any other reproductive effects. 

16. OTHER INFORMATION 

Latest Revision(s): 

Reference number: 000000047038 

Date of Revision : 07/25/2008 

Date Printed: 09/18/2008 
p. 25 

Material Safety Data Sheet 

1 . PRODUCT AND COMPANY IDENTIFICATION 

Transportation : CHEMTREC : (800) 424-9300 

(24 hrs., 7 days a week) 

Medical: Rocky Mountain Poison Center : (303) 623-5716 

(24 hrs., 7 days a week) 

Product Information 

Product name: 

Synonyms: FA 

Molecular formula: 

Chemical family: 

Product use: j in coa mgs, composities, and thermoplastic polymers. 

2. HAZARDS IDENTIFICATION 

Emercaency Overview 

Color: black 

Physical state: solid 

Form : powder 

Odor: none 

WARNING! 

MAY FORM COMBUSTIBLE DUST AIR MIXTURES. 

MAY CAUSE EYE, SKIN AND RESPIRATORY TRACT IRRITATION . 

Potentia l Health Effects 

Primary routes of exposure : 

Inhalation and skin contact. 

Signs and symptoms of acute exposure : 

Dust: May cause eye irritation . May cause irritation of respiratory tract . Prolonged or repeated contact may dry skin 

and cause irritation . 
p. 26 

Material Safety Data Sheet 

3. COMPOSITION/INFORMATION ON INGREDIENTS 

Chemical Name CAS-No. wtlWt OSHA 

Hazardous 

The substance(s) marked with a "Y" in the Hazard column above, are those identified as hazardous chemicals under 

the criteria of the OSHA Hazard Communication Standard (29 CFR 1910.1200) . 

This material is classified as hazardous under Federal OSHA regulation . 

4. FIRST AID MEASURES 

Inhalation : 

If inhaled, remove victim to fresh air. If not breathing, give artificial respiration . If breathing is difficult, give oxygen . 

Get medical attention. 

Skin: 

In case of contact, immediately flush skin with plenty of water. Remove contaminated clothing and shoes. Get 

medical attention if irritation persists . Wash clothing before reuse. Thoroughly clean shoes before reuse. 

Eyes: 

Immediately flush eye(s) with plenty of water. Get medical attention if irritation persists . 

Ingestion: 

If swallowed, DO NOT induce vomiting . Get medical attention. Never give anything by mouth to an unconscious 

person . 

5. FIRE-FIGHTING MEASURES 

Flash point: no data available 

Auto-ignition temperature: no data available 

Lower flammable limit (LFL): no data available 

Upper flammable limit (UFL): no data available 

Extinguishing media (suitable) : 

Water spray, Carbon dioxide (C02), Dry powder 

Protective equipment: 

Fire fighters and others who may be exposed to products of combustion should wear full fire fighting turn out gear 

(full Bunker Gear) and self-contained breathing apparatus (pressure demand I NIOSH approved or equivalent). 

Fire fighting equipment should be thoroughly decontaminated after use. 

Further firefighting advice : 

Version 1 .1 Issued on: 0712512008 Page: 21 10 
p. 27 

Material Safety Data Sheet 

A solid stream of water can cause a dust explosion. 

Do not use a solid stream of water. 

Fire and explosion hazards: 

Dust clouds generated during handling and/or storage can form explosive mixtures with air. Dust explosion 

characteristics vary with the particle size, particle shape, moisture content, contaminants, and other variables. 

Note : Check that all equipment is properly grounded and installed to satisfy electrical classification requirements . As 

with any dry material, pouring this material or allowing it to free-fall or to be conveyed through chutes or pipes can 

accumulate and generate electrostatic sparks, potentially causing ignition of the material itself, or of any flammable 

materials which may come into contact with the material or its container. 

6. ACCIDENTAL RELEASE MEASURES 

In case of spill or leak: 

Sweep up and shovel into suitable containers for disposal . Use only non-sparking tools . Wet down dust with water 

spray jet . Spills should be contained and placed in suitable containers for disposal . Consult a regulatory specialist to 

determine appropriate state or local reporting requirements, for assistance in waste characterization and/or 

hazardous waste disposal and other requirements listed in pertinent environmental permits. 

7. HANDLING AND STORAGE 

Handling 

General information on handling : 

Keep away from heat, sparks and flames . 

Avoid contact with the skin, eyes and clothing. 

Avoid breathing dust . 

Keep container closed . 

Avoid creating dust in handling, transfer or clean up. 

Prevent dust accumulation . 

Check that all equipment is properly grounded and installed to satisfy electrical classification requirements. 

Use only with adequate ventilation . Wash thoroughly after handling . 

Container hazardous when empty. 

Emptied container retains product residue. 

Follow label warnings even after container is emptied . 

RESIDUAL DUSTS MAY EXPLODE ON IGNITION . 

DO NOT CUT, DRILL, GRIND, OR WELD ON OR NEAR THIS CONTAINER. 

Improper disposal or reuse of this container may be dangerous and/or illegal . 

Storaae 

General information on storage conditions: 

Store in well ventilated area away from heat and sources of ignition such as flame, sparks and static electricity . 

Ensure that all storage and handling equipment is properly grounded and installed to satisfy electrical classification 

requirements . Static electricity may accumulate when transferring material . All storage containers, including drums, 

cylinders and IBCs, must be bonded and grounded during filling and emptying operations . 

Storage incompatibility - General: 

Store separate from oxidizers. 

Version 1 .1 Issued on: 0712512008 Page: 3110 
p. 28 

Material Safety Data Sheet 

Storage incompatibility-Segregation (specific) : 

Strong oxidizing agents 

8. EXPOSURE CONTROLS/PERSONAL PROTECTION 

Only those components with exposure limits are printed in this section . Limits with skin contact designation above have skin contact 

effect . Air sampling alone is insufficient to accurately quantitate exposure . Measures to prevent significant cutaneous absorption 

may be required . Limits with a sensitizer designation above mean that exposure to this material may cause allergic reactions. 

Engineering controls : 

Investigate engineering techniques to reduce exposures below airborne exposure limits . Provide ventilation if 

necessary to control exposure levels below airborne exposure limits (see above) . If practical, use local 

mechanical exhaust ventilation at sources of air contamination such as open process equipment. Consult 

ACGIH ventilation manual or NFPA Standard 91 for design of exhaust systems. 

Respiratory protection : 

Avoid breathing dust. Where airborne exposure is likely or airborne exposure limits are exceeded (if applicable, 

see above), use NIOSH approved respiratory protection equipment appropriate to the material and/or its 

components (full facepiece recommended) . Consult respirator manufacturer to determine appropriate type 

Version 1 .1 Issued on: 07/25/2008 Page: 4 110 
p. 29 

Material Safety Data Sheet 

equipment for a given application. Observe respirator use limitations specified by NIOSH or the manufacturer. 

For emergency and other conditions where there may be a potential for significant exposure or where exposure 

limit may be significantly exceeded, use an approved full face positive-pressure, self-contained breathing 

apparatus or positive-pressure airline with auxiliary self-contained air supply . Respiratory protection programs 

must comply with 29 CFR § 1910.134 . 

Skin protection : 

Wear appropriate chemical resistant protective clothing and chemical resistant gloves to prevent skin contact. 

Consult glove manufacturer to determine appropriate type glove material for given application. Rinse 

immediately if skin is contaminated . Wash contaminated clothing and clean protective equipment before reuse. 

Wash thoroughly after handling . 

Eye protection : 

Where eye contact may be likely, wear chemical goggles and have eye flushing equipment available. 

9. PHYSICAL AND CHEMICAL PROPERTIES 

Color: black 

Physical state : solid 

Form: powder 

Odor: none 

pH: no data available 

Density: no data available 

Specific Gravity (Relative no data available 

density) : 

Bulk density : 100 - 400 kg/m3 

Vapor pressure : no data available 

Vapor density : no data available 

Boiling pointlboiling no data available 

range: 

Freezing point: no data available 

Melting point/range: no data available 

Solubility in water: insoluble 

4UNNIO-A 

Version 1 .1 Issued on: 0712512008 Page: 6 ! 10 
p. 30 

Material Safety Data Sheet 

10. STABILITY AND REACTIVITY 

Stability: 

This material is chemically stable under normal and anticipated storage, handling and processing conditions . 

Materials to avoid: 

Strong oxidizing agents 

Conditions I hazards to avoid: 

To avoid thermal decomposition, do not overheat. 

Hazardous decomposition products: 

Thermal decomposition giving toxic products : 

Carbon oxides 

11 . TOXICOLOGICAL INFORMATION 

Data on this material and/or its components are summarized below. 

40000MUM~ Acute toxicity 

Oral: 

Slightly toxic. (rat) LD50 = 1,600 mg/kg . 

EANNImm I 

Dermal : 

No more than slightly toxic. (rabbit) LD50 > 1,000 mg/kg. 

Carcinoaenicitv 

Chronic inhalation, intratracheal injection administration to rat, mouse, hamster, guinea pig / No increase in tumor incidence was reported . 

Carcinogenicitv 

Repeated inhalation, intratracheal injection administration to rat / affected organ(s) : lung / signs: fibrosis 

Human experience 

Inhalation : 

Respiratory tract: Epidemiology studies of workers indicate pulmonary function is not affected ; no significant increases in cancer. 

Acute toxicitv 

Oral : 

No more than slightly toxic. (rat) LD50 > 2,000 mg/kg. 

Dermal: 

No more than slightly toxic. (rat) LD50 > 2,000 mg/kg. 
p. 31 

Skin Irritation : 

Slightly irritating . (rabbit) 

Eye Irritation : 

Moderately irritating . (rabbit) 

Other routes : 

Skin Sensitization: 

Not a skin sensitizer . (mouse) 

Genotoxicity 

Assessment in Vitro: 

No genetic changes were observed in laboratory tests using: bacteria, human cells 

12. ECOLOGICAL INFORMATION 

Chemical Fate and Pathway 

No data are available . 

Ecotoxicology 

Data on this material and/or its components are summarized below. 

Aquatic toxicity data: 

Practically nontoxic . Leuciscus idus (Golden orfe) 48 h LCO = 1,000 mg/I 

Microorganisms : 

Practically nontoxic . Bacteria ECO = 5,000 mg/I 

13. DISPOSAL CONSIDERATIONS 

Waste disposal : 

Where possible recycling is preferred to disposal or incineration . If recycling is not an option, incinerate or dispose of in accordance with federal, state, and local regulations. Consult a regulatory specialist to determine appropriate state or local reporting requirements, for assistance in waste characterization and/or hazardous waste disposal and other requirements listed in pertinent environmental permits. Note : Chemical additions to, processing of, or otherwise altering this material may make this waste management information incomplete, inaccurate, or otherwise inappropriate. Furthermore, state and local waste disposal requirements may be more restrictive or otherwise different from federal laws and regulations. 
p. 32 

14. TRANSPORT INFORMATION 

US Department of Transportation (DOT): not regulated 

International Maritime Dangerous Goods Code (IMDG) : not regulated 

15. REGULATORY INFORMATION 

Chemical Inventory Status 

EU. EINECS EINECS Conforms to 

US. Toxic Substances Control Act TSCA The components of this product are all on 

the TSCA Inventory . 

Australia . Industrial Chemical (Notification and AICS Conforms to 

Assessment) Act 

Canada. Canadian Environmental Protection Act DSL All components of this product are on the (CEPA). Domestic Substances List (DSL). (Can . Gaz. Canadian DSL list . 

Part II, Vol. 133) 

published by ERMA New Zealand 

United States - Federal Reaulations 

SARA Title III - Section 302 Extremely Hazardous Chemicals : 

SARA 302: No chemicals in this material are subject to the reporting requirements of SARA Title III, Section 302. 

SARA Title III - Section 311/312 Hazard Categories : 

Acute Health Hazard 

SARA Title 111- Section 313 Toxic Chemicals: 

SARA 313: This material does not contain any chemical components with known CAS numbers that exceed the threshold (De Minimis) reporting levels established by SARA Title 111, Section 313. 

Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) - Reportable Quantity (RQ): 

Japan. Kashin-Hou Law List ENCS (JP) Conforms to 

Korea. Toxic Chemical Control Law (TCCL) List KECI (KR) Conforms to 

Philippines. The Toxic Substances and Hazardous PICCS (PH) Conforms to 

and Nuclear Waste Control Act 

China. Inventory of Existing Chemical Substances IECSC (CN) Conforms to 

New Zealand. Inventory of Chemicals (NZIoC), as NZIOC Conforms to 

#

. _ 

l] 

Version 1 .1 Issued on: 0712512008 Page: 8 l 10 
p. 33 

Material Safety Data Sheet 

The components in this product are either not CERCLA regulated, regulated but present in negligible concentrations, or regulated with no assigned reportable quantity. 

OSHA Regulated Carcinogens (NTP, IARC, OSHA Listed): 

NTP: 

No component of this product present at levels greater than or equal to 0.1 % is identified as a known or anticipated carcinogen by NTP. 

IARC: 

No component of this product present at levels greater than or equal to 0.1 % is identified as probable, possible or confirmed human carcinogen by IARC. 

OSHA: 

No component of this product present at levels greater than or equal to 0.1 % is identified as a carcinogen 

or potential carcinogen by OSHA. 

United States - State Regulations 

Massachusetts Right to Know 

New Jersey Right to Know 

Chemical .Name._ _ AN* 

Pennsylvania Right to Know 

Chemical Name CAS-NQ._ .- 

California Prop. 65 

This product does not contain any chemicals known to the State of California to cause cancer, birth defects, or any other reproductive effects. 

16. OTHER INFORMATION 

Latest Revision(s): 

Reference number: 000000054675 

Date of Revision : 07/25/2008 

Date Printed: 07/25/2008 

p. 35 

Material Safety Data Sheet 

1 . PRODUCT AND COMPANY IDENTIFICATION 

Transportation : CHEMTREC: (800) 424-9300 

(24 hrs ., 7 days a week) 

Medical : Rocky Mountain Poison Center: (303) 623-5716 

(24 hrs ., 7 days a week) 

Product Information 

Product name: 

Synonyms: 

Molecular formula: 

Chemical family : 

Product use: 

2. HAZARDS IDENTIFICATION 

Emergency Overview 

Color: black 

Physical state: liquid 

Odor: slight 

WARNINGI 

MAY CAUSE SKIN IRRITATION . 

MAY CAUSE ALLERGIC SKIN REACTION. 

Potential Health Effects 

Primary routes of exposure : 

Inhalation and skin contact. 

Signs and symptoms of acute exposure : 

May cause skin irritation . Allergic skin reaction : redness, rash . 

Skin: 

Practically nontoxic . Moderately irritating . (based on components) 

Eyes: 

Slightly irritating . (based on components) 
p. 36 

Material Safety Data Sheet 

Ingestion : 

Practically nontoxic . (based on components) 

3. COMPOSITION/INFORMATION ON INGREDIENTS 

Chemical Name CAS-No. wtlwt OSHA 

Hazardous 

The substance(s) marked with a "Y" in the Hazard column above, are those identified as hazardous chemicals under 

the criteria of the OSHA Hazard Communication Standard (29 CFR 1910.1200) . 

This material is classified as hazardous under Federal OSHA regulation . 

4. FIRST AID MEASURES 

Inhalation : 

If inhaled, remove to fresh air. 

Skin : 

In case of contact, immediately flush skin with soap and plenty of water. Remove contaminated clothing and shoes. 

Get medical attention if symptoms occur. Wash clothing before reuse. Thoroughly clean shoes before reuse. 

Eyes: 

Immediately flush eye(s) with plenty of water. 

Ingestion: 

If swallowed, DO NOT induce vomiting . Get medical attention. Never give anything by mouth to an unconscious 

person. 

5. FIRE-FIGHTING MEASURES 

Flash point: > 392 °F (> 200 °C) (closed cup) 

Auto-ignition temperature: Not determined 

Lower flammable limit (LFL): Not determined 

Upper flammable limit (UFL): Not determined 

Extinguishing media (suitable) : 

Carbon dioxide (C02), Dry powder, Water spray 

Protective equipment: 

Version 1 .1 Issued on: 0712512008 Page: 2 l 10 
p. 37 

Material Safety Data Sheet 

Fire fighters and others who may be exposed to products of combustion should wear full fire fighting turn out gear (full Bunker Gear) and self-contained breathing apparatus (pressure demand / NIOSH approved or equivalent). 

Further firefighting advice : 

Fire fighting equipment should be thoroughly decontaminated after use. 

Fire and explosion hazards: 

When burned, the following hazardous products of combustion can occur: 

Carbon oxides 

6 . ACCIDENTAL RELEASE MEASURES 

In case of spill or leak: 

Spills should be contained and placed in suitable containers for disposal . Consult a regulatory specialist to 

determine appropriate state or local reporting requirements, for assistance in waste characterization and/or 

hazardous waste disposal and other requirements listed in pertinent environmental permits . 

7. HANDLING AND STORAGE 

Handling 

General information on handling: 

Wash thoroughly after handling . 

Avoid contact with the skin, eyes and clothing . 

Emptied container retains vapor and product residue. 

Observe all labeled safeguards until container is cleaned, reconditioned or destroyed. 

Storaa e 

General information on storage conditions : 

Keep containers tightly closed in a dry, cool and well-ventilated place. 

Storage stability - Temperature:-

36 - 104 °F (2 - 40 °C) 

Storage incompatibility - General : 

Store separate from strong acids, strong bases, and strong oxidizing agents . 

8. EXPOSURE CONTROLS/PERSONAL PROTECTION 

Version 1 .1 Issued on: 0712512008 Page: 3 I 10 
p. 38 

Only those components with exposure limits are printed in this section. Limits with skin contact designation above have skin contact 

effect . Air sampling alone is insufficient to accurately quantitate exposure . Measures to prevent significant cutaneous absorption 

may be required . Limits with a sensitizer designation above mean that exposure to this material may cause allergic reactions. 

Engineering controls : 

Investigate engineering techniques to reduce exposures . Provide ventilation if necessary to control exposure 

levels below airborne exposure limits (see above). If practical, use local mechanical exhaust ventilation at 

sources of air contamination such as open process equipment. 

Respiratory protection : 

Avoid breathing vapor or mist . Where airborne exposure is likely or airborne exposure limits are exceeded (if 

applicable, see above), use NIOSH approved respiratory protection equipment appropriate to the material 

and/or its components (full facepiece recommended). Consult respirator manufacturer to determine appropriate 

type equipment for a given application . Observe respirator use limitations specified by NIOSH or the 

manufacturer. For emergency and other conditions where there may be a potential for significant exposure or 

where exposure limit may be significantly exceeded, use an approved full face positive-pressure, self-contained 

breathing apparatus or positive-pressure airline with auxiliary self-contained air supply. Respiratory protection 

programs must comply with 29 CFR § 1910.134. 

Skin protection : . 

Wear appropriate chemical resistant protective clothing and chemical resistant gloves to prevent skin contact. 

When handling this material, gloves of the following type(s) should be worn: 

Neoprene 

Nitrile rubber 

Impervious butyl rubber gloves 

Wear face shield and chemical resistant clothing such as a rubber apron when splashing may occur. Rinse 

immediately if skin is contaminated . Wash contaminated clothing and clean protective equipment before reuse. 

Wash thoroughly after handling . 

Eye protection : 

Use good industrial practice to avoid eye contact. 

9. PHYSICAL AND CHEMICAL PROPERTIES 

Color: black 

Physical state: liquid 

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p. 39 

Material Safety Data Sheet 

Odor: slight 

pH: not determined 

Density: 1,150 - 1,200 kg/m3 77 °F (25 °C) 

Vapor pressure: not determined 

Vapor density: not determined 

Boiling point/boiling > 392 'F (> 200 *C) 

range: 

Melting pointlrange: > 392 °F (> 200 °C) 

Solubility in water: < 1 mg/I 

10. STABILITY AND REACTIVITY 

Stability: 

This material is chemically stable under normal and anticipated storage, handling and processing conditions . 

Materials to avoid: 

Strong oxidizing agents 

Strong acids 

strong bases 

Conditions I hazards to avoid: 

Keep away from heat and sources of ignition . To avoid thermal decomposition, do not overheat . 

Hazardous decomposition products : 

Thermal decomposition giving toxic products : 

Carbon oxides 

11 . TOXICOLOGICAL INFORMATION 

Oral : 

Practically nontoxic. (rat) LD50 > 5,000 mg/kg. 

Dermal: 

Practically nontoxic . (rabbit) LD50 > 6,000 mg/kg . 

Skin Irritation : 

Moderately irritating . (rabbit) 

Version 1 .1 Issued on: 07/2512008 Page: 6 I 10 
p. 40 

Eye Irritation : 

Slightly irritating . (rabbit) 

Material Safety Data Sheet 

Skin Sensitization: 

Repeated skin exposure. (laboratory animal) Skin allergy was observed . 

Genotoxicity 

Assessment in Vitro: 

Both positive and negative reponses for genetic changes were observed in laboratory tests using: 

bacteria 

Genotoxicitv 

Assessment in Vivo: 

No genetic changes were observed in laboratory tests using: animals, rodent 

Developmental toxicity 

Exposure during pregnancy. (rat and rabbit) / No birth defects were observed. 

Human experience 

Skin contact: 

~ Skin : Skin allergy was observed. 

Carcinoaenicitv 

Repeated inhalation, intratracheal injection administration to rat / affected organ(s) : lung / signs: fibrosis 

Human experience 

Inhalation : 

Respiratory tract: Epidemiology studies of workers indicate pulmonary function is not affected ; no significant 

increases in cancer. 

Acute toxicity 

Oral : 

No more than slightly toxic. (rat) LD50 > 2,000 mg/kg . 

Dermal : 

No more than slightly toxic. (rat) LD50 > 2,000 mg/kg. 

Skin Irritation : 

Slightly irritating . (rabbit) 

Eye Irritation : 

Moderately irritating . (rabbit) 

Other routes : 
p. 41 

Material Safety Data Sheet 

Not a skin sensitizer. (mouse) 

Skin Sensitization: 

Genotoxicity 

Assessment in Vitro: 

No genetic changes were observed in laboratory tests using: bacteria, human cells 

12. ECOLOGICAL INFORMATION 

Chemical Fate and Pathway 

Data on this material and/or its components are summarized below. 

Biodegradation : 

Not readily biodegradable . (28 d) biodegradation 2 - 3 % 

Octanol Water Partition Coefficient: 

log Pow ca. 3.8 

Ecotoxicolopy 

Data on this material and/or its components are summarized below. 

Aquatic toxicity data : 

Practically nontoxic . Oncorhynchus mykiss (rainbow trout) LC50 > 1,000 mg/I 

13. DISPOSAL CONSIDERATIONS 

Waste disposal : 

Where possible recycling is preferred to disposal or incineration . If recycling is not an option, incinerate or dispose of in accordance with federal, state, and local regulations. Consult a regulatory specialist to determine appropriate 

state or local reporting requirements, for assistance in waste characterization and/or hazardous waste disposal and other requirements listed in pertinent environmental permits. Note: Chemical additions to, processing of, or otherwise altering this material may make this waste management information incomplete, inaccurate, or otherwise inappropriate. Furthermore, state and local waste disposal requirements may be more restrictive or otherwise different from federal laws and regulations. 

14. TRANSPORT INFORMATION 

US Department of Transportation (DOT): not regulated 

International Maritime Dangerous Goods Code (IMDG) 

UN Number : 3482 

Proper shipping name : ENVIRONMENTALLY HAZARDOUS SUBSTANCE, LIQUID, N.O .S . 
p. 42 

Material Safety Data Sheet 

Technical name : (EPOXY RESIN) 

Class : g 

Packaging group : III 

Marine pollutant : no 

Flash point : > 392 °F (> 200 °C) closed cup 

15. REGULATORY INFORMATION 

Chemical Inventory Status 

EU. EINECS EINECS Conforms to 

US. Toxic Substances Control Act TSCA The components of this product are all on 

the TSCA Inventory. 

Australia. Industrial Chemical (Notification and AICS Conforms to Assessment) Act 

(CEPA)

Canada. Canadian Environmental Protection Act DSL All components of this product are on the . Domestic Substances List (DSL). (Can. Gaz. Canadian DSL list. Part II, Vol. 133) 

Japan. Kashin-Hou Law List ENCS (JP) Conforms to 

Korea. Toxic Chemical Control Law (TCCL) List KECI (KR) Conforms to 

Philippines. The Toxic Substances and Hazardous PICCS (PH) Conforms to and Nuclear Waste Control Act 

China. Inventory of Existing Chemical Substances IECSC (CN) Conforms to 

United States - Federal Regulations 

SARA Title 111- Section 302 Extremely Hazardous Chemicals: 

SARA 302: No chemicals in this material are subject to the reporting requirements of SARA Title III, Section 302. SARA Title III - Section 311/312 Hazard Categories : Acute Health Hazard 

SARA Title III - Section 313 Toxic Chemicals: 

SARA 313: This material does not contain any chemical components with known CAS numbers that exceed the threshold (De Minimis) reporting levels established by SARA Title 111, Section 313. 

Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) - Reportable Quantity (RQ): 

The components in this product are either not CERCLA regulated, regulated but present in negligible concentrations, or regulated with no assigned reportable quantity . 

Version 1 .1 Issued on: 07/2512008 Page: 8110 
p. 43 

Material Safety Data Sheet 

--J 

OSHA Regulated Carcinogens (NTP, IARC, OSHA Listed): 

NTP: 

No component of this product present at levels greaterthan or equal to 0.1% is identified as a known or 

anticipated carcinogen by NTP. 

IARC: 

No component of this product present at levels greater than or equal to 0.1% is identified as probable, 

possible or confirmed human carcinogen by IARC. 

OSHA: 

No component of this product present at levels greater than or equal to 0.1 °1o is identified as a carcinogen 

or potential carcinogen by OSHA. 

United States - State Regulations 

Massachusetts Right to Know 

CAS-No. 

New Jersey Right to Know 

No components are subject to the New Jersey Right to Know Act. 

Pennsylvania Right to Know 

California Prop. 65 

This product does not contain any chemicals known to the State of California to cause cancer, birth 

defects, or any other reproductive effects. 

16. OTHER INFORMAT10N 

Latest Revision(s): 

Revised Section(s) : 

Reference number: 

Date of Revision: 07/25/2008 

Date Printed : 09/18/2008 

CANKUNI 

Version 1 .1 Issued on: 07125/2008 Page: 9110 
p. 44 

Material Safety Data Sheet 
SPONSOR . 

STUDY TITLE 

ACUTE ORAL TOXICITY IN RATS 

"ACUTE TOXIC CLASS METHOD" 

STUDY DIRECTOR 

Catherine Pelcot 

DATE OF ISSUE ,9s r~;~ 2_~' 

TEST FACILITY 

CIT 

BP 563 - 27005 Evreux - France 

IFM Recherche BY 563 27005 Evreux Cedex 

S.N.C au Copital de 846 .092 ¬ T61 : +33 (0)2 32 29 26 26 

788 060 465 R .C.S. Evreux Fax : +33 (0)2 32 67 87 OS 

VAT : FR I 9 788 060 465 E-mail : CIT@citox.com 

Agrement Credit Impot Recherche Web site : www.citox .com 

CONFIDENTIAL 

France 
CONTENTS 

STATEMENT OF THE STUDY DIRECTOR 4 

OTHER CIT SCIENTIST INVOLVED IN THE STUDY 5 

STATEMENT OF QUALITY ASSURANCE UNIT -~`-

< 

6 

SUMMARY 7 

RESUME 

1 . INTRODUCTION 

2. MATERIALS AND METHODS 

2.1 TEST ITEM 

2.1 .1 Identification 

2.1 .2 Vehicle 

2.1 .3 Dosage form preparation 

2.2 TEST SYSTEM 

2 .2 .1 Animals 

2 .2 .2 Environmental conditions 

2 .2 .3 Food and water 

2.3 TREATMENT 

2.3 .1 Fasting of the animals 

2.3 .2 Study design 

2.3 .3 Administration of the test item 

2.4 CLINICAL EXAMINATIONS 

2.4 .1 Clinical signs and mortality 

2.4 .2 Body weight 

2.5 PATHOLOGY 

2.5 .1 Sacrifice 

2.5 .2 Macroscopic necropsy examination 

2.5 .3 Preservation of tissues 

2.6 

2.7 

2 .8 

2.9 

3. 

3.1 

3.1 .1 

3.1 .2 

3 .2 

4. 

EVALUATION AND INTERPRETATION OF THE RESULTS 

ARCHIV ING 

CHRONOLOGY OF THE ST. ULY 

STUDY PLAN ADHERENCE 

RESULTS 

CLINICAL EXA$IINATIQNS 

a: 

Clinical signs and mortality (table 1) 

Body weight (figure 1, tables 2 and 3) 

PATHOLOGY (table 4) 

CONCLUSION 

9 

11 

11 

11 

11 

11 

12 

12 

12 

12 

12 

13 

13 

13 

13 

14 

14 

14 

14 

14 

14 

14 

14 

15 

15 

15 

16 

16 

16 

16 

16 

16 
Figure 1 : Mean body weight of the treated animals 1 7 

Table 1 : Individual clinical signs and mortality 18 

Table 2: Individual and mean body weight and weekly body weight change of treated rats 

(g) 19 

Table 3 : Body weight - CIT historical data of control animals dosed by oral route 20 

Table 4 : Individual macroscopic examinations at necropsy 21 

APPENDICES 22 

1 . Test article description and analytical certificate 23 

2. Diet formula 26 

3 . CIT GLP certificate 28 and 29 
STATEMENT OF THE STUDY DIRECTOR 

The study was performed in compliance with CIT's standard operating procedures and the 

following principles of Good Laboratory Practice: 

. OECD Principles of Good Laboratory Practice (as revised in 1997), ENV/MC/CHEM 

(98) 17 and all subsequent OECD consensus documents. 

. Directive 2004/10/EC of the European Parliament and of the Council of 11 February 2004 on 

the harmonization of laws, regulations and administrative provisions relating to the 

application of the Principles of Good Laboratory Practice and the verification of their 

applications for tests on chemical substances (OJ No. L50 of 20.2 .2004) . 

. Decret N° 2006-1523 du 04 decembre 2006 concernant les Bonnes Pratiques de Laboratoire 

(Journal Officiel du 06 decembre 2006), Ministere de 1'Economie, des Finances et de 

1'Industrie . 

However, no chemical analysis of the dosage forms was performed as part of this study. 

This exception is not considered to impact the overall GLP status of the study. 

The study was also conducted in compliance with the following Animal Protection regulations : 

. Council Directive 86/609/EEC of 24th November 1986 on the harmonization of laws, 

regulations or administrative provisions relating to the protection of animals used for experimental or other scientific purposes . 

. Guidance document on the recognition, assessment, and use of clinical signs as humane 

endpoints for experimental animals used in safety evaluation, OECD Environmental Health and Safety Publications, No . 19. 

I declare that this report constitutes a true and faithful record of the procedures undertaken and 

the results obtained during the performance of the study. 

f ., 

This study was performed at CIT, BP 563, 27005 Evreux, Franc¢: 

Y 

Toxicology 

C. Pelcot Study completion date: ~-1 A pk . L J,a ~ 

Study Director 
OTHER CIT SCIENTIST INVOLVED IN THE STUDY 

Director of Toxicology 

CIT Management 

J.J . Legrand Date: 

Doctor of Veterinary Medicine 
STATEMENT OF WALITY ASSURANCE UNIT 

Inspections performed at CIT: 

The CIT Quality Assurance Unit conducted the inspections detailed below: 

Dates 

Type of inspection Inspection Reported to the Reported to 

Study Director Management 

p. 50 

6 

Study plan 27 July 2007 27 July 2007 02 August 2007 

Report 12 February 2008 14 February 2008 10 March 2008 

In addition, at about the same time, process-based and facility inspections relevant to this study 

were carried out by the Quality Assurance Unit. 

The inspections; were performed in compliance with CIT Quality Assurance Unit procedures and 

the principles of Good Laboratory Practices . 

The final report is considered to constitute an accurate and complete reflection of the study raw 

data. 

_- 

. caL7- c.GCC.~ Date: -2S ~-tt,, ~ 

C1T Quality Assurance Unit 
SUMMARY 

~_ . ., . 

guidelines. 

, 17th December 2001) and EC (2004/73/EC, B.1 tris, 29th Apri12004) 

The study was conducted in compliance with the principles of Good Laboratory Practice 

Regulations . 

Methods 

The test item was prepared in 0.5% methylcellulose and was administered by oral route 

(gavage), to groups of three fasted female Sprague-Dawley rats . 

The study design was as follows : 

Dose-level Volume Female 

(mg/kg) (mL/kg) 

300 10 3 

2000 14.3 3 

2000 14.3 3 

Clinical signs, mortality and body weight gain were checked for a period of up to 14 days 

following the single administration of the test item. 

All animals were subjected to necropsy. 

Results 

Dose-level of 300 mg/kg (three females) 

No deaths and no clinical signs were noted at this dose-level. 

Dose-level of 2000 mg/kg (three females then confirmation on three other females_) 

No deaths occurred. 

Hypoactivity, piloerection and dyspnea were noted in 3/6 animals on day 1 . 

When compared to CIT historical control animals, a slightly lower body weight gain was noted 

between day 1 and day 8 in 1/3 females treated at the dose-level of 300 mg/kg, without any 

relevant consequence at the end of the observation period, and in 1/6 females treated at the 

dose-level of 2000 mg/kg, between day 8 and day 15. The body weight gain of the other animals 

treated at the dose-level of 300 or 2000 mg/kg was not affected by treatment with the test item. 

At necropsy, no apparent abnormalities were observed in any animal . 
Conclusion 

itions of this study, the oral LDo of the test item 

According to the classification criteria laid down in Council Directive 67/548/EEC (and subsequent adaptations), concerning the potential toxicity by oral route, the test item should not be classified. 
RESUME 

~ ignes irec ~ces e CDE (n° 423, 17 decem re e e a EE (2004/73/CEE, B. 

tris, 29 avril 2004). 

L'dtude a ete realisee conform6ment aux regles de Bonnes Pratiques de Laboratoire . 

Methode 

Le produit a etd prepar6 clans de la methylcellulose a 0,5 % et administr6 par voie orale 

(gavage), a des groupes de 3 femelles rats Sprague-Dawley mis a la diete hydrique. 

Le schema de 1'etude a ete le suivant : 

Dose Volume Femelle 

(mg/kg) (mL/kg) 

300 10 3 

2000 14,3 3 

2000 14,3 3 

Les signes cliniques, la mortalite et 1'evolution ponderale des animaux ont ete suivis pendant une 

pgriode de 14 jours apres 1'administration unique du produit . 

Un examen anatomopathologique a ete effectue sur tous les animaux. 

Resultats 

Dose de 300 mg/kg (3 femelles) 

Aucune mortalite et aucun signe clinique ne sont observes a cette dose. 

Dose de 2000 my/kg (3 femelles puis confirmation sur 3 autres femelles) 

Aucune mortalite West enregistree. 

Hypoactivite, pilogrection et dyspnee sont notees chez 3/6 animaux au Jour 1 . 

En comparaison avec celle des temoins historiques du CIT, 1'evolution ponderale est legerement reduite entre le Jour 1 et le Jour 8 chez 1/3 femelle traitee a la dose de 300 mg/kg, sans aucune consdquence notable a la fin de la p6riode d'observation, et chez 1/6 femelle traitde a la dose de 2000 mg/kg entre le Jour 8 et le Jour 15. L'evolution ponderale des autres animaux traitgs aux 

doses de 300 ou 2000 mg/kg n'est pas influencde par le traitement avec le produit. 

L'examen macroscopique des principaux organes realisd chez tous les animaux ne met en evidence aucune anomalie apparente. 
Conclusion 

p. 54 

10 

--- m---conditions ex 6riment l d- cette etude la DL 0 orale du produ' 

- - ------- --- 

Selon les criteres de classification d6crits dans la Directive 67/548/CEE (et ses adaptations), 

concernant la toxicite aigue par voie orale, le produit ne devrait pas etre classe . 
l. INTRODUCTION 

! is study was to evaluate the toxicity of the test ite 

~llowing a single oral administration in rats . 

p. 55 

11 

In the assessment of the toxic characteristics of a test item, determination of acute oral toxicity is 

an initial step. It provides information on health hazards likely to arise following a short-term 

exposure by the oral route in humans and enables the test item to be ranked in different 

classification systems. 

The acute toxic class method is a stepwise procedure . 

The test item is administered orally to one group of animals at one of the defined dose-levels, 

each step using three animals of one sex. 

Absence or presence of compound-related mortality of the animals dosed at one step determines 

the next step, i. e. : 

. no further testing is needed, 

. the next step is performed with the same dose-level, 

. the next step is performed at the next higher or the next lower dose-level. 

The study was conducted in compliance with: 

. EC Directive 2004/73/EC, B.1 tris, 29th April 2004, 

. OECD Guideline No . 423, 17th December 2001 . 

2. MATERIALS AND METHODS 

2.1 TEST ITEM 

e: 

batch 

CAS number 

Description at re" ce'°ip~' 

container: one plastic flask 

date of receipt: 29 June 2007 

storage conditions: at room temperature and protected from humidity 

composition : see analytical certificate 

expiry date: June 2008. 

Data relating to the characterization of the test item are documented in a test article description 

and an analytical certificate (presented in appendix 1) provided by the Sponsor. 

2.1 .2 Vehicle 

The vehicle used was 0.5% methylcellulose: methylcellulose, batch No. 017K0052 (Sigma, 

Saint-Quentin-Fallavier, France) in purified water (prepared at CIT by reverse osmosis). 
2.1 .3 Dosage form preparation 

p. 56 

12 

The test item was not soluble at the concentration of 100 mg/mL in purified water, 0.5% 

methylcellulose and corn oil. At the maximal concentration of 70 mg/mL, a suspension was 

obtained in purified water or in 0.5% methylcellulose. As the suspension was more 

homogeneous in 05% methylcellulose, this vehicle was retained. 

On the day of treatment, the test item was ground to a fine powder using a mortar and pestle, 

then was prepared at the chosen concentration in the vehicle . 

The test item preparation was made freshly on the morning of administration by the CIT 

Pharmacy and any unused material was discarded that same day. 

2.2 TEST SYSTEM 

2 .2 .1 Animals 

Species, strain and sex: nulliparous and non-pregnant female rat, Sprague-Dawley Rj : SD (IOPS 

Han). 

Reason for this choice: rodent species generally accepted by regulatory authorities for this type 

of study. 

Breeder : Janvier, Le Genest-Saint-Isle, France . 

Number and sex: 3 groups of three females were used. 

Age/weight: on the day of treatment, the animals were approximately 8 weeks old, and had a 

mean body weight ::E standard deviation of 189 ± 6 g. 

Acclimation : at least 5 days before the beginning of the study . 

Allocation to study: before the beginning of the study, on day 1, the required number of animals 

was selected according to body weight and clinical condition . 

Identification : individually by earnotches . 

2.2 .2 Environmental conditions 

The conditions in the animal room were set as follows : 

. temperature : 22 ± 2°C 

. relative humidity : 30 to 70% 

. light/dark cycle: 12 h/12 h 

. ventilation : approximately 12 cycles/hour of filtered, non-recycled air . 

The temperature and relative humidity were under continuous control and recording. The records were checked daily and filed . In addition to these daily checks, the housing conditions 

and corresponding instrumentation and equipment are verified and calibrated at regular 

intervals . 

The animals were housed in polycarbonate cages with stainless steel lid (48 cm x 27 cm 

x 20 cm). Each cage contained one to seven animals during the acclimation period and three rats 

of the same group during the treatment period . 

Each cage contained autoclaved sawdust (SICSA, Alfortville, France) . 

Sawdust is analyzed by the supplier for composition and contaminant levels. 

2 .2 .3 Food and water 

All the animals had free access to SsniffR/M-H pelleted diet (SSNIFF Spezialdiaten GmbH, 

Soest, Germany), except as noted in "2.3 .1 Fasting of the animals" . 

Each batch of food is analyzed by the supplier for composition and contaminant levels. 

The diet formula is presented in appendix 2 . 

Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum . Bacteriological and chemical analyses of water are performed regularly by external laboratories. 
p. 57 

13, 

These analyses include the detection of possible contaminants (pesticides, heavy metals and 

nitrosamines) . 

No contaminants were known to have been present in the diet, drinking water or bedding 

material at levels which may be expected to have interfered with or prejudiced the outcome of 

the study. 

2.3 TREATMENT 

2.3 .1 Fasting of the animals 

The animals were fasted for an overnight period of approximately 18 hours before dosing, but had free access to water. 

Food was given back approximately 4 hours after administration of the test item (dose-level of 

300 mg/kg) or after the first administration of the test item (dose-level of 2000 mg/kg). 

2.3 .2 Study design 

Three females were used for each step. 

The dose-level used as the starting dose-level was selected from one of four fixed levels, 5, 50, 300 or 2000 mg/kg body weight . 

As no information on the toxic potential of the test item was available, for animal welfare 

reasons, the starting dose-level of 300 mg/kg was chosen. 

After the first assay, as no deaths occurred, another assay was carried out on three animals at the 

next higher dose-level, i.e . 2000 mg/kg. 

After the second assay, as no deaths occurred, the results were then confirmed in three other females at the dose-level of 2000 mg/kg. 

The study design was as follows : 

Dose-level Volume Female 

(mg/kg) (mL/kg) 

300 10 3 

2000 14.3 3 

2000 14.3 3 

2.3 .3 Administration of the test item 

The dosage form preparations were administered to the animals under a volume of 10 mL/kg 

(dose-level of 300 mg/kg) or 14 .3 mL/kg (dose-level of 2000 mg/kg). 

The administration was performed once (dose-level of 300 mg/kg) or twice at a 2-hour interval (dose-level of 2000 mg/kg) by oral route using a metal gavage tube fitted to a 2 or 5 mL plastic syringe (0.1 or 0.2 mL graduations) . 

The volume administered to each animal was adjusted according to body weight determined on 

the day of treatment. 
2.4 CLINICAL EXAMINATIONS 

p. 58 

14 

The administration was performed in the morning of day 1 ; it was followed by a 14-day observation period. 

2 .4 .1 Clinical signs and mortality 

The animals were observed frequently during the hours following administration of the 

test item, for detection of possible treatment-related clinical signs. Thereafter, observation of the animals was made at least once a day. 

Type, time of onset and duration of clinical signs were recorded for each animal individually . 

2.4.2 Body weight 

The animals were weighed individually just before administration of the test item on day 1 and then on days 8 and 15. 

The body weight gain of the treated animals was compared to that of CIT control animals with the same initial body weight. 

2.5 PATHOLOGY 

2.5 .1 Sacrifice 

On day 15, all animals were killed by carbon dioxide asphyxiation . 

2.5 .2 Macroscopic necropsy examination 

All study animals were subjected to a macroscopic examination as soon as possible after death. After opening the thoracic and abdominal cavities, a macroscopic examination of the main 

organs (digestive tract, heart, kidneys, liver, lungs, pancreas, spleen and any other organs with 

obvious abnormalities) was performed. 

2.5 .3 Preservation of tissues 

No organ samples were taken. 

2.6 EVALUATION AND INTERPRETATION OF THE RESULTS 

The interpretation of results was based on the flow charts of Annex 2 of the OECD Guideline No. 423, 17th December 2001 and of Annex 3 of the Directive 2004/73/EC, B.1 tris, 29th April 2004. 
2.7 ARCHIVING 

p. 59 

15 

The following study materials are archived by CIT, 27005 Evreux, France, for 10 years after the end of the in vivo phase of the study : 

. Study plan and possible amendments, 

. raw data, 

. correspondence, 

. final report and possible amendments. 

On completion of this period, the archived study materials will be returned to the Sponsor, or 

may be archived at CIT for a further period (at additional cost) . The total duration of archiving (depending on regulations) will be the responsibility of the Sponsor. 

In addition, raw data not specific to the study including, but not limited to, certificates of analyses for food, water and bedding (if applicable) and records of environmental data and equipment calibration, are also archived by CIT for at least 30 years. 

2.8 CHRONOLOGY OF THE STUDY 

The chronology of the study was as follows : 

Procedure Date 

Experimental starting date (first treatment) 

Experimental completion date (necropsy of the last animal) 

2.9 STUDY PLAN ADHERENCE 

The study was performed in accordance with 

07 August 2007 

11 September 2007 

--- -- amendments, with the following deviations from the agreed Study plan: 

. the temperature and relative humidity recorded in the animal room were sometimes the target ranges specified in the Study plan, 

. on day l, the body weight of one animal (female No. 

(177 g), 

. the actual administered dose was 2000 mg/kg (instead 

amendment to the Study plan). 

ubsequent 

outside of 

01) was slightly lower than 180 g 

of 2002 mg/kg as specified in the 

These minor deviations were not considered to have compromised the validity or integrity of the study. 
3. RESULTS 

3.1 CLINICAL EXAMINATIONS 

3 .1 .1 Clinical signs and mortality (table 1) 

Dose-level of 300 mg/kg (three females) 

No deaths and no clinical signs were noted at this dose-level. 

Dose-level of 2000 mz/kg (three females then confirmation on three other females) No deaths occurred . 

Hypoactivity, piloerection and dyspnea were noted in 3/6 animals on day 1 . 

3.1 .2 Body weight (figure l, tables 2 and 3) 

p. 60 

16 

When compared to CIT historical control animals, a slightly lower body weight gain was noted between day 1 and day 8 in 1/3 females treated at the dose-level of 300 mg/kg, without any relevant consequence at the end of the observation period, and in 1/6 females treated at the dose-level of 2000 mg/kg, between day 8 and day 15. The body weight gain of the other animals treated at the dose-level of 300 or 2000 mg/kg was not affected by treatment with the test item. 

3 .2 PATHOLOGY (table 4) 

Macroscopic examination of the main organs of the animals revealed no apparent abnormalities . 

4. CONCLUSION ~ 

n_tal_ _- conditions . of_ .t_-ig _ : .,q . 1,ju rhP ,1a.t _ My of the test item 

_ __ ._.. ._. . ._ . 

ODD mg/kg in rats . 

According to the classification criteria laid down in Council Directive 67/548/EEC (and subsequent adaptations), concerning the potential toxicity by oral route, the test item should not be classified. 
Figure 1 : Mean body weight of the treated animals 

320 

300 

280 

260 

240 

220 

200 

180 

160 

140 

(g) 

® 

days 

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 

p. 61 

17 

--4w- 300 mg/kg -o--2000 ng/kg (first assay) -,A -2000 mg/kg (confirmatory assay) 
Table 1 : Individual clinical signs and mortality 

Dose-level Time Females Mortality Clinical signs 

(mg/kg) 

300 30 min 

1 h - 2 h - 4 h ~ 01-02-03 No None 

D2toDl5 J 

p. 62 

18 

-------------------------------------------------------------------------------------------------------------------- 

2000 15 min - 1 h - 2 h 04-05-06 No None 

(first assay) 

2 h 30 05-06 No Hypoactivity, 

piloerection, dyspnea 

04 No Piloerection, dyspnea 

4 h - 6 h 04-05-06 No Hypoactivity, 

piloerection, dyspnea 

D 2 to D 5 04-05-06 No None 

-------------------------------------------------------------------------------------------------------------------- 

2000 30 min -1 h 

(confirmatory 2 h - 2 h 30 - 4 h ~ 07-08-09 No None 

assay) D 2 to D 15 J 

min : minutes 

h : hour 

D :day 
Table 2: Individual and mean body weight and weekly body weight change of treated rats (g) 

Dose-level Volume 

Sex Animals 

Days 

mg/kg mL/kg 1 (1) 8 (1) 

300 10 Female 01 177 

02 195 

03 192 

15 

35 212 12 224 

37 232 13 245 

26 218 18 236 

M 188 33 221 14 235 

SD 10 6 10 3 11 

2000 14 .3 Female 04 185 

(first assay) 05 193 

06 190 

50 235 17 252 

40 233 15 248 

51 241 4 245 

M 189 47 236 12 248 

SD 4 6 4 7 4 

2000 14.3 Female 07 

(confumatory 08 

assay) 09 

190 41 231 22 253 

186 46 232 21 253 

197 44 241 9 250 

M 191 44 235 17 252 

SD 6 3 6 7 2 

(1) = body weight gain 

M = mean 

SD = standard deviation 
Table 3 : Body weight - CIT historical data of control animals dosed by oral route 

Check of the body weight (g) of CIT historical control animals 

Method : Acute oral administration 

CIT Studies / Experimental : CIT/Study Nos. 27218 RDR / February 2004 

Completion date 29691 RDR / April 2005 

31103 RDR / January 2006 

Reference item : Purified water 

Volume of administration : 10 mL/kg 

Animals 

Species, strain : Rat, Sprague-Dawley Rj : SD (IOPS Han) 

Breeder : Janvier, Le Genest-Saint-Isle, France 

Age on day 1 : 8 weeks 

Volume 

(mL/kg) 1 

10 Female M 

SD 

n 

M :mean 

(1) : body weight gain 

SD : standard deviation 

n : number of animals 

Sex 

(1) 

Days. 

8 (1) 15 

200 38 240 15 257 

8 5 7 6 8 

30 30 30 30 30 

p. 64 

20 
Table 4 : Individual macroscopic examinations at necropsy 

Dose-level Time Females Macroscopic 

(mg/kg) abnormalities 

p. 65 

21 

300 D 15 01-02-03 No apparent abnormalities 

-------------------------------------------------------------------------------------------------------------------- 

2000 D 15 04-05-06-07-08-09 No apparent abnormalities 

D: day 
APPENDICES 
1 . Test article description and analytical certificate 
TEST ARTICLE DESCRIPTION 

p. 68 

24 

IDENTITY 

Test article name 

Chemical name 

CAS number 

EINECS number 

Origin 

Batch number 

Arkema filing number 

PHYSICAL AND CHEMICAL PROPERTIES 

Appearance : black powder 

Part.icule size : 30 gm. (approximately) 

Specific Gravity : 2,1 kg/m3 at 20°C 

Autoignition temperature : > 400 °C (standard : NF EN 50281-2-1) 

Solubility : in water : insoluble 

TOXICOLOGICAL INFORMATIONS AND USE SAFETY 

See Safety Data Sheet. 

STORAGE AND DISPOSAL 

Storage 

Expiry date 

Disposal 

: Keep hermetically closed in a dry, cool and well-ventiled 

place . 

: June 2008 

: Incineration 
Nom du responsable du taboratoirel Laboratory Director : 

DETERMINATION ! ITEM 

Reference de la methode 

RESULTAT l RESULT d'analyse 1 Analysis 

reference 

Powder characteristics 

Ash content °k 7.6% ATG 

r 

~

"'°' 

70 . 
2. Diet formula 
SsniffR/M-H 

Complete diet for rats/mice - maintenance 

Crude proteins 

Crude fat 

Crude fiber 

Crude ash 

Amino Acids 

Lysine 

Methionine 

Cystine 

Glycine 

Leucine 

Isoleucine 

Arginine 

Phenylalanine 

Tryptophan 

Histidine 

Tyrosine 

Aspartic acid 

Glutaminic acid 

Valine 

Threonine 

Constituents 

Calcium 1 .00% 

Phosphorus 0.70% 

Sodium 0.25% 

Magnesium 0.20% 

Potassium 0 .90% 

19.00% 

3 .30% 

4.90% 

6.70% 

1 .00% 

0.30% 

0,30% 

0.90% 

1 .30% 

0.70% 

1 .20% 

0.90% 

0.25% 

0.50% 

0.60% 

1 .70% 

3 .80% 

0 .90% 

0 .70% 

Vitamins go kg) 

A 

D3 

E 

B1 

B2 

B6 

B12 

Biotin 

Pantothenic acid 

Choline 

Folic acid 

Nicotic acid 

K3 

Inositol 

Trace elements (je kg) 

Manganese 

Copper 

Zinc 

Iodine 

Iron 

Selenium 

Cobalt 

ME (je kg) 

Item numbers 

V1530 Meal 

V1531 Micromeal 

V 1534 Pellets 10 mm 

V1535 Pellets 15 mm 

90 mg 

12 mg 

75 mg 

2 mg 

220 mg 

0.2 mg 

2 mg 

15,000 IE 

1,000 IE 

100 mg 

10 mg 

20 mg 

12 mg 

80 pg 

400 pg 

30 mg 

1,600 mg 

4 mg 

60 mg 

5 mg 

50 mg 

12.2 MJ 
3 . CIT GLP certificate 
!lSertt- " 4eHft - F'40-8f 

Eiirusr tcWe FraN(;&rse 

GROUPE INTERMINISTERIEL DES PRODUITS CHIMIQUES 

Paris, le 

Objet : Evaluation de la conformite aux Bonnes Pratiques de Laboratoires (BPL) selon les directives 

2004/9/CE et 2004/10/CE du 11 f6vrier 2004 . 

Subject : Assessment of compliance with Good Laboratory Practices (GLP) under the EC directives 

2004/9 and 2004/10 of II February 2004. 

ConsBcutivement ii votre engagement vis-a-vis du GIPC et du COFRAC et en application du d6cret n° 2006-

1523 du 4 decembre 2006 concernant les bonnes pratiques de laboratoires et modifiant le ddcret n° 81-278 du 25 

mars 1981 portant creation d'un groupe interministeriel des produits chimiques, je vous confirme que le GIPC, 

au vu des r6sultats du contr8le exerc8 par le Comite frangais d'accr6ditation (COFRAC) - Section Laboratoires a 

d8cid8 pour votre installation du statut suivant : 

Following your engagement vis-a-vts the GIPC and COFRAC and in application of the decree 11' 2006-1523 of 

4 December 2006 relating to the good laboratory practices and modifying the decree n° 81-278 of 25 March 

1981 giving birth to an interministerial group of chemical products (GIPC), I confirm to you that the GIPC, 

given the results of the inspection realised by the French Committee of accreditation (COFRAC) - Laboratory 

Section has taken the following decision relating to your installation: 

Respect des principes de BPL Date d'inspection : 7-8 mars 2007 

Respect of the GLP principles Date of inspection ., 7-8 mars 2007 

Domaines de reconnaissance: Inspection de renouvellement (i .r) 

1 - essais physico-chimiques Renewal inspection (i.r) 

2 - etudes de toxicite 

3 - etudes de mutag8nicife Date de decision du GIPC : 29 juin 2007 

4 - etudes 8cotoxicologiques sur les Date of GIPC decision: 29 juin 2007 

organismes aquatiques et terrestres 

8 - methodes de chimie analytique et Date de prise d'effet : 8 mars 2007 

clinique Date of implementation: 8 mars 2007 

Areas of expertise : Ann6e de premiere conformitd : 1989 

1 = Physico-chemical testing Year of the first conformity: 1989 

2 = Toxicily studies 

3 = Mutagenicity studies Dur6e de validite : 18 mois 

4 = Environmental toxicity studies on aquatic or Time of validity: 18 months 

terrestrial organisms 

8 =Analytical and clinical chemistry 

:r_ _ . ._ . 

CENTRE INTERNATIONAL DE TOXICOLOGIE 

(CIT) 

MISEREY - BP 563 

27005 EVREUX CEDEX 

- s JUli . 2007 

Pierre CkEYSSEL 

Conseiller d'Etat h 

Secr6tariat general du GIPC - DGE- Simap - 12, rue Villiot - 75572 Paris cedex 12 

Telephone : 0153 44 96 10 - T61ecopic : 0 153 44 9172 
Figure 1 : Mean body weight of treated rats 15 

Table 1 : Individual clinical signs and mortality 16 

Table 2: Cutaneous reactions 1 7 

Table 3 : Individual and mean body weight and weekly body weight change (g) 18 

Table 4: Body weight - CIT historical data of control animals dosed by dermal route 19 

Table 5 : Individual macroscopic examinations at necropsy 20 

APPENDICES 21 

1 . Test article description and analytical certificate 22 

2. Diet formula 25 

3. CIT GLP certificate 27 and 28 
:~s----' Safety& Health 

Research Laboratories 

STUDY TITLE 

ACUTE DERMAL TOXICITY 

IN RATS 

STUD YDIRECTOR 

Catherine Pelcot 

DATE OF ISSUE 

TEST FACILITY 

CIT 

BP 563 - 27005 Evreux - France 

LABORATORY STUDY NUMBER 

IFM Recherche BY 563 27005 Evreux Cedex France 

S .N.C au Capital de 846.092 ¬ T6I. : +33 (0)2 32 29 26 26 

788 060 465 RC.S. Evreux Fax : +33 (0)2 32 67 87 05 

UAT : FR19 788 060 465 E-maif : CfT@citox.com 

Agrement Crddit fmpdt Recherche Web site : vdww.citox_com 

p. 75 

CONFIDENTIAL 

I 
CONTENTS 

STATEMENT OF THE STUDY DIRECTOR 4 

OTHER CIT SCIENTIST INVOLVED IN THE STUDY 5 

STATEMENT OF QUALITY ASSURANCE UNIT 6 

SUMMARY 7 

RESUME 8 

1 . INTRODUCTION 9 

2. MATERIALS AND METHODS 9 

2.1 TEST ITEM 9 

2.1 .1 Identification 9 

2.1 .2 Dosage form preparation 9 

2.1 .3 Other item 9 

2 .2 TEST SYSTEM 10 

2 .2 .1 Animals 10 

2 .2 .2 Environmental conditions 10 

2.2 .3 Food and water 10 

2.3 TREATMENT 11 

2.3 .1 Study design 11 

2.3 .2 Preparation of the animals 1 1 

2.3 .3 Administration of the test item 11 

2.4 CLINICAL EXAMINATIONS 11 

2.4 .1 Clinical signs and mortality 11 

2.4 .2 Body weight 11 

2.5 PATHOLOGY 12 

2.5 .1 Sacrifice 12 

2.5 .2 Macroscopic necropsy examination 12 

2.5 .3 Preservation of tissues 12 

2 .6 DATA EVALUATION 12 

2.7 ARCHIVING 12 

2.8 CHRONOLOGY OF THE STUDY 13 

2.9 STUDY PLAN ADHERENCE 13 

3 . RESULTS 14 

3 .1 CLINICAL EXAMINATIONS 14 

3 .1 .1 Mortality (table 1) 14 

3 .1 .2 Clinical signs and cutaneous reactions (tables 1 and 2) 14 

3 .1 .3 Body weight (figure 1, tables 3 and 4) 14 

3.2 PATHOLOGY (table 5) 14 

4. CONCLUSION 14 
STATEMENT OF THE STUDY DIRECTOR 

The study was performed in compliance with CIT's standard operating procedures and the 

following principles of Good Laboratory Practice: 

. OECD Principles of Good Laboratory Practice (as revised in 1997), ENV/MC/CHEM 

(98) 17 and all subsequent OECD consensus documents. 

. Directive 2004/10/EC of the European Parliament and of the Council of 11 February 2004 on 

the harmonization of laws, regulations and administrative provisions relating to the 

application of the Principles of Good Laboratory Practice and the verification of their 

applications for tests on chemical substances (OJ No. L50 of 20.2.2004). 

. D6cret N° 2006-1523 du 04 d6cembre 2006 concernant les Bonnes Pratiques de Laboratoire 

(Journal Officiel du 06 d6cembre 2006), Ministere de 1'Economie, des Finances et de 1'Industrie . 

The study was also conducted in compliance with the following Animal Protection regulations: 

. Council Directive 86/609/EEC of 24th November 1986 on the harmonization of laws, 

regulations or administrative provisions relating to the protection of animals used for 

experimental or other scientific purposes. 

. Guidance document on the recognition, assessment, and use of clinical signs as humane 

endpoints for experimental animals used in safety evaluation, OECD Environmental Health 

and Safety Publications, No. 19 . 

I declare that this report constitutes a true and faithful record of the procedures undertaken and the results obtained during the performance of the study. 

This study was performed at CIT, BP 563, 27005 Evre; , France . 

Toxicology 

C. Pelcot Study cor}ipletion date: 31 rj.,rej, 

Study Director 
OTHER CIT SCIENTIST INVOLVED IN THE STUDY 

Director of Toxicology 

CIT Management 

J.J . Legrand Date: ~ Z- kt,  ,t ~,1 too Doctor of Veterinary Medicine 
STATEMENT OF OUALITY ASSURANCE UNIT 

Inspections performed at CIT: 

The CIT Quality Assurance Unit conducted the inspections detailed below: 

Dates 

Type of inspection Inspection Reported to the Reported to 

Study Director Management 

Study plan 27 July 2007 27 July 2007 02 August 2007 

Report 13 February 2008 14 February 2008 12 March 2008 

In addition, at about the same time, process-based and facility inspections relevant to this study 

were carried out by the Quality Assurance Unit. 

The inspections were performed in compliance with CIT Quality Assurance Unit procedures and the principles of Good Laboratory Practices . 

The final report is considered to constitute an accurate and complete reflection of the study raw data. 

C'OccC~ Date: -A 5 _70 p, s?czs 

CIT Quality Assurance Unit 
SUMMARY 

-

o .402.,._24t~ 

_ __ . _ _ --m 

- eruaiy 1987) _and EC (92/69%EEC, B.3, 31st July 1992) gui'delines . 

The study was conducted in compliance with the principles of Good Laboratory Practice 

Regulations . . 

Methods 

The test item was applied to the skin of one group of ten Sprague-Dawley rats (five males and 

five females). 

The application was performed with the test item in its original form at the dose-level of 

2000 mg/kg. 

The test site was then covered by a semi-occlusive dressing for 24 hours. 

Clinical signs, mortality and body weight gain were checked for a period of 14 days following 

the single application of the test item. 

All animals were subjected to necropsy. 

Results 

No deaths and no clinical signs were observed during the study. 

A black coloration of the skin was noted in all the animals from day 2 until day 10 and in all the 

males and in one female between day 11 and day 15 (end of the observation period). This 

coloration masked the evaluation of cutaneous reactions in all the animals from day 2 until 

day 6. 

Crusts were observed in 1/5 males and 2/5 females between day 11 and day 15 (end of the 

observation period) . 

When compared to CIT historical control animals, the overall body weight gain of the animals 

was not affected by treatment with the test item. 

No apparent abnormalities were observed at necropsy in any animal . 

Conclusion 

mental - conditions of-this - study, the dermal LDo of 

According to the classification criteria laid down in Council Directive 67/548/EEC (and 

subsequent adaptations), concerning the potential toxicity by dermal route, the test item should 

not be classified. 
RESUME 

senformement aux lignes direcfrices de "1'OCDE~'402, 24 f6vrier 1987) et -e a CEE 

(92/69/CEE, B.3, 31 juillet 1992). 

L'etude a ete realis6e conformement aux r6gles de Bonnes Pratiques de Laboratoire. 

Methode 

Le produit a ete applique sur la peau d'un groupe de 10 rats Sprague-Dawley (5 males et 

5 femelles) . 

L'application a ete effectuee avec le produit tel quel a la dose de 2000 mg/kg. 

Le site de traitement a ensuite dte recouvert d'un pansement semi-occlusif pendant 24 heures. 

Les signes cliniques, la mortalite et 1'evolution ponderale des animaux ont ete suivis pendant une periode de 14 jours apres 1'application unique du produit . 

Un examen anatomopathologique a ete effectue sur tous les animaux . 

Resultats 

Aucun signe clinique ni aucune mortalite ne sont observds pendant 1'etude . 

Une coloration noire de la peau est notee chez tous les animaux du Jour 2 au Jour 10 et chez 

tous les males et 1 femelle entre le Jour 11 et le Jour 15 (fin de la periode d'observation) . Cette coloration a masque l'evaluation des reactions cutanees chez tous les animaux du Jour 2 au Jour 6. 

Des croutes sont observdes chez 1/5 male et 2/5 femelles entre le Jour 11 et le Jour 15 (fin de la periode d'observation) . 

Comparee a celle des animaux temoins historiques du CIT, 1'evolution ponderale des animaux 

West pas influenc6e par le traitement. 

L'examen macroscopique des principaux organes des animaux ne met en gvidence aucune anomalie apparente. 

Conclusion 

cette etude,, . _la -DL 0 dermale du produit 

Selon les criteres de classification dgcrits dans la Directive 67/548/CEE (et ses adaptations) 

concernant la toxicite aigue par voie dermale, le produit ne devrait pas etre classe . 
1 . INTRODUCTION b 

p. 82 

9 

' 

e objective of this study was to evaluate the toxicity of the test item GRAPHISTRENGTH 

Cl 00 micronised following a single dermal application to rats . J 

In the assessment of the toxic characteristics of a test item, determination of acute dermal 

toxicity is an initial step. It provides information on health hazards likely to arise following a 

short-term exposure by the dermal route in humans. 

The study was conducted in compliance with: 

. OECD guideline No. 402, 24th February 1987, 

. EC Directive No. 92/69/EEC, B.3, 31st July 1992. 

2. MATERIALS AND METHODS 

2.1 TEST ITEM 

2.1 .1 Identification 

suppli 

name: 

batch num 

CAS number: 

Description at 

container: one plastic flask 

date of receipt: 29 June 2007 

storage conditions : at room temperature and protected from 

composition : see analytical certificate 

expiry date: June 2008. 

humidity 

Data relating to the characterization of the test item are documented in a test article 

and in an analytical certificate (presented in appendix 1) provided by the Sponsor. 

2.1 .2 Dosage form preparation 

The test item was administered in its original form. 

2.1 .3 Other item 

description 

Purified water (prepared at CIT by reverse osmosis) was used in order to moisten the test item 

and ensure a good contact with the skin. 
2.2 TEST SYSTEM 

2.2 .1 Animals 

p. 83 

10 

Species, strain : rat, Sprague-Dawley Rj : SD (IOPS Han). 

Reason for this choice: rodent species generally accepted by regulatory authorities for this type 

of study. 

Breeder: Janvier, Le Genest-Saint-Isle, France. 

Number and sex: one group of ten animals (five males and five females) . 

Females were nulliparous and non pregnant. 

Age/weight: on the day of treatment, the animals were approximately 8 weeks old and had a 

mean body weight f standard deviation of 287 ± 6 g for the males and 214 f 5 g for the females. 

Acclimation: 4 days before the beginning of the study. 

Identification : individually by earnotches. 

2.2 .2 Environmental conditions 

The conditions in the animal room were set as follows: 

. temperature: 22 f 2°C 

. relative humidity: 30 to 70% 

. light/dark cycle: 12 h/12 h 

. ventilation : approximately 12 cycles/hour of filtered, non-recycled air. 

The temperature and relative humidity were under continuous control and recording. The 

records were checked daily and filed . In addition to these daily checks, the housing conditions 

and corresponding instrumentation and equipment are verified and calibrated at regular 

intervals . 

During the acclimation period, one to seven animals of the same sex were housed in 

polycarbonate cages with stainless steel lid (48 cm x 27 cm x 20 cm). 

During the treatment period, the animals were housed individually in polycarbonate cages with 

stainless steel lid (35.5 cm x 23 .5 cm x 19.3 cm). 

Each cage contained autoclaved sawdust (SICSA, Alfortville, France) . 

Sawdust is analyzed by the supplier for composition and contaminant levels. 

2.2 .3 Food and water 

All the animals had free access to SsniffR/M-H pelleted diet (SSNIFF Spezialdiaten GmbH, 

Soest, Germany). Each batch of food is analyzed by the supplier for composition and 

contaminant levels. 

The diet formula is presented in appendix 2. 

Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum. 

Bacteriological and chemical analyses of water are performed regularly by external laboratories . 

These analyses include the detection of possible contaminants (pesticides, heavy metals and 

nitrosamines). 

No contaminants were known to have been present in the diet, drinking water or bedding 

material at levels which may be expected to have interfered with or prejudiced the outcome of the study. 
2.3 TREATMENT 

2.3 .1 Study design 

As the test item was anticipated to be non-toxic at the dose-level of 2000 mg/kg, a limit test was performed by application of 2000 mg/kg of the test item to one group of ten animals (five males and five females). 

2.3 .2 Preparation of the animals 

On the day before treatment, the dorsal area of each animal was clipped (i.e . approximately 

5 cm x 7 cm for males and 5 cm x 6 cm for females) using an electric clipper. Only animals with healthy intact skin were used for the study. 

2.3 .3 Administration of the test item 

A single dose of 2000 mg/kg of the test item in its original form was placed on a hydrophilic 

gauze pad (pre-moistened with 2 mL of purified water) and then applied to an area of the skin representing approximately 10% of the total body surface of the animals, calculated according to Meeh's formula (1) (i.e . approximately 5 cm x 7 cm for the males and 5 cm x 6 cm for the 

females) . The test item and the gauze pad were held in contact with the skin for 24 hours by 

means of an adhesive hypoallergenic aerated semi-occlusive dressing and a restraining bandage . 

This dressing prevented ingestion of the test item by the animal. On removal of the dressing, any residual test item could not be removed using a moistened cotton pad. 

The dose applied to each animal was adjusted according to the body weight determined on the day of treatment. 

2.4 CLINICAL EXAMINATIONS 

2.4 .1 Clinical signs and mortality 

The animals were observed frequently during the hours following administration of the test item, for detection of possible treatment-related clinical signs. Thereafter, observation of the animals was made at least once a day until day 15. 

Type, time of onset and duration of clinical signs were recorded for each animal individually. 

From day 2, any local cutaneous reaction was recorded. 

2.4 .2 Body weight 

The animals were weighed individually just before administration of the test item on day 1 and then on days 8 and 15 . 

The body weight gain of the treated animals was compared to that of CIT control animals with a similar initial body weight. 

(1) Meeh's formula 

S = k3 p2 

S = total surface of the animal in dm2 

p = weight of the animal in grams 

k = 0.09 for the rat 
2.5 PATHOLOGY 

2.5 .1 Sacrifice 

On day 15, all animals were killed by carbon dioxide asphyxiation. 

2.5 .2 Macroscopic necropsy examination 

p. 85 

12 

All study animals were subjected to a macroscopic examination as soon as possible after death. After opening the thoracic and abdominal cavities, a macroscopic examination of the main organs (digestive tract, heart, kidneys, liver, lungs, pancreas, spleen and any other organs with obvious abnormalities) was performed . 

2 .5 .3 Preservation of tissues 

No organ samples were taken. 

2.6 DATA EVALUATION 

Evaluation of the toxicity of the test item following a single dermal application in rats should include the relationship, if any, between the animals' exposure to the test item and the incidence and severity of all abnormalities including behavioural and clinical abnormalities, macroscopic lesions, body weight changes, mortality and any other toxic effects. 

2.7 ARCHIVING 

The following study materials are archived by CIT, 27005 Evreux, France, for 10 years after the end of the in vivo phase of the study : 

. Study plan and possible amendments, 

. raw data, 

. correspondence, 

. final report and possible amendments. 

On completion of this period, the archived study materials will be returned to the Sponsor, or 

may be archived at CIT for a further period (at additional cost). The total duration of archiving (depending on regulations) will be the responsibility of the Sponsor. 

In addition, raw data not specific to the study including, but not limited to, certificates of analyses for food, water and bedding (if applicable) and records of environmental data and equipment calibration, are also archived by CIT for at least 30 years. 
2.8 CHRONOLOGY OF THE STUDY 

The chronology of the study was as follows : 

Procedure Date 

Experimental starting date (treatment) 21 August 2007 

Experimental completion date (necropsy of the last animal) 04 September 2007 

2.9 STUDY PLAN ADHERENCE 

The study was performed in accordance with Study plat'l~~J!~!~111U~~ subsequent 

amendments, with the following deviations from the agreed Study plan : 

. the temperature and relative humidity recorded in the animal room were sometimes outside of the target ranges specified in the Study plan, 

. the acclimation of the animals was 4 days (instead of at least 5 days) before the beginning of 

the study. 

These minor deviations were not considered to have compromised the validity or integrity of the study. 
3. RESULTS 

3.1 CLINICAL EXAMINATIONS 

3.1 .1 Mortality (table 1) 

No deaths occurred during the study. 

3 .1 .2 Clinical signs and cutaneous reactions (tables 1 and 2) 

p. 87 

14 

No clinical signs were observed during the study. 

A black coloration of the skin was noted in all the animals from day 2 until day 10 and in all the 

males and in one female between day 11 and day 15 (end of the observation period) . This 

coloration masked the evaluation of cutaneous reactions in all the animals from day 2 until day 6. 

Crusts were observed in 1/5 males and 2/5 females between day 11 and day 15 (end of the 

observation period) . 

3 .1 .3 Body weight (figure 1, tables 3 and 4) 

When compared to CIT historical control animals, the overall body weight gain of the animals 

was not affected by treatment with the test item. 

3.2 PATHOLOGY (table 5) 

Macroscopic examination of the main organs of the animals revealed no apparent abnormalities . 

4. CONCLUSION 

Under the experimental conditions of this study, the dermal LDo of the test item 

II~~ 000 mg/kg in rats . 
. . ~. --~. 

Figure 1 : Mean body weight of treated rats 

400 . 

380 

360 

340 

320 

300 

280 

260 

240 

220 

200 

(9) 

O 

days 

4 5 6 7 8 9 10 11 12 13 14 15 

-s-Male -O-Female 

p. 88 

15 
Table 1 : Individual clinical signs and mortality 

Dose-level Time Animals Mortality Clinical signs 

(mg/kg) Males Females 

2000 30 min 

2 h - 5 h ~ 01-02-03-04-05 06-07-08-09-10 No None 

D2toD15J 

min : minutes 

h :hour 

D : day 
Table 2: Cutaneous reactions 

Dose-level Time Animals Cutaneous reactions 

(mg/kg) Males Females 

2000 D 2 to D 6 01-02-03-04-05 06-07-08-09-10 Black coloration of the skin 

which could have masked a 

possible erythema 

D 7 to D 10 01-02-03-04-05 06-07-08-09-10 Black coloration of the skin 

No cutaneous reactions 

D 11 06-10 Crusts 

01-02-03-04-05 07 Black coloration of the skin 

No cutaneous reactions 

08-09 None 

D 12 and D 13 03 Crusts, black coloration of the 

skin 

06-10 Crusts 

01-02-03-04-05 07 Black coloration of the skin 

No cutaneous reactions 

08-09 None 

D 14 01-02-03-05 07 Black coloration of the skin 

No cutaneous reactions 

06-10 Crusts 

04 08-09 None 

D 15 01-02-03-05 07 Black coloration of the skin 

No cutaneous reactions 

06 Crusts 

04 08-09-10 None 

D: day 
Table 3 : Individual and mean body weight and weekly body weight change (g) 

Dose-level Days 

Sex Animals 

mg/kg 1 (1) 8 (1) 15 

2000 Male 01 295 52 347 53 400 

02 280 49 329 44 373 

03 287 47 334 53 387 

04 288 50 338 49 387 

05 284 50 334 51 385 

M 287 50 336 50 386 

SD 6 2 7 4 10 

2000 Female 06 214 17 231 19 250 

07 220 16 236 21 257 

08 206 19 225 20 245 

09 214 15 229 14 243 

10 218 19 237 26 263 

M 214 17 232 20 252 

SD 5 2 5 4 8 

(1) = body weight gain 

M = mean 

SD = standard deviation 
Table 4: Body weight - CIT historical data of control animals dosed by dermal route 

Check of the body weight (g) of CIT historical control animals 

Method : Acute dermal administration 

CIT Studies / Experimental : CIT/Study Nos. 27220 RDR / February 2004 

Completion date 29692 RDR / April 2005 

31105 RDR / January 2006 

Reference item : Purified water 

Volume of administration : 5 mL/kg 

Animals 

Species, strain : Rat, Sprague-Dawley Rj : SD (IOPS Han) 

Breeder : Janvier, Le Genest-Saint-Isle, France 

Age on day 1 : 8 weeks 

Volume 

(mL/kg) Sex 1 (1) 

Days 

8 (1) 15 

5 Male M 333 46 380 48 428 

SD 36 14 32 9 34 

n 29 29 29 29 29 

5 Female M 216 24 241 19 260 

SD 9 11 14 9 16 

n 29 29 29 29 29 

M : mean 

(1) : body weight gain 

SD : standard deviation 

n : number of animals 

p. 92 

19 

460 

440 

(420 ' 

400 

380 

360 

340 

320 

300 

280 

260 

240 

220 days 

200 

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 

+ -Male --A--Female 
Table 5 : Individual macroscopic examinations at necropsy 

Dose-level Time Animals Macroscopic 

(mg/kg) Males Females abnormalities 

2000 D 15 01-02-03-04-05 06-07-08-09-10 No apparent abnormalities 

D: day 
APPENDICES 
1 . Test article description and analytical certificate 
TEST ARTICLE DESCRIPTION 

PHYSICAL AND CHEMICAL PROPERTIES 

Appearance : black powder 

Particule size : 30 jim ( approximately) 

Specific Gravity : 2,1 kg/m3 at 20°C 

Autoignition temperature : > 400 °C (standard : NF EN 50281-2-1) 

Solubility : in water : insoluble 

TOXICOLOGICAL INFORMATIONS AND USE SAFETY 

I See Safety Data Sheet. 

I STORAGE AND DISPOSAL 

Storage : Keep hermetically closed in a dry, cool and well-ventiled 

place . 

Expiry date : June 2008 

~ Disposal : Incineration 

p. 96 

23 
p. 97 

24 

Reference de la m6thode 

d'analyse l Analysis 

reference 

Nom du responsable du laboratoirel Laboratory Director : 

Signature 
2. Diet formula 
SsniffRIM-H 

Complete diet for rats/mice - maintenance 

Crude proteins 

Crude fat 

Crude fiber 

Crude ash 

Amino Acids 

Lysine 

Methionine 

Cystine 

Glycine 

Leucine 

Isoleucine 

Arginine 

Phenylalanine 

Tryptophan 

Histidine 

Tyrosine 

Aspartic acid 

Glutaminic acid 

Valine 

Threonine 

Trace elements (je 

Manganese 

Copper 

Zinc 

Iodine 

Iron 

Selenium 

Cobalt 

19.00% 

3.30% 

4.90% 

6.70% 

1 .00% 

0.30% 

0.30% 

0.90% 

1 .30% 

0.70% 

1 .20% 

0.90% 

0.25% 

0.50% 

0.60% 

1 .70% 

3 .80% 

0 .90% 

0.70% 

Constituents 

Calcium 

Phosphorus 

Sodium 

Magnesium 

Potassium 

Vitamins Qe kg) 

A 

D3 

E 

B1 

B2 

B6 

B12 

Biotin 

Pantothenic acid 

Choline 

Folic acid 

Nicotic acid 

K3 

Inositol 

ME (je kg) 

1 .00% 

0.70% 

0.25% 

0 .20% 

0.90% 

kg) 

90 mg 

12 mg 

75 mg 

2 mg 

220 mg 

0.2 mg 

2 mg 

Item numbers 

V1530 Meal 

V1531 Micromeal 

V1534 Pellets 10 mm 

V1535 Pellets 15 mm 

15,000 IE 

1,000 IE 

100 mg 

10 mg 

20 mg 

12 mg 

80 ltg 

400 pg 

30 mg 

1,600 mg 

4 mg 

60 mg 

5 mg 

50 mg 

12 .2 MJ 
3 . CIT GLP certificate 
"&.u - Auaaa . rMWrAuE 

Ittiruar1CWe FxamJ~ 

GROUPE I1VTERIYIINISTERIEL DES PRODUITS CHIMIQUES 

Paris, le 

Objet : Evaluation de la conformite aux Bonnes Pratiques de Laboratoires (BPL) selon les directives 

2004/9/CE et 2004/10/CE du 11 f&vrier 2004. 

Subject : Assessment of compliance with Good Laboratory Practices (GLP) under the EC directives 

2004/9 and 2004/10 of 11 February 2004. 

ConsBcutivement A votre engagement vis-A-vis du GIPC et du COFRAC et en application du d6cret n° 2006-

1523 du 4 d6cembre 2006 concernant les bonnes pratiques de laboratoires et modifiant le d6cret n° 81-278 du 25 

mars 1981 portant creation d'un groupe interministbriel des produits chimiques, je vous confirme que le GIPC, 

au vu des rdsultats du controle exere6 par le Comite frangais d'accr6ditation (COFRAC - Section Laboratoires a 

d8cid6 pour votre installation du statut suivant : 

Following your engagement vis-a-vis the GIPC and COFRAC and in application of the decree n'2006-1523 of 

4 December 2006 relating to the good laboratoiy practices and modifying the decree n° 81-278 of 25 March 

1981 giving birth to an interministerial group of chemical products (GIPC), I confirm to you that the GIPC, 

given the results of the inspection realised by the French Committee of accreditation (COFRAC - Laboratory 

Section has taken the following decision relating to your installation : 

Respect des principes de BPL Date d'inspection : 7-8 mars 2007 

Respect of the GLP principles Date of inspection : 7-8 mars 2007 

Domaines de reconnaissance ; Inspection de renouvellement (i.r) 

1- essais physico-chimiques Renewal inspection (i.r) 

2 - 6tudes de toxicit6 

3 - 6tudes de mutagbnicite Date de decision du GIPC : 29 juin 2007 

4 - 6tudes 6cotoxicologiques sur les Date of GIPC decision : 29 juin 2007 

organismes aquatiques et terrestres 

8 - m6thodes de chimie analytique et Date de prise d'effet : 8 mars 2007 

clinique Date of implementation: 8 mars 2007 

Areas of expertise : Annee de premi8re conformitd : 1989 

1=Physico-chemical testing Year of the first conformity: 1989 

2 = Toxicity studies 

3 = Mutagenicity studies Durde de validit6: 18 mois 

4 = Environmental toxicity studies on aquatic or Time of validity: 18 months 

terrestrial organisms 

8 =Analytical and clinical chemistry 

: 

CENTRE INTERNATIONAL DE TOXICOLOGIE 

(CIT) 

MISEREY - BP 563 

27005 EVREUX CEDEX 

- 9 JUIi. 2007 

Pierre CkEY'SSEY, 

Conseiller d'Etat h 

Secrbtariat general du GIPC - DGE- Simap - 12, rue Villiot - 75572 Paris cedex 12 

Telephone : 0153 44 96 10 -T616copie : 01 53 44 9172 
p. 102 

CONFIDENTIAL 

STUDY TITLE 

ACUTE EYE IRRITATION 

IN RABBITS 

STUDYDIRECTOR 

Catherine Pelcot 

DATE OF ISSUE 

"Q3,- pcecle 

TEST FACILITY 

CIT 

BP 563 - 27005 Evreux - Prance 

IFM Recherche 

S .N.C au Capita! de 846,092 ¬ 

788 060 465 R.C.S . Evreux 

VAT ER ! 9 788 060 465 

Agremenz Credit Impet Recherche 

LAB O UMBER 

B.P 563 27005 Evreux Cedex France 

T6. : +33 (0)2 32 29 26 26 

Fax : +33 (0)2 32 67 87 05 

E-mail : CIT@citox.com 

Web site : www.citox .com 

T6j 
CONTENTS 

STATEMENT OF THE STUDY DIRECTOR 3 

OTHER CIT SCIENTIST INVOLVED IN THE STUDY 4 

STATEMENT OF QUALITY ASSURANCE UNIT 5 

SUMMARY 

RESUME 

6 

7 

1 . INTRODUCTION 8 

2. MATERIALS AND METHODS 8 

2.1 TEST ITEM 8 

2.1 .1 Identification 8 

2.1 .2 Dosage form preparation 8 

2.2 TEST SYSTEM 

2 .2 .1 Animals 

2 .2 .2 Environmental conditions 

2.2 .3 Food and water 

2.3 TREATMENT 10 

2.3 .1 Selection of the animals 10 2.3 .2 Administration of the test item 10 

2.4 OCULAR EXAMINATIONS 10 

2.4 .1 Duration of the observation period 10 

2.4 .2 Description and evaluation of ocular reactions 10 

2 .5 BODY WEIGHT 11 

2.6 INTERPRETATION OF RESULTS 11 

2.7 ARCHIVING 12 

2.8 CHRONOLOGY OF THE STUDY 12 

2.9 STUDY PLAN ADHERENCE 12 

3. RESULTS 13 

4. DISCUSSION 13 

5 . CONCLUSION 13 

Table 1 : Individual ocular examinations and mean values of the scores recorded for each 

animal (24, 48 and 72 hours) 14 

Table 2 : Individual body weight (g) 16 

APPENDICES 17 

1 . Test article description and analytical certificate 18 

2. Diet formula 21 

3. CIT GLP certificate 23 and 24 
STATEMENT OF THE STUDY DIRECTOR 

The study was performed in compliance with the principles of Good Laboratory Practice as 

described in: 

. OECD Principles of Good Laboratory Practice (as revised in 1997), ENV/MC/CHEM 

(98) 17 and all subsequent OECD consensus documents. 

. Directive 2004/10/EC of the European Parliament and of the Council of 11 February 2004 on 

the harmonization of laws, regulations and administrative provisions relating to the 

application of the Principles of Good Laboratory Practice and the verification of their 

applications for tests on chemical substances (OJ No. L50 of 20.2.2004) . 

. Decret N° 2006-1523 du 04 d6cembre 2006 concernant les Bonnes Pratiques de Laboratoire 

(Journal Officiel du 06 d6cembre 2006), Ministere de 1'Economie, des Finances et de 

1'Industrie. 

The study was also conducted in compliance with the following Animal Protection regulations : 

. Council Directive 86/609/EEC of 24th November 1986 on the harmonization of laws, regulations or administrative provisions relating to the protection of animals used for 

experimental or other scientific purposes. 

. Guidance document on the recognition, assessment, and use of clinical signs as humane endpoints for experimental animals used in safety evaluation, OECD Environmental Health 

and Safety Publications, No. 19. 

I declare that this report constitutes a true and faithful record of the procedures undertaken and the results obtained during the performance of the study. 

This study was performed at CIT, BP 563, 27005 Evreux, Fran 

Toxicology 

/ 

- 

C. Pelcot Study completion/date : J1 [A Fn L j Study Director , 
OTHER CIT SCIENTIST INVOLVED IN THE STUDY 

Director of Toxicology 

CIT Management 

J.J. Legrand Date: 

Doctor of Veterinary Medicine 
STATEMENT OF QUALITY ASSURANCE UNIT 

Inspections performed at CIT: 

The CIT Quality Assurance Unit conducted the inspections detailed below: 

Type of inspection 

Dates 

Inspection Reported to the Reported to 

Study Director Management 

Study plan 27 July 2007 27 July 2007 02 August 2007 

Report 25 March 2008 27 March 2008 08 April 2008 

In addition, at about the same time, process-based and facility inspections relevant to this study 

were carried out by the Quality Assurance Unit. 

The inspections were performed in compliance with CIT Quality Assurance Unit procedures and the principles of Good Laboratory Practices . 

The final report is considered to constitute an accurate and complete reflection of the study raw data. 

A . ac- e.e: c~cc.> Date: .2 3 TIP, 2,:7c-,a 

CIT Quality Assurance Unit 
SUMMARY 

p. 107 

6 

g 

29th April 2004) guidelines. 

The study was conducted in compliance with the principles of Good Laboratory Practice 

Regulations . 

Methods 

The test item was first administered to a single male New Zealand White rabbit. Since the 

test item was irritant on this first animal, but without persistence until the end of the observation 

period, it was then evaluated sequentially in two other animals. 

A single dose of the test item in its original form was introduced into the left conjunctival sac. 

The right eye was not treated and served as control . 

The eyes were not rinsed after administration of the test item. 

Ocular reactions were observed approximately 1 hour, 24, 48 and 72 hours after the 

administration and then daily until complete reversibility of the ocular reactions (day 9 at the 

latest) . 

The mean values of the scores for chemosis, redness of the conjunctiva, iris lesions and corneal 

opacity were calculated for each animal. 

Results 

A slight to marked chemosis was noted in all the animals from day 1 until day 7 (1/3 animals) or 

8 (2/3 animals) . A slight to severe redness of the conjunctiva was observed in all the animals 

from day 1 until day 5 (1/3 animals) or 8 (2/3 animals) . A clear discharge was observed in all 

the animals from day 1 until day 2, 5 or 6. 

An iritis was noted in all the animals on day 2 ; it persisted in one animal until day 4. 

A slight or moderate corneal opacity was recorded in all the animals on days 2 and 3; it persisted 

until day 5 in 2/3 animals. This corneal opacity covered the whole area of the cornea of 

1/3 animals on day 2. 

Mean scores calculated for each animal over 24, 48 and 72 hours were 2.7, 2.3 and 2.7 for 

chemosis, 2.0, 1 .7 and 2.7 for redness of the conjunctiva, 0.3, 0.3 and 1 .3 for iris lesions and 2 .0, 

1 .0 and 2.0 for corneal opacity . 

Conclusion 

According to the classification criteria laid down in Council Directive 67/548/EEC (and 

subsequent adaptations), the test item should be considered as irritant to eyes. 
p. 108 

7 

RESUME 

e Lapin ont ete dva uees se 

CEE (92/69/CEE, B.5, 31 juillet 1992). 

L'etude a et6 realisee conformgment aux regles de Bonnes Pratiques de Laboratoire . 

Methode 

Le produit a d'abord dte applique sur un seul Lapin male New Zealand White. Le produit ayant montre des proprietgs irritantes sur ce premier animal mais sans persistance jusqu'a la fin de la periode d'observation, il a ensuite ete teste sdquentiellement sur deux autres animaux. 

Une dose unique de produit tel quel a ete introduite dans le cul-de-sac conjonctival de Neil gauche. L'oeil droit a servi de temoin. 

Aucun ringage des yeux n'a ete realise apres 1'administration du produit. 

Les reactions oculaires ont ete observees environ l, 24, 48 et 72 heures apres I'administration puis quotidiennement jusqu'a reversibilite complete des lesions oculaires observees (jour 9 au plus tard). 

La moyenne des scores pour le chemosis, la rougeur de la conjonctive, les lesions de 1'iris et l'opacite de la cornee a etd calculde pour chaque animal. 

Resultats 

Un chemosis leger A marque est note chez tous les animaux du jour 1 aux jours 7 (1/3 animal) ou 8 (2/3 animaux). Une rougeur de la conjonctive legere a grave est observee chez tous les animaux du jour 1 aux jours 5 (1/3 animal) ou 8 (2/3 animaux). Un larmoiement clair est observe chez tous les animaux du jour 1 aux jours 2, 5 ou 6. 

Une inflammation de 1'iris est notee chez tous les animaux au jour 2 ; elle persiste chez 1 animal jusqu'au jour 4. 

Une opacite corngenne legere ou moderge est observee chez tous les animaux aux jours 2 et 3 ; elle persiste jusqu'au jour 5 chez 2/3 animaux. Cette opacite corneenne a couvert la totalite de la cornee d'1/3 animal au jour 2. 

La moyenne des scores enregistres pour chaque animal apres 24, 48 et 72 heures est de 2,7 ; 2,3 et 2,7 pour le chemosis, 2,0 ; 1,7 et 2,7 pour la rougeur de la conjonctive, 0,3 ; 0,3 et 1,3 

pour les lesions de 1'iris et 2,0 ; 1,0 et 2,0 pour 1'opacite corneenne. 

Conclusion 

Selon les criteres de classification decrits dans la Directive 67/548/CEE (et ses adaptations), le produit est considere comme irritant pour les yeux. 
1. INTRODUCTION 

In the assessment of the toxic characteristics of a test item, determination of the irritant effects 

on the eyes of mammals is an important initial step . Information derived from this test serves to 

indicate the possible hazards likely to arise from exposure of the eyes, ~ and associated mucous 

membranes, to the test item. 

This study was conducted in compliance with: 

. OECD guideline No. 405, 24th April 2002, 

. EC Directive No. 2004/73/EC, part B .5, 29th April 2004. 

2. MATERIALS AND METHODS 

2.1 TEST ITEM 

2.1 .1 Identification 

Description at receipt: -blacZ powder 

container: one plastic flask 

date of receipt : 29 June 2007 

storage conditions: at room temperature and protected from humidity 

composition : see analytical certificate 

expiry date: June 2008. 

Data relating to the characterization of the test item are documented in a test article description 

and in an analytical certificate (presented in appendix 1) provided by the Sponsor. 

2.1 .2 Dosage form preparation 

The test item was used in its original form. 

The pH of the test item at the concentration of 1% in purified water was not measurable (non soluble and colored test item). 
2.2 TEST SYSTEM 

2.2 .1 Animals 

p. 110 

9 

Sex, species, strain: male New Zealand White rabbits. 

Reason for this choice: species generally accepted by regulatory authorities for this type of study. 

Breeder: Grimaud freres selection S .A.S., La Corbiere, Roussay, France 

Number: three animals were used, as recommended by the international guidelines. 

Age/weight: on the day of treatment, the animals were 2 to 4 months old and had a mean body weight f standard deviation of 2.9 f 0.1 kg. 

Acclimation : at least 5 days before the beginning of the study. 

Identification : individual metal ear tag . 

2.2 .2 Environmental conditions 

The conditions in the animal room were set as follows : 

. temperature: 18 ± 3°C 

. relative humidity: 30 to 70% 

. light/dark cycle: 12 h/12 h 

. ventilation : approximately 12 cycles/hour of filtered, non-recycled air. The temperature and relative humidity were under continuous control and recording. The records were checked daily and fled. In addition to these daily checks, the housing conditions and corresponding instrumentation and equipment were verified and calibrated at regular intervals. 

The animals were housed individually in Techniplast (64 cm x 63 cm x 30 cm) or Pajon (50 cm 

x 57 cm x 75 cm) cages. 

Each cage was equipped with a food container and a water bottle. 

2.2 .3 Food and water 

During the study, the animals had free access to 110C pelleted diet (SAFE, Villemoisson, Epinay-sur-Orge, France). 

Food is analyzed regularly by the supplier for composition and contaminant levels. The diet formula is presented in appendix 2. 

Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum . Bacteriological and chemical analyses of water are performed regularly by external laboratories . These analyses include the detection of possible contaminants (pesticides, heavy metals and nitrosamines) . 

No contaminants were known to have been present in the diet or drinking water at levels which 

may be expected to have interfered with or prejudiced the outcome of the study. 
2.3 TREATMENT 

2.3 .1 Selection of the animals 

p. 111 

10 

Just before treatment, the eyes of each animal were examined in order to check the absence of 

any signs of ocular irritation, ocular defects or pre-existing corneal injury . 

2.3 .2 Administration of the test item 

The test item was first administered to a single animal (No. 474). Since the test item was irritant, but without persistence at the end of the observation period, on this first animal, it was then evaluated sequentially on other animals (No. 483 then No. 484). 

A single dose (the maximal quantity of the test item administrable into the eye was approximately 1/3 of the 100 mg required quantity) of the test item in its original form was introduced into the conjunctival sac of the left eye after gently pulling the lower lid away from the eyeball . 

The lower and upper eyelids were held together for about one second to avoid any loss of 

test item. The right eye, which remained untreated, served as control . 

The eyes were not rinsed after administration of the test item . 

2.4 OCULAR EXAMINATIONS 

2.4 .1 Duration of the observation period 

The eyes were examined approximately 1 hour, 24, 48 and 72 hours after administration of the 

test item. 

Since there were persistent ocular reactions at 72 hours, the observation period was extended up to their complete reversibility (day 9 at the latest) . 

2 .4 .2 Description and evaluation of ocular reactions 

Conjunctival reactions, iritis and corneal opacification were evaluated daily for each animal. For the evaluation of corneal opacification (presence or absence, affected area), the eyes were examined under a UV lamp after instillation of one or two drops of 0.5% sodium fluorescein solution (a clear fluorescence is visible in the areas of opacification) . This evaluation was performed on day 2 and repeated thereafter whenever necessary. 

Ocular reactions were scored according to the following numerical scale: 

Coniunctival lesions and discharge 

Chemosis (lids and/or nictitating membranes) 

. no swelling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0 

. any swelling above normal (includes nictitating membranes). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . _ . . . . . . . . . . . . . 1 

. obvious swelling with partial eversion of lids . . . . . . . . . . . . . . . . . . . . . . . . ., . . . ., ., . ., . . . . ., .,_ . . . . . . . . . . . . . . ., . . . . . . .2 

. swelling with lids about half-closed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3 

. swelling with lids more than half-closed . . . . . . . . . . . . . . . . . . . . . ., . . ., . ., . . _ , . . . . . ., ., . . . . .,_ . ._ . ., . . . . ., . . . . . . .4 

Redness (refers to palpebral and bulbar conjunctivae, cornea and iris) 

. blood vessels normal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .0 

. a number of blood vessels definitely hyperemic (injected). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 

. diffuse, crimson colour, individual vessels not easily discernible . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 

. diffuse, beefy red . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3 
p. 112 

11 

Discharge 

. absence of discharge . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .0 

. slight discharge (does not include small amounts normally found in 

inner canthus) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1 

. discharge with moistening of lids and hairs adjacent to lids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 

. discharge with moistening of lids and hairs on wide area around the eye . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 

Iris lesions 

. normal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .0 

. markedly deepened rugae, congestion, swelling, moderate circum-corneal 

hyperemia, or injection, any of these or combination of any thereof, iris still 

reacting to light (sluggish reaction is positive) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1 

. no reaction to light, haemorrhage, gross destruction (any or all of these) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2 

Corneal lesions 

Degree of opacity (area most dense taken for reading) 

. no ulceration or opacity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .0 

. scattered or diffuse areas of opacity (other than slight dulling or normal lustre), 

details of iris clearly visible . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1 

. easily discernible translucent area, details of iris slightly obscured . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2 

. nacreous areas, no details of iris visible, size of pupil barely discernible . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3 

. opaque cornea, iris not discernible through the opacity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4 

Area of opacity 

. one quarter (or less) but not zero . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . l 

. greater than one quarter but less than a half. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2 

. greater than one half but less than three quarters . . . . . . . . . . . . . . . . . . . . . . . . . . . . ., . . . . . . . . . . . . . . . . . . . . . . ., . . . . . . . . . . . . . . .3 

. greater than three quarters up to whole area. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4 

Any other lesions observed were noted. 

2.5 BODY WEIGHT 

Each animal was weighted at the beginning (day of treatment) and at the end of the observation 

period. 

2.6 INTERPRETATION OF RESULTS 

The results obtained were evaluated in conjunction with the nature and the reversibility of the 

scores observed, whilst taking into account all the reactions of the treated animals. 

Criteria for irritation 

A substance or a preparation is considered irritant for the eyes if, when applied to the eye of the 

animal, significant or severe ocular lesions are caused within 72 hours after exposure and which 

persist for 24 hours or more after treatment with the test item. 

All the scores at each reading time (24, 48 and 72 hours) and for an effect are used for 

calculating the respective mean values. 
2.7 ARCHIVING 

The following study materials are archived by CIT, 27005 Evreux, France, for 10 years after the end of the in vivo phase of the study: 

. Study plan and possible amendments, 

. raw data, 

. correspondence, 

. final report and possible amendments. 

On completion of this period, the archived study materials will be returned to the Sponsor, or may be archived at CIT for a further period (at additional cost) . The total duration of archiving (depending on regulations) will be the responsibility of the Sponsor. In addition, raw data not specific to the study including, but not limited to, certificates of analyses for food, water and bedding (if applicable) and records of environmental data and equipment calibration, are also archived by CIT for at least 30 years. 

2.8 CHRONOLOGY OF THE STUDY 

The chronology of the study is summarized as follows : 

Procedure 

Experimental starting date 

(day of treatment of the first animal) 

Experimental completion date 

(end of the observation period) 

Date 

14 August 2007 

07 September 2007 

2.9 STUDY PLAN ADHERENCE 

The study was performed in accordance with 

amendments, with the following deviation from trn-~-agreed Study plan: 

. the temperature and relative humidity recorded in the animal room were sometimes outside of the target ranges specified in the Study plan. 

This minor deviation was not considered to have compromised the validity or integrity of the study . 
3. RESULTS 

The observations recorded during the study are presented in table 1 . 

The body weight is presented in table 2. 

A slight to marked chemosis (grades 1 to 3) was noted in all the animals from day 1 until day 7 (1/3 animals) or 8 (2/3 animals). A slight to severe redness of the conjunctiva (grades 1 to 3) was observed in all the animals from day 1 until day 5 (1/3 animals) or 8 (2/3 animals). A clear discharge was observed in all the animals from day 1 until day 2, 5 or 6. An iritis of grade 1 was noted in 2/3 the animals on day 2. In the third animal, an iritis of grade 2 was noted on day 2; an iritis of grade 1 persisted until day 4. 

A slight or moderate corneal opacity (grade 1 or 2) was recorded in all the animals on day 2; it persisted until day 3 (1/3 animals) or 5 (2/3 animals) . This corneal opacity covered the whole area of the cornea of 1/3 animals on day 2 . 

Mean scores calculated for each animal over 24, 48 and 72 hours were 2.7, 2.3 and 2.7 for chemosis, 2.0, 1 .7 and 2 .7 for redness of the conjunctiva, 0.3, 0.3 and 1 .3 for iris lesions and 2 .0, 1 .0 and 2.0 for corneal opacity. 

4. DISCUSSION 

The grade 2 observed in one animal only on day 2 seems doubtful and not coherent with the grades 1 noted the next days. 

5. CONCLUSION 
Table 1 : Individual ocular examinations and mean values of the scores recorded for each animal 

(24, 48 and 72 hours) 

Rabbit Region Description Scores Mean Interpretation 

number of eye of ocular irritation (+) 

reactions Ih 24h 48h 72h score (1) (-) 

D1 D2 D3 D4 

474 Conjuncfivae Chemosis 3 3 3 2 2.7 (+) 

Redness 2 2 2 2 2.0 (-) 

Discharge 1 2 0 0 0 .7 

Iris 0 1 0 0 0 .3 (-) 

Corneal opacity Intensity 0 2 2 2 2.0 (+) 

Area 0 4 3 1 2.7 

Other Su Su Su Su 

Fluoresoein / U U U 

483 Conjunctivae Chemosis 3 3 2 2 2 .3 (+) 

Redness 2 2 2 1 1,7 (_) 

Discharge 1 2 1 1 1 .3 

Iris 0 1 0 0 0.3 (-) 

Corneal opacity Intensity 0 2 I 0 1 .0 (-) 

Area 0 3 1 0 1 .3 

Other Su Su * * 

Fluorescein / U U U 

484 Conjunctivae Chemosis 3 3 3 2 2.7 (+) 

Redness 2 3 3 2 2.7 (+) 

Discharge 1 1 1 I 1 .0 

Iris 0 2 1 1 1 .3 (+) 

Corneal opacity Intensity 0 2 2 2 2.0 (+) 

Area 0 3 3 2 2.7 

Other Su Su * * 

Fluorescein / U U U 

(1) mean of scores on days 2, 3 and 4 

h=hour 

D =day 

(+) = irritant according to E.E .C. criteria 

(-) =non-irritant according to E.E .C. criteria 

* = none 

U = fluorescein batch Nos. L239 and L653 

/ = fluorescein not used 

Su = residual test item 
Table 1 (continued) 

Rabbit Region Description Scores 

number of eye of ocular 

reactions 

D5 D6 D7 D8 D9 474 Conjunctivae Chemosis 2 2 1 1 0 

Redness 2 2 1 1 0 

Discharge 0 0 0 0 0 

Iris 0 0 0 0 0 

Corneal opacity Intensity 1 0 0 0 0 

Area 1 0 0 0 0 

Other * * * * 

Fluorescein U U 

483 Conj unctivae Chemosis I I 1 0 

-

Redness 1 0 0 0 - 

Discharge 1 1 0 0 

Iris 0 0 0 0 

Corneal opacity Intensity 0 0 0 0 

Area 0 0 0 0 

Other * * * * _ 

Fluorescein 

484 Conjunctivae Chemosis 2 2 2 1 0 

Redness 1 I 1 l 0 

Discharge 1 0 0 0 0 

Iris 0 0 0 0 0 

Corneal opacity Intensity 1 0 0 0 0 

Area 1 0 0 0 0 

Other * * * * * 

Fluorescein U U 

D =day 

* = none 

U = fluorescein batch Nos. L239 and L653 

/ = fluorescein not used 

- = ocular examination not performed 
Table 2: Individual body weight (g) 

Sex Animals Day of End of the 

treatment observation period 

Male 474 2778 2993 

483 2780 2914 

484 3015 3140 

M 2858 3016 

SD 136 115 

M = mean 

SD = standard deviation 
p. 118 

17 

APPENDICES 
1 . Test article description and analytical certificate 
T~t article name 

Chemical narne 

CAS number 

UNECS number 

Qrigill 

Batch number 

ftling n 

F 

eci tic C`armFi.ty 

ASIt!f31gn1ttC3'Ll tei31pelfUt7%2 

: black powd2t 

; 30p,m ( apprnximataly ) 

: 2,1~ kOjM3 at 2(}'C 

`%' 400 °C (Standard : 'e``IF E't~ .'r>E}7$1-2-1 ) 

Solubility . in wawC : !I?54l1iblo 

: Keep hermetically closed in e dry, coot and wtil-ventited 

place . I JUDO 

Incineration 
DETERMINATION I ITEM 

Reference de la m6thode 

RESULTAT 1 RESULT d'analyse 1 Analysis 

reference 

Powder characteristics 

Ash content % 7.6% ATG 

76 WW ei h-in ._ ., . 

Nom du responsable du laboratoirel Laboratory Director : 
2. Diet formula 
p. 123 

22 

Ref 110 

COMPLETE DIET 

RABBIT BREEDING DIET 

Appearance: 3 mm diameter granules 

Conditioning: bags of 20 kgs 

Daily portion : Rabbits 150 g, water ad libitum . 

FORMULA % 

Cereals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Grain biproducts and 

leguminous plants . . . . . . . . . . . . . . . . . . . . . 

Vegetable protein (soya bean 

meal, yeast) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Vitamin and mineral mixture . . . 

AVERAGE ANALYSIS % 

Calorific value (Kcal/kg) . . . . . . . . . . 

Moisture. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Lipids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Carbohydrates (N.F.E.) . . . . . . . . . . . . 

Fibre. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Minerals (ash) . . . . . . . . . . . . . . . . . . . . . . . . . . . 

AMINO ACID VALUES 

(calculated in mg/kg) 

Arginine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Cystine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Lysine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Methionine. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Tryptophan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Glycine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

FATTY ACID VALUES 

(calculated in mg/kg) 

Palmitic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Palmitoleic acid . . . . . . . . . . . . . . . . . . . . . . . . 

Stearic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Oleic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Linoleic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Linolenic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . 

33.8 

48 

14 

4.2 

3100 

10 

15 

2.3 

48.2 

17 

7.5 

11300 

3400 

9300 

2800 

2400 

8700 

6400 

0 

600 

6400 

12100 

2400 

MINERALS (calculated in mg/kg) 

Nat. CMV 

val. val. Total 

----------- ------ P . . . . . . . . . . . . . . . . . . . . . 3600- ------ 

2900 6500 

Ca . . . . . . . . . . . . . . . . . . 4200 5800 10000 

K . . . . . . . . . . . . . . . . . . . . 12000 0 12000 

Na . . . . . . . . . . . . . . . . . . 400 2000 2400 

Mg . . . . . . . . . . . . . . . . . . 2500 100 2600 

Mn . . . . . . . . . . . . . . . . . 50 40 90 

Fe . . . . . . . . . . . . . . . . . . . 150 150 300 

Cu . . . . . . . . . . . . . . . . . . Traces 15 15 

Zn . . . . . . . . . . . . . . . . . . 30 45 75 

Co . . . . . . . . . . . . . . . . . . 0.1 1 .5 1 .6 

1 . . . . . . . . . . . . . . . . . . . . . 0.1 0 0.1 

Cl . . . . . . . . . . . . . . . . . . . 300 3000 3300 

VITAMINS (calculated per kg) 

Nat. CMV 

--- val. _------val.----- .-- Total _ .-- 

Vitamin A Traces 10000 IU 10000 IU 

Vitamin D3 0 Ii 1000 IU 1000 fu 

Vitamin Bl 5 mg 0 mg 5 mg 

Vitamin 132 4 mg 0 mg 4 mg 

Vitamin 133 20 mg 0 mg 20 mg 

Vitamin 136 1 mg 1 mg 2 mg 

Vitamin B12 0 mg 0 mg 0 mg 

Vitamin E 15 mg 25 mg 40 mg 

Vitamin K3 0 mg 1 mg 1 mg 

Vitamin PP 60 mg 5 mg 65 mg 

Folic acid 0 mg 0 mg 0 mg 

Biotin 0 mg 0 mg 0 mg 

Choline 1000 mg 1000 mg 2000 mg 

Available under quality "Control Ref. : I 10" 

SAFE, 7 rue Gallidni, Villemoisson, 91360 Epinay-sur-Orge 

Tel: 01 .69 .04.03 .57 - Fax : 01 .69.04.81 .97 

(Ref. Doe. UAR: 2000) 
3 . CIT GLP certificate 
fdLerM - l3gilPtF - Frmataltd 

RArusr.eQFte FsAxC~ 

GROUPE INTERMINISTERIEL DES PRODUI'I'S C$IMIQUES 

Paris, le - 9 JU1L, 2007 

Objet : Evaluation de la conformM aux Bonnes Pratiques de Laboratoires (BPL) selon les directives 

2004/9/CE et 2004/10/CE du 11 fevrier 2004. 

Suhjeet : Assessment of compliance with Good Laboratory Practices (GLP) under the EC directives 

2004/9 and 2004/10 of 11 February 2004. 

Cons ¬cutivement a votre engagement vis-a-vis du GIPC et du COFRAC et en application du d6cret n° 2006-

1523 du 4 decembre 2006 concemant les bonnes pratiques de laboratoires et modifiant le decret n° 81-278 du 25 

mars 1981 portant cr6ation d'un groupe interminist6riel des produits chimiques, je vous confirme quo le GIPC, 

au vu des r6sultats du controle exere8 par le Comite franqais d'accr6ditation (COPRAC) - Section Laboratoires a 

d6cid¬ pour votre installation du statut suivant : 

Following your engagement vis-k-vis the GIPC and COFRfIC and in application of the decree n° 2006-1523 of 

4 December 2006 relating to the good laboratory practices and modifying the decree n° 81-278 of 25 March 

1981 giving birth to an intermlnisterial group of chemical products (GIPC), I confirm to you that the GIPC, 

given the results of the inspection realised by the French Committee of accreditation (COFR,4C) - Laboratory 

Section has taken the following decision relating to your installation; 

Respect des principes de BPL Date d'inspection : 7-8 mars 2007 

Respect of the GLP principles Date of inspection : 7-8 mars 2007 

Domaines de reconnaissance : Inspection de renouvellement (i.r) 

1- essais physico-chimiques Renewal inspection (i.r) 

2 - 6tudes de toxicite 

3 - 8tudes de mutagenicite Date de d¬ cision du GIPC : 29 juin 2007 

4 - 6tudes ecotoxicologiques sur les Date of GIPC decision.- 29 juin 2007 

organismes aquatiques et terrestres 

8 - mbthodes de chimie analytique et Date de prise d'effet : 8 mars 2007 

clinique Date of implementation : 8 mars 2007 

Areas of expertise : AnnBe de premiere conformite : 1989 

1= Pltysico-chemical testing Year of lhe first conformity: 1989 

2 = Toxicity studies 

3 =Mutagenicity studies Durt;e de validit6 : 18 mois 

4 =Environmental toxicity studies on aquatic or Time of validity: 18 months 

terrestrial organisms 

8 =Analytical and clinical chemistry 

I 

CENTRE INTERNATIONAL DE TOXICOLOGIE 

(CIT) 

MISEREY - BP 563 

27005 BVREUX CEDEX 

Pietre CkE-YSSEL. 

Conseiller d'Etat h 

Secr6tariat general du GIPC - DGE- Simap - 12, rue Villiot - 75572 Paris codex 12 

Tzfl~,phone : 0153 44 96 10 - Tblecopie : 0153 44 9172 
p. 126 

CONFIDENTIAL 

STUDY TITLE 

EVALUATION OF SKIN SENSITIZATION POTENTIAL IN MICE 

USING THE LOCAL LYMPH NODE ASSAY (LLNA) 

STUDY DIRECTOR 

Guillaume Sire 

DATE OF ISSUE 

(D~ ~o-c-)An- ~ 

TEST FACILITY 

Crr 

BP 563 - 27005 Evreux - France 

LA.B ~TO ,~IMBER 

IFM Recherche S.R 563 27005 Evreux Cedex France 

SAS au Capital de 846.000 ¬ 

Te1 : +33 (0)2 32 29 26 26 

788 060 465 R.CS. Evreux Fax : +33 (0)2 32 67 87 05 

VAT : FR 19 788 060 465 E-mail : CIT@citox.com 

Agrememt Credit imp6t Recherche Web site : www.citox .com 

Vy 
CONTENTS 

STATEMENT OF THE STUDY DIRECTOR 4 

OTHER CIT SCIENTIST INVOLVED IN THE STUDY 5 

STATEMENT OF QUALITY ASSURANCE UNIT 6 

SUMMARY 7 

RESUME 9 

1 . INTRODUCTION 11 

2. MATERIALS AND METHODS 11 

2.1 TEST MATERIALS 11 

2.1 .1 Test item 11 

2.1 .2 Vehicle 11 

2.1 .3 Dosage form preparation 11 

2.1 .4 Other materials 12 

2.1 .4 .1 Reference item (positive control) 12 

2.1 .4 .2 Reagent used for the proliferation assay 12 

2.2 TEST SYSTEM 12 

2.2 .1 Animals 12 

2.2 .2 Environmental conditions 12 

2.2 .3 Food and water 13 

2.3 TREATMENT 13 

2.3 .1 Preliminary test 13 

2 .3 .2 Main test 13 

2.3 .2.1 Study design 13 

2.3 .2.2 Administration of the test materials 14 

2.4 CLINICAL EXAMINATIONS 14 

2.4 .1 Clinical signs, morbidity and mortality 14 

2.4 .2 Body weight 14 

2.4 .3 Ear thickness measurements and recording of local reactions 14 

2.5 PROLIFERATION ASSAY 15 

2.5 .1 Intravenous injection of 3H-TdR and sampling of auricular lymph nodes 15 

2.5 .2 Preparation of auricular lymph node cell suspensions and determination 

of proliferation 15 

2.5 .3 Interpretation of results 15 

2.6 ARCHIVING 15 

2.7 CHRONOLOGY OF THE STUDY 16 

2.8 STUDY PLAN ADHERENCE 16 
3. RESULTS 17 

3 .1 CHOICE OF THE VEHICLE 17 

3 .2 PRELIMINARY TEST 17 

3 .3 MAIN TEST 17 

3 .3 .1 Systemic clinical signs and mortality 17 

3 .3 .2 Body weight (table 2) 17 

3 .3 .3 Local irritation (tables 3 and 4) 17 

3 .3 .4 Proliferation assay 17 

4. CONCLUSION 17 

Table 1 : Results of the preliminary test (ear thickness measurements) 18 

Table 2: Individual body weight and body weight gain (g) 19 

Table 3 : Evaluation of cutaneous reactions 21 

Table 4: Ear thickness measurements (mm) 22 

Table 5: Study results 24 

APPENDICES 25 

1 . Test article description and analytical certificate 26 

2. Diet formula 29 

3 . Historical data 31 

4. CIT GLP certificate 33 and 34 
STATEMENT OF THE STUDY DIRECTOR 

The study was performed in compliance with CIT's standard operating procedures and the following principles of Good Laboratory Practice : 

. OECD Principles of Good Laboratory Practice (as revised in 1997), ENV/MC/CHEM (98) 17 and all subsequent OECD consensus documents. 

. Directive 2004/10/EC of the European Parliament and of the Council of 11 February 2004 

on the harmonization of laws, regulations and administrative provisions relating to the application of the Principles of Good Laboratory Practice and the verification of their applications for tests on chemical substances (OJ No. L50 of 20.2 .2004). 

. Decret N° 2006-1523 du 04 decembre 2006 concernant les Bonnes Pratiques de Laboratoire (Journal Officiel du 06 decembre 2006), Ministere de 1'Economie, des Finances et de 1'Industrie. 

However, no chemical analysis of the dosage forms was performed as part of this study. This exception is not considered to impact on the overall GLP status of the study. 

The study was also conducted in compliance with the following Animal Protection regulations : 

. Council Directive 86/609/EEC of 24th November 1986 on the harmonization of laws, regulations or administrative provisions relating to the protection of animals used for experimental or other scientific purposes. 

. Guidance document on the recognition, assessment, and use of clinical signs as humane endpoints for experimental animals used in safety evaluation, OECD Environmental Health and Safety Publications, No. 19. 

I declare that this report constitutes a true and faithful record of the procedures undertaken and the results obtained during the performance of the study. 

This study was performed at CIT, BP 563, 27005 Evreux, France. 

Toxicology 

G. Sire Study completion date: U(~ 

Study Director 

DESS Cellular Engineering 
OTHER CIT SCIENTIST INVOLVED IN THE STUDY 

Director of Toxicology 

CIT Management 

J.J. Legrand Date: 0 S 

Doctor of Veterinary Medicine 
STATEMENT OF OUALITY ASSURANCE UNIT 

Inspections performed at CIT: 

The CIT Quality Assurance Unit conducted the inspections detailed below: 

Dates 

Type of inspection Inspection Reported to the Reported to 

Study Director Management 

Study plan 12 February 2008 12 February 2008 14 February 2008 

Report 19 August 2008 19 August 2008 21 August 2008 

In addition, at about the same time, process-based and facility inspections relevant to this study 

were carried out by the Quality Assurance Unit. 

The inspections were performed in compliance with CIT Quality Assurance Unit procedures 

and the principles of Good Laboratory Practices . 

The final report is considered to constitute an accurate and complete reflection of the study raw 

data. 

Marie-Christine Ottini-Bertin Date: 0 S ' ~ -P, ~ ~ 

CIT Quality Assurance Unit 
T 

Lymph Node Assay (LLNA . Evaluation 

_ 

of local irrita_ . .,__ . .  ~. u, ,u ~ This study was conducted in compliance with the principles of Good Laboratory Practice Regulations . 

Methods 

A preliminary test was first performed in order to define the concentrations of test item used in the main test. 

In the main test, twenty-eight female CBA/J mice were allocated t 

ups of four animals receiving the test ite 

e concentration of 0.1, 0.25, 0.5, 1 or 2.5%, 

to be 

ga~control group of four animals receiving the vehicle (propylene glycol), 

. one positive control group of four animals receiving the reference a-hexylcinnamaldehyde (HCA), a moderate sensitizer, at the concentration of 25%. 

item, 

During the induction phase, the test item, vehicle or reference item was applied over the ears (25 pL per ear) for 3 consecutive days (days 1, 2 and 3). After 2 days of resting, the proliferation of lymphocytes in the lymph node draining the application site was measured by incorporation of tritiated methyl thymidine (day 6). The obtained values were used to calculate stimulation indices (SI). 

The irritant potential of the test item was assessed in parallel by measurement of ear thickness on days 1, 2, 3 and 6. 

Results 

The test item was not soluble in the recommended vehicles, consequently, a homogenous suspension, obtained in propylene glycol, was used for the study. The maximum practicable concentration in this vehicle was 2.5%. 

Therefore, the concentrations selected for the preliminary test were 0.25, 0.5, 1 and 2.5%. 

for 

Since the test item was non-irritant in the preliminary test, the highest concentration retained the main test was the maximal practicable concentration (2.5%). 

Systemic clinical signs and mortality 

No mortality and no clinical signs were observed during the study. 

Local irritation 

A black coloration of the skin of the ears was noted on days 2 and 3 in all animals treated at the concentrations of 1 and 2.5% and on day 6 in 2/4 animals treated at the concentration of 2.5%. No noteworthy increase in ear thickness was observed in the animals of the treated groups. 
p. 133 

8 

Proliferation assay 

A significant lymphoproliferation was noted in the positive control group given HCA at 25%, 

the study was therefore considered valid. 

No noteworthy lymphoproliferation was noted at any of the tested concentrations. 

The results are presented in the following table: 

Treatment Concentration Irritation level Stimulation Index 

(%) (SI) 

Test item 0 .1 non-irritant 1 .13 

Test item 0.25 non-irritant 0.72 

Test item 0.5 non-irritant 0.84 

Test item 1 non-irritant 0.97 

Test item 2.5 non-irritant 0.76 

---------- - -------- - --------- - ---------- ---------- ------------------ -------------- 

HCA 25 - 8 .59 

Conclusion 

ions of this study, the test ite 

not induce delayed contact hyper' ensi 
RESUME 

e-une~ -ny- erseiisibilite e co a-ire - ee 771 77!77 F Lymph Node Assay" (LL°tslA) . L'irritation locale a ete egalement dvaluee en parallele. L'etude a ete rdalisee conformdment aux r6gles de Bonnes Pratiques de Laboratoire. 

Methode 

Dans un premier temps, un essai pr6liminaire a ete realisd afin de determiner les concentrations de produit a utiliser dans 1'essai principal. 

Dans I'essai principal, 28 souris femelles CBA/J ont ete 

. 5 groupes traites de 4 animaux recevant le produi 

la concentration de 0,1 ; 0,25 ; 0,5 ; 1 ou 2,5 % 

. 1 groupe t6moin n6gatif de 4 animaux recevant le vehicule (propylene glycol), 

. 1 groupe temoin positif de 4 animaux recevant le produit de r6ference, a-hexylcinnamaldehyde (HCA), un sensibilisant modere, d la concentration de 25 %. 

Pendant la phase d'induction, le produit, le vehicule ou le produit de refdrence a etd appliqud sur les oreilles (25 pL par oreille) pendant 3 jours consecutifs (jours 1, 2 et 3). Apres 2 jours de repos, la proliferation lymphocytaire dans le noeud lymphatique drainant le site d'application a ete mesure par incorporation de methyl thymidine tritiee (jour 6) . Les valeurs obtenues ont ete utilisees pour calculer 1'indice de stimulation (IS). 

Le potentiel irritant du produit a 6t6 evalue en parall6le en mesurant 1'epaisseur de 1'oreille aux jours 1, 2, 3 et 6. 

Resultats 

Le produit n'a pas ete soluble dans les vehicules recommand6s, par consequent, une suspension homogene, obtenue dans le propylene glycol, a ete utilisee pour 1'6tude. La concentration maximale utilisable dans cc vehicule a ete 2,5 %. 

Les concentrations retenues pour 1'essai preliminaire ont ete 0,25 ; 0,5 ; 1 et 2,5 %. 

Le produit n'ayant pas ete irritant lors de 1'essai preliminaire, la concentration maximale retenue pour 1'essai principal a ete la concentration maximale utilisable (2,5 %). 

Signes clinigues systemicgues et mortalit6 

Aucune mortalite ni aucun signe clinique Wont et6 observ6s pendant 1'etude . 

Irritation locale 

Une coloration noire de la peau des oreilles a ete notee aux jours 2 et 3 chez tous les animaux traites aux concentrations 1 et 2,5 % et au jour 6 chez 2/4 animaux traitds a la concentration de 2,5 %. Aucune augmentation notable de 1'epaisseur de 1'oreille n'a et6 notde chez les animaux trait6s avec le produit. 

Test de proliferation 

Une lymphoprolifdration significative a ete not6e avec le groupe temoin positif ayant requ le HCA a 25 %, 1'etude est done consid6ree valide . 

Aucune lymphoproliferation n'a et6 notee aux concentrations testees . 
Les resultats sont prdsentes dans le tableau suivant : 

Traitement Concentration Niveau Indice de Stimulation 

(%) d' irritation (IS) 

Produit 0,1 non-irritant 1,13 

Produit 0,25 non-irritant 0,72 

Produit 0,5 non-irritant 0,84 

Produit 1 non-irritant 0,97 

Produit 2,5 non-irritant 0,76 

---- ------------------------- ------ 

------- ------- 

HCA 25 

Conclusion 

8,59 

Dans nos conditions experimentales, le produi 

induit une hypersensibilite de contact retardee ns Me test du LLNA. 
p. 136 

1 . INTRODUCTION 

ll 

s study was to evaluate the potential of the test ite~~~~ 

induce delayed contact hypersensitivity, using the murine oca ymp o e 

ssay (LLNA). 

This study was based on the design adopted by ICCVAM (Interagency Coordination 

Committee on the Validation of Alternative Methods, ICCVAM 1999) and ECETOC 

(Monograph No. 78 Skin sensitization Testing for the Purpose of Hazard Identification and 

Risk Assessment, September 2000), with the addition of the evaluation of local irritation . 

The study has been designed to comply with the following guidelines : 

OECD Guideline No. 429, 24th April 2002, 

EC Directive No. 2004/73/EC, part B.42, 29th April 2004. 

2. MATERIALS AND METHODS 

2.1 TEST MATERIALS 

2.1 .1 Test item 

description : black powder 

container : one plastic flask 

date of receipt : 29 June 2007 

storage conditions : at room temperature and protected from humidity 

purity/composition : see analytical certificate 

expiry date: June 2008. 

Data relating to the characterization of the test item are documented in an analytical certificate 

and a test item date sheet (presented in appendix 1) provided by the Sponsor. 

2 .1 .2 Vehicle 

The vehicle used was propylene glycol, batch No. S35142-236 (Aldrich, 

Saint-Quentin-Fallavier, France) . 

2 .1 .3 Dosage form preparation 

The concentrations were expressed in % (w/v). 

For the dosage form preparation at 2.5%, the test item was ground to a fine powder using a 

mortar and pestle, before to add the vehicle . The lower concentrations were obtained by 

dilution in the vehicle . 

All dosage form preparations were made freshly on the morning of administration and any 

unused material was discarded that same day. 
p. 137 

2.1 .4 Other materials 

2.1 .4 .1 Reference item (positive control) 

12 

The reference item (positive control) was a-hexylcinnamaldehyde (RCA), batch No. 04012JE 

(Aldrich, Saint-Quentin-Fallavier, France), dissolved in a mixture acetone/olive oil (4/1, v/v) at 

the concentration of 25% (v/v). 

The preparation was made freshly on the morning of administration and any unused material 

was discarded that same day. 

2.1 .4 .2 Reagent used for the proliferation assay 

The reagent used for the proliferation assay was 

batch No . B505 (Amersham, Les Ulis, France) . 

13H] methyl-thymidine (3H-TdR), 

Three days before the injections, the required quantity of 3H-TdR was diluted in 0.9% NaCI 

(20 pCi in 250 pL of 0.9% NaCl per animal). The obtained solution was stored at +4°C and 

protected from light before use. 

2.2 TEST SYSTEM 

2.2 .1 Animals 

Species, strain and sex: 

Reason for this choice: 

Age/weight: 

Number: 

Breeder: 

Acclimation: 

Allocation to groups : 

Identification : 

CBA/J mouse, nulliparous and non-pregnant females. 

species generally accepted by regulatory authorities for this type of 

study. Females have been chosen since this sex is recommended by 

regulatory authorities for this type of study. 

on the first day of the treatment period, the animals of the preliminary 

test were approximately 10 weeks old and the animals of the main test 

were approximately 9 weeks old and had a mean body weight ± 

standard deviation of 21 .2 ± 1 .1 g. 

4 females for the preliminary test, 28 females for the main test . 

Janvier, Le Genest-Saint-Isle, France. 

at least 5 days before the beginning of the study. 

animals were assigned to the treatment groups by hand procedure . 

individually by a number on the tail . 

2.2 .2 Environmental conditions 

The conditions in the animal room were set as follows : 

. temperature: 22 f 2°C 

. relative humidity: 30 to 70% 

. light/dark cycle: 12 h/12 h (7 :00-19:00) 

. ventilation : approximately 12 cycles/hour of filtered, non-recycled air. 

The temperature and relative humidity are under continuous control and recording. The records 

are checked daily and filed . In addition to these daily checks, the housing conditions and 

corresponding instrumentation and equipment are verified and calibrated at regular intervals . 

The animals were housed individually in disposable crystal polystyrene cages 

(22.00 cm x 8.50 cm x 8.00 cm). Each cage contained autoclaved sawdust (SICSA, Alfortville, 

France) . Sawdust is analyzed by the supplier for composition and contaminant levels. 
p. 138 

13 

2.2 .3 Food and water 

All animals had free access to SSNIFF R/M-H pelleted maintenance diet (SSNIFF 

Spezialdiaten GmbH, Soest, Germany) and tap water (filtered using a 0.22 micron filter) 

contained in bottles. 

Each batch of diet is analyzed for composition and contaminant level by the supplier. 

The diet formula is presented in appendix 2. 

Bacteriological and chemical analyses of water, including the detection of possible 

contaminants (pesticides, heavy metals and nitrosamines) are performed regularly by external 

laboratories. The results of these analyses are archived at CIT. 

No contaminants are known to be present in the diet, drinking water or sawdust at levels which 

may be expected to interfere with or prejudice the outcome of the study . 

2.3 TREATMENT 

2.3 .1 Preliminary test 

To assess the irritant potential of the test item (through ear thickness measurement), a 

preliminary test was performed on a small number of animals, as follows : 

. the test item was prepared at the concentrations of 2.5, 1, 0.5 and 0.25%, 

. for 3 consecutive days, the animals received applications of 25 pL of the dosage form 

preparations to the external surface of both ears (one concentration per ear), 

. measurement of the ear thickness (using a micrometer) was performed each day before 

treatment and 72 hours after the last application . 

2.3 .2 Main test 

2.3 .2.1 Study design 

The concentrations of test item were selected according to the criteria specified in the 

International Guideline and on the basis of the results of the solubility and preliminary assays. 

The study design was as follows : 

Groups Number of animals Treatment Concentration (%) 

1 4 females Vehicle 0 

2 4 females Test item 0.1 

3 4 females Test item 0.25 

4 4 females Test item 0.5 

5 4 females Test item 1 

6 4 females Test item 2.5 

7 4 females HCA 25 
p. 139 

2.3 .2 .2 Administration of the test materials 

14 

On days 1, 2 and 3, a dose-volume of 25 p,L of the control or dosage form preparations was 

applied to the dorsal surface of both ears, using an adjustable pipette fitted with a plastic tip . 

In order to avoid licking and to ensure an optimized application of the test materials, the 

animals were placed under light isoflurane anesthezia during the administration. 

No massage was performed but the tip was used to spread the preparation over the application 

sites . No rinsing was performed between each application . 

2.4 CLINICAL EXAMINATIONS 

2.4 .1 Clinical signs, morbidity and mortality 

The animals were observed at least once a day during the study for clinical signs, signs of 

morbidity or mortality . 

2.4 .2 Body weight 

The animals were weighed individually on the first day of the study (day 1) and on the day of 

sacrifice (day 6). 

2.4 .3 Ear thickness measurements and recording of local reactions 

Ear thickness measurements and recording of local reactions were performed in order to assess 

any possible irritant effect of the test item, as possible irritancy may be involved in false 

positive lymphoproliferative responses. 

On days 1, 2 and 3 (before each cutaneous application) and on day 6 (after sacrifice), the 

thickness of the left ear of each animal of the vehicle control and treated groups was measured 

using a micrometer. 

No measurement of ear thickness was performed for the animals of the positive control group. 

Any irritation reaction (erythema and edema) was recorded in parallel . Any other observation 

(coloration, presence of residual test item, . . .) was noted. 

The irritation level of the test item was determined according to the following table: 

% increase in ear thickness 

between day 1 and day 3 or 6 

Irritation level Interpretation 

< 10% I Non-irritant 

10-30% lI Slightly irritant 

> 30% 111 Irritant 
2.5 PROLIFERATION ASSAY 

2.5 .1 Intravenous injection of 3H-TdR and sampling of auricular lymph nodes 

p. 140 

15 

Lymph node cell proliferative responses were measured as described by Kimber and Dearman (1991). On day 6, all animals of all groups received a single intravenous injection of 250 pL of 0.9% NaCl containing 20 pCi of 3H-TdR (specific activity of 25 Ci/mmol) via the tail vein. Approximately 5 hours later, the animals were killed by cervical dislocation and the auricular lymph nodes were excised . The lymph nodes were pooled for each experimental group. 

2.5 .2 Preparation of auricular lymph node cell suspensions and determination of proliferation 

For each experimental group, a single cell suspension of auricular lymph node cells (ALNC) 

was prepared by mechanical dissagregation in Petri dishes with the plunger of a syringe. Cell 

suspensions were washed with 15 mL of 0.9% NaCl and pellets obtained were re-suspended in 0.9% NaCl for numeration of lymphocytes (cellularity) and determination of their viability by 

exclusion of trypan blue. Each cell suspension was then centrifuged and pellets were 

precipitated with 3 mL of 5% (w/v) trichloroacetic acid (TCA) in purified water at +4°C 

overnight. After a last centrifugation, the pellets were precipitated with 1 mL of 5% TCA. 

Three mL of Ultima Gold "R scintillation fluid (Packard) were added in order to measure 

incorporation of 3H-TdR usirng (3-scintillation counting . 

The results were expressed as disintegration's/mn (dpm) per group and per node. Stimulation Indices (Sl) were calculated according to the following formula: 

SI = 

dpm of treated group 

dpm of control group 

2.5 .3 Interpretation of results 

The test item was considered as a skin sensitizer when the SI for a dose group is >_ 3 . Other relevant criteria such as cellularity, radioactivity levels and ear thickness were also taken into 

account for the interpretation of results . 

2.6 ARCHIVING 

The following study materials are archived by CIT, 27005 Evreux, 

the end of the in vivo phase of the study : 

. Study plan and possible amendments, 

. raw data, 

. correspondence, 

. final report and possible amendments. 

France, for 10 years after 

On completion of this period, the archived study materials will be returned to the Sponsor, or 

may be archived at CIT for a further period (at additional cost). The total duration of archiving (depending on regulations) will be the responsibility of the Sponsor. 

In addition, raw data not specific to the study, including, but, not limited to, certificates of analyses for food, water and bedding (if applicable) and records of environmental data and equipment calibration, are also archived by CIT for at least 30 years. 
p. 141 

2.7 CHRONOLOGY OF THE STUDY 

The chronology of the study is summarized as follows : 

Procedure Date 

Experimental starting date (solubility assay) 13 March 2008 

First day of treatment of the preliminary test 18 March 2008 

First day of treatment of the main test 26 March 2008 

Day of injection of 3H-TdR and ALNC collection 31 March 2008 

Experimental completion date 04 September 2008 

2.8 STUDY PLAN ADHERENCE 

16 

The study was performed in accordance with the d subsequent 

amendments . There were no deviations from the agreed Study plan. 
3 . RESULTS 

3.1 CHOICE OF THE VEHICLE 

p. 142 

17 

The test item was not soluble in the recommended vehicles, consequently, a homogenous 

suspension, obtained in propylene glycol, was used for the study. The maximum practicable 

concentration in this vehicle was 2.5%. 

3.2 PRELIMINARY TEST 

The results of the preliminary test are presented in table l . 

A black coloration of the skin of the ears was observed at the concentrations of 1 and 2.5% on 

days 2 and 3 . 

The test item was non-irritant in the preliminary test, whatever the concentrations tested. 

The highest concentration retained for the main test was therefore the maximal practicable 

concentration (2 .5%). 

3 .3 MAIN TEST 

3 .3 .1 Systemic clinical signs and mortality 

No clinical signs and no mortality were observed during the study. 

3 .3 .2 Body weight (table 2) 

The body weight change of treated animals was similar to that of control animals. 

3.3 .3 Local irritation (tables 3 and 4) 

A black coloration of the skin of the ears was noted on days 2 and 3 in all animals treated at the 

concentrations of I and 2.5% and on day 6 in 2/4 animals treated at the concentration of 2.5%. 

No noteworthy increase in ear thickness was observed in the animals of the treated groups . 

3.3 .4 Proliferation assay 

Results of proliferation assay are presented in table 5 . 

The quantity of cells obtained in each group was satisfactory and the cellularity correlated with 

incorporation of 3H-TdR. The cell viability was higher than 70% in each group. 

In the positive control group given HCA at the concentration of 25%, a moderate increase in cellularity and a stimulation index exceeding the threshold value of 3 (SI - 8.59) were noted. The study was therefore considered valid. 

No noteworthy lymphoproliferation was noted with the test item at any of the tested 

concentrations . 

4. CONCLUSION 

itions of this study, the test item~~ 

Local Lymph Node Assay. 

id not induce delayed contact hypersens~tivity - Fill e murme 
Table 1 : Results of the preliminary test (ear thickness measurements) 

p. 143 

18 

DI D2 D3 D6 

Number Concentration Ear Local Ear Local Ear Local Far Local % Animal % Thickness reaction Thickness reaction Thickness reaction Thickness reaction 

Female 301 2.5 RE 0.24 0 0.25 0/C 0.26 0/C 0.26 0 8 33 Female 301 1 LE 0.25 0 0.25 0/C 0.25 0/C 0.27 0 

. 

8 00 Female 302 2.5 RE 0.24 0 0.24 0/C 0.24 0/C 0.24 0 . 

0.00 Female 302 1 LE 0.24 0 0.24 0/C 0.24 0/C 0.23 0 4.17 Female 303 0.5 RE 0.24 0 0.24 0 0.24 0 0.25 0 4.17 Female 303 0.25 LE 0.25 0 0.25 0 0.25 0 0.25 0 0.00 Female 304 0.5 RE 0.25 0 0.25 0 0.25 0 0.25 0 0.00 Female 304 0.25 LE 0.25 0 0.24 0 0.25 0 0.26 0 4.00 

RE= tight ear 

LE= left ear 

D=day 

0 =no cutaneous reaction 

C=black coloration of the ears 

% = percentage of earth ickness increase compared today I 
Table 2: Individual body weight and body weight gain (g) 

Days 

Groups Animal 

No . L 6 (1) 

1 01 18 .9 19.4 0.5 

02 21 .3 20.9 -0 .4 

03 21 .0 21 .4 0.4 

04 18 .8 19 .7 0.9 

M 20.0 20.4 0.3 

SD 1 .3 1 .0 0.5 

2 05 21 .1 21 .9 0.8 

06 20 .4 21 .2 0.8 

07 22 .0 21 .9 -0 .1 

08 21 .6 22.0 0.4 

M 21 .3 21 .8 0 .5 

SD 0.7 0.4 0 .4 

3 09 22.6 22.0 -0.6 

10 19.4 20.2 0 .8 

11 21 .8 21 .5 -0 .3 

12 20.2 21 .3 1 .1 

M 21 .0 21 .3 0.3 

SD 1 .5 0 .8 0 .8 

4 13 21 .4 19.1 -2.3 

14 20.6 21 .0 0 .4 

15 19.7 19 .5 -0 .2 

16 21 .3 22.3 1 .0 

M 20.8 20.5 -0.3 

SD 0.8 1 .5 1 .4 

(1) = body weight gain 

M = mean 

SD=standard deviation 
Table 2 (continued) 

Days 

Groups Anirrral 

No . 1 6 (1) 

5 17 20.6 21 .0 0.4 

18 20.5 21 .5 1 .0 

19 22.1 22.2 0.1 

20 21 .5 21 .4 -0 .1 

M 21 .2 21 .5 0.3 

SD 0.8 0.5 0 .5 

6 21 21 .7 22.6 0.9 

22 22.0 22.3 0.3 

23 23 .8 22.5 -1 .3 

24 22.6 21 .2 -1 .4 

M 22.5 22.2 -0 .4 

SD 0.9 0.6 1 .2 

7 25 21.6 22.5 0.9 

26 22.0 22.9 0.9 

27 22.1 19.5 -2 .6 

28 21.1 21 .7 0.6 

M 21 .7 21.7 -0 .1 

SD 0 .5 1 .5 1 .7 

(1)=body weight gain 

M = mean 

SD = standard deviation 
Table 3 : Evaluation of cutaneous reactions 

Days 

Groups Dosage Animal 

form No. 1 2 3 6 

1 vehicle 01 0 0 0 0 

02 0 0 0 0 

03 0 0 0 0 

04 0 0 0 0 

2 test item 05 0 0 0 0 

0.1% 06 0 0 0 0 

07 0 0 0 0 

08 0 0 0 0 

3 test item 09, 0 0 0 0 

0.25% 10 0 0 0 0 

11 0 0 0 0 

12 0 0 0 0 

4 test item 13 0 0 0 0 

0.5% 14 0 0 0 0 

15 0 0 0 0 

16 0 0 0 0 

5 test item 17 0 0/C 0/C 0 

1% 18 0 0/C 0/C 0 

19 0 0/C 0/C 0 

20 0 0/C 0/C 0 

6 test item 21 0 0/C 0/C 0 

2.5% 22 0 0/C 0/C 0/C 

23 0 0/C 0/C 0/C 

24 0 0/C 0/C 0 

0=no cutaneous reaction 

C=black coloration of the ears 

vehicle =propylene glycol 
Table 4: Ear thickness measurements (mm) 

Groups Dosage 

form 

1 vehicle 

2 test item 

0 .1% 

3 test item 

0.25% 

Days 

Animal 

No. 1 2 d 1 3 d2 6 d3 

01 0.26 0.25 -0 .01 0.25 -0.01 0.25 -0.01 

02 0.26 0.26 0.00 0.26 0 .00 0.25 -0 .01 

03 0.26 0.27 0.01 0.26 0 .00 0.25 -0 .01 

04 0.25 0.25 0.00 0.24 -0 .01 0.25 0.00 

M 0.26 0.26 0.00 0.25 -0 .01 0 .25 -0.01 

SD 0.01 0.01 0.01 0.01 0.01 0.00 0 .01 

% (*) 0.00 -1 .94 -2.91 

05 0.25 0.25 0.00 0.24 -0 .01 0 .24 -0 .01 

06 0 .26 0 .26 0.00 0.25 -0 .01 0 .25 -0 .01 

07 0 .26 0.25 -0.01 0.25 -0 .01 0 .25 -0 .01 

08 0 .25 0.24 -0.01 0.26 0.01 0.27 0 .02 

M 0.26 0.25 -0 .01 0 .25 -0.01 0.25 0 .00 

SD 0.01 0.01 0.01 0 .01 0 .01 0.01 0.02 

% (*) -1 .96 -1 .96 -0 .98 

09 0 .27 0.26 -0.01 0.26 -0 .01 0.26 -0 .01 

10 0.25 0.24 -0.01 0.24 -0 .01 0.24 -0.01 

11 0.26 0.25 -0.01 0.25 -0.01 0.25 -0.01 

12 0.26 0.25 -0 .01 0.26 0.00 0.25 -0.01 

M 0.26 0 .25 -0 .01 0.25 -0.01 0.25 -0 .01 

SD 0 .01 0.01 0.00 0.01 0.01 0.01 0.00 

% (*) -3 .85 -2 .88 -3 .85 

M = mean 

SD =standard deviation 

(*)=percentage ofearthickness increase compared to day 1 

d 1 =difference of ear thickness between day 2 and day 1 

d2=difference ofearthickness between day 3 and day 1 

0 =difference of ear thickness between day 6 and day 1 

vehicle --pro~ylene glycol 

--- 
Table 4 (continued) 

Groups Dosage 

form 

4 test item 

0.5% 

5 test item 

1% 

6 test item 

2.5% 

AnimalNo. 1 

13 

14 

15 

16 

Days 

2 dl 3 d2 6 d3 

0 .25 0.24 -0.01 0.25 0.00 0.25 0.00 

0 .25 0.25 0.00 0.25 0.00 0.25 0.00 

0 .25 0.26 0.01 0.25 0.00 0.26 0 .01 

0 .25 0.25 0.00 0.25 0.00 0.25 0 .00 

M 0.25 0 .25 0.00 0 .25 0.00 0.25 0.00 

SD 0.00 0 .01 0.01 0 .00 0 .00 0.01 0.01 

% (*) 0.00 0 .00 1 .00 

17 

18 

19 

20 

0 .25 0.26 0.01 0.25 0.00 0.26 0.01 

0 .26 0.25 -0.01 0.26 0.00 0.25 -0 .01 

0 .27 0.25 -0 .02 0.26 -0 .01 0.26 -0.01 

0 .25 0.25 0.00 0.25 0.00 0.24 -0.01 

M 0.26 0 .25 -0.01 0.26 0.00 0.25 -0 .01 

SD 0.01 0.01 0.01 0.01 0.01 0.01 0.01 

% (*) -1 .94 -0.97 -1 .94 

21 

22 

23 

24 

0 .25 0.25 0.00 0.26 0.01 0.26 0.01 

0 .25 0.25 0.00 0.25 0.00 0.25 0.00 

0 .25 0.25 0.00 0.24 -0.01 0.24 -0.01 

0.25 0.25 0.00 0.26 0.01 0.26 0.01 

M 0.25 0.25 0.00 0.25 0.00 0.25 0.00 

SD 0.00 0.00 0.00 0.01 0.01 0.01 0.01 

% (*) 0.00 1 .00 1 .00 

M = mean 

SD =standard deviation 

(*) = percentage of ear thickness increase compared to day 1 

d I =digerence of ear thickness between day 2 and day 1 

d2=difference ofearthickness between day 3 and day 1 

d3 =difference of ear thickness between day 6 and day 1 
p. 149 

Table 5: Study results 

Groups and 

Cell count 

Viability Arnomt 

u d 7 -Y

Nunub 

r 

dpm dpm Stinnulation 

Incre&sein 

carthickness 

oncentrations iable dead fcells x 

of 0 

n des, ergroup, ernode ndex(SI) I/ bct~veen 

: 7 

ev 

Fel 

-- 

per 

group 

day 1nd day 6) 

-- -- 

I Vehicle 103 8 92 .79 5.15 ~ i 8 576.81 72 .10 -2 .91 i 

Test rtem - - - -- - 1 

0 .1% 

183 1 99 .46 9.15 1 .78 8 654 .34 81 .79 1 .13 -0 .98 1 

Test rtem 

3 

0.25% 

81 3 96.43 4 .05 0 .79 8 416 .37 52 .05 0 .72 -3 .85 1 

Test item 

0 .5% 

82 7 92 .13 4.10 0 .80 8 484 .86 60,61 0.84 1 .00 1 

Test item 

I % 

69 18 79 .31 3 .45 0.67 8 558 .54 69.82 0.97 -1 .94 1 

Test item 

2 .5% 

88 6 93 .62 4 .40 0 .85 8 436 .90 54 .61 0 .76 1 .00 I 1 I 

HCA ------ 

25% 

214 18 92 .24 21 .40 4 .16 8 4956 .76 619.60 8 .59 

I 

viable cells 

. viability - x 100 

viable cells + dead cells 

dpm=disintegrations per ninute 

HCA =a-hexylcinnamaldehyde 

amount of cells (x 10° cells) in the treated group I =non-hritant (Bncrease in car thicFaess < loo/) 

. cellularity index ECi value =theoretical concentration resulting in a SI value of 3 amount of ce0s (x 10° cells) in the vehicle group 

dpm o£treated group 

. stinulation index 

24 

3 value 

NA 

dpm of control group 
APPENDICES 
1 . Test article description and analytical certificate 
PHYSICAL AND CHEMICAL PROPERTIES 

Appearance : black powder 

Particule size : 30 gm ( approximately) 

Specific Gravity : 2,1 kg/m3 at 20°C 

Autoignition temperature : > 400 °C (standard : NF EN 50281-2-1) 

Solubility : in water : insoluble 

TOXICOLOGICAL INFY3RiVIATIOiVS AND USE SAFETY 

See Safety Data Sheet. 

I STORAGE AND DISPOSAL 

Storage : Keep hermetically closed in a dry, cool and well-ventiled 

place . 

Expiry date : June 2008 

Qisposal : Incineration 

p. 152 

27 
Date 5 juin 2007 

r 

Nom du responsable du laboratoirel Laboratory Director : 

Signature 

RWrence de la m6thode 

d'analyse / Analysis 

reference 

p. 153 

28 
2 . Diet formula 
SSNIFF RJM-H 

Complete diet for rats/mice - maintenance 

Constituents 

Crude proteins 19 .00% Calcium 1 .00% 

Crude fat 3 .30% Phosphorus 0.70% 

Crude fiber 4.90% Sodium 0.25% Crude ash 6 .70% Magnesium 0.20% 

Potassium 0.90% 

Amino Acids Vitamins (je kg) 

Lysine 1 .00% A 15,000 IE Methionine 0.30% D3 1,000 IE 

Cystine 0.30% E 100 mg 

Glycine 0.90% B1 10 mg 

Leucine 1 .30% B2 20 mg 

Isoleucine 0.70% B6 12 mg 

Arginine 1 .20% B12 80 pg 

Phenylalanine 0.90 .% Biotin 400 fig 

Tryptophan 0 .25% Pantothenic acid 30 mg Histidine 0.50% Choline 1,600 mg 

Tyrosine 0.60% Folic acid 4 mg 

Aspartic acid 1 .70% Nicotic acid 60 mg 

Glutaminic acid 3.80% K3 5 mg 

Valine 0.90% Inositol 50 mg 

Threonine 0.70% 

Trace elements (je kg) ME (je kg) 12.2 MJ 

Manganese 90 mg 

Copper 12 mg 

Zinc 75 mg Item numbers 

Iodine 2 mg V 1530 Meal 

Iron 220 mg V1531 Micromeal 

Selenium 0 .2 mg V 1534 Pellets 10 mm 

Cobalt 2 mg V1535 Pellets 15 mm 
3 . Historical data 
Historical data 

Skin sensitization potential in miee (CBA!J straln) 

using the Local Ly-mph Node Assay (LLhFA) 

Vehicle. control Propy lone Glycol 

from December 2004 to December 2006 

Amount dpm/node increase, in 

of cells ear thickness 

x106 cel:ls 

minimum 2.75 33.28 -4.63 

maximum 7.90 76.64 1 .02 

Moan 5.03 47.93 -0.98 

SD 1 .65 10.60 1 .78 

Nb 17 

SD - Standard deviation 

Nb = number of values 
4. CIT GLP certificate 
La-4 " dd.ref -x ternue 

Urosxrqye Pxu+ec-use 

GROUPE AVTERMINISTERIEL DES PRODUTI'S CHI&IIQUES 

Paris, le - 9 JUIL 2001 

Objet : Evaluation de la conformit6 aux Bonnes Pratiques de Laboratoires (BPL) selon les directives 

2004/9/CE et 2004/10/CE du 1 l Wrier 2004 . 

Subject : Assessment of compliance with Good Laboratory Practices (GLP) under the BC directives 

200419 and 2004110 of 11 February 2004. 

Consdcutivement n votre engagement vis-it-vis du GIPC et du COFRAC et en application du ddcret n° 2006-

1523 du 4 d8cembie 2006 concernant les bonnes pratiques de laboratoires et modifiant le dccret n° 81-278 du 25 

mars 1981 portant crdation d'un groupe interminist¬riel des produits chimiques, je vous confurne que le GIPC, 

au vu des rdsultats du contr6le exerc8 par le ComitB tranpais d'accrdditation (COFRAC) - Section Laboratoires a 

d8cid6 pour votre installation du statut suivant : 

Following your engagement vis-d-vis the GIPC and COFRAC and in application ofthe decree n° 2006-1523 of 

4 December 2006 relating to the good laboratory practices and modifying the decree n° 81-278 of 25 March 

1981 giving birth to an interministerial group of chemical products (GIPC), I confirm to you that the GIPC, 

given the results of the inspection realised by the French Committee of accreditation (COFR.4C) - Laboratory 

Section has taken the following decision relating to your installation : 

Respect des principes de BPL Date d'inspection : 7-8 mars 2007 

Respect of the GLP principles Date of inspection : 7-8 mars 2007 

Domaines de reconnaissance : Inspection de renouvellement (i .r) 

1- essais physico-chimiques Renewal inspection (P.r) 

2 - 6tudes de toxicitd 

3 - etudes de mutagsnicite Date de d6cision du GIPC : 29 juin 2007 

4- etudes 6cotoxicologiques sur les Date of GIPCdecision: 29juin 2007 

organismes aquatiques et terrestres 

8 - m6thodes de chimie analytique et Date de prise d'effet : 8 mars 2007 

clinique Date of implementation : 8 mars 2007 

Areas of expertise ; Ann6e de premiere conformit8 : 1989 

1=Physico-chemical testing Year of the first conformity: 1989 

2 = Toxicity studies 

3 =Mutagenicity studies Duree de validite : 18 mois 

4 = Environmental toxicity studies on aquatic or Time of validity: 18 months 

terrestrial organisms 

8 =Analytical and clinical chemistry 

- 

CENTRE INTERNATIONAL DE TOXICOLOGIE 

(CIT) 

MISEREY - BP 563 

27005 EVREUX CEDEX 

Pierre C 

Conseiller d'Etat h 

Secretariat g6ndral du GIPC - uGE- Simap - 12, rue Villiot - 75572 Paris codex 12 

Te18phone : 0153 44 96 10 - T ¬ 1ecopie : 01 53 44 91 72 
STUDY TITLE 

ACUTE DERMAL IRRITATION 

IN RABBITS 

STUDYDIRECTOR 

Catherine Pelcot 

DATE OF ISSUE 

C,qs f~Dw q:~s 

TEST FACILITY 

CTT 

BP 563 - 27005 Evreux - France 

--_ -~- . " Safety & Health" 

Research Laboratories 

fFM Recherche BY 563 27005 Evreux r e dex France 5 .N.C au Capital de 846 .092 ¬ T61 . : +33 (0)2 32 29 26 26 788 060 465 R .C.S . Evreux Fax : +33 (0)2 32 67 87 05 VAT : FR! 9 788 060 465 E-mail : C(T@citox.com Agremenr Credit Imp6t Recherctie Web site : www.citox .com 

p. 160 

CONFIDENTIAL 

1 ~ 
CONTENTS 

STATEMENT OF THE STUDY DIRECTOR 4 

OTHER CIT SCIENTIST INVOLVED IN THE STUDY 5 

STATEMENT OF QUALITY ASSURANCE UNIT 6 

SUMMARY 7 

RESUME 9 

1 . INTRODUCTION 11 

2. MATERIALS AND METHODS 11 

2.1 TEST ITEM 11 

2.1 .1 Identification 11 

2.1 .2 Dosage form preparation 11 

2.2 TEST SYSTEM 12 

2.2 .1 Animals 12 

2.2 .2 Environmental conditions 12 

2.2 .3 Food and water 12 

2.3 TREATMENT 13 

2.3 .1 Preparation and selection of the animals 13 

2 .3 .2 Application of the test item 13 

2 .4 CUTANEOUS EXAMINATIONS 13 

2.4 .1 Duration of the observation period 13 

2.4 .2 Description and evaluation of cutaneous reactions 13 

2.5 BODY WEIGHT 14 

2.6 INTERPRETATION OF RESULTS 14 

2.6 .1 Interpretation of results 14 

2.6 .1 .1 Criteria for irritation 14 

2.6 .1 .2 Criteria for corrosion 14 

2.7 ARCHIVING 15 

2.8 CHRONOLOGY OF THE STUDY 15 

2.9 STUDY PLAN ADHERENCE 15 

3 . RESULTS 16 

4. CONCLUSION 16 

Table l : 3-minute exposure - Cutaneous examinations and mean values of the scores 

recorded for the first animal (24, 48 and 72 hours) 17 

Table 2: 1-hour exposure - Cutaneous examinations and mean values of the scores 

recorded for the first animal (24, 48 and 72 hours) 18 

Table 3: 4-hour exposure - Individual cutaneous examinations and mean values of the 

scores recorded for each animal (24, 48 and 72 hours) 19 

Table 4: Individual body weight (g) 21 
APPENDICES 22 1 . Test article description and analytical certificate 23 2. Diet formula 2( 3 . CIT GLP certificate 28 and 29 
STATEMENT OF THE STUDY DIRECTOR 

The study was performed in compliance with the principles of Good Laboratory Practice as 

described in: 

. OECD Principles of Good Laboratory Practice (as revised in 1997), ENV/MC/CHEM 

(98) 17 and all subsequent OECD consensus documents. 

. Directive 2004/101EC of the European Parliament and of the Council of 11 February 2004 on 

the harmonization of laws, regulations and administrative provisions relating to the 

application of the Principles of Good Laboratory Practice and the verification of their 

applications for tests on chemical substances (OJ No. L50 of 20.2 .2004). 

. Ddcret N° 2006-1523 du 04 decembre 2006 concemant les Bonnes Pratiques de Laboratoire 

(Journal Officiel du 06 decembre 2006), Minist&e de 1'Economie, des Finances et de 

1'Industrie. 

The study was also conducted in compliance with the following Animal Protection regulations: 

. Council Directive 86/609/EEC of 24th November 1986 on the harmonization of laws, 

regulations or administrative provisions relating to the protection of animals used for 

experimental or other scientific purposes. 

. Guidance document on the recognition, assessment, and use of clinical signs as humane 

endpoints for experimental animals used in safety evaluation, OECD Environmental Health 

and Safety Publications, No. 19. 

I declare that this report constitutes a true and faithful record of the procedures undertaken and 

the results obtained during the performance of the study. 

This study was performed at CIT, BP 563, 27005 Evreux, France,( ! 

Toxicology 

C. Pelcot Study completion d0e: J1 (A~i; t a~-? 

Study Director ,I 
OTHER CIT SCIENTIST INVOLVED IN THE STUDY 

Director of Toxicology 

CIT Management 

J.J . Legrand Date: 

Doctor of Veterinary Medicine 

r, 

fj ~tfGf 
STATEMENT OF QUALITY ASSURANCE LTNIT 

Inspections performed at CIT" 

The CIT Quality Assurance Unit conducted the inspections detailed below: 

Type of inspection 

Dates 

Inspection Reported to the Reported to 

Study Director Management 

Study plan 27 July 2007 27 July 2007 02 August 2007 

Report 21 March 2008 27 March 2008 08 April 2008 

In addition, at about the same time, process-based and facility inspections relevant to this study were carried out by the Quality Assurance Unit. 

The inspections were performed in compliance with CIT Quality Assurance Unit procedures and the principles of Good Laboratory Practices . 

The final report is considered to constitute an accurate and complete reflection of the study raw data. 

a . D C cc~_e~~~> Date: 

CIT Quality Assurance Unit 
SUMMARY 

r s acc g o OECD (N : , 2 th April 2002) and EC (2004/73/EC B .4; 

29th April 2004) guidelines. 

The study was conducted in compliance with the principles of Good Laboratory Practice 

Regulations . 

Methods 

The test item was first applied for periods of 3 minutes, 1 hour and 4 hours to a single male 

New Zealand White rabbit . Since the test item was neither severely irritant nor corrosive on this 

first animal, it was then applied simultaneously for 4 hours to two other animals. 

A single dose of 500 mg of the test item in its original form was applied to the closely-clipped 

skin of one flank. The test item was held in contact with the skin by means of a semi-occlusive 

dressing. 

Cutaneous reactions were observed approximately 1 hour, 24, 48 and 72 hours after removal of 

the dressing and then daily until reversibility of cutaneous reactions (day 8). 

The mean values of the scores for edema were calculated for each animal. 

Results 

After a 3-minute exposure (one animal) 

No cutaneous reactions were observed. 

A grey coloration of the skin was noted from day 1 until day 4. 

After a 1-hour exposure (one animal) 

A grey coloration of the skin was noted from day 1 until day 4. This coloration could have 

masked a possible very slight erythema on day 1 . No cutaneous reactions were observed from 

day 2 until day 4. 

After a 4-hour exposure (three animals) 

A grey coloration of the skin was noted in all the animals all over the observation period . This 

coloration could have masked a possible very slight or well-defined erythema from day 1 until 

day 2, 3 or 4. 

A very slight erythema was noted in 1/3 animals on days 4 and 5 . A dryness of the skin was 

noted in 2/3 animals between day 4 and day 7 . 

Due to the coloration of the skin, the mean scores over 24, 48 and 72 hours for each animal were 

not calculable for erythema. For edema, the mean scores over 24, 48 and 72 hours for each 

animal were 0.0, 0.0 and 0.0 . However, taking into account the possible erythema masked by the 

coloration, the maximal mean values over 24, 48 and 72 hours for erythema could be: 0.3, 

1 .0 and 1 .3 . 
ible maximal mean values for erythema, the test item 

mg-7o 

subsequent adaptations), the test item should not be classified as irritating to the skin. 
RESUME 

e Lapm se 

(2004/73/CEE, B .4, 29 avril 2004). 

L'etude a ete realisee conformement aux regles de Bonnes Pratiques de Laboratoire . 

Methode 

Le produit a d'abord ete applique pendant 3 minutes, 1 heure et 4 heures sur 1 seul Lapin male New Zealand White. Le produit n'ayant pas montre de proprietes severement irritantes ou corrosives sur ce premier animal, il a ensuite ete applique simultanement pendant 4 heures sur 2 autres Lapins. 

Une dose unique de 500 mg de produit tel quel a ete appliquee sur une surface de peau tondue 

au niveau d'un des flancs . Le produit a ete maintenu en contact avec la peau au moyen d'un pansement semi-occlusif. 

Les reactions cutanees ont ete observees environ 1, 24, 48 et 72 heures apres I'enlevement du pansement puis quotidiennement jusqu'a reversibilite complete des reactions cutanees (jour 8). La moyenne des scores pour 1'oedeme a ete calculee pour chaque animal. 

Res ultats 

Apres une exposition de 3 minutes (1 animal) 

Aucune reaction cutanee n'est observee. 

Une coloration grise de la peau est notee du jour 1 au jour 4. 

Apres une exposition de 1 heure (1 animal) 

Une coloration grise de la peau est notee du jour 1 au jour 4. Cette coloration a pu masquer un eventuel erytheme tres leger au jour 1 . Aucune reaction cutanee n'est observee du jour 2 au jour 4. 

Apres une exposition de 4 heures (3 animaux) 

Une coloration grise de la peau est notee chez tous les animaux sur toute la periode 

d'observation. Cette coloration a pu masquer un eventuel erytheme tres leger ou bien defini du jour 1 au jour 2, 3 ou 4. 

Un erytheme tres leger est note chez 1/3 animal aux jours 4 et 5. Une secheresse cutanee est notee chez 2/3 animaux entre le jour 4 et le jour 7. 

En raison d'une lecture masquee de 1'erytheme He a la coloration de la peau par le produit, les 

scores moyens pour chaque animal apres 24, 48 et 72 heures ne sont pas calculables pour 1'erytheme. Pour 1'oedeme, les scores moyens calcules sont de 0,0, 0,0 et 0,0 . Cependant, en tenant compte de t'eventuel erytheme masque par la coloration, les scores moyens maximum 

apres 24, 48 and 72 heures pour 1'erytheme pourraient etre : 0,3, 1,0 et 1,3. 
Conclusion 

les conditions experimentales de cette etu de par voie cutanee chez le Lapin, le produit 

-- --- mpechant de c uire sur son potentiel irrita 

es criteres de classification decrits dans 1a Directive 67%5 

ne devrait pas etre classe comme irritant pour la peau. 
1. INTRODUCTION 

s study was to evaluate the potential of the test ite 

induce skin irritation following a single topical applicahoin to-ra ~ i s . 

In the assessment of the toxic characteristics of a test item, determination of the irritant and/or 

corrosive effects on the skin of mammals is an important initial step. Information derived from 

this test serves to indicate the possible hazards likely to arise from exposure of the skin to the 

test item. 

This study was conducted in compliance with: 

. OECD guideline No. 404, 24th April 2002, 

. EC Directive No. 2004/73/EC, B.4, 29th April 2004. 

2. MATERIALS AND METHODS 

2.1 TEST ITEM 

2.1 .1 Identification 

Description at receipt:~Tack powder 

container: one plastic flask 

date of receipt: 29 June 2007 

storage conditions : at room temperature and protected from humidity 

composition : see analytical certificate 

expiry date : June 2008. 

Data relating to the characterization of the test item are documented in a test article description 

and in an analytical certificate (presented in appendix 1) provided by the Sponsor. 

2.1 .2 Dosage form preparation 

The test item was used in its original form. 

Fresh dosage form preparations were made by the CIT Pharmacy on the 

administration and any unused material was discarded that same day. 

morning of 

The pH of the test item at the concentration of 1% in purified water was not measurable (non 

soluble and colored test item). 
2.2 TEST SYSTEM 

2 .2 .1 Animals 

Sex, species, strain : male New Zealand White rabbits . 

Reason for this choice: species generally accepted by regulatory authorities for this type of 

study. 

Breeder: Grimaud freres selection S.A.S ., La Corbi6re, Roussay, France. 

Number: three animals were used, as recommended by the international guidelines . 

Age/weight: on the day of treatment, the animals were 2 to 4 months old and had a mean body 

weight t standard deviation of 2 .5 ± 0.1 kg. 

Acclimation : at least 5 days before the beginning of the study. 

Identification : individual metal ear tag . 

2.2 .2 Environmental conditions 

The conditions in the animal room were set as follows : 

. temperature : 18 ± 3°C 

. relative humidity: 30 to 70% 

. light/dark cycle: 12 h/12 h 

. ventilation : approximately 12 cycles/hour of filtered, non-recycled air . 

The temperature and relative humidity were under continuous control and recording. The 

records were checked daily and filed. In addition to these daily checks, the housing conditions 

and corresponding instrumentation and equipment were verified and calibrated at regular 

intervals. 

The animals were housed individually in Techniplast (64 cm x 63 cm x 30 cm) or Pajon (50 cm 

x 57 cm x 75 cm) cages. 

Each cage was equipped with a food container and a water bottle. 

2.2 .3 Food and water 

During the study, the animals had free access to 110C pelleted diet (SAFE, Villemoisson, 

Epinay-sur-Orge, France). 

Food is analyzed regularly by the supplier for composition and contaminant levels . 

The diet formula is presented in appendix 2. 

Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum . 

Bacteriological and chemical analyses of water are performed regularly by external laboratories . 

These analyses include the detection of possible contaminants (pesticides, heavy metals and 

nitrosamines) . 

No contaminants were known to have been present in the diet or drinking water at levels which 

may be expected to have interfered with or prejudiced the outcome of the study. 
2.3 TREATMENT 

2.3 .1 Preparation and selection of the animals 

p. 172 

13 

The day before treatment, both flanks of each animal were clipped using electric clippers and 

just before treatment, the skin of each animal was examined in order to check the absence of any 

signs of skin irritation . 

2.3 .2 Application of the test item 

The test item was first evaluated on a single animal (No. 513/474). The durations of exposure 

were 3 minutes, 1 hour and 4 hours. 

Since the test item was neither severely irritant nor corrosive on this first animal, it was then 

applied simultaneously for 4 hours to two other animals (Nos . 483 and 484). 

Doses of 500 mg of the test item in its original form were placed on a gauze pad moistened with 

purified water, which was then applied to an area of approximately 6 cm 2 of the anterior 

left flank (application for 3 minutes), the anterior right flank (application for 1 hour) or the 

posterior right flank (application for 4 hours) of the animals . 

The gauze pad was held in contact with the skin by means of an adhesive hypoallergenic aerated 

semi-occlusive dressing and a restraining bandage. 

The untreated skin served as control . 

After removal of the dressing, any residual test item was wiped off by means of a dry cotton pad 

or a cotton pad moistened with acetone then with water. 

2.4 CUTANEOUS EXAMINATIONS 

2.4 .1 Duration of the observation period 

The skin was examined approximately 1 hour, 24, 48 and 72 hours after removal of the dressing . 

Since there were persistent irritation reactions at 72 hours, the observation period was extended 

up to their complete reversibility (day 8). 

2.4.2 Description and evaluation of cutaneous reactions 

Dermal irritation was evaluated for each animal according to the following scoring scale : 

Erythema and eschar formation : 

. no erythema . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 0 

. very slight erythema (barely perceptible) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 

. well-defined erythema. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 

. moderate to severe erythema . . . . . . . . . . . . . . . . . . . . . . . . . . . ._ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 

. severe erythema (beet redness) to slight eschar formation (injuries in depth) . . . . . . 4 

Edema formation 

. no edema . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . :. . . . . . . . . . . . . . . . . . . . . 0 

. very slight edema (barely perceptible) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 

. slight edema (edges of area well-defined by definite raising) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 

. moderate edema (raised approximately 1 millimeter) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 

. severe edema (raised more than 1 millimeter and extending beyond area 

of exposure) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 

Any other lesions were noted . 
2.5 BODY WEIGHT 

p. 173 

14 

Each animal was weighted at the beginning (day of treatment) and at the end of the observation 

period. 

2.6 INTERPRETATION OF RESULTS 

The results obtained were evaluated in conjunction with the nature and the reversibility of the 

findings observed. 

2.6 .1 Interpretation of results 

2.6 .1 .1 Criteria for irritation 

A substance or a preparation is considered to be irritating to the skin if, when it is applied to 

healthy intact animal skin for up to 4 hours, significant inflammation is caused and which 

persists for 24 hours or more after the end of the exposure period . 

2.6 .1 .2 Criteria for corrosion 

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 of skin tissue on at least one animal during the 

test for skin irritation, or if the result can be predicted (for example : from strongly acid or 

alkaline reactions) . 

All scores obtained at each reading time (24, 48 and 72 hours) for an effect are used for 

calculating the respective mean values. 
2.7 ARCHIVING 

p. 174 

15 

The following study materials are archived by CIT, 27005 Evreux, France, for 10 years after the 

end of the in vivo phase of the study: 

. Study plan and possible amendments, 

. raw data, 

. correspondence, 

. final report and possible amendments. 

On completion of this period, the archived study materials will be returned to the Sponsor, or 

may be archived at CIT for a further period (at additional cost). The total duration of archiving 

(depending on regulations) will be the responsibility of the Sponsor. 

In addition, raw data not specific to the study including, but not limited to, certificates of 

analyses for food, water and bedding (if applicable) and records of environmental data and 

equipment calibration, are also archived by CIT for at least 30 years. 

2.8 CHRONOLOGY OF THE STUDY 

The chronology of the study is summarized as follows : 

Procedure 

Experimental starting date 

(day of treatment of the first animal) 

Experimental completion date 

(end of the observation period) 

2.9 STUDY PLAN ADHERENCE 

The study was performed in accordance w 

Date 

07 August 2007 

16 August 2007 

amendments, with the following deviation fro - -agr-7-~y an: 

nd subsequent 

. the temperature and relative humidity recorded in the animal room were sometimes outside of the target ranges specified in the Study plan. 

This minor deviation was not considered to have compromised the validity or integrity of the study. 
3. RESULTS 

The observations recorded during the study are presented in tables 1, 2 and 3 . 

The body weight is presented in table 4. 

After a 3-minute exposure (one animal) 

No cutaneous reactions were observed. 

A grey coloration of the skin was noted from day 1 until day 4. 

p. 175 

16 

After a 1-hour exposure (one animal) 

A grey coloration of the skin was noted from day 1 until day 4. This coloration could have masked a possible very slight erythema (grade 1) on day l . No cutaneous reactions were observed from day 2 until day 4. 

After a 4-hour exposure (three animals) 

A grey coloration of the skin was noted in all the animals all over the observation period. This 

coloration could have masked a possible very slight or well-defined erythema (grade 1 or 2) 

from day 1 until day 2, 3 or 4. 

A very slight erythema (grade 1) was noted in 1/3 animals on days 4 and 5 . A dryness of the 

skin was noted in 2/3 animals between day 4 and day 7 . 

Due to the coloration of the skin, the mean scores over 24, 48 and 72 hours for each animal were 

not calculable for erythema. For edema, the mean scores over 24, 48 and 72 hours for each 

animal were 0 .0, 0.0 and 0.0. 

However, taking into account the possible erythema masked by the coloration, the maximal 

mean values over 24, 48 and 72 hours for erythema could be: 0.3, 1 .0 and 1 .3 . 

4. CONCLUSION 

rimental conditions of this study, when applied topicall 

IM 
p. 176 

17 

Table 1 : 3-minute exposure - Cutaneous examinations and mean values of the scores recorded 

for the first animal (24, 48 and 72 hours) 

Rabbit Dermal Scores Mean 

number Irritation irritation 

Ih 24h 48h 72h score (1) 

D1 D2 D3 D4 

513/474 Erythema 0 0 0 0 0.0 

Edema 0 0 0 0 0.0 

Other C C C C 

(1) mean of scores on days 2, 3 and 4 

h = hour 

D =day 

C = grey coloration of the skin 
p. 177 

18 

Table 2: 1-hour exposure - Cutaneous examinations and mean values of the scores recorded for 

the first animal (24, 48 and 72 hours) 

Rabbit Dermal Scores Mean 

number Irritation irritation 

Ih 24h 48h 72h score (1) 

D1 D2 D3 D4 

513/474 Erythema C I 0 0 0 0.0 

Edema 0 0 0 0 0.0 

Other * C C C 

(1) mean of scores on days 2, 3 and 4 

h = hour 

D =day 

* = none 

C = grey coloration of the skin 

CI = grey coloration of the skin which could have masked a possible very slight erythema 
Table 3 : 4-hour exposure - Individual cutaneous examinations and mean values of the scores 

recorded for each animal (24, 48 and 72 hours) 

Rabbit Dermal Scores Mean 

number Irritation irritation 

lh 24h 48h 72h score (1) 

D1 D2 D3 D4 

513/474 Erythema C2 C1 0 0 (2) 

Edema 0 0 0 0 0.0 

Other * * C C 

483 Erythema C1 C1 C1 C1 (2) 

Edema 0 0 0 0 0 .0 

Other * * * 

484 Erythema C2 C2 C1 1 (2) 

Edema 0 0 0 0 0.0 

Other * * * C/S 

(1) mean of scores on days 2, 3 and 4 

(2) = not calculable 

h =hour 

D = day 

(+) = irritant according to E.E.C. criteria 

(-)=non-irritant according to E.E.C. criteria 

* =none 

S=dryness of the skin 

C=grey coloration of the skin 

C1 =grey coloration of the skin which could have masked a possible very slight erythema 

C2 grey coloration of the skin which could have masked a possible well-defined erythema 
Table 3 (continued) 

Rabbit Dermal Scores 

number Irritation 

513/474 Erythema 

Edema 

Other 

483 Erythema 0 0 0 0 

Edema 0 0 0 0 

Other C/S C/S C/S C 

484 Erythema 1 0 0 0 

Edema 0 0 0 0 

Other C/S C/S C/S C 

D = day 

S = dryness of the skin 

C = grey coloration of the skin 

- = cutaneous examination not performed 
Table 4: Individual body weight (g) 

Sex Animals Day of End of the 

treatment observation period 

Male 513/474 2607 2763 

483 2440 2560 

484 2351 2557 

M 2466 2627 

SD 130 118 

p. 180 

21 

M = mean 

SD = standard deviation 
p. 181 

22 

APPENDICES 
1 . Test article description and analytical certificate 
WENTITY 

Test artitle name 

Cbenimal name 

CAS numlrcr 

EINECS number 

Origin 

ch nuniber 

Arkerr.1a filing number 

~'STCALANI) CHE.',,Nl'fC-AL PROPERTIES 

Azrpcananc.e 

Partic"Ile. s 

Specific Gravity 

Antotgnition temperature 

Solub-111q, 

: black powder 

: 30 gin ( appmx:it.i9trl.y } 

: 2,1 kgleO at. 2VC 

: > 400 °C (standard : NT EN 502& 1-2-1) 

: in waier -. insoluble 

: Keep hcrni,etically closed in a dry, coo( avid well-ventiled 

place . 

: Jane 2003 

Incineration 
Reference de la m6thode 

DETERMINATION I ITEM RESULTAT I RESULT d'analyse I Analysis 

reference 

Powder characteristics 

Ash content % 7.6% ATG 

70 - 

. a,,.~. w 

Nom du responsable du taboratoirel Laboratory Director : 

Signature 

p. 184 

25 
2. Diet formula 
p. 186 

27 

Ref: 110 

COMPLETE DIET 

RABBIT BREEDING DIET 

Appearance: 3 mm diameter granules 

Conditioning: bags of 20 kgs 

Daily portion : Rabbits 150 g, water ad libitum . 

FORMULA % 

Cereals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Grain biproducts and 

leguminous plants . . . . . . . . . . . . . . . . . . . . . 

Vegetable protein (soya bean 

meal, yeast) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Vitamin and mineral mixture . . . 

AVERAGE ANALYSIS % 

Calorific value (Kcal/kg) . . . . . . . . . . 

Moisture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Lipids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Carbohydrates (N.F .E.) . . . . . . . . . . . . 

Fibre. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Minerals (ash) . . . . . . . . . . . . . . . . . . . . . . . . . . . 

AMINO ACID VALUES 

(calculated in mg/kg) 

Arginine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Cystine. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Lysine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Methionine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Tryptophan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Glycine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

FATTY ACID VALUES 

(calculated in mg/kg) 

Palmitic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Palmitoleic acid . . . . . . . . . . . . . . . . . . . . . . . 

Stearic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Oleic acid. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Linoleic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 

Linolenic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . 

33.8 

48 

14 

4.2 

3100 

10 

15 

2.3 

48.2 

17 

7.5 

11300 

3400 

9300 

280.0 

2400 

8700 

6400 

0 

600 

6400 

12100 

2400 

MINERALS (calculated in mg/kg) 

Nat. CMV 

val. val . Total 

P . . . . . . . . . . . . . . . . . . . . . 3600 2900 6500 

Ca . . . . . . . . . . . . . . . . . . 4200 5800 10000 

K . . . . . . . . . . . . . . . . . . . 12000 0 12000 

Na . . . . . . . . . . . . . . . . . 400 2000 2400 

Mg . . . . . . . . . . . . . . . . . . 2500 100 2600 

Mn . . . . . . . . . . . . . . . . 50 40 90 

Fe . . . . . . . . . . . . . . . . . . 150 150 300 

Cu . . . . . . . . . . . . . . . . . . Traces 15 15 

Zn . . . . . . . . . . . . . . . . . . 30 45 75 

Co . . . . . . . . . . . . . . 0.1 1 .5 1 .6 

I . . . . . . . . . . . . . . . . . . . . . 0.1 0 0.1 

Cl . . . . . . . . . . . . . . . . . . . 300 3000 3300 

VITAMINS (calculated per kg) 

Nat. CMV 

val. val. Total 

-------------------------------- Vitamin A Traces 10000 lU 10000 IU 

Vitamin D3 0 IU 1000 IU 1000 fu 

Vitamin 131 5 mg 0 mg 5 mg 

Vitamin 132 4 mg 0 mg 4 mg 

Vitamin 133 20 mg 0 mg 20 mg 

Vitamin 136 1 mg 1 mg 2 mg 

Vitamin B12 0 mg 0 mg 0 mg 

Vitamin E 15 mg 25 mg 40 mg 

Vitamin K3 0 mg 1 mg 1 mg 

Vitamin PP 60 mg 5 mg 65 mg 

Folic acid 0 mg 0 mg 0 mg 

Biotin 0 mg 0 mg 0 mg 

Choline 1000 mg 1000 mg 2000 mg 

Available under quality "Control Ref. : 110" 

SAFE, 7 rue Galli6ni, Villemoisson, 91360 Epinay-sur-Orge 

Tel : 01 .69 .04.03.57 - Fax : 01 .69.04.81 .97 

(Ref. Doc. UAR: 2000) 
3 . CIT GLP certificate 
i,a ..rlf - Ill~na - rroreraai 

Wum-eQpe FcaVCP"e 

GROUPE IIVTEIt1YIINISTERIEL DES PRODUITS CHIMIQUES 

Paris, le - s JuIL. 2007 

OObjet : Evaluation de la conformite aux Bonnes Pratiques de Laboratoires (BPL) selon les directives 

200419/CE et 20041101CE du 11 f8vrier 2004. 

Subject : Assessment of compliance with Good Laboratory Practices (GLP) under the EC directives 

2004/9 and 2004/10 of 11 February 2004. 

Cons6cutivement i3 votre engagement vis-6-vis du GIPC et du COFRAC et en application du d8cret n° 2006-

1523 du 4 dccembre 2006 concernant les bonnes pratiques de laboratoires et modifiant le d6cret n° 81-278 du 25 

mars 1981 portant creation d'un groupe interminist6riel des produits chimiques, je vous confirme quo le GIPC, 

au vu des rdsultats du contr8le exere6 par le Comite fran~ais d'accr8ditation (COFRAC) - Section Laboratoires a 

d6cid8 pour votre installation du statut suivant : 

Following your engagement vis-k-vis the GIPC and COFRAC and in application of the decree n° 2006-1523 of 

4 December 2006 relating to the good laboratory practices and modifying the decree n° 81-278 of 25 March 

1981 giving birth to an interministerial group of chemical products (GIPC, 1 confrrm to you that the GIPC, 

given the results of the inspection realised by the French Committee of accreditation (COFRAC) - Laboratory 

Section has taken the following decision relating 10),01V installation . 

Respect des principes de BPL Date d'inspection : 7-8 mars 2007 

Respect of the GLP principles Date of inspection : 7-8 mars 2007 

Domaines de reconnaissance : Inspection de renouvellement (i .r) 

1- essais physico-chimiques Renewal inspection (4 r) 

2 - etudes de toxicit8 

3 - 8tudes de mutag6nicit6 Date de d¬cision du GIPC : 29 juin 2007 

4 - rstudes icotoxicologiques sur les Date of GIPC decision : 29 juin 2007 

organismes aquatiques et terrestres 

8 - m8thodes de chimie analytique et Date de prise d'effet : 8 mars 2007 

clinique Date of implementation: 8 mars 2007 

Areas of expertise : Annee de premiere conformitd : 1989 

1 =Physico-chemical testing Year of the first conformity: 1989 

2 = Toxicity studies 

3 =Mutagenicity studies Dur¬e de validite : 18 mois 

4 = Environmental toxicity studies ort aquatic or Time of validity : 18 months 

terrestrial organisms 

8 =Analytical and clinical chemistry 

CENTRE INTERNATIONAL DE TOXICOLOGIE 

(CIT) 

MISEREY - BP 563 

27005 EVREUX CEDBX 

( 

IrA\A~ 

Pierre CPEYSSEL 

Conseiller d'Etat h 

Secr6tariat gdneral du GIPC - DGE- Simap - 12, me Villiot - 75572 Paris codex 12 

Telephone : 0153 44 96 10 - Telecopie : 0153 44 9172