# TSCA NON-CONFIDENTIAL BUSINESS INFORMATION DOCUMENT DESCRIPTION DOCUMENT CONTROL NUMBER DATE RECEIVED 

# COMMENTS:DE 

# L S

DOES NOT CONTAIN CBI

## UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON D.C., 20460 OFFICE OF 

### ~PRO' CHEMICAL SAFETY AND POLLUTION PREVENTION 

# Notice of Change inCBI Chemical Identity Status Pre-Manufacture Notice (PMN) P-03-0645 Notice of commencement (NOC) received: 216/2004 Please be advised that the Confidential Business Information (CBI) claim forthe above referenced substance was dropped bythe information submitter inthe NOC. The identified non-confidential chemical name for this substance as provided in the NOC is: 1,3-BENZENEDIMETHANAMINE, POLYMERS WITH EPICHLOROHYDRINPOLYETHLENE GLYCOL REACTION PRODUCTS* The current TSCA Inventory name is 1,3-Benzenedimethanamine, polymers with epichlorohydrin-polyethylene glycol reaction products CASRN: 652968-34-8
This notice as submitted tothe NCIC files on 9/1/201 1 Name of staffer: Jeffrey Santacroce Position: Asst. NOC Coordinator Branch: RDMB/IMD/OPPT/OCSPP 

# CONTAINS NO OIPMN Page 1 

### ; 

## Page 1 of 101

____________________________________________________ Form Approved. O.M.B. No. 2070-0012. Approval Exp s 10-31-96. U.S. ENVIRONMENTAL PROTECTIONAGENCV, - -~ 

### GNY S NY_ 

## PREMANIJFACTURE NOTICE 

## HOJ M, 1 8 AIN 101: 56 

# IL ~ 

# 111 ~ 

# III Iii ompanY Sanitized

inthe Premanufacture Notice

* Before youcomplete 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). * Ifa user feehas been remitted for this notice (40 CFR 700.45), indicate inthe boxes above the TS-user fee identification number youhave generated. Rememher, your user fee ID numbermust also appear on your corresponding fee remittance, which is sent to EPA,Washington Financial Management Center (3303), P 0 360399M, Pittsburgh, PA 15251-6399, Attn. TSCA User fee. Part I- GENERAL INFORMATION TEST DATA AND OTHER DATA You mustprovide the currently correct Chemical Abstracts (CA) Name ofthe new You are required to submit all test data in yourpossession or control and t chemical substance, evenifyou claim she identity as confidential. You may authorize provide a description ofall other data known to or reasonably ascertainable by another personi to submit chemical idensity information for you, but your submission will you, ifthese data are related to thehealth and envirornental effects on the sot he complete and the review will notbegin until EPA receives this information. A letter mauctrpoesndtibininomresoripslofhe w insupport ofyour submuission should reference yourTS userfee identification number. mnfcue rcsig itiuini omre so ipslo h e You must submit an original and twocopies ofthis notice including all test data. Ifyou ceia usac.Sadr ieauecttosmyh umte o ai h claimed any information asconfidential, a single sanitized copy must also be sbmittedl. open scientific literature. Complete test data (written in English). not surnaics ofdata, must be submitted ifthey do not avnear inthe open literature. Youshould clearly identify whether test data is on the substance oron an analog. Als ,the Pars 11 - HUMAN EXPOSURE AND ENVIRONMENTAL RELEASE chemical composition ofthe testedmaterial should he characterized. Foilcwing are examples oftest data and otherdata. Data should be submitted according to ifthere are several manufacture, processing, or use operations tobe described inPant 11, the requirements of§720.50 ofthe Premainufacture Notification Rule (40 GFR sectionsA and Boftbis notice, repreduce the sections as needed. Part 720).
Part Ill - LISTOF ATI'ACITMENTS Test Dats (Check Below any included inthis notice) Attach additional sheets ifthere is not enough space so answer aquestion fully. Label each *Environmental fate data 

## [) Yes *Other data 

## [] continuation sheet with shecorresponding section beading. in Pant ill, listthese attachments, any test data or other data and any optional information included inthe notice. * Health effectsdata 

## [] Yes Riskassessments * Environmental effects data 

## 5 Yes Structure/activity rela oshipsf OPTIONAL INFORMATION Physical/Chemical Properties* 

### ~ Yes Test data not inthe po ssion orcontrol ofthesubmi ter

## [] Youmay include any information that you want EPA soconsider inevaluating the new substance. On page IIofthis,form, space has beenprovided for you so describe 'A physical and chemical propeities worksheet islocated on the lastpage ofthi fonrs. pollution prevention and recycling information you may have regarding the new substance. TYPE OF NOTICE (Check Only One) So-called "binding" boxes are included throughout thisform foryou to indicate your willingness to he bound so certain statements you make inthis section, such as use, PMN (Premanufacture Notice) production volume, protective equipment ..This option isintended to reduce delays that 

## lJ routinely accompany shedevelopment ofconsent orders or Significant New Use Rules. Except in the case ofexemption applications (suchas TMEA, LVE, LOREX) where 

## [J INTERMEDIATE PMN (submitted insequence with final product certain information provided in suchnotification isbinding on the submitter when the Agency approves the exemption application, checking abinding box inthis notice does not SNUN (Significant New Use Notice) by itselfprohibit the submitterfrom later deviating from the information (except chemical E idetiy) eprte i th frm ThIEA (Test Marketing Exemption Application) CONFIDENTIALITY CLAIMS 

# [] LYE (Low Volume Exemption) @40 CFR 723.50(c))r You may claim any information in this notice as confidential. To assert a claim on the= form, markc (X) the confidential box nest tothe information that you claim as confidential. 

## Li LOREX (LowRelease/Lo Exposure Exemption) @40 CFR 7 

### c)22 

### "I To assert aclaim in an attachment, circle or bracket the information you claim as 

# E lI V)>C: confidential. Ifyou claim information inthe notices asconfidential.you mus also nrvde LE oiiainLRXMdfcto asnitized version ofshe notice. ( including attachments). For additional instructions o LEMdicaon[] LRX oifCato claiming information as confidential, rmad the Instructions Manual. ISTI ICNOIAEDPN e 

# [I Mark (x)ifany information inthis notice isclaimed asconfidential. # ofchemicals orpolymers 1 (Prenotice Communication #I required, enter#on page 3)___ EPA FORM 7710-2 Rv -5 Replaces previous editions ofEPA Form, 710-25. 

# -25 (Rev S-5

## Page 2 of 101 Public reporting burden for thiscollection of information isestimated toaverage 110hours per response, including timeforrviwg instructions, searching existing data sources, gathering and maintainingthe data needed, and completing and reviewing the coleion o information. Send comments regarding the burden estimate or any other aspect ofthis collection ofinformation, including suggestions Or reducing this burden, to Director,Collection Strategies Division (2822), U.S. Environmental Protection Agency, 1.200 Pennsylvania A e., N.W., Washington, D.C. 20460; and to the Office ofManagement andBudget, Paperwork Reduction Act (2070-0012), Washington, D.C. 20 03. 

### CERTIFICATION
- A Printed copy of this signature page, with original signature, must be submittedIcertify that to the bestofmyknowledge and beliefI. The company named in Part 1,section A, subsection Ilaofthis notice form intends tomanufacture or import for a -commercial purpose, other than in small quantities solely for research and development, the substance identified in Iart 1,

Section B. 2. All information provided in thisnotice iscomplete and truthful as ofthe date ofsubmission. 3. 1am submitting with this notice all test data inmy possession orcontrol and adescription ofall other data known to or reasonably ascertainable by me asrequired by 

### § 720.50 ofthe Premanufacture Notification Rule. Additional Certification Statements: Ifyou are submitting a PMN, Intermediate PM4N, Consolidated PMN, or SNUTN, check the following user fee certiflcati )n statement that applies: SThe Company named in Part 1,Section Ahas remitted the fee of$2500 specified in40 CFR 700.45(b), or El The Companynamed in Part I, SectionA has remitted the fee of$1000 foran Intermediate PMN (defined 

### @40 CF 700.43) in accordance with 40 CER 700.45(b), or Dl The Company named in Part ISection Ais a small business concemn under 40 CFR 700.43 andhas remitted a fee o~$100 in accordance with 40 CFR 700.45(b). Ifyou are submritting alow volume exemption (LVE) application inaccordance with 40 CER 723.50(c)(1) or a Low re ease and low exposure exemption (LoRex) application in accordance with 40 CER 723.50(c)(2),check the following certi 

### ~ation statements: 

# F The manufacturer submitting thisnotice intends to manufacture or import the new chemical substance for commerc al purposes, other than insmall quantities solely forresearch and development, underthe terms of40CFR 723.50. SThemanufacturer is familiar with the terms ofthis section andwill comply with those terms; and 

## F- Thenew chemical substance for which thenotice is submitted meets all applicable exemption conditions. SIfthis application is for an LVE in accordance with40 CFR 723.50(c)(1), the manufacturer intends to commence

manufacture ofthe exempted substance for commercial purposes within Iyear ofthe date ofthe expiration ofthe 3 day review period. The accuracy ofthe statements you make in this notice should reflect your best prediction oftheanticipated facts regarding the chemical substance described herein. Any knowing and willfulmisinterpsretationIssubjectto criminalpenalty pursuantto 1s usc 1001. ____cfidential Signa~tu ditie ofA tborizoi Official (Ongina] signature Requwred) Date Signat-6agh t- (isf plicabi

- Date

EPA FORM 7710-25 (Rev. 5-95) Page2PMN Page 3 

## Page3of 101 PartI - GENERAL INFORMATION Section A - SUBMITTER ILDENTIFICATION ConfiMark ()the "Confidential" box next toany subsection you claimas confidential dential_ Ia. Person Nameofauthorized official Position NuitingU.. Mr.. 1(zuki Puj imoto Mathager -ChemicalsE 

# ] Notce(inU .) CompanyF 

# 1 Mitsubishi Gas Chemical America, Inc. 520 M~adison Avenue

b. Agent (if Name ofauthorized official Psto applicable) Robert 7. .Reynolds consultant Company Regulatory Compiliance servicea Mailing address (number and street) 22312 colony Woods Drive City, State Postal Code Telephone (include area code) Apex, NC 27523 (919) 303-5018 c. if you are submitting this notice aspart ofajoint submission, mark (X)this box._________________________ Joint Submitter (if Name ofauthorized official Psto applicable)[I Company

2. Technical Name ofauthorized official Psto Contact (in Robert J.. Reynolds Consultant U.S.)

2312 Colony Woods Drive

3. Ifyou have had aprenotice communication (PC) concerningthis notce and EPA asaigned aPCNumber to the notice, enterthe number. Mr X 4. Ifyou previously submitted an exemption application for the chemical substance covered bythis notice, enter the exemption number assigned by EPA Ifyou previouslysubmitted aPMN fur this substance enter the PMN fin

number assigned byEPA (i.e. withdrawn orincomplete). - ifr none 

# E: 5. Ifyou havesubmitted anotice ofBona fide intent tomanufacture or import forthe chemical substance covered bythis notice, enterthe notice number Mr X assigned by EPA. if none 

# I 6.. Typofotce-as(X tr :2. I mf 

# El ot 6.Tp fNtc ak()Only IOnly 1 3. Both 

# [] Binding Option Lit Binding Option Mark (X) IMark (X)

____
EPA FORM7710-25 (Rev. 5-95) Page 3

### PMN Page4 

## Page4of 101 PartI - GENERAL INFORMATION - Continued____ Sectin B-CHEMIAL iD TIT iNFOMATIO: YoumustproIde acurrently conrect Chemical Abstracts (CA) name ofthe substance based othninth ColcieIdx(9C0) ofCA nomenclature rules and conventions.

________ Mark (X)the "Confidential" boxnext to any item you claimas confidential Compilete either item 1(Class I or2substances) or2(Polymers) as appropriate. Complete all other items. If another person willsubmit chemical identity information for you (for either Item I or2), mark (X)the box atthe right. ConfiIdentifythe name, company, and addressofthat person in acontinuation sheet.L dential 1.Class 1 or 2chemical substances (fordefiniitions ofclass I and class2substances, see the Istructions Manual) a. Class of substance- Mark (X) I []CasI or 2 E ls b. Chemical name (Currently correct Chemical Abstracts (CA) Name that isconsistent with TSCA Inventory listings for similar substances. For Class I substances aCA Index Name must beprovided For Class 2substanceseither a CA Index Name or CA Preferred Namne muttibe provided which ever isappropriate based on CA 9C0 nomenclature rules and conventions).

c. Please identify which method you usedto develop or obtainthe specified chemical identit inormation reported inthis notice: (cheek one). 

# [IMethod I (CAS Inventory Expert Service - acopy ofthe Identification Method 2(Other Source)

report obtained fromthe CAS Inventory Expert Services mustbe submitted as an attachment tothis notice) exists forthesubstanceI e. For aclass I substance, provide acomplete and correct chemical structurediagram. For aclass2substance, provide acorrect representative or partial chemical structure diagram, as complete as can beksnown, ifonecan be reasonably ascertained. Please seethe E-PMN Instruction Manual For discussionof "native format" diagram software which can be helpfu~l in reviewingyour substance.X



# Q Mark (X) this box if youattach acontinuation sheet._____
EPA FORM 7710-25 (Rev. 5-95) Page 4

### PMNPage 4a 

## Page5of101 Foraclass 2 substance - (1)Listtheimmediate precursor substances withtheir respective CAS Registry Numbers. (2)Describe the nature ofthe recion orprocess. (3)Indicate the rangeofcomposition and the typical composition (where appropriate)._____

e-(1) Listthe immediate precursor substances withtheir respective CAS Registry Numbers. GonfiName (CAS dential

e.(2) Describe the nature ofthe reaction or process. This is an.imported material. The process isunknown.II

e. (3) Indicate the range ofcomposition and thetypical composition (where appropriate). There is no detectable residual monomer in the PMN substance[I

10Markc (X) this box ifyou attach acontinuation sheet._______ EPA FORM 7710-25 (Rev. 5-95) Page 4a

### PMNPage 5 

## ;Page6of 101 PartI - GENERAL INFORMATION - Continued Section B - CHEMICAL IDENTITY INFORMATION - Continued 2. Polymers (For a definition ofpolymer, see the Instructions Manual.) Confidential a. Indicate the number-average weight ofthe lowest molecular weight composition ofthepolymer you intend to manufacture. Indicate maximum weightpercent oflow molecular weight species (not includingresidual monomners, reactants, or solvents) below 500 and below 1,000 absolute molecular weight ofthatcomposition. Describethe methodsofmeasurement orthe basis for your estimates: GPC

### Li Other 

## 1 ]: (Specify below)_____ (i) lowestnumber average molecular weight: 1,831 See Attachment 8 (ii) maximum weight %below 500 molecular weight:0 (iii)maximum weight %below 1000 molecular weight: 0 Mark (X)this boxifyou attach a continuation sheet.______ b. You mustmake separate confidentiality claims formonomer or otherreactant identity, composition information, and residual information. Mi(X)the "Confidential" box next to any item youclaim as confidential (I)

- Provide the specific chemical name and CAS Registry Number (ifa number exists) ofeach monomer orotherreactant used inthe maufacture of the polymer. (2) - Mark (X)this column ifentry incolumn (1)isconfidential. (3) - Indicate thetypical weightpercent ofeachmonomer orotherreactant inthe polymer. (4) - Choose "yes" from drop down menu ifyou want amonomer or otherreactant used attwo weight percent or less to be listed as part ofthe polymer description on the TSCA Chemical Substance Inventory. (5) - Mark (X)this column ifentries in columns (3)and (4)are confidential. (6) - Indicate the maximum weight percent ofeachmonomer or other reactant thatmay be presentas a residual inthepolymer as manufa tured for comamercial purposes. (7) - Mark (X) this column ifentry incolumn (6) isconfidential.___________

_____ Monomer or otherreactant andCAS Registry Number Confi- Typical Include in Confi- Maximum Confidential composition identity dential residual dential

_ _ _ _ _ _ _ _ _ _ _ _ _(4) (_) (6) (7) 

# % 

# T._.......

______ _[___EiOL EZ [Ii

_ _ 

# L

_ 

### V0i 

# LI %] _L 0 ak X ti bxifyu tac

cninaio het E ]____ EPA ORM771025 Rev 5-9) Pge% 

# 6%PMN Page 5a; 

## Page 7 

## of 101
c. Please identify which method you used to develop or obtain the specified chemical identity information reported in this notice (check one). CBI 

# [E] Method I (CAS Inventory Expert Service - a copy ofthe identification report [] Method 2 (other source) obtained from CAS Inventory ExpertService must be submitted as an attachment to this notice) 

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

CAS Registry Number (ifanumber already exists for the tubstance)li 

# ]

e. Provide a correct representative or partial chemical structure diagram, as complete as can be known, ifone can be reasonably ascertained.. Plea!e see the E-PMN Instruction Manual for discussion of"native format" diagram software which can be helpful in reviewing your substance.[ 

# J

ElMark (X) this box ifyou attach acontinuation sheet.

EPA FORM 7710-25 (Rev. 5-95) Page SaPMN Page 6 

## Page8ofl101 Part I - 

### GENERAL INFORMATION - 

### Continued

___ SectionB - CHEMICAL IDENTITY INFORMATION - Con~tinued 3. Impurities (a)

- Identify each impurity thatmay be reasonably anticipated to be presentus the chemical substance asmanufactured forcommercial purpose. Provide the CAS Registry Number ifavailable. Ifthere are unidentified impurities, enter "unidentified." (b)

- Estimate the maximum weight% ofeach impurity. Ifthere are unidentified impurities, estimate their total weight%.4 Impurity and CAS Registry Number Maximum Confipercent dential (a) _b

0

0 

# [J Mark (X)thisbox ifyouattach acontinuation sheet. 4. Synonyms - Enterany chemical synonyms for the new chemical identified insubsection Ior 2. Confidential 

# L~x] E] Mar. (X) this box ifyouattach acontinuation sheet. 5. Trade identification -List trade names for thenew chemical substance identified in subsection 1or2. Memory Green Crystal[I 

# [] Mark (X) this boxifyouattach a continuation sheet. 6. Generic chemical name - Ifyou claim chemical identify asconfidential, youmust provide a generic name for your substance thatreveals thespecific chemical identity ofthe new chemical substance tothe maximun extentpossible. Refer to the TSCA Chemical Substance Inventory, 1985 Edition, Appendix B for guidance on developing generic names. Polyether polyol 

# fl Mark (X)this box ifyou attach acontinuation theet_

____ 7. Byproducts - Describe any byproducts resulting fromthe rmufacture, processing, use, ordisposal ofthe newchemical substance. Provide the CAS Registry Number ifavailable.

_______________ ____ Byproduct CAS Registry Number Confi- (1) (2) dential None_ 

# [] Mark (X) this box ifyou attach acontinuation sheet. EPAFORM 7710-25 (Rev. 5-95) Page 6

### PMVN Page 7 

## Page9of 101 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 ofproduction. Also estimate the max nurn production volume for any consecutive 12-month period during the first three years ofproduction. Estimates should be on 100% nei chemical substance basis. For aLow Volume Exemption application, ifyouchoose to have your notice reviewed at a lower production volun than 10,000 kg/yr, specify the volume and mark (x) in the binding box. Ifgranted, you are bound tothisvolume Maximum first 12-month production (kg/yr) Maximum 1 2-month production (kglyr) Cot i- Binding ( 0 %n wc e i a su sa c ba i)(100% new chemical substance basis) den ial Option~ 2. UseInformation - Youmust make separate confidentiality claims for the description ofthe category ofuse, the percent ofproduction volume devotdd toeach category, the formulation of the new substance, and otheruse information. Mark (X) the "Confidential" Boxnext to any itemyou claim as confident a]. a. (1)
--Describe each intended category ofuse ofthe new chemical substance by funtction and application. (2) -Mark (X)this column ifentry col mn(I) is ,onlldential business information (CB1). (3)
--Indicate your willingness to have the infonmation provided incolumn (1)binding. (4)
--Estimate the percent of total rToduction forthe first three years devoted toeach category ofuse. (5)-Mark (X) this column ifentry incolumn (4)is confidential business information QRBI). (6) -- Estimate thepercent ofthe new substance as formulated inmixtures, suspensions, emulsions, solutions, or gels as manufactured for commercial purposes al sites under (our control associated with each category ofuse. (7)
-- Mark (X) this column ifentry in column (6) is confidential business information (CB1). (8)
--Indickte %of 3roduct volume expected for the listed "use" sectors. Mark more than one box ifappropriate. Mark (X) to indicate your willingness tohave the use type p ovided in (8) iinding. (9)
--Mark (X) thiscolumn ifentry(ies) in column (8) is (ae)onfidential business information (C 1) Category ofuse (1) (by function and application i.e. a CR1 &-ng Prod- CBI %in CBI % ofsubstance expected peru!e CR! dispersive dye for finishing polyester fibers) Mark~ %uc tion Fon(8 (2) (3) (4) (5) (6) (7) S11. i-ldn cors- liding (9) boartd mmrtot mria S0on

__ water retention Aid for orrianental %]F~ plAnts 

# [ 1 10010 

## EL 1 100%00 

# _______[II[II[]%[IHI

*If youhave identified a "consumer" use,please provide on a continuation sheetadetailed description ofthe use(s) ofthischemical substance in consui e products.

Inaddition include estimates oftheconcentration ofthe new chemical substance as expected inconsumer products and describe the chemical reactions by which this substance loses its identityin the consumer product. 

### [X1Mark (X) thisbox ifyou attach a continuation sheet. b.Generic use Ifyou claim any ctitegory ofuse description in subsection2a asconfidential, enter a generic description ofthat category. Read t eInstruction description Manual forexamples ofgeneric use descriptions. 

# DMark (X) thisbox ifyou attach acontinuation sheet. 3. Hazard Information - Include in thenotice acopy ofreasonable facsimile ofany hazard warning statement, label, material safety data sheet, or othei Biding

information which will be provided to any personwho isreasonably likely tobe exposed to this substance regarding protective equipment orpractices for Mwkto) the safehanding, transport, use, ordisposal ofthenew substance. Listinpart IIIhazard information you include. M~ Mark (X) thisbox ifyou attach hazard information.[1 

# 1 EPA FORM 7710-25 (Rev. 5-95) Page 7PMN Page8 

## Pagel10of 101 Part11- HUMAN EXPOSURE AND ENVIRONMENTAL RELEASE Section A - INDUSTRIAL SITES CONTROLLED BY THE SUBMITTER Mark (X)the "Confidential" box nextto ary item you claim as confidential Complete section A for each type ofmanufacture, processing, oruse operation involving the new chemical substance atindustrial 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 Confla. Identity -- Enter the identity ofthe site at which the operation will occur. dential 

### Name[I Siteaddress (number and street) City, County, State,ZIP code Ifthe same operation will occur at more than one site, enter the number ofsites. Identify the production rates oroperations, include all the information requested in this section for those[X 

### Mak()0 Manufacturing 

### 0I Processing 

### 1 

# [Zs]

c. Amount and Duration -- Complete 1 or 2 as appropriate_______________

_____________ Maximum kg/batch tiooA.cwc ciicai Hours/batch Batches/year 1. Batch btac)[i] Maximum kg/day (IOVA amchanical Hours/day Days/year 2. Continuous substancc) 

# [ii
d. Process description 

# [] Mark (X)to indicate yourwillingness to have your process description binding. 

# [ I (1) Diagramthe majorunit operation steps andchemical conversions. Include interim storage andtransport containers (specify- e.g. 5gallon pails, !5 gallon drum, rail car, tank truck, etc.). (2) Provide the identity , the approximate weight (by kg/day or kg/batch ona 100%new chemnical substance basis), and entry point ofall starting mtrials and feedstocks (including reactants, solvents, catalysts, etc.), andof all products, recycle streams,and wastes. Include cleaning chemicals (note frequency ifnot used daily orper batch.). (3) Identity bynumber the points ofrelease, including small orintermittent releases, to theenvironment ofthe new chemical substance. Ifreleasing otwo media atthe same step, assign a second release number forthe second medium. Product will be imported to the U.S. and stored at strategically located public warehouses fr distribution to retail outlets. Submitter may import tIhe PM'N substance in bulk, and contrac with

adomestic facility for repackaging None of these facilities 3.s under control of the subsj ea'.



# ElMark (X)this box ifyou attach a continuation sheet. EPA FORM 7710-25 (Rev. 5-95) Page8

### PMN Page 8a; 

## Page 11 of 101 Confidential

EPAFORM 7710-25 (Rev. 5-95) Page 8aPMN Page9 ; 

## Page12 ofl101 Part11- HUMAN EXPOSURE AND ENVIRONMENTAL RELEASE - Continued Section A - INDUSTRIAL SITES CONTROLLED BY THE SUBMHTTER - Continued 2. Occupational Exposure - You mustmake separate confidentiality claims forthe description ofworker activity, physical form ofthenew chemical SubaCe, number of works exposed, and duration ofactivity. Mark (X) the "Confidential" box next toany itemn you claim as confidential. (1)- Describe the activities (i.e. bag dumping, totefilling, unloading drums, sam~pling, cleaning, etc.) inwhich workers may be exposed to the substance (2)- Mark (X)this column ifentryin column (1)is confidential business information (CBI). (3) - Describe any protective equipment andengineering controls usedtoprotect workers. (4) and (6)- Indicate yourwillingness tohave the information provided incolumn (3)or(5) binding. (5) - Indicate the physical form(s) ofthe new chemical substance (e.g., solid: crystal, granule,powder, or dust) and %new chemical substance (ifpart a mixture) atthe time ofexposure. (7)

# -- Mark (X)this column ifentryincolunm (5)isconfidential business information (CBI). (8) - Estimate the maximum number ofworkers involved in eachactivity for all sites combined. (9) -Mark (X)this column ifentry in column (8) is confidential business information (C131). (10) and (11) -- Estimatethe maximum duration ofthe activity forany worker inhours perday and days per year. (12) -- Mark (X) this colwnmin ifetries in columns (10) and (11) are confidential business infrmtion_(C131).

____ ____ __ Worker activity CBI Protective Equipment/ Bindig Physical Bawidg C~l #of CB1 Maximum duration CBI forms(s) (i.e., bag dumping, filling drums) Engineering Controls Op t ion and%new Option Workers Hrs/day Days/yr Mark WX substance MarkW Exposed (2) (4) (5) (6) (7) (8) (9) (10) (1I) (12)


# Mark 

# L ZXE

_ -_s 

# _Hfyuatahacniuainset

_____A []R Zr12 

# (Rv 5-95 9PMNPage 9a; 

## Page 13of 101 3. Environmental Release and Disposal -- You mustmake separate confidentiality claims for therelease numnber and the amount ofthenew chemical sub -,ace released andother release and disposal information. Mark (X)the "Confidential" box nextto each item you claim as confidential. (1) --Enterthe number ofeach release point identified in theprocess description, partIt,section A, subsection Ild(3). (2) - Estimate the amount ofthenew substance released (a) directly to the environment or (b)into control technology (in kglday or kg/batch). (3).-- Mark (X) this column ifentries incoluims (1)and (2) are confidential business information(C131). (4) - Identify'themedia (stack air, fugitive air(optional-see Instruction Manual), surface water, on-site or off-site land or incineration, POTW, or other (!peci1f')) to which the new substance willbe released from thatrelease point. (5) --a. Describe control technology, ifany, andcontrol efficiency thatwill heused to limit therelease ofthenew substance to the environment. Forrel-ases disposed ofonland, characterize the disposalmethod and statewhether itisapproved fordisposal ofRCRA hazardous waste. On a continuation seet, for each site describe any additional disposal methods that will be used and whether the waste issubject to secondary or tertiary on-site treatment b. Estimate the amount releasedtothe environment after control technology (in kg/day). (6) - Mark (X) this column ifentries incolumns (4) and (5)are confidential business information (CBI). (7) -I1dentify the destination(s) ofreleases towater. Please supplyNPDES (National Pollutant Discharge Elimination System) numbers for direct dischages or NPDES numbers ofthe POT (Publicly Owned Treatment Works). Mak(X) ifthe POTWname orNPDES #is confidential business informatio (C2BI). Release Amount ofnew C131 Medium of Control technology andefficiency (you may wishto optionally attach Cal Number substance released release efficiency data)

___ _______

(1) (2a) (2b) cg.9. Binding (3) (4) (5a) Mr()(5b) (6) 

# El IL I

_E 

# D _E (7)[I Mak_)th_

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# eesstowtr N D S B 

# El_________Li]d Li __Li] FII

()Mark(X) the desf.ata otinuation sheeto t. N D S AFM1 

### (evs) 5-5)Pge9PMN Page 10; 

## Page 14 of 101 Part11- HUMAN EXPOSURE AND ENVIRONMENTAL RELEASE - Continued Section B - INDUSTRIAL SITES CONTROLLED BY OTHERS Complete section B for typical processingoruse operations involvingthe newchemical substance at sites you do not control. importers do not haveto comp]-e this section for operations outsidethe U.S.; however, you must report any processing or use activities afterimpost Seethe Instructions Manual. Complete aseparate sectionBfor eachtype of processing,ormve operation involvingthe newchemical subsiance. Ifthe same operation isperformed at more than one sitedescribethe typical operation counon to these sites. Identifyadditional siteson acontinuation sheet. 1(a). OperationDescription -- To claim information in thissection asconfidential, circle or bracket the specific information thatyou claim as confidential. (I) -- Diagram the major unit operation steps and chemical conversions, including interim storage and transport containers (specify - e.g. 5gallon pails, 55 gallon drums, rail cars, tank trucks, etc). On the diagram, identifyby letter andbriefly describe each workeractivity. (2)--Either in the diagram or inthe textfield 1(b) below, provide the identity, the approximate weight (bykg/day or kg/batch. on an 100% new chemical substancebasis), andentry pointof aUl feedstocks (including reactant solvents and catalysts, etc) and allproducts, recyclesteams, and wastes. Include cleaningchemicals (note frequency if not used daily or perbateh). (3)--Either inth, diagram or inthe text field I1(b) below, identifyby numberthe points ofrelease, including smallor intermittent releases, to theenvironment ofthe newchemical substance. (4) Please enter the l# ofsites (remember toidentify the locations ofthese sites on acontinuation sheet):

______ #I

ofsitesCB



## Diagram the major unit operation steps and chemical conversions...

(b). (Optional) This space is foratextdescription to clarifythe diagram above. 

# [E1 ]

UMark (X)this box ifyou attach acontinuation sheet. EPA FORM 7710-25 (Rev. 5-95) Page 10 

# /YPMN Page 10a 

## Page15of 101 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 ofworkers exposed for all sites combined. (4)- Estimate the typical duration ofexposure per worker in (a) hours per day and (b) days peryear. (6) - Describe physical fonm ofexposure and %new chemical substance (ifin mixture), and any protective equipment and engineering controls, ifany, usCd to protect workers.

- (7) - Estimate the percent ofthe ne~w substance as formulated when packaged or used as a final product. (9) --From the process diagram above, enter the numnber ofeach release point. Complete 9-13 for each release point identified. (10) - Estimate the amount ofthe new substance released (a) directly to the environment or (b) into control technology to the environent (in kg/day or kgbteh). (12) - Describe media ofrelease i.e. stack air, fuigitive air (optional-see Instructions Manual), surface water, on-site or off-site land orincineration, POTW, o ther (specify) and control technology, ifany, that will be used to limit the release ofthe new substance to the environmsent. (14) -- Identify byproducts; which mayresult from the operation.

_______ 

### ~(3), (5), (8) (11), (13) and (15) --Mark (X) this column ifany ofthe proceeding entries are confidential business information (CBI)____ Lett

#o CR1 DuraionrcOf CB1 Protective Equip. /Engineering Controls/Physical Formand! %new substance i B of Workers Epsur FormActi- Exposed ulation vity (1) (2) (3) (a (4) (5) (6) (7) (8)

Release Amiount of BIMedia ofRelease &Control Technology CR! Number New Substance Reteased (9)L (10a) (10b) (i)(12) (13)

(14)- Byproducts: (15) 

# EJ Mark (X) this box ifyou attach a continuation sheet,_______ 

## OPTIONAL POLLUTION PREVENTION INFORMATION To claim information inthe following section asconfidential circle orbracket the specific information that you 

### claimasconfidential. In this section you mayprovide information notreported elsewhere inthis form regarding your efforts toreduce orminimize patetial risks associated with activities surrounding manufacturing, processing, use and disposal ofthePMN substance. Please include new* formation pertinent to pollution prevention, including source reduction, recycling activities and safer processes orproducts available due t4 the new chemical substance. Source reduction includes the reduction in the amount or toxicity ofchemical wastes by technological mad fcation, process and procedure modification, product reformulation, raw materials substitution, and/orinventory control. Recycling refe stothe reclamation ofuseful chemical companents from wastes thatwould otherwise be treated orreleased as air emissions or water & -harges, or land disposal. Descriptions ofpollution prevention, sourcereduction and recycling should emphasize potential riskreduction su )Sequent to compliance with existing regulatory requirements and can beeither quantitative orqualitative. The EPA is interested in the infoi ation to assess over-all netreductions intoxicity orenvironmental releases and exposures, not the shifting ofrisksto other environmental media ornonenvironmental areas (e.g., occupational orconsumer exposure). In addition, information on therelative cost or performance cha acteristics of the PMN substance topotential alternatives may beprovided. All information provided in this section will be taken intoconsideration duringthereview of this substance. See PMN Instructions 1ManualandPollution Prevention Guidance manualforguidance andexamples.

__ ____

EPA FORM 7710-25 (Rev. 5-95) Page 10aPMN Page 11 

### ; 

## Page 16 of 101 OptionalPollution Prevention Information(Continued) Describe the expected net benefits, such as (1)an overall redudion in riskto human health orthe environment (2) areduction in the volume manufactured; (3) a reduction in the generation ofwaste mn terials through recycling, source reduction or other means; (4) a reduction in potential toxicity or human exposure and/or environn ental release; (5)an increase in product performance, a decrease in the cost ofproduction and/or improved operation efficiency othe new chemical substance in comparison to existing chemical substances used in similar application; or (6) the extent to which tht new chemical substance may be a substitute for an existing substance that poses a greater overall risk tohuman health or the en% ironment.



# F] Mark (X this box ifyou attach acontinuation sheet. EPA FORM 7710-25 (Rev. 5-95) Page I1I

### PMN Page 12 ;Page 17 

### of 19 

### PartIII -- 

### LIST OF ATTACHMENTS___ Attach continuation sheets for sections ofthe form and test data and other data (including physical/chemical properties and structure/activity infornation), and optional information after this page. Clearly identify the attachment andthe section ofthe formto which itrelates, if appropriate. Number consecutively the pages ofany paper attachments. In the colurmi below, enter the inclusive page numbers ofeach attachment. Electronic attachments can be identified by filename. 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 ofthe notice form a sanitized version ofany attachment in which you claim information as confidential.

___________ # Attachment name Attachment Filename Attachment Confi- ________________________________________________page_ number(s) dential 1 Consumer Use continuation (19 

# L 2 

# 1 11
3 Material Safety Data Sheet 

# I

________ 

### 21-25 

# 11
4 Bacterial Reverse Mutation Study 

# j_ ________j26-47jj 5 Skin Sensitization Study j_ ________j48-64 _ 

# El 6 Primary Skin Irritation Study j

________ 65-79 

# L 7 Acute Oral Toxicity Study 

# [B__________ O-100 0I 

## 811 

# L L L L El n () hibxfouatacacotiuaio

see. ntrheatacmet am adumer
EPA_

_ _ _ _ _ _ FOM71-5(ev -5 ae1PMN PageI13 ; 

## Page18ofl101 PHYSICAL AND CHEMICAL PROPERTIESWORKSHEET__ To assist EPA'sreview ofphysical and chemical properties data, please complete the following worksheetfor data you provide and include it inthe notice. Identify theproperty measured, the page ofthenotice on which the property appears, the value ofthe property, the units in whichthe property is measured (as necessary), and whether ornot the property isclaimed as confidential. Ifthe attachments are electronic, give the attachment number (found onpage 12) at (b). Thephysical state ofthe neat substance should be provided. These measured properties should be for the neat(100/6 pure) chemical substance. Properties that are measured for mixtures or formulations shouldbe sonoted (%/PMN substance in -J. You are notrequired to submit this worksheet; however, EPA strongly recommends thatyou do so, as itwill simplify'review and ensure that confidential informnation isproperlyprotected. You should submit this worksheet as a supplement toyour submissionoftestdata. Thisworksheet is not a substitute for submission oftest data. Property Mark(X PaeValue Measured ConSiif jnumer Euiate dendal (a) provided (b) (c) ( NrE) 

### Physical state ofneat substance 

# [Ii[J (s) L( 

# Ig M 

## [j] Vapor pressure[I @ Temperature 0

## C 

# ElTonr E:_] Density/relative density 

# [E ] g/cm3

____ 

# I Solubility 

### @ Temperature ______C[] 

# ED

Solvent___

__ __ g/L

_ Solubility in water @Ternperature

_____ 

# 111.1/L ::

Melting_ _ _ tepeatr _ o Boiling / sublimation temperature@_ -_tonf pressure 

# [I1I0 

# E::] Spectra 

# EI] [I Dissociation constant[ 

# ] ] Particle size distribution 

# L ]__ I Octanol / waterpartition coefficient 

# FI] I]J Henry's L.aw constant E l

____________ 

# i Volatilization fromwater EI] 

# [Fi1 Volatilization fromsoil 

# 0 1] [I pH @ concentration 

# [ED

____________ __ Flammability 

# LI E:1_ L Explodability 

# 17] ____________3 ___ 

# I Adsorption /coefficient 

# Lii 

# ED11
Other - Specify 

### EZ________________ 

# I]- Other- Specify[I_________

_ ___1 

# ] [J Mark~ (X)this boxNf youattach a continuation sheet Enter theattachment name andnumber. ATTACHMENT PAGE 13 

# 49

## Page 19 of 101 Continued From: Part 1, Section C2a, pg 

### 7 Continuation Title: Consumer Use continuation Insert narrative continuation here Cni 

### The PMN substance will be supplied as small, irregularly-shaped pellets that are about 0.1-1 cT 

# L in diameter. The pellets,though dry inappearance and feel, will be about 80% water. They will be packaged incontainers upto20 kg capacity perpackage and sold to consumers through garden supply outlets. The PMN substance may also be shipped in bulk or large capacity containers to nurseries. The PMN substance may also be shipped inbulkto adomestic, contract packaging facility. The PMN substance will be mixed 1:1 with potting soil for use. The PMN substance does not react, dissolve, degrade ordecompose during normal use.

Insert any picture here (PDF) Confiiential

Check here to add another continuation sheet 

### Page 19 of 19

### ---- --- - ------ Page 20 of 1011wVJ 

# CI~3 InventoryExpertSe ice Adivision oftheAmerican Chemical Sociecy Phone: 800-631-1884, 614-447-36 (e60-2308) FaX:6 4-447-3747* PoWstj Fax Note 7671 DaeE-mail: ans rs@casorg Phy I 

### 0 

### 5/8 jPhane Fax*
- #__ _ _ _ _ _ _ _ _ INVENTORY EXPERT SERVICE REPORT

IES-ORTIER NUMBER: 69691 REGISTRY NUMBER.- Not Registered

CA INDEX NAME

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

Ifthis box is checked, CAS has made correction(s) marked in red to your IES ordex. -Pleasemake the same correction(s) to your PXM1 before submitting it to the EPA.

2W406lnngy "dvrRoad PO.09 3343 Caleanb"g Ohio43210-034

FEB-4-1999 THU 12:55AM ID: PAIE: 1

## Page 21 of 101 

# MITSUBISHI GAS CHEMICAL COMPANY, INC. Page 1 f5 MATERIAL SAFETY DATA SHEET Product Name: Memory Green Crystal Preparation date: 2/2C/03 _SECTION 1 -CHEMICALPOUTADCMAYIETFATN 

# ] MSDS REVISION #:New PRODUCT NAME: Memory Green Crystal SYNONYMS: None DISTRIBUTED BY: Mitsubishi Gas Chemical America, Inc. 520 Madison Avenue, 17th Floor NewYork, NY 10022, USA PHONE NUMBERS: Inquiries - 

## (212) 752-4620 (business hours) CHEMTREC - 

## (800) 424-9300 (transportation emergencies, 

# ON 

# b

h'R.E.01E .-

,] Component % CAS No. Exposure Limits Polyether polyol 20 None Not established Water 80 7732-18-5 Not established

***********************EMERGENCY 

### OVERVIEW * ***' *V 

## Product consistsof odorless, yellow-green, irregularly-shaped beads. There are no known orexpected hazards associatedwith handling this product. Avoid contactwith eyes. Wash thoroughly after handling. POTENTIAL HEALTH EFFECTS EYES: No adverse health effects known or expected, although small particles may cause mechanical irritation. SKIN: No adverse health effects known or expected. Section 3continued on next page

## Page 22 of 101 

# MITSUBISHI GAS CHEMICAL COMPANY, INC. Page 2cf 

## 5 MATERIAL SAFETY DATA SHEET Product Name: Memory Green Crystal Preparation date: 2/2C/03



## INGESTION (swallowing): No adverse health effects known or expected. INHALATION (breathing): No adverse health effects known or expected. CHRONIC EFFECTS/CARCINOGENICITY: This product (or component) is not listed in IARC Monographs, the NTP Nnth Annual Report orthe ACGIH TLVs as a carcinogen or potential carcinogen. 0,%- HA does not regulate itas acarcinogen.
EYE CONTACT: Flush with water, lifting upper and lower lids occasionally.
FLASH POINT: Not applicable FLASH POINT METHOD: Not applicable UPPER EXPLOSION LIMIT: Unavailable LOWER EXPLOS ION LIMIT: Unavailable AUTOIGNITION TEMPERATURE: Unavailable EXTINGUISHING MEDIA: Water spray, carbon dioxide, dry chemical orfoam FIRE AND EXPLOSION HAZARDS: None known FIRE-FIGHTING EQUIPMENT: Wear full protective clothing and self-contained breathing apparatus with full facepiece. Combustion products may include toxicfumes.

## Page 23 of 101 

# MITSUBISHI GAS CHEMICAL COMPANY, INC.Pae3 5 MATERIAL SAFETY DATA SHEET Product Name: 

## Memory Green Crystal Preparation date: 2/20)03

Shovel spilled material into containers. Thoroughly sweep up residual material. Keep inclosed or covered containers when not inuse. Store incool dry place with adequate ventilation. SCTIO 8

- 

## EXPOSURE CONTROLSIPROA PRTC -N RESPIRATORY PROTECTION: Not required under normal conditions ofuse. SKIN PROTECTION: Not required under normal conditions of use. EYE PROTECTION: Protective eye wear or safety glasses are recommended. ENGINEERING CONTROLS: Not required under normal conditions of use.
APPEARANCE: Yellow-green beads 

## @770

## F (250 C) SPECIFIC GRAVITY: Unavailable VAPOR PRESSURE (mm Hg): Nil VAPOR DENSITY (Air =1): Heavierthan air INITIAL BOILING POINT: Decomposes EVAPORATION RATE: Nil (Ethyl Ether = 

## 1) SOLUBILITY inWATER: Insoluble, although product absorbs significant amoL nts of water BULK DENSITY: Unavailable VOLATILE %: 80% SOFTENING POINT: Unavailable

## Page 24 of 101 

# MITSUBISHI GAS CHEMICAL COMPANY, INC. Page 4of 5 MATERIAL SAFETY DATA SHEET Product Name: Memory Green Crystal Preparation date: 2/20,103

STABILITY (conditions to avoid): Stable under normal conditions. INCOMPATIBILITIES (materials to avoid): Avoid contactwith acids, and oxidizing agents. HAZARDOUS DECOMPOSITION PRODUCTS: Thermal decomposition products include: carbon monoxide, carbon dioxide rnd noxious gases HAZARDOUS POLYMERIZATION: Not known to occur
Acute Oral LD 5



## o(rat) - 

## greater than 2,000 mg/kg Skin irritation
- 

## Not irritating Mutagenicity (Ames test)
- 

## negative
No data available
Incineration isthe recommended disposal method for all chemical wastes; however, this material may be deposited inalandfill inaccordance with applicable local, slate and Federal regulations. This material isnot considered ahazardous waste under current RCRA definitions.
Not regulated under current DOT, IMO or ICAO regulations.

## Page 25 of 101 

# MITSUBISHI GAS CHEMICAL COMPANY, INC. Page 5 f5 MATERIAL SAFETY DATA SHEET Product Name: 

## Memory Green Crystal Preparation date: 2/20/03 

## SECTIO 15 -REG1JATORY777777"nN 

## TSCA INFORMATION: This product isnot, to the best of our knowledge, listed on the TSCA Inventory. se of this material is restricted to technically qualified individuals for research ind development purposes only. SARA 313 INFORMATION: SARA requires submission of annual reports of release of toxic chemicals hat appear in 40 CFR 372. This information must be included in all MSDS that are copied and distributedfor this material. Components present inthis product at alevel that could require reporting under the statute are: None
HAZARD RATING: HEALTH 0 0-LEAST FIRE 1 1 - 

## SLIGHT REACTIVITY 0 2- MODERATE OTHER -3 -HIGH 4- EXTREME HAZARD RATING METHOD: HMIS REASON FOR REVISION: New MSDS
The product information contained herein isbelieved to be accurate as of the date of the Material Safety )ata Sheet, and isprovided without warranty, expressed or implied, as to the results of use of this information o-the producttowhich itrelates. Recipient assumes all responsibility for the use of this information and the use 

### (E lone or incombination with any other product), storage or disposal of the product, including any resultant personal injury or property damage. 

### ~END 

## OF REPORT

## Page 26 of 101 B020514 

## Confirmation Statement

General Testing Research Center, Japan Oistuff Inspectors' Corporation

Sponsor Mitsubishi Gas Chemical Company, Inc. Title Bacterial Reverse Mutation Study of MEMORY GREEN CRYSTAL Study No. B020514

The study had been conducted in the General Testing Research Center, Japan Oilstuff Inspectors' Corporation. The original study report was written in Japanese and translated into English. I hereby declare that this report faithfully raflects the original study report as accurately as possible to my knowledge.

Translator

_________ _____ Date: Masato Nakamura, M. S. Scientist of Mutagenicity Testing Group

2

### Page 27 of 101 B020514

Submitted to: Mitsubishi Gas Chemical Company, Inc.



## Re p or t
Bacterial Reverse Mutation Study of MEMORY GREEN CRYSTAL (Study No. :B020514)

June 25, 2002
GENERAL TESTING RESEARCH CENTER, JAPAN OILSTUFF INSPECTORS' CORPOZATION

-I3

## Page 28 of 101
B020514 

## Statement

General Testing Research Center, Japan Oistuff Inspectors' Corpor tion

Sponsor Mitsubishi Gas Chemical Company, Inc. Title :Bacterial Reverse Mutation Study of MEMORY GREEN CRYSTAL Study No. B020514

The study in this report was conducted in compliance with the followirg Good Laboratory Practice Standards.

Japanese GLP standards Applied to Industrial Chemicals (Kanpogyo No. 39, Yakuhatsu No.229, 59 Kikyoku No.85, 1984; revised in 1988 and 2000) GLP Standards under the Japanese Industrial Safety and Health Law (N tification No.76, Ministry of Labor, 1988 revised in 2000)

Management Masakatsu Usami sealed Date: June 25, 2002 Study Director : Eiichi Machigaki sealed Date: June 25, 2002

4

## Page 29 of 101 B020514 

## Quality Assurance Statement

General Testing Research Center, Japan Oilstuff Inspectors'I Corporation

Sponsor Mitsubishi Gas Chemical Company, Inc. Title :Bacterial Reverse Mutation Study of MEMORY GREEN CRYSTAL Study No. B 020514
By the inspect ions and audit s i s

ted belIow, I hereby certif y that the studrwas carried out in accordance with its protocol and with our Standard Operating Pro- edures, and that the report faithfully describes the methods and procedures applied, and that the reported results reflect the raw data of the study accurately.

Inspctin Dte o inpecion Date of reporting to InspecitionrDate ofi ns e t o the study director or aditor aditand to the management Prtocol May 8, 2002 May 8, 2002 Srtdyl Procdur June 4, 2002 June 4, 2002 Draft June 19, 2002 June 19, 2002 Report Report June 25, 2002 June 25, 2002

Quality Assurance Unit: Sadayoshi Ohtubo sealedDate: June 25, 2002

## Page 30 of 101 B0205 14 

## Outline of the Study
1. Title :Bacterial Reverse Mutation Study of MEMORY GREEN CRYSTAL (Study No. : B020514)
2. Purpose: To assess the mutagenicity of the test substance in bacter al mutation assays, using Salmonella typhimui-iuv and Eschez-ichia coli strains.
3. Guideline: Guidelines under the Japanese Industrial Safety and Heal ;h Law (1997) and Guidelines under the Japanese Industrial Safetj and Health Law (1988 and revised in 1997).

4.GLP :GLP under the Japanese Industrial Safety and Health Law 1984; revised 1988 and 2000) and GLP under the Japanese Industria LSafety and Health Law (1988 and revised in 2000).
5. Sponsor: Study ordered by Mitsubishi Gas Chemical Company, Inc. 5-2, Marunouchi 2-chome Chiyoda-ku, Tokyo, Japan (Responsible person) Masanori Hataya Study ordered

Mitsubishi Chemical Safety Institute Ltd. 1-30, Shiba 2-chome, Minato-ku, Tokyo
6. Organization in: Testing Facility Contract General Testing Research Center, Japan Oilstuff Inspectors' Corporation 10-4, 1-Chome, Mikagetsukamachi, Higashinada-ku, Kobe-shi, Hyogo 6

## Page 31 of 101 B020514 7. Study Contributors Management Masakatsu Usami Data Recorder :Etsurou Fujiwara Study Director :Ejichi Machigaki sealed Date: Jn 2,O00 Other Contributors Masato Nakamura, Osamu Iwaihara, Chikako Tsutsufti, Eriko Iwasaki, Naomi Ikeda
8. Study Date :(Initiation of the Study) May 8, 2002 (Initiation of the Test) May 27, 2002 (Submission of the Test) June 6, 2002 (Submission of the Final-Report) June 25, 2002
9. Data analysis: No statistical methods were used. 10. There were no unforeseeable circumstances that had affected t st results undesirably. 11. Retention The protocol, all raw data, the test substance sample, documents and the final report will be retained in the safekeeping facility of

-General Testing Research Center. The materials described hbove except

for the test substance sample will be retained for ten years after submission of the application for Japanese Industrial Safety and Health Law, section 57-3-1 or the notification of coming urder section 4-1-1, 2, or 3 of Industrial Chemicals. The test substance and specimen will be retained for a shorter period of the following two periods: 10 years after receiving notification according to the Industrial Chemicals or making notification according to the Industrial Safety and Health Law, or a period that allows stable storage without deterioration in quality. Further retention will be discus 3ed with the sponsor.
7

## Page 32 of 101 B020514 

## Contents Page Summary ...... ............................................................ 9
Materials and Methods............................................................10 1. Test materials ............................................................ 10 2. Bacterial tester strains. ............................................. 11 3. Media.... ............................................................... 12 4. S9 mix ................................................................. 13 5. Test method ......................... I...................... ............ 13
Results and Discussion ......................................................... 16
References .................................................................. 17
Tables...... ................................................................ 18 Figures ................................................................... 20 Background data .............................................................. 23

8

## Page 33 of 101 B0205 14 

## Summary
MEMORY GREEN CRYSTAL was examined for the mutagenicity in bacterial assa s using five different strains, Salmonella typhimur-ium TA98, TA1QO, TA1535, TA1537 and Esche-ric-hia coli WP2 uvrA. In both the dose-finding test and the main test, the number of revertant colonies induced by the test substance was less than twice that inthe corresponcling solvent control value for all tester strains both the presence and absence of $9 mix.

Based on these findings, it was concluded that MEMORY GREEN CRYSTAL is nt mutagenic under the test conditions employed.

9

## Page 34 of 101 B020514 

## Materials and Methods 1. Test materials 1

. 1 Test substance

Name of the new chemical substance (IIJPAC nomenclature) Trade name MEMORY GREEN CRYSTAL Structural formula or rational formula

(or outline of manufacturing method, incase both are unknown) Purity of the new Impossible Lot No. of the new chemica1 chemical substance to measure substance to be tested MG-R02 tested Name and concentration

of impurities

________ ____ CAS No. Vprpesr Molecular weight Impossible Partition____ Melting point Appearance at Lgtylo lsi Boiling point ambenLihtyelowelati Solubility in Sovn ouiiyS ability in several solvents SovnSolblt lvent Water (water holding ability) tbe2 Water suspended at 50 mg/mL1)tbe) DMSO suspended at 50 mg/mL ' Acetone suspended at 100 mg/niL others (-) DMS0: diniethyl sulfoxide Storage condition: kept at a room temperature and in desiccator 1): Based on the results of the vehicle selection test conducted in the testing facility. 2): Heat, discoloring and foam formation were not observed in the test substance preparation.

10

## Page 35 of 101 B020514 1. 2 Control Substance The negative control and the positive controls were as follows. Name Abbreviation Supplier Lot No. Purity(%) Negative control Wk ueCeia W92 Sterilized distilled DW Wand ures Ltd.a DG72 Water IdsreLd Positive controls Wk ueCeia 2-(2--Furyl)-3-(5-nitro AF-2 WaoPr hmcl WTH1046 8. 1 -2-furyl)acrylamide Industries, Ltd. Sodium azide NaN3 Wako Pure Chemical DM78 9 Sodium az___________ Industries, Ltd. DW45 98 9-Aminoacridine 9-A Aldrich Chemical 03024JR 8. 8 hydrochloride CompanyII NAEthyl-jN-nitro-AN- ENNG Aldrich Chemical 12219AS 97 nitrosoguanidine Company_____ 2-Aminoanthracene 2-AA Wake Pure Chemical ELJ6826 7.4 _______________________________Industries, Ltd.

_____ __ 2. Bacterial tester strains 2. 1 Strains1 Five genetically different strains of Salmonella typhimui-iumTA98, TA .00, TA1535, TA1537 and Esche-richia coi WP2 uvrA- were employed. These strains wre received from Japan Bioassay Research Center on the date of May 13, 1998. rhe genetic properties of the strains are listed inthe following table. Gene Additional mutations Stan eai emrne Rfatenetai tp Affectedmutation TA98 his D uvi- B rfa pkMl01 Frameshift TA1535 his G uv2-B .rfa pk11 Base-pair change TAIS37 his C uP--B rfa Frameshift WP2 uvrA- ti-p E uvi- A + Base-pair change 2. 2 Reason for selection The tester strains were widely used in a bacterial reverse mutatiar study and were recommen~ded in Guideline the Japanese Industrial Safety and iealth Law (Notification No.77, Ministry of Labor, September 1, 1988; revised Jume 2, 1997) and Japanese Guidelines on Industrial Chemicals (Kanhoan No. 287, Eis i NO, 127, Heisei 09. 10. 31 Kikycku No. 2, October 31 1997) and Guideline th Japanese

## Page 36of 101 B020514 IndustrialI Safety and Health Law (Not if ication No. 77, Ministry of Labo ,September 1, 1988; revised June 2, 1997). 2.3 Check of genotypic characteristics The tester strains were checked for their genotypic characteristics such as amino acid requirement, UV sensitivity, membrane mutation(rfa) and R-factor before the study. 2.4 Storage Eight milliliters of fresh bacterial suspension (cultured with shaking at 37 0C for 8 hours in a liquid growth medium) was mixed with 0.7 mL of DMSO and quickly frozen in a dry ice-acetone mixture. The stocks of the tester Strains were stored below -80*C. 2.5 Bacterial suspension Twenty microliters of stock culture of each tester strain was momulated into 10 mL liquid growth medium and grown with shaking at 37 "C for 8 hours. Densities of the fresh bacterial cultures were measured with a spectrometer. (ell numbers for the suspensions were calculated by the 10-fold dilution method.
3. Media 3. 1 Liquid growth medium Oxoid Nutrient Broth 'No. 2 (Unipath Ltd. lot No. 02859365) was dissolved in distilled water (25 g/L) and autoclaved at 121 'C for 15 minutes. 3.2 Minimal glucose agar plate. The agar plate (TESMEDIADAN MEDIUM, lot No. ANI190DR prepared on Ap -il 2, 2002) used for this study was purchased from Oriental Yeast Co. , Ltd. 3.3 Top agar Aqueous amino acid solution for S. typhimurium strains, which contained 0. 5 mmol/L each of L-histidine and D-biotin was prepared and sterilized by filtration (0. 45 um pore size), and then stored in a refrigerator. Aqueous amino acid solution for E.coli, which contained 0.5 mmol/L L-tryptophane Vas prepared and sterilized by filtration (0. 45 g mpore size), and then stored in a refrigerator. 12

## Page 37 of 101 B020514 Powdered agar (0.6g Bacto-Agar; Difco Laboratories) and 0.5 g of sod urn chloride were dissolved in 100 mL of distilled water and autoclaved. The abo- e-mentioned amino acid solution was added to this agar solution in the ratio of l.to 10 and kept at about 45 00 until use.
4. S9 mix 4.1 S9 The S9, which was prepared from the liver of rats treated with phenobarbital (30 mg/kg i.p. on the first day and 60 mg/kg i.p. on the second to fourth day) and 5,6-benzoflavone (80 mg/kg i.p. at the third phenobarbital injection), was purchased from Kikkoman Corporation. The S9 was stored below -80 'C and was used in both the dose-finding test and the main test.

Details of the S9 used were as follows: Lot No. RAA-459 Prepared on February 22, 2002 Using animal SrgeDwe a Species, Strain SrgeDwe a Sex, Age (inweeks) Male, 7 weeks old Weight 214 - 239 g
4. 2 S9 mix The composition of the S9 mix 'is listed in the following table. Th( S9 mix was freshly prepared for each assay, and was stored in ice. Components Amount in 1 mL S9 0. 1 mL MgClz 6H 20 8 umol KCl 33 Atmol D-glucose-6-phosphate 5 A mol /3-NADPH 4 gmol 

### /-NADH 4 umol Na-phosphate buffer (pH 7.4) 100 'Umol Sterilized distilled water Remainder13

## Page 38 of 101
B020514

5. Test method2 5.1 Preparation of the test substance solutions and the positive contro s According to the information from the sponsor, the test substance inc uded water. The examination of vehicle was performed with sterilized distilled wat r(DW), DM30 and acetone. The test substance suspended in DW(S0mg/mL), DMSO(50 mg/ni) and acetone (100 mg/mL).

Thus, DiV was selected as the vehicle for the test substance. The test substance was suspended in DiV and was diluted into each concentration by 2-fold dilution. The test substance suspensions were prepared just before use. As for positive control, NaN3 was dissolved in DW, and others were issolved in DM30. One milliliter aliquots of these solutions were dispensed into tubes for freezing and stored below -20 00. Froze aliquots were thawed at room temperature before use. 5. 2 Test substance concentrations In the dose-finding test, six dose levels ranging from 5 to 5000 Ug /plate were graded ata common ratio of 4.

Microbial growth inhibition was not observed in any tester strains Precipitation of the test substance was observed on the agar plate at 313 u g/plate or more in the absence of 39 mix and at 1250 g g/plate or more in the presence of 39 mix In the main test, five dose levels ranging 313 to 5000 pg/plate were graded at the common ratio of 2 based on the results of the dose-finding test. 5. 3 Rev~rse mutation test The reverse mutation test was done by the preincubation method. For one concentration, 0. 1 mL,of the test substance solution, 0. 1 mL of each bacterial suspension and 0. 5 aL, of either 0. 1 mol/L sodium phosp iate buffer (PH 7. 4) for the assay without metabolic activation or 39 mix for thE assay with metabolic activation were mixed in a sterilized test tube and incubated by gentle shaking at 37 'C for 20 minutes. After preincubation, 2 mL of molten op agar was 14

## Page 39 of 101 B020514 added to this mixture and poured onto a minimal glucose agar plate. len the agar overlay solidified, these plates were incubated at 37 'C for 48 hourt. The level of microbial toxicity was confirmed by observation of backgrounc lawn with stereoscopic microscope. Revertant colonies were counted manually or with an automatic colony counter.

Two plates per dose level were used for each assay. The test was perf rmed twice. Negative and positive control tests listed below were also done in each assay. Strain -S9smix (Mug! plate) +S9mix (gug/plate) 

### TA98 AF-2 0.1 

# 1 2-AA 0.5 TA100 AF-2 0.01 2-AA 1 TA1535 NaN3 0.5 2-AA

- 2 TAIS37 9-AA 80 2-AA 2 WP2 uvrA- ENNG 2 2-AA 10
5. 4 Contamination test The highest dose solution and S9 mix was mixed with molten top agar and the mixture was overlaid on a minimum glucose agar plate. No bacterial contamination was observed. 5.6 5judgment criteria The test substance was judged to be positive in this assay when the mean number of revertant colonies is more than double of the negative control va ue and when it showed significantly dose-dependent increase in a reproduci le manner. Otherwise, the mutagenicity of the test substance was judged to be negative.

No statistical methods were used for data analysis.

## Page 40 of 101
B020514 

## Results and Discussion
The results are shown in Tables 1, 2 and Figs. 1 - 5. In the main test, the number of revertant colonies induced by the test substance was less than twice of that in the corresponding solvent control value for all test strains both the presence and absence of S9 mix. The same results were obtained in the dose-finding test. The reproducibility of these negative iesults was confirmed.

Neither bacterial nor fungal contamination were observed in the resalts of the contamination test.

Microbial growth inhibition was not observed. Precipitated test substance was observed on the agar plate. The viable bacteria counts were as follows. Baceril tste Stain -Base pair exchange type Frameshift type TAlOO TA1535 WP2uvrA- TA98 TA1537 Number of dose-finding 4. 46452412 viable test 4 . . . . bacteria Main test 29 4
. . .

(X10 9 /mL)2. 47492613 In the positive control tests using specific mutagens with and with ut S9 mix, significant revertant colonies more than double of the solvent control value were induced. For solvent-control and positive control, the number of reverta. t colonies in all the tester strains were within the normal range of our 1aboratory' s background data. These results confirmed that the tests were performed correctly. Based on these results, it was concluded that MEMORY GREEN GRYSrAL is not mutagenic under the test conditions employed.

16

## Page 41 of 101 B020514 

## References 1) Maron, D. M. and Ames, B. N. (1983) Revised methods for the Salmon( la mutagenicity test, Mut. Res. , 113, 173-215. 2) Mutagenicity test in Industrial Safety and Health Law (1991) The t st guideline and the GLP, edited by the Chemical Substance Investigat: on

17

## Page 42 of 101 B020614 Table 1 Results of the Dose-finding Test Test substance :MEMORY GREEN CRYSTAL

Testing periods From May 27, 2002 toMay 30,2002 With (+)or Test material Number of revertants (number of clonies/plate) without -) concentration Base-pair change type craiseshift type S9 nix (Mg/plate) TA 100 TA 1535 WP2uvrA- TA 98 TA 1537 75 9 27 11 4 Solvent control 88( 82) 5 7) 25( 26) 10( 1) 0 7) 89 9 24 13 8 5 79( 84) 8( 9) 25( 25) 8( 1 5( 7) 87 7 30 8 6 S

9nix 20 97 ( 92) 5 6 ) 30( 30) 8( 8) 8( 7) 79 6 29 13 3 -)78 78 ( 79) 7( 7) 33( 31) 11( 12) 4( 4) go9 5 18 9 6 313 89(90) 4( 5) 22(20) 5 7) 8( 7) t104 8 18 15 10 1250 78( 91) 7( 8)28(23) 9(12) 5 8) t 90 5 30 16 6 5000 84( 87) 7 6) 26 (28) 9(13) 3 5) 92 7 26 27 9 Solvent control 106( 99) 10 9) 25 ( 26) 26( 27) 7 8) 100 29 15 10 5 97(101) 9 7) 34(32) 21(18) C 9) 97 6 32 15 9 S9aix 20 124(111) 9 8 8) 32(32) 21(18) 5 7) 98 8 29 16 7 ()78 134 (116) 9 9 ) 28( 29) 19( 18) 6( 7) 105 6 31 20 6 313 115 ( 110) 6( 6 ) 26(29) 13C 17) 9 8) t100 7 28 16 4 1250 112 ( 106) 6 C 7) 24( 26) 14( 15) 7 6) t 98 9 24 13 5 5000 97 (98) 8( 9) 29(27) 16(15) 5 5) Name and A F- 2 NaN 3 ENNG AF-2 9 -AA Positive concentration 0. 01 0.5 2 0. 1 80 control (u g/plate)

_________________ not requiring Number of revertant 418 351 633 569 164 S9 ii colonies per plate 457 ( 438 ) 354 ( 353 ) 648 C 641 ) 585 ( 577 )170 C167 Nameand 2-AA 2-AA 2-AA 2-AA 2-AA Positive concentration 12 10 0. 5 2 control (m a/plate)

______________________________________ requiring Number of revertant 633 350 564 532 161 S9 mix colonies per plate ,664 C649 )359 ( 355 )665 C 615 ) 51 ( 517 )L 92 C177 AF-2 2-(2-furyl)-3-(5-nitro-2-furyl)acrylaaide, NaN3 sodium azide, ENNG A-ethyl-V' -nitro-N-ntorosoguani dine, 9-AA :9-aminoacridine hydrochloride, 2-AA 2-aninoanthracene ):Mean Precipitated test substance was observed on the agar plate.

## Page43 of 101B051

Table 2 Results of the Main Test Test substance: MEMORY GREEN CRYSTAL Testing periods From June 3, 2002 to June 6, 2002 With (+) or Test material Number of revertants (number of colonies/plate)

________ without H- concentration Base-pair change type Frameshift type S9 mix (g g/plate) TA 100 TA 1535 WP2uvrA- TA 98 TA 1537 85 11 12 12 6 Solvent control 98( 92) 5( 8)26(19) 18 15) 9 8) t 78 8 21 14 8 313 117( 98) 5( 7)20(21) 13( 14) 10 9) ill1 12 19 11 9 S9mix 625 92(102) 5 9)23(21)14(13) 5( 7) t100 10 18 23 6 C)1250 91( 96) 8( 9) 18( 18 ) 10( 17) 5 6) t 75 10 27 15 7 2500 99( 87) 6 8) 15( 21) 13( 14) 5 6) T 101 10 17 17 9 5000 117(109) 5 .8)23(20) 16( 17) 7 8) 132 9 20 18 12 Solvent controll109(121 8( 9)21(21)31(25-)] 10( 11) T 130 14 30 24 15 313 128(129) 11( 13) 22( 26) 23 24) 13( 14) T 128 7 23 24 16 S9mix 625 115(122) 8( 8)22(23)26(25) 116(16) t124 10 26 18 11 ()1250 110(117) 14( 12) 26( 26) 28 23) 9(10) '118 11 35 15 10 2500 105(112) 8(10) 19(27)24(20) 16(13) t114 4 22 18 13 5000 102(108) 6( 5)28(25)29(24) 7(10) Name and AF-2 NaN3 ENNG AF-2 9 -AA Positive concentration 0. 01 0. 5 2 0. 1 s0 control Cua/plate) not requiring Number of revertant 378 378 658 508 144 S9 mix colonies per plate 377 ( 378 )329 ( 354 )625 ( 642 )494 ( 501 )160 (152 Name and 2 -AA 2-AA 2-AA 2 -AA 2-AA Positive concentration 1 2 10 0. 5 2 control(u/ t)

________ _______ ________ _______ ____ ___

reuring Number of revetant 609 361 6014528 S9 mix colonies per plate 584 ( 597 )342 ( 352 )635 (621 )404 ( 430 1 74 (151 AF-2 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide, NaN3 sodium azide, ENNG A-ethyl-A" -nitro-N-ntorooguanidine, 9-AA : -aminoacridine hydrochloride, 2-AA 2-aminoanthracene C :Mean t:Precipitated test substance was observed on the agar plate.

19

## Page 44 of 101B051 .200 

## [TA 100] 150

0. 0. 

### 5 0
0



### 40

40

020

## Page 45 of 101 B020514 *40
30 aT CL

.~20 0 0 C 

### *~10
0 O 313 625 1250 2500 5000 Consentration 

### ( gg/plate) Fig.3 Dose-response curve of MEMORY GREEN CRYSTAL (Main Test) 0 

### -S9mix . 

### 0- .+s9rix


### 40 

## CTA981 30

0.E

0
0



### ~ 10

## Page 46 of 101 B020614 

## [TA15371 30
CL
LD 

### 20 C 0 0



### 00 313 625 1250 2500 5000 Consent ration (pu g/p Iate) Fig.5 Dose-response curve of MEMORY GREEN CRYSTAL (Main Test)

--S9mix -0- 

### .S9mix

22

## Page 47 of 101 B020514 

## Background Data WlBackground data of the negative (solvent) control value Strain

______ -S9mix _____+S9mix
___ AMerage Average± Number Average Average:± umber

______________ 2SD _____2SD

___ TA100 90 72 - 107 60 ill 89 - 133 59 TA1535 8 4 -12 58 8 4 -12 _56 VWP2uvzA- 20 14 - 26 53 22 16 - 28 _52 TA98 16 11 - 21 61 21 16 - 27 60 TA1537 7 5 -10 58 10 7 -13 58 (2)Background data of the positive control value Strain -S9mix _____+S9mix

___ Average Average±t Number Average Average-± Number

_______ 2SD

_____ _____ 2SD

___ TA100 468 346 - 589 48 666 542 - 789 __48 TA1535 382 337 - 427 43 313 266 - 359 42 WP2uvrA- 614 471 - 756 40 723 600 - 846 __40 TA98 552 458 - 645 49 447 349 - 546-] 49 TA1537 193 91 -294 43 158 134 - 181 43 Background data was collected from September 4, 2001 to, March 27, 2002

23

## Page 48 of 101 PPLFINAL REPORT: P020071

VERIFICATION OF TRANSLATION
This is to certify that the English translation presented here conforms to the origina. Japanese report. Study No.: P020071 Title: Skin Sensitization Study ofMEMORY GREEN CRYSTAL in Guinea Pigs According to Buehler Test Method

Masami Kokubu Date Panapharm Laboratories Co., Ltd.

## Page 49 

## of 101 PPLFINALREPOF T:P020071

Skin Sensitization Study ofMEMORY GREEN CRYSTAL in Guinea Pigs According to Buehler Test Method

FINAL REPORT -



### August 16, 2002

Sponsor: Mitsubishi Gas Chemical Company, Inc. 2-5-2, Marunouchi, Chiyoda-ku, Tokyo 100-8324, Japan Organization under Contractor: Mitsubishi Chemical Safety Institute Ltd. 2-1-30, Shiba, Minato-ku, Tokyo 105-0014, Japan Test Facility: Safety Assessment Laboratory Panapharm.Laboratories (PPL) Co., Inc. 1285, Kurisaki-machi, Uto-shi, Kumamoto 869-0425, Japan

## Page 50 

## of 101 PPL FINAL REPORT: P020071



### SIGNATURE PAGE

Study Director
Signed inthe original Seal affixed August 16, 2002 Masarni Kokubu Date Panapharnn Laboratories Co., Ltd.

## Page 51 of 101 PPL FINAL REPORT: P 20071



### STUDY SCHEDULE 1. Study inception: May2, 2002 2. Animal receipt: May 9, 2002 3. Sensitization period: May 21, 2002 - 

### June 4, 2002 4. Challenge date: June 18, 2002 5. Observation period: June 19, 2002 - 

### June 20, 2002 6. Study completion: August 16, 2002 RETAINED MATERIALS AND THEIE REPOSITORY 1.- Records concerning the protocol 2. Records concerning the animals 3. Records concerning the animal room conditions 4. Records concerning the test article 5. Records concerning the results ofthe study 6. Final reports 7. Other materials concerning the study The above materials shall be kept inthe archives ofPanapharm,LaboratorieE Co., Ltd. for 5years after completion ofthe study (from August 16, 2002 toAugust 16, 2007). The handling ofthe above materials after 5years shall be determined through discussion with the sponsor.

## Page 52 of 101 PPL FINAL REPORT: P( 2007 1



### CONTENTS Page SIGNATURE PAGE............... STUD SCHDULE...........................................i RETAINED MATERIALS AND THEIR REPOSITORY
SUMMARY...........................................1 OBJECTIVE...........................................2 

### MATERIALS AND METHODS....................................2 RESULTS............................................6 

### DISCUSSION...........................................6 REFERENCES..........................................7 

## Table 1 Body weights ofguinea pigs in skin sensitization study of 

## MEMORY GREEN CRYSTAL ............................ 8 Table 2 Skin scores ofguinea pigs in skin sensitization study of MEMORY GREEN CRYSTAL ............................ 9 Photographs Iand2....................................... 10 Appendix (individual data)

## Page 53 

## of 101 PPL FINAL REPORT: P02 071



### SUMMARY The skin sensitivity ofMEMORY GREEN CRYSTAL was investigated using H-artley guinea pigs according to the Buehler Testmethod. In

### the case that the MEMORY 

### GREEN CRYSTAL sensitized group was elicited by 

### MEMORY GREEN CRYSTAL, no skinresponses 

### were seen. The positive rates were 0 % 

### and the mean scores ofskinresponse were 0 ateach observation time after 24 and 48 ho respectively. In the case thatnon-sensitized group was applied MEMORY GREEN CRYSTAL, no s in responses were seen on any ofthe applied sites, and no skinirritation couldberecogniize From the above mentioned, skin sensitivity forMEMORY GREEN CRYSTAL could o 

## berecognized underthepresent studyconditions. 

# f

## Page 54of 101PPL FINALREPORT: P 20071

OBJECTIVE As apart ofnon-clinical studies ofthe safety evaluation ofMEMORY GREEN CRY TAL, the skin sensitivity in guinea pigs was investigated according to the Buehler Test methol

. 

### MATERIALS AND METHODS 1. Test Article and the Vehicle MEMORY GREEN CRYSTAL 

## (LotNo. MG-R02) provided by Mitsubishi Gas Chemical Company, Inc. was used as the test article. MEMORY GREEN CRYST was yellow powder. Japanese pharmnacopoeia water forinjection (LotNo. IE90, 0 suka Pharmaceutical Factory, Inc.) was used as the vehicle. The test article was preserved in a desiccator in the test article room at roomtemperature. The vehicle was preserved ina lockerin thetest article room atroom temperature. 2. Preparationof TestArticle MEMORY GREEN CRYSTAL and water for injection was mixed atthe rate of05gof MEMORY GREEN CRYSTAL to 3ml,ofwater forinjection, and the mixture was ellkneaded to prepare the gel like material immediately before use. 0.05 gofthe mix eto be usedwas weighed ona piece oflint cloth of2.0 X

### 2.0 cm. 3. Animals and Husbandry Eighteen female guinea pigs (Cij:Hlartley, Charis River, Japan, Inc.) were pur hased at an age of5weeks. During the quarantine and acclimation periods for 11I days, all animals were examined for clinical signs and weighed to confirm that theywere healthy. Fifteen animals at an age of7 weeks were used for the study. Body weights on the first day ofapplication were 373.1-432.6g. The animals were h used in an aluminum cage (Wx H xD: 430x360x700 mm) with 1or 2 animals per ca e in the animal room (No. 16) in the rabbit and guinea pig area ofthe pharmacology laboratory, which was controlled at a temperature of24±2'C (permissible range: 21- 27'C), relative humidity 55±10% (permissible range: 35-75%), 12-hour light/da -k cycle (lighting from 7:00A.M. to 7:00P.M.) and 13-15 changes/hour ventilation. The animals were allowedto receive apellet diet (RC4, Oriental Yeast Co., Ltd.) ani to drink well water supplemented with NaCIO (about 0.2 ppm) adlibitum. The aimal cages were changed at frequency ofonce or more pertwo weeks. The animalioom was washed with water, drained and cleaned with a disinfectant-soaked mop every day.
-2-

## Page 55 

## of 101 PPL FINAL REPORT 0~7



### 4. ConstitutionofTest Groups, Reason for Selection ofDose Levels and Selection of Animals Constitution ofthe test groups is as shown below. 

### Sensitization Challenge Am sf Group Sex Article for DoeArticle for Dose (Animal os.) sensitization Doechallenge MEMORY MEMORY 10

- 

### I GREEN 0.05 g/site X 

### 3times RSEN 0.05 glsite 

### (Nos.286 295) CRYSTALCRSA MEMORY5 TI Non-treated GREEN 0.05 glsite 

### (Nos. 29 -300) _______________CRYSTAL

____ 

### 0.05 glsite x 3 

### times ofMEMORY GREEN CRYSTAL was applied to the sensitiz tion 

### group and 0.05 glsite ofMEMORY GREEN 

## CRYSTAL was applied to the challenge group. The usual dosage level fortheBuheler Test method was selected. After th health condition andbody weight ofthe animals a daybefore the start ofthe sensitiz tion was confirmed, 15 animals were selected. A study animal number was given to the individual animals, in the order ofyounger quarantine numbers. The study animals were identified byear tagnumber, and a label was attached each cage bearing the study n mber, animal number, study group number and sex. After group assignment, the remaini g animals were excluded from the study. 5.Route andMethod forDosing The method, frequency andperiod ofdosing were determined according to the B ehler Testmethod'), ofwhichis described in the guidelines for toxicity studies ofskin sensitization to drugs. 5.1. Sensitization Method The hair ofthe animals was clipped (about 4 x 

### 5cm) on the back ofthe ne k region with a clipper (DC-5, Shimizu Denki Kogyo Co. Ltd.) andwas shaved ith an electric shaver (BS 5597, BRAUN). The clipped skin area was wiped of with a piece of sanitary cotton containing ethanol. 0.05 g ofarticle for sensitizatio was loaded onto a 2 x,

### 2 cm.piece oflint cloth (Yamato Kogyo Co. Ltd.). The ii t cloth was affixed on the skin area and covered with a sheet oflaboratory film (PARAFILM'M, American National Can TM). 

### Elastic adhesive bandage (NICHIBAN -ELASTOPORE, No.50-5) was bound around the neck to hold he skin
-3-

## Page 56 of 

### 101 PPL FINAL REPORT: P0 0071
application for 6hours. The first dosing day was determined as-day 0.- The procedure of6 hour occlusive application a day wasperformed atotal ofthree t 

### mes on day 0, day 7and day 14 for two weeks at frequency ofonce perweek (at the interval of7 days). The non-sensitized group was free from this sensitization procedure. 5.2. Challenge Two weeks after the final sensitization (day 28), the hair ofthe animals was clipped on the back ofthe neck region with a clipper and was shaved with an el ctric shaver. One square part measuring 2 x

### 2 cmn was obtained from theletfak A piece of2 x

### 2 cm lint cloth loaded with 0.05 g ofarticle for challenge,was affix. d on the square part, and was covered with a sheet oflaboratory film. After 6 hour , 

### the animals were free from the occlusive application, and the article was wiped off.. Group 1, 11 Anterior 

# LMEMORY
GREEN CRYSTAI,

Posterior

*2x2 

### cm

-4-

## Page 57 of 101 PPL FINAL REPORT: P02 071



### 6. Method andFrequency ofObservation andEvaluation oftheApplied Skin The applied skin was scored for the grade oferythema or edema 24 and 48 hours after the removal ofthe occlusive application according to the standards for evalua ion ofDraize') as shown below. (1) Erythema and eschar formation 0 No erythema I Very slight erythema (barely perceptible) 2 Well defined erythema 3 Moderate to severe erythema 4 Severe erythema (beet redness) to slight eschar formation (injuries in depth) (2) Edema formation 

### 0 No edema 1 Very slight edema (barely perceptible) 2 Slight edema(edges of area well defined by definite raising) 3 Moderate edema (raised approximately I mm) 4 Severe edema (raised more than 1 mm and extending beyond area ofexposure) Mean score =X~)() 

### Number oftested animals Positive rate _Number 

### ofpositive animals*X10 M% Number oftested animals *Number 

### ofanimals showing skin response ofa score ofone ormore in (1) r (2) items. The mean score ofskin response and the positive rate were calculated at each observation time of24 and 48 hours after the removal ofthe occlusive applicatio The skin sensitivity oftest article was evaluated based on these parameters. 7. Clinical Observation and Body Weight Measurement Clinical observation was performed twice at pre- andpost-application ofthe sensitization and the challenge, respectively. The animals were weighed on th first sensitization day, the challenge day and the observation day 48 hours after remo al of the occlusive application.

-5 -

## Page 58 of 101 PPL FINAL REPORT: P0 0071

8. Photography The photography was performed 24 and 48 hours after the challenge. 9. Statistical Analysis Mean value and standard deviation ofbodyweight for each group were calculate Statistical analysis was not performed.
RESULTS 1. Clinical Observation No abnormal clinical signswere observed in each group during the sensitization eriod. 2. Body Weight Body weights ofeach group are shown in Table 1. No abnormal changes to the body weight were observed in any group. 3. Skin Reaction The score ofskinresponse and thepositive rate are shown in Table 2and Appen 

### ~x1, andphotographs ofthe skin are shown in Photographs Iand 2. InMEMORY GREEN CRYSTAL sensitized animals (Group I), no skin reaction inducedby MEMORY GREEN CRYSTAL challenge was seen on any ofthe applie sites of10 animals during the observation times of24 and48 hours afterthe challenge. e positive rate was 0 %,and themean scores ofskin response were 0at each observat on time, respectively. Innon-sensitized animals(Group HI), no skinreaction induced by MEMORY G EN CRYSTAL application was seen on any ofthe applied sites of5animals during the observation times of24 and 48 hours after the challenge. The positive rate was 0 0,and 

### the mean score ofskin response was 0at each observation time, respectively.
DISCUSSION Thepotency ofskin sensitivity for MEMORY GREEN CRYSTAL was investigated using guineapigs according to the Buehler Testmethod '). 

### In the case that MEMORY GREEN CRYSTAL was applied on the skin ofthe non- -6-

## Page 59 

## of 101 PPL FINAL REPORT: P-02 71



### sensitized group, no skin reactions were seen on any ofthe applied sites at each observatio time and the skin response by irritation 

### ofMEMORY GREEN CRYSTAL could notbe recognized. In the case that MEMORY GREEN CRYSTAL was challenged on the skin f the MEMORY GREEN CRYSTAL sensitized group, no skinreaction was seen on any of he applied sites at each observation time andthe skin response by sensitivity ofMEMORY GREEN CRYSTAL could notbe recognized. From the above mentioned, the skin sensitivity ofMEMORY GREEN CRYSTAL coul notbe recognizedunder the present studyconditions.
REFERENCES 1) Buehler E.V. (1965) :Delayed contact hypersensitivity in the guinea pig. Arch. Dermatol., 91, 171. 2) Draize, J.H. (1959): "Dermal toxicity", Appraisal ofthe safety ofchemicals infoo s, drugs and cosmetics, p4.6-59, Association ofFood &Drug Officials ofthe United St tes, Topeka, Texas.

-7 -

## Page 60 of 101 Study &o.P020071 Table I Body weights ofguinea pigs in skin sensitization study ofMEMORY GREEN CRYSTAL

Group Animal Bodyweights (g) number Day 0') Day 28 b) Day 30 c

286 397.1 592.5 593.2 287 374.5 516.6 516.4 288 412.4 582.8 582.9 289 432.6 666.9 667.2 290 374.3 530.4 531.6 1291 402.7 578.6 577.2 292 390.1 559.6 562.1 293 392.9 603.1 601.9 294 377.9 583.3 583.4 295 373.1 546.9 546.6 Mean 392.8 576.1 576.3 ±S.D. 19.4 42.3 42.1

296 357.9 569.8 567.2 297 388.9 531.5 534.7 298 391.0 605.0 607.1 UI 299 400.5 636.4 638.5 300 393.3 553.4 555.4 Mean 386.3 579.2 580.6 :±S.D. 16.5 41.7 41.8 a) Day ofthe start ofthe sensitization b) Day ofthe challenge c)Day ofthe final observation

## Page 61 of 101

C00

C>C

Cl

Cc a) E)a E

0 

### ~ 0 CZ

E a80 t= on C C a)ar)
.

.u .0 w I C 43 C La S z~ .a)- tc)) E E

- a)) o C 0££

-

## Page 62 of 101 PPLFINALREPORT :P020071
Photograph 1 The skin response ofthe sensitized guineapigs 24hours after application ofMEM )RY GREEN CRYSTAL. AnimalNo. :288 24hr MEMORY GREEN CRYSTAL I eft side No.2S8No 

### skinreaction isseen.

Phoorp2

The skinresponse ofthenon-sensil zed guinea pigs 24hours after MEMORY GREEN CRYSTAL application. Animal No. :300 P020071 MEMORY GREEN CRYSTAL /Left side 24hr No skinreaction is seen. No.300

10-

## Page 63 of 101 PPL FINAL REPORT: P 20071

Skin Sensitization Study ofMEMORY GREEN CRYSTAL in Guinea Pigs according to Buehler Test Method
(Individual data)

Appendix I Individual skin scores ofguinea pigs in skin sensitization study ofMEM RY GREEN CRYSTAL

## Page 64 of 101 SuyNo.P020071
Appendix 1 Individual skin scores ofguineapigs in skin sensitization study ofMEMORY GREEN CR)STAL Score ofskinresponse

____ Article for Article for Animal 24 h?) 48 hr? GOPsensitization challenge number E em d Edema E~hm d Ed-ma eschar c) eschar

frainb) formation fomtinb formation'c 286 0 0 0D 287 0 0 0 288 0 0 0 289 0 0 0 MEMORY MEMORY 290 0 0 0

GREEN GREEN CRYSTAL CRYSTAL 291 0 0 0 292 0 0 0 293 0 0 0 294 0 0 0 295 0 0 0

____ 296 0 0 0 MEMORY 297 0 0 00 HI GREEN 298 0 0 0 0 CRYSTAL 299 0 0 0 0 300 0 0 0 0 a) Time after removal ofocclussive application. b) Score oferythema formation: 0;No erythema 1; Very slight erythemna (barely perceptible) 2; Well defined erythema 

### 3;Moderate to severe erythema 

# i 4; Severe erythema (beet redness) to slight escharformation (injuried i depth)' c) Score ofedema formation: 0-; No edema I1;

Very slight edema (barely perceptible) 2; Slight edema (edges ofarea well defined by definite raising) 3; Moderate edema (raised approximately I mm) 4; Severe edema (raised more than I mm and extending beyond area of exposure)

## Page 65 

## of 101 PPL FINAL REPORT: P20070



### VERIFICATION OF TRANSLATION
This is to certify that the English translation presented here conforms to the ogna Japanese report. Study No.: P020070 Title: Primary Skin Irritation Study ofMEMORY GREEN CRYSTAL in Rabbits



## A/' Masami Kokubu Date Panapharm Laboratories Co., Ltd.



# /& /A2 1

## Page 66 

## of 101 PPLFINAL REPORT: P)20070

PrimarySkin IrritationStudy of MEMORY GREEN CRYSTAL in Rabbits
FINAL REPORT -



### June 27, 2002

Sponsor: Mitsubishi Gas Chemical Company, Inc. 2- 5-2, Marunouchi, Chiyoda-ku, Tokyo 100-8324, Japan Organization under Contractor: Mitsubishi Chemical Safety Institute Ltd. 2-1-30, Shiba, Minato-ku, Tokyo 105-0014, Japan Test Facility: Safety Assessment Laboratory Panapharm Laboratories (PPL) Co., Ltd. 1285, Kurisaki-machi, Uto-shi, Kumamoto 869-0425, Japan

## Page 67 of 101 PPL FINAL 

### REPORT: P(20070

SIGNATURE PAGE

Study Director
Signed in the original Seal affixed June 27, 2002 Masami Kokubu Date Panapharmn Laboratories Co., Ltd.

## Page 68 

## of 101 PPL FINAL REPORT: P(20070

STUDY SCHEDULE 1. Study inception: May 2,2002 2. 

### Animal receipt: May 7,2002 3. 

### Administration: May 14, 2002 4. 

### Observation period: May 15, 2002 - 

### May 17, 2002 5. 

### Study completion: June 27, 2002 RETAINED MATERIALS AND THEIR REPOSITORY 1. 

### Records concerning the protocol 2. 

### Records concerning the animals 3. Records concerning the animal room conditions 4. 

### Records concerning the test article 5. 

### Records concerning the results ofthe study 6. Final reports 7. 

### Other materials concerning the study The above materials shall be kept in the archives ofPanapharm Laboratorie Co., Ltd. for 10 years after completion ofthe study (from June 27, 2002 to June 26,-2012). The handling ofthe above materials after 10 years shall be determined throug discussion with the sponsor.

## Page 69of 101 PPL FINAL REPORT: 1020070



### PARTICIPATING PERSONNEL IN THE STUDY 1. Study director: Masami Kokubu 2. 

### Responsible technician: Shigenari Okumura 3. 

### Person responsible for test article management: Hiroyuki Izumi 4. 

### Person responsible for quarantine (veterinarian): Emiko Kuwasaki 5. Preparatioii oftest article, etc.: Chikako Tokumitsu and Naoko Kimura 6. Animal receipt: Shigenari Okumura 7. 

### Animal care: Shigenari Okumura, Kazuaki Sasaki, Tatsumi In i, Hironori Shigekane and Misuzu Ueba 8. 

### Group assignment: Shigenari Okumura 9. Clinical observation: Shigenari Okumura, Toshihiro Katsuta, Kazuaki Sasaki and Tatsumi Inui 10. Body weight measurement: Shigenari Okumura, Toshihiro Katsuta ai d Hironori Shigekane 11. Administration: Toshihiro Katsuta 12. 

### Scoring: Masami Kokubu.and Toshihiro Katsuta 13. Photography: Shigenari Okumura and Toshihiro Katsuta

## Page 70 

## of 101 PPL FINAL REPORT: P20070

CONTENTS Page SIGNATURE PAGE............... STUDY SCHEDULE.......................................i RETAINED MATERIALS AND THEIR REPOSITORY

.... .. .... .... .... ... PARTICIPATING PERSONNEL IN THE STUDY

SUMMARY 1.............. OBJECTIVE ........... .... 2 MATERIALS AND METHODS....................................2 RESULTS ............ ... 6 DISCUSSION .......... ..... 6 REFERENCES .......... ..... 6 Table 1 Body weights ofrabbits in primary skin irritation study of MEMORY GREEN CRYSTAL

.... ... ... .... ... ... ... 7 Table 2 Skin scores ofrabbits inprimary skinirritation study of MEMORY GREEN CRYSTAL Photograph I......................................... .9

## Page 71 

## of 101 PPL FINAL REPORT: P(20070

SUMMARY The primary skin irritation forMEMORY GREEN CRYSTAL was investigated using ma e New Zealand White rabbits. No skinreaction was observed onany ofintact and abraded skin from 24 hours to 72 hours 

### after MEMORY GREEN CRYSTAL occlusive application. Theprimary irritation index 

## (... was 0. MEMORY GREEN CRYSTAL was evaluated as mildlyirritants. From the abovementioned, the primary irritation ofMEMORY GREEN CRYSTAL coulInot berecognized under the present study conditions.

## Page 72 of 101 PPLFINAL REPORT: P20070

OBJECTIVE As a part ofnon-clinical studies ofthe safety evaluation ofMEMORY GREEN CRYSTAL, the primary skin irritation studywas carried outusing 

## rabbits. MATERIALS AND METHODS 

## 1. TestArticle MEMORY GREEN CRYSTAL (LotNo.MG-R02) provided by Mitsubishi Gas Chemical Company, Inc. was used as the test article. Japanese pharmacopoeia wat r for injection (Lot No.1IE90, Otsuka Pharmaceutical Factory, Inc.) was used as the vehicle The test article was preserved in adesiccator in the test article room atroom temper Lture. The vehicle was preserved in the test article room atroom temperature. 2. PreparationofTest Article MEMORY GREEN CRYSTAL and water for injection was mixed at the rate of05g of MEMORY GREEN CRYSTAL to 3mL ofwater for injection, and the mixture was ellkneaded to prepare the gel like material. These preparations were conducted immediatelybefore use. 3. Animals andHusbandry Five male New Zealand White (Kbs:NZW) Rabbits aged 4 months were purc ased from Kitayama Labes Co., Ltd. The weights ofsome animals at thereceipt were less than the minimum value (3.0 kg) ofweights at the time ofreceipt in aprotocol. The reduction in body weights ofthe animals,wasjudged tobe due to the stress durivgthe transportation, because no abnormal clinical signs were observed in any ofthe animals at the receipt ofanimals. The deviation ofprotocol wasjudged not to affect the study, since there were no clinical sign ofchange in the animals and thebody weights increased smoothly after the receipt. The animals were quarantined and acclim ted for six days. In this period, all animals were examined for clinical signs and weighed to confirm thatthey were healthy. Three animals were selected and used for the study. Body weights on the application day were 2.497-2.986 kg. The animals were hused in an aluminum cages (Wx H xD: 430x360x700 mm) at one per cage in the animal room (No. 14) in the rabbit and guinea pig area ofthe metabolism-pharmacology research institute, which was controlled at a temperature of24±2'C (permissible range: 21-27C), relative humidity 55±10% (permissible range: 35-75%), 12-hour lighqdark cycle (lighting from 7:00A.M. to 7:00P.M.) and 13-15 changes/hour ventilation.
-2-

## Page73 o 101PPL FINAL REPORT: P)20070

Actual ranges oftemperature andrelative humidity in the animal room during th study 

## were 23-25 

## 'Cand 54-65 %,respectively. The animals were allowed to receiv a pellet diet (RC4, 

### Oriental Yeast Co., Ltd.) and to drink well water supplemented with NaCIO (about 0.2 ppm) adlibitum. The animal cages were changed once at gr up assignment. The animal room was washed with water, drained andcleaned wit a disinfectant-soaked mop every day. 4. ConstitutionofTest Group, Reason for Selection ofDose Levels and Selection f Animals Three animals ofone group were used in this study. Test article Dose Sex Number of Animal Nos.

_____________(glsite) animals 

### MEMORY

GREEN 0.5 6" 3 No. 1-3 CRYSTAL

_________ ____ 

### The hair ofthe animals on the back oftheir necks was gently clipped with a clipper (DC-6, Shimiizu Denki Kogyo Co. Ltd.) a daybefore the start ofapplication. The ;kin conditionwas observed, andthree healthy animals, which had slight island skin and good skin condition forthe application, were selected, and a study animal number was givento the individual animals, in the order ofyounger quarantine numbers. The animals ere identified by numbers onthe inside oftheir left ear,marked with ablack oily felt-ti pen, and each cagehad cage cards attached with study number, animal number and sex. The remaining animals after the animal selection were excluded from the study. The dose of0.5 glsite MEMORY GREEN CRYSTAL was selected asthe genera dose level fortheprimary skin irritation study. 5. Route and Method forDosing Percutaneous route by occlusive application was selected because ofthe usua administration route ofprimary skin irritation study. The hair clipped on their backs was divided into two parts and a 2.5 X 

### 2.5 cm square dotted by means ofablac oily felt-tip pen was marked in each part. A # -shaped abrasion was inflicted with 23G needle in one ofthe square sites on eitherthe right or left side. Non-injured (iritact) skin and injured (abraded) skin was one set, andthe test article was applied on I ese squares.

-3-

## Page 74 of 101 PPL FINAL REPORT: P)20070

A 0.5 

### g ofthe gel-like test article was loaded uniformly onto apiece 

### of2.5 X2.5 -m square lint cloth. The lint cloth was affixed to each square ofthe clipped skin ith a piece of4X4 

### cm mesh bandage (NICHIBAN -MESHPOREO, No.50-10). The mnal was bound around the body with an netbandage (NABIS -NET BANDAGE No f4, AO-2424-04) to maintain the 24 hour occlusive application. The animal was equipped with a protective collar. The lint cloth was removed from the applied skin after 24 hours. At the same time, the dosing article was removed and the skin sjuare was wiped with apiece ofsanitary cotton containing distilled water. <Application pattern> Animal No. Animal No. 1,3 2 

### Anterior Anterior Memory Memory Memory 

## I Memory

Green I Green Green Green Crystal Crystal Crystal 

## I Crystal 2.5 

### I2.5 cm cm 2.T5 cm 2.5cm 

### Posterior Posterior
6. Observation, Evaluation and Measurement The skin reaction on the applied skin ofall animals was scored according to t ie procedure for evaluation ofskin reactions by Draize') 24hours, 48 hours and 72 ours after the application. The scores ofreactions (erythema, eschar and edema formation) ofthe intact kin and abraded skin in each animal were summed up 24hours and 72 hours after t application. The sum ofthe scores was divided with 4 to obtain mean score.

-4-

### JPage 

## 75 of 101 PPL FINAL REPORT: P(20070

6.1. Erythema and eschar formation Score Skin reaction

O No erythema 1 Very slight erythema (barelyperceptible) 2 Well defined erythema 3 Moderate to severe erythema 4 Severe erythema (beet redness) to slight eschar ____formation 

### (injuries in depth) 6.2. Edema formation Score Skin reaction



### 0 No edema 1 Very slight edema (barelyperceptible) 2 Slight edema (edges ofarea well defined by 2 definite raising) 3 Moderate edema (raised approximately I mm) 4 Severe edema (raised more than 1mmn and extending beyond area ofexposure) The mean score of all animals was calculated to obtain primary irritation index (P.1.1., The maximum 8 points). The irritation grade was evaluated on the b sis of scale shown below. Photography of the application skin was carried out r the typical sample at each following time. 

## o : P.M. 5 2 Mildlyirritants 2 < P.M.. :_ 5 Moderately irritants 5 < P.M.. !- 8 Severe irritants The animals were observed once a day and weighed at pre-application on the application day and the final observation day. 7. Statistical Analysis Mean value and standard deviation ofbodyweight were calculated. Statistical analysis was not performed.

-5-* PPL FINAL REPORT: Pi120070 

## Page 76 of 101

RESULTS 1. Clinical Observation No abnormal clinical signs were observed in any animal. 2. Body Weight Body weights are shown in Table 1. All animals showed favorable weight transition. 3. Skin Reaction Skinreactions are shown inTable 2. For MEMORY GREEN CRYSTAL occlusive application, no skin reactions on either the intact orabraded skin were observed atobservation times of24hours, 48 hours and 2 hours afterthe occlusive application. The P.1.I. was 0. DISCUSSION Afterthe gel-like article and water forinjection was absorbed in MEMORY GREEN CRYSTAL, itwas applied occiusivelly for 24 hours, the skin condition ofthe applied are was observed. No skinreaction on anyofthe sites ofthe intact or abraded skin area was observed at any observation time. The P.1.1. was 0. MEMORY GREEN CRYSTAL was evaluated as amildly irritant from the scale ofP.1.1. by Draize. From the above mentioned, primary skinirritation ofMEMORY GREEN CRYSTAI could notrecognized.
REFERENCES 1)Draize, J.H. (1959) :"Dermal toxicity", Appraisal of the safety of chemicals in foods,

drugs and cosmetics, p46-59, Association ofFood &Drug Officials ofthe United States, Topeka, Texas.

-6-

## Page 77 of 101

-0 cn W) C

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I-.4

## Page 78 of 101

0.

ccl c,0 00

*00 00 z 0 'd I .0C- e C<

. 

### ~- .~cc 

### ~ ~0c0
~ cc
cuU 

### ~ 0

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### ~~8ray%= Ivui iuu YrL rNALKJrUKI: ruz/-UU 

## Page 79 of 101

Photogrph I 

### ~ Skin of24 hours afterthe application :r >~'Animal No.2.



### ~ MEMORY GREEN CRYSTAL: 24wNo reactionatsitesofmitactand abraded skins.

9

### Page80 o 101PPL FINAL REPORT: P 0069



### VERIFICATION OF TRANSLATION

This is to certify that the English translation 

## presented here conforms to the orignal Japanese report.
Study No.: P020069
Title: Single Oral Administration Toxicity Study ofMEMORY GREEN CRYSTAL in Rats (Limit Test)

Kumiko Honda Date Panapharm Laboratories Co., Ltd.

## Page 81 of 101 PPL FINAL REPORT: PC20069



### Single Oral Administration Toxicity Study of MEMORY GREEN CRYSTAL in Rats (Limit Test)

FINAL REPORT -



### June 25, 2002



### Sponsor: Mitsubishi Gas Chemical Company, Inc. 2-5-2, Marunouchi, Chiyoda-ku, Tokyo 100-8324, Japan Organization under Contract: Mitsubishi Chemical Safety Institute Ltd. 2-1-30, Shiba, Minato-ku, Tokyo 105-0014, Japan Test Facility: Safety Assessment Laboratory Panapharm Laboratories (PPL) Co., Ltd. 1285, Kurisaki-machi, Uto-shi, Kumamoto 869-0425, Japan

## Page 82 of 101 PPLFINAL REPORT: P0' 0069


### SIGNATURE PAGE

Study Director
Signed in the original Seal affixed June 25, 2002 Kumiko Honda Date

Panapharm Laboratories Co., Ltd.

## Page 83 of 101 PPL FINAL REPORT: P0K0069
STUDY SCHEDULE 

### 1. Study 

### inception: April 25, 2002 2. Animal receipt: May 7, 2002 3. Administration: May 14, 2002 4. Necropsy: May 28, 

### 2002 

## 5. Study completion: June 25, 2002 

### ADVENTITIOUS CIRCUMSTANCES AFFECTING THE RELIABILITY 0 THLE STUDY AND DEVIATION FROM THE PROTOCOL There were no adventitious circumstances affecting the reliability ofthe stud and no deviation from the protocol. RETAINED MATERIALS AND THEIR REPOSITORY 1. Records concerning the protocol 2. Records concerning the animals 3. Records concerning the animal room conditions 4. Records concerning the test article 5. Records concerning the results ofthe study 6. CD-R 7. Final reports 8. Other materials concerning the study The above materials shall be kept inthe archive ofPanapharm Laboratodes Co., Ltd. for 10 years after completion ofthe study (from June 26, 2002 to. une 25, 2012). The handling ofthe above materials after 10 years shall be determined through discussion with the sponsor. 

### PARTICIPATING PERSONNEL IN THE STUDY 1. Study director: Kumiko Honda 2. Responsible technician: Eiji Murata 3. Person responsible for pathological examination: Masao'Hamamura 4. Person responsible 

### for test article management: Hiroyuki Izumi 5. Person responsible for quarantine (veterinarian): Emiko Kuwasaki 6. Preparation oftest article mixture, etc.: Noriko Kojima,and Naoko Kimura

## Page 84 of 101 PPL FINAL REPORT: PO 0069


### 7. Animal receipt: Yasuo Nakaoka 8. Animal care: EijiMurata, Teruhiko Enoshima, Yasuo Nakaok Tomohiro Ikeda, Hidehiro Ogata, Kumiko Hond Sayumi Tominaga, Misako Ueba, Noriko,Kojima. Akemi Goto and Yukari Douki 9. Group assignment: Eiji Murata 10. Administration: Eiji Murata 11. Clinical observation: Eiji Murata, Teruhiko Enoshima, Yasuo Nakaok;t Tomohiro Ikceda, Hidehiro Ogata, Kumiko Hond;t Sayumi Tominaga, Misako Ueba and Noriko Ko.inia 12. Body weight measurement: Eiji Murata, Teruhiko Enoshima, Yasuo Nakaokk Kumiko Honda, Akemi Goto and Yukari Douki 13. Necropsy: Takafumi Oshikata, Yutaka Nakahara and Hideyaki Watanabe 14. Management ofcomputer system: Ryoichi Tagami and Miho Watanabe

## Page 85 of 101 PPL FINAL REPORT: P 0069 

## CONTENTS SIGNATURE PAGE ........ STUDY SCHEDULE......... i ADVENTITIOUS CIRCUMSTANCES AFFECTING THE RELIABILITY OF

THE STUDY AND DEVIATIONS FROM THE PROTOCOL

....... ... ... RETAINED MATERIALS AND THEIR REPOSITORY...................iii PARTICIPATING 

### PERSONNEL IN THE STUDY......................iii SUMMARY ......... 1 OBJECTIVE ......... 

### 2 MATERIALS AND METHODS ......... 2 RESULTS ......... 4 

### DISCUSSION ......... 5 Figure 1 Body weight changes.................................6 Table 1 Clinical signs 7 Table 2 Body weights .. .......... 9 Table 3 Necropsy findings . ........... 10 Appendices

-iv -

## Page 86 of 101 PPL FINAL REPORT: PO 069 

### SUMM4ARY MEMORY GREEN CRYSTAL was orally administered to SD-IGS rats 

### (5male ind 

### 5female 

### Crj:CDrats) at a dose level of2000 mg/kg. No death occurred in any animal during the observation period. There were no changes ascribable to administration ofthe test article in any ofthe clinical observa -ion, body weight measurement or necropsy. In conclusion, under the conditions ofthis study, it was considered that the approximate lethal dose in rats ofboth sexes was above 2000 mg/kg.

## Page 87 of 101 PPL FINAL REPORT: P02 069 

### OBJECTIVE As a part ofnon-clinical studies ofthe safety evaluation, MEMORY GREEN CRYSTAL was orally administered to rats to evaluate its acute toxicity.
MATERIALS AND METHODS 1. Test Article MEMORY GREEN CRYSTAL (lotNo. MG-R02) provided by Mitsubishi Gas Chemical Company, Inc. was used as the test article. Corn oil (lot No. V1P5345 Nacalai Tesque Co., Ltd.) was used as a vehicle. The test article was stored ina desiccator, and the corn oil was stored in a locker in the test article room at room temperature. 2. Animals and Husbandry Seven male and female rats (Crj :CD(SD)IGS) aged 5weeks old were purchas d from Charles River Japan, Inc. The animals were quarantined and acclimatize for 6 days. During this period, all the animals were examined for clinical signs anl weighed in order to confirm the in health status. Finally, 5 males and5 femnales were selected by the randomization method on the day before dosing and emplo red in this study at the age of6 weeks. The body weights on the day ofdosing wer 165.5-169.6 gfor the males and 117.3-121.5 g for the females. The animals were housed in an animal room (No. 09) in barrier system area a controlled at a temperature of 24t2 0

### C (permissible range: 21-27'C) and relative humidity of 55±10% (permissible range: 35-75%) with a 12-hour light/dark cycle (lighting fom 7:00 A.M. to 7:00 P.M.) and 13-15 ventilation changes/hour. The animals wer. housed individually in stainless steel wire bottom cages (WxHxD: 260xH200xD380 mm). The actual temperature and relative humidity ofthe animal room during the study were 22-26'C and 53-64%, respectively. The animals were given access to pellet diet (CRF-1, Oriental Yeast Co., Ltd., Tokyo, Japan) and well water supplemented with sodium hypochlorite (approximately 2 

### ppm) adlibitumfro an automatic dispenser or feeding bottle. Diet containing impurities below the permissible limit, as verified by Japan Food Research'Laboratories (ajuridical foundation, Tokyo, Japan) at the request of Oriental Yeast Co. Ltd., was purchased. The water was analyzed at Kakujo Nankyukaken Center Ltd,. (Kumamoto, Japan) and verified as satisfying the requirements ofthe Water Quality Standard (Waterworks Law). All housing equipment and diet were sterilized by 

### autoel ving -2-

## Page 88 of 101 PPL FINAL REPORT: P 069 before use. Cages and cage racks were changed once at group assignment, and the sanitary trays under the cages were changed 2times aweek. The animal room was cleaned and wiped with a disinfectant-soaked mop every day. 

### 3.. Constitution ofTest Group,Reason for DeterminationofDose Levels and GroupAssignment Constitution ofthe test groups istabulatedbelow. Group Dose Concentration Dose volume Sex No. of Anij al (mg/kg) (mg/niL) (niL/kg) animals numer M 5 

### 101-05 2000mg/kg 2000 100 20F 5 20-5

Based on the information on the test article analysis provided by the sponsor, the test article was considered to be hypotoxic. The dose level was set at 2000 mgkg, which was the highest dose ofthe toxicity guidelines. The numbers of animals were 5males and 5females. On the day before do ing, the animals were divided into groups by the randomization method onthe basis f body weights. After the animal assignment, the animals were identified individually by tail number with an oily felt pen and ear punch. A cage card 11 ting the study number, animal number, dose level and sex was attached to each cage. The animals that remained after grouping were excluded from the study. 

### 4. Route andMethod for Administration The oral route was chosen based on the expected route ofhuman exposure. All the animals were fasted overnight (for about 18 hours). The test article mixtui e was administered to the rats once by oral gavage. The amount ofthe dosing solution was calculated based in the individual body weight on the day of administration. The rats were given food about 3 hours after the administraticn 

### 5.Method ofPreparationofTheTest Article Mixture withThe Vehicle and Frequency ofPreparation Just before dosing, the test article was suspended in corn oil at a concentrati n of 100 mg/mL. The containers ofthe test article mixture were labeled with the udy number, test article name, date ofpreparation and name ofthe preparing perso nel.

-3-

## Page 89 of 101 PPL FINAL REPORT: P920069 

### 6. Method andFrequency of Observation,Examination and Measurement 1) Clinical Observation The 

### observation period was for 14 days after administration. All the animals were observed for clinical signs andmortality until 6hours after administration on the day ofadministration (Day 0) and once daily thereafter. 2) Body Weight Measurement The body weights were measured before administration (Day 0) and on D.LYS 1, 3, 7, 10 and 14. 3) Necropsy All the animals were anesthetized with either and necropsied at the end ofthe observation period for 14 days. 7. Statistical Analysis The mean and standard deviation ofbody weights were calculated.
RESULTS 1. Clinical Observation The clinical signs are presented in Table 1andAppendix 1. Diarrheal stool was observed in 2 males and 3 females from 1to 3 hours alter administration. In addition, soiled periproctal was observed in 1female at 1.hour after administration. No abnormal signs were observed in any animals thereafter. 2. Body Weight The body weight changes are presented in Figurel, Table 2 and Appendix 2. The body weights ofall the animals increased normally during the observation period. 3. Necropsy The results are presented in Table 3and Appendix 3. No abnormal findings were observed in any animal.

-4-

## Page 90 of 101REOT 206
DISCUSSION MEMORY GREEN CRYSTAL was orally administered to male and female rats at a dose level of 2000 mg/kg. No death occurred in any animal during the observation period. Diarrheal stool was 

## observed inboth sexes from 1to 3hours after administration. Soiled periproctal ias 

### observed in 1female from 1to 3hours after administration. These signs had 

### disappeared by 6 hours after administration. In addition, there were no changes ascribable to administration ofthe test article in the body weight measurement or necropsy. The diarrheal stool and soiled periproctal observed inthis study were considered to have been induced by corn oil and not related to administration ofth( test article. In conclusion, under the conditions ofthis study, it was considered that the approximate lethal dose inrats ofboth sexes was above 2000 mg/kg.

-5-

## Page 91 

## of 101 Study No P020069

-- 

### 2000 mg/kg M~


### 300-

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### 0 1 3 7 10 14 

### Days

400 

### F

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### 2000 mg/kg

'~300-

200-

0 1 3 7 10 14 

### Days Figue 1 Body weight changes 6

## Page 92 of 101

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## Page96 o 101PPL FINAL REPORT: F020069 

### CONTENTS Appendix 1 Individual clinical signs - 

### Male, Female.......................1 Appendix 2 Individual body weights - 

### Male, Female.......................3 Appendix 3 Individual necropsy findings - 

### Male, Female, Day 14............... 4

## Page 97 of 101

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## Page 101 of 101