Form A roved. O.M.B. No. 2070-0012. A ires 10-31-96. 

## u.s. ENVIRONMENTALPROTECTIONAGENCY 

### AGENCYUSE ONLY**~D~.~te~o~f~r~~i~Pt~-----=~~~~~~--~~--~~~~----------------~** r:/

....A. 

### PREMANUFACTURE C , '-) 

### , ft 

### .., ;ojr:'! 

### NOTICE " It::­ ,j 

## Company Sanitized

nterthe tOtal nUll''''', v<**,,~o-­** inthe Premanufacture Notice 

### GENERALINSTRUCTIONS You mustprovide all infonnation requested in this fonn to the extentthat it is known to orreasonably ascertainable byyou. Makereasonable estimates ifyou do not have actual data. Beforeyou completethis fonn, you should read the "Instructions Manual for Premanufacture Notification" (the InstructionsManual is available from the Toxic SubstancesControl Act (TSCA) Infonnation Service by calling202-554-1404,orfaxing 202-554-5603). Ifa userfee hasbeen remitted for this notice (40 CFR700.45), indicate in the boxes abovetheTS-userfee identification numberyou have generated. Remember, youruserfee ID numbermust alsoappearonyourcorresponding fee remittance, which is senttoEPA,HQAccountingOperations Branch (PM-226), P.O. 360399M, Pittsburgh, PA 15251-6399,Attn. TSCA Userfee.
PartI
-- GENERAL INFORMATION TESTDATA ANDOTHERDATA You must provide the currentlycorrectChemicalAbstracts (CA)Name ofthe new You arerequired to submitall testdata in yourpossession orcontrol and to provide chemical substance, even ifyou claim the identity as confidential. You may authorize a description ofall otherdataknown toorreasonably ascertainablebyyou, ifthese anotherpersonto submit chemical identity information for you, but your submissionwill data arerelatedtothehealth and environmental effects on themanufacture, not be complete and the reviewwill not begin until EPAreceives this information. A processing, distribution in commerce, use, ordisposal ofthe newchemical letterin support ofyour submission shouldreference your TS userfee identification substance. Standard literature citationsmaybesubmitted for data in theopen number. You must submit an original and two copies ofthis notice including all test data. scientific literature. Completetest data (written in English) not summariesofdata Ifyou claimed any information as confidential, a single sanitized copy must also be mustbe submitted ifthev donotappearin the openliterature. You should clearly submitted. identifY whethertestdata is on the substanceoronan analog. Also, thechemical composition ofthe tested material should becharacterized. Followingare examplesoftestdataand otherdata. Datashould be submitted accordingtothe PartII
-- HUMANEXPOSURE AND ENVIRONMENTAL RELEASE requirements of§nO.50ofthePremanufactureNotificationRule (40 CFRPart Ifthere are several manufacture, processing, oruse operationsto be described in Part II, nO). sections A and B ofthis notice,reproduce the sections as needed.
Part 1II

-- LIST OF ATTACHMENTS TestData (CheckBelow any included in this notice) Attach additional sheetsifthere is not enough space to answera question fully. Label Environmental fate data 181 Yes Otherdata D Yes each continuation sheetwiththe corresponding section heading. InPart 1II, listthese attachments, any test data orotherdata and any optional information included in the Health effectsdata 181 Yes Risk assessments notice.
. Environmental effects data 181 Yes Structure/activityrelationships Physical/Chemical Properties· 181 Yes Testdata not in the possession OPTIONAL INFORMATION orcontrol ofthe submitter You may include any informationthatyou want EPA to consider in evaluating the new Aphysical and chemical properties worksheet is located on the last page ofthis form. substance. On page II ofthis form, space has been provided for you to described pollution prevention and recycling information you may have regardingthe new TYPE OF NOTICE (Check OnlyOne) substance. 

### [8] PMN (Premanufacture Notice) So-called "binding"boxes are includedthroughout this form for youto indicate your willingness to be bound to certain statementsyou make in this section, such as use, production volume, protective equipment.
. . This option is intended to reduce delays 

## D INTERMEDIATEPMN (submitted in sequence't'it!J final product PMN) thatroutinely accompanythe development ofconsent ordersorSignificantNewUse 

### = . = Rules. Except in the case ofexemption applications (such as TMEA, LVE, LOREX) ..L.­ SNUN(Significant New Use Notice) where certain information provided in such notification is binding on the submitterwhen 

## D :r­ the Agency approves the exemption application, checkinga binding box in this notice \:J Cl-ry --::) F;' doesnot by itselfprohibit the submitterfrom laterdeviating from the information (except TMEA(TestMarketing Exemption Application)-;':; 

## D**-~Jn** chemical identity) reported in the form.**~** '-Ir'q 

## D LVE (Low Volume Exemption)@40CPR723.50(cXI)
. _: •._ CONFIDENTIALITYCLAIMS**~** 

### (')r;; 

## D LOREX (LowReleaselLowExposureExemption)@40C:riiW.50(cX2) You may claim any information in this notice as confidential. To assert a claim on the CO 

# .. form, mark (X) the confidential box next to the information that you claim as confidential. To assert aclaim in an attachment, circle orbracket the information you claim as confidential. Ifyou claim information in the notices as confidential you must also provide a sanitized version ofthe notice (including attachments). Foradditional instructions on claiming information as confidential, readthe Instructions Manual. 

# D Mark (x) ifany information in this notice is claimed as confidential. 

## D
IS TH LVE Modification IS ACONSOLIDATED PMN? 

## D D LOREx..J)1odification U1 Yes # ofchemicals (Prenotice Communication # required, enter # on page 3) EPA FORM 7710-25 {Rev. 5-95) Replaces previous editions ofEPAFonn 7710-25. 

## IPublicreporting burden for this collection ofinformation is estimated to average) 10hoursperresponse, includingtime for reviewing instructions, searchingexistingdata sources, gatheringand maintainingthedataneeded, and completing and reviewingthe collection of information. Send commentsregardingthe burden estimate or any otheraspect ofthiscollection ofinformation, including suggestionsfor reducingthis burden, to Chief, InformationPolicy Branch,PM-223, U.S. Environmental ProtectionAgency, 401 M. St., S.W.,Washington, D.C. 20460; and to theOfficeofManagementand Budget,Paperwork ReductionAct (2070-0012), Washington, D.C. 20503. CERTIFICATION I certifythattothe bestofmyknowledge andbelief: I. The company named in PartI, section A, subsection I a ofthis notice form intends to manufacture orimportfor a commercial purpose, otherthan in small quantities solelyfor research and development, the substance identified in PartI, Section B. 2. All informationprovided inthis notice is complete andtruthful as ofthe date ofsubmission. 3. I am submittingwiththis notice all testdata in mypossession orcontrol and a description ofall other dataknowntoor reasonably ascertainable byme asrequired by §720.50ofthe PremanufactureNotificationRule.
AdditionalCertificationStatements: Ifyou are submittinga PMN, IntermediatePMN,ConsolidatedPMN, orSNUN, checkthe following userfee certification statementthatapplies: 

## IZI TheCompany named in Part I, Section A hasremitted the fee of$2500 specified in40CFR700.45(b), or 

# o TheCompanynamed in PartI, Section A hasremittedthe fee of$1000 foran Intermediate PMN (dermed @40CFR 700.43)in accordance with40CFR700A5(b), or 

# o The Company named in PartI Section A is a small business concernunder40 CFR700.43 and hasremitted a fee of$100 in accordance with40CFR700.45(b). lfyouare submittinga lowvolumeexemption (LVE)application in accordance with40CFR723.50(cXI)ora Lowrelease and low exposure exemption (LoRex) application in accordancewith40CFR723.50(c)(2), checkthe following certification statements: 

# o Themanufacturer submittingthis notice intendsto manufacture orimportthe new chemical substance for commercial purposes, otherthan in small quantities solelyfor research and development, underthe terms of40CFR723.50. 

# oThe manufactureris familiar with the terms ofthis sectionandwill complywiththose terms; and 

# oThe newchemical substance for which the notice is submittedmeets all applicable exemptionconditions. 

# o Ifthis application is for an LVEin accordance with 40CFR723.s0(cXl),the manufacturer intendsto commence manufacture ofthe exempted substance for commercial purposes within 1year ofthe date ofthe expiration ofthe 30day reviewperiod. Tbe accuracy ofthestatementsyou makeinthis noticeshould reflectyourbestpredictionoftheanticipated facts regardingtbechemicalsubstance described herein. Anyknowinl! andwillful misinterpretationis subiecttocriminal penaltvpursuantto 18USC 1001. Confidential Signature and title ofAuthorized Official (Original SignatureRequired) Date
Signature ofagent
- (ifapplicable) Date

EPA FORM 7710-25 (Rev. 5-95) Page 2 

## x

# x

you the chemical
PMN numberassignedby EPA (i.e. withdrawnor incomplete). Ifyou for the

Type ofNotice

- Mark (X) J. Binding Option Telephone

Position 

# .. Name Submitting Notice (in U.S.)

b. Agent(if applicable)

c. Ifyou are Company

City, State,ZIP Code

Company Mailing address and street)

mark box. Confidential

Joint Submitter(if Nameof applicable)

street) City,

Contact (in U.S.) Company Mailing address (numberand street) City, State, Code

Mark (X) ifnone substancecovered by this notice,enterthe exemptionnumberassignedby Mark (X) EPA. Ifyoupreviously submitted aPMNforthis substanceenterthe ifnone 5. Mark (X) 2. Import 6. Only 3. 

# o Both 

# o BindingOption Mark Mark
EPA FORM 7710-25 (Rev. 5-95) Page 31. a. b. name correct Abstracts that consistent listings similar ForClass I substances aCAIndexName mustbe provided. ForClass2 substances eithera CAIndexNameorCAPreferred Namemust beprovided, which everis appropriatebasedonCA9CI nomenclaturerules and conventions).

c. Please identifYwhichmethod you used todevelop orobtainthe specifiedchemical identity information in this notice: (check 

# o Method 1 (CAS InventoryExpertService - a copyofthe Identification 

## 0 Method 2 (OtherSource) report obtained from the CAS InventoryExpert Services mustbe d.

e. Fora class I a complete correct precursor substances theirrespectiveCAS RegistryNumbers. (2) thenatureofthe reaction orprocess. (3) Indicatethe range ofcomposition andthetypical composition (where appropriate). (4) Providea correctrepresentativeorpartial chemical structure diagram. as comolete as canbe known. ifone canbereasonablv ascertained. based on



## x X

X X X

attach a continuation sheet.
EPA FORM 7710-25 (Rev. 5-95) Page 4a. Indicate the number-average weightofthe lowestmolecularweight composition ofthe polymeryou intend to manufacture. Indicate maximum weightpercentoflowmolecularweight species (notincluding residual monomers, reactants, orsolvents)below 500 and below 1,000 absolute molecularweight ofthatcomposition. Describe the methods ofmeasurementorthebasis foryourestimates: GPC

## 0 Other 

## 0: (Specify) i) lowestnumberaverage molecularweight: ii) maximumweight % below 500 molecularweight: iii) maximumweight %below 1000 molecularweight:
You must make separate monomerorotherreactant composition information, and residual "Confidential"box nextto any item you claimas confidential (1)· Providethe specific chemical name and CAS RegistryNumber(ifanumberexists) ofeachmonomerorotherreactantused in the manufacture of thepolymer. Mark (X)this column ifentry in column (1) is confidential. Indicate the typical weightpercentofeach monomer or otherreactantin the polymer. (2) - (3) • (4) • Mark (X)the identitycolumn ifyou wantamonomeror otherreactantused at two weightpercentorless to belistedas part ofthe polymer description on theTSCAChemical Substance Inventory. Mark (X)this column ifentriesin columns (3)and (4)are confidential. (5)

- (6)

- Indicate the maximum weightpercentofeach monomerorotherreactantthatmaybe present as aresidual in thepolymeras manufactured for commercial purposes. Mark this column if

(CAS Inventory ExpertService- acopy ofthe identification report obtained from CAS InventoryExpert Servicemust be submitted as as to this d. The currentlycorrect name for the polymer % % %

e. Provide acorrectrepresentativeorpartial chemical structure diagram, as complete as can beknown, ifone can % % %

polymers.

attach a continutation sheet EPA FORM 7710-25 (Rev. 5-95) Page 5;2812757385 # L,/ 4-15-04; 3:00<"M;BaKer <"e1:rOII1:e HSE

(n) Identify each impurity that may be reasonably**~nticipated** to be prescnt in the chemical subsmnce as manufactured forcorrunercial purpose. Provide the CAS Registry Number ifavailable. Ifthcrearc unidentified impurities, enter "unidentified." %ofeach
. estimatetheir total 1m

%
%
%
%
% ConfiConfidential 

# o Mark (X) this box ifyou attach acontinuation sheet. 5. identified in subsection I or 2. 

# o Mark (X) this box ifyou aunch acontinuation sbeet. 6. you claim chemical liS con you mUSl provide ageneric name for yoursubst:mce (hu( reveals the specific chemical identity ofthenew chemical substance to the ma)(imum extentpossible. Refer to (he TSCA Chemical Substance Inventory, 1985 &lilion. Appendix Bfor guidance on developing generic names. Quaternary ammonium hydroxide

resulting from the manufacture. processing. use, ordispOSllIofthe new chemical ct CAS .lIIach acontinuation sheet.
antiCipated

EPA FORM 7710-25 (Rell.5-95) I)age 6

### PartI
-- GENERAL INFORMATION

-- Continued Section C

-- PRODUCTION,IMPORT,AND USE INFORMATION: Mark (X)the "Confidential"boxnexttoanyitem you claim as confidential. 1. Production volume

­ Estimatethe maximum production volume duringthe first 12months ofproduction. Also estimate the maximum production volume for any consecutive 12-monthperiodduringthe firstthree years ofproduction. Estimates should be on 100%new chemical substance basis. For aLow Volume EXemption application, ifyou choose to haveyournotice reviewed at a lowerproduction volumethan 10,000 kg/yr, specifYthe volume and mark (x) in thebindingbox. Ifgranted, you are bound to this volume Maximum fust 12-monthproduction (kglyr) Maximum 12-monthproduction (kg/yr) Conti- Binding (100%new chemical substancebasis) (100%newchemical substancebasis) dential Option Mark (x) X X 2. Use Information

-- You must make separate confidentialityclaims forthedescription ofthecategoryofuse, the percentofproduction volume devoted to each category, theformulation of thenew substance, and otheruse information. Mark (X)the "Confidential"Boxnextto any item you claim as confidential. a. (1)

-­ Describeeach intended categoryofuse ofthe new chemical substanceby function and application
.. (2)

-­ Mark (X)this column ifentrycolumn (l) is confidential business information (CBI). (3)

-­ Indicateyour willingness to have the information provided in column (1)binding. (4)

-­ Estimate the percent oftotalproduction for the first three years devoted to each category ofuse. (5)

-­ Mark (X)this column ifentry in column (4) is confidentialbusiness information (CBI). (6)

-­ Estimate the percentofthenew substanceas formulated in mixtures, suspensions, emulsions, solutions, orgelsas manufactured for commercialpurposes at sitesunderyourcontrol associated with each category ofuse. (7)

-­ Mark (X) this column ifentry in column (6) is confidentialbusiness information (CBI). (8)

-­ Indicate%ofproductvolumeexpected forthe listed "use" sectors. Markmorethan one box ifappropriate. Mark (X) to indicate yourwillingnessto have the usetype provided in (8) binding. (9)

-­ Mark (X)this column ifentry(ies) in column (8) is (are)confidential business information (CBI). Categoryofuse (I) CBI Binding Produc- CBI %in CBI %ofsubstanceexpectedperuse CBI Option tion% Form­ (8) (byfunction and application i.e. adispersive dye Mark(x) uIation Site- 

# E Indus- Com- Binding forfinishing polyesterfiberS) (2) (3) (4) (5)

# ± limited trial mercia! Option (9) X X X X % % % % % % % % % % % 

# = % %

.. Ifyouhave identified a"consumer" use, pleaseprovide onacontinuation sheetadetailed description ofthe use(s) ofthis chemical substance in consumerproducts.

In addition includeestimatesoftheconcentrationofthenew cllemical substanceasexpected in consumerproducts and describethechemical reactions bywhichthis substanceloses its identityintheconsumerproduct. DMark(X)thisboxifyouattach acontinuation sheet. b. Generic Ifyouclaimanycategoryofuse description in subsection2aas confidential, enteragenericdescription ofthatcategory. Read the use InstructionsManual forexamples ofgeneric usedescriptions. description Destructive use as a fuel additive.

D Mark (X)this box ifyou attach acontinuation sheet. 3. Hazard Information

- Include in the noticeacopy ofreasonable facsimile ofanyhazard warning statement, label,material safetydatasheet, orother Binding

information wllich will be providedto any personwllo is reasonably likely to beexposedtothis substanceregardingprotectiveequipmentorpractices Option forthe safehanding, transport, use, ordisposal ofthe new substance. List inpart IIIhazard information you include. Mark (x) D Mark (X)tllis box ifyou attachhazard information. 

## X Part11-- HUMANEXPOSURE AND ENVIRONMENTAL RELEASE
EPAFORM 7710-25 (Rev. 5-95) Page7

# x you Section A -- INDUSTRIAL SITES CONTROLLED BY THE SUBMITTER Complete section A for each type ofmanufacture, processing, oruse operation invo new at mdustrial sitesyou control. Importers do nothave to complete this section for operations outside the U.S.; however, you may still have reporting requirements if there are further industrial after' You must describethese See instructions manual 1. Confiofthe site at which the nnF'r"t,nn will occur. dential

City, County, State, ZIP code Ifthe same operation will occurat more than one site, enter the number ofsites. Identifythe additional sites on a continuation sheet, and ifany ofthe siteshave significantly different production rates oroperations, include all the information requested in this section for those sites as attachments. Mark (X) this box ifyou attach a continuation sheet. Mark (X) b. Type-­ 

# x DProcessing Duse 

# x I. Batch Maximum kglbatch (100%newchemical Hourslbatch Batches/year substance) 2. Continuous d. Process description Mark (X) to indicate yourWillingness to have your process description binding. (I) Diagramthe major unit operation steps and chemical conversions. Include interim storage and transportcontainers (specify- e.g. 5gallon pails, 55 drum, rail car, tank truck, etc.). (2) Provide the identity, the approximateweight (by kg/day or kglbatch on a 100"10 new chemical substancebasis), and entry pointofall starting materials and feedstocks (including reactants, solvents, catalysts, etc.), and ofall products, recycle streams, and wastes. Include cleaningchemicals (note frequency ifnot used daily or per batch.). (3) Identify by numberthe pointsofrelease, includingsmall or intermittentreleases, to the environment ofthe new chemical substance.

this box ifyou attach a continuation sheet.



### Part 11-- HUMANEXPOSUREAND ENVIRONMENTAL RELEASE -- Continued EPA FORM 7710-25 (Rev. 5-95) Page 8 

# xSection A

-- INDUSTRIAL SITESCONTROLLEDBY THESUBMITTER

­ Continued 2. OccupationalExposure
­ You must make separate confidentiality claimsfor the description ofworkeractivity, physical form ofthe new chemical substance, numberofworks exposed, and duration ofactivity. Mark (X) the "Confidential" box nextto any item you claim as confidential. (1) --Describethe activities (i.e. bag dumping, tote filling, unloading drums, sampling,cleaning, etc.) in which workers may beexposed to the substance. (2)

--Mark (X)this column ifentry incolumn (I)is confidential business information (CBI). (3)

- Describe any protective equipment and engineeringcontrolsused to protectworkers. (4) and (6)

- Indicateyourwillingnessto have the information provided in column (3) or (5) binding. (5)

-­Indicatethe physical form(s) ofthe new chemical substance (e.g., solid: crystal, granule, powder, ordust) and %new chemical substance (ifpartofamixture) atthe time ofexposure. (7)

- Mark (X) this column ifentry in column (5) is confidential business information (CBI). (8)

- Estimate the maximum numberofworkers involved in each activity for all sites combined. (9)

- Mark (X) this column ifentry in column (8) is confidential business information (CBl). I (10) and (11)

-- Estimatethe maximum duration ofthe activity for any worker in hours perday and days peryear. (12)

- Mark (X)this column ifentries in columns (IO) and (II)are confidential business information (CBI). i Workeractivity eBl Protective Equipment! Binding Physical forms(s) Binding eBI #of eBI Maximu duration eBI m 

### I (Le., bagdumping, filling drums) Engineering Controls Option and %new Option Workers Rrs/day Days/yr Mark (x) substance Marl< (x) Exposed (I) (2) (3) (4) (5) (6) (7) (8) (9) (IO) (II) (12) i Quality control sampling Gloves, face shield X X X X

Drumming product Gloves, face shield X X X X Unitwashout Gloves, safety X X X X aoaales 

## D Mark (X) this boxifyou attach acontinuation sheet. 3. Environmental Release andDisposal

- You mustmake separate confidentialityclaims for therelease number and the amountofthe new chemical substance released and otherrelease and disposal information. Mark (X) the "Confidential"box nexttoeach item you claim as confidential. (1)

--Enterthe numberofeach release point identified inthe process description, part 11, section A, subsection Id(3). (2)

--Estimatethe amountofthe new substance released (a) directly to the environmentor(b) into control technology (inkg/day orkglbatch). (3)

- Mark (X) this column ifentries incolumns (1) and (2) are confidential business information (CBI). (4) -Identifythe media (stackair, fugitive air (optional-see Instruction Manual), surfacewater, on-site oroff-site landorincineration, POTW, orother(specify» to whichthe new substancewill be released from thatrelease point. (5)

- a. Describecontrol technology, ifany, andcontrol efficiencythatwill be used to limit the release ofthe new substanceto the environment. Forreleases disposed ofon land, characterizethe disposal method and state whether it is approved for disposal ofRCRAhazardous waste. On acontinuation sheet, for each sitedescribe any additional disposal methods thatwill be used and whetherthewaste is subject to secondary or tertiary on-sitetreatment. b. Estimate the amount released to theenvironmentaftercontrol technology (in kg/day). (6)

- Mark (X) thiscolumnifentries incolumns (4) and (5) are confidentialbusiness information (CBl). (7)

--Identifythe destination(s) ofreleases to water. Please supplyNPDES (National Pollutant Discharge Elimination System)numbers fordirect discharges or NPDES numbers ofthe POTW (Publici OwnedTreatmentWorks). Mark (X) ifthe POTW name orNPDES # is confidential business information (CBI). Release Amountofnew substance CBI Mediaof Control technology and efficiency (you may wishtooptionally attachefficiency data) CB] Number released release

(1) (2a) (2b) e.g. stack air Binding (3) (4) (Sa) Mark(X) (5b) (6) X X X X X X

(7) Mark(X) the 

## U POTW provide name(s) below: CBI 

## D Navigable 

# D Other

- Specify provideNPDES# CBI destination(s) of 

## 7 waterway X releases to water. 

## D Mark (X) thisbox ifyou attach acontinuation sheet.

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

### Part 11-- HUMAN EXPOSUREAND ENVIRONMENTALRELEASE

-- Continued Section B
-- INDUSTRIALSITESCONTROLLEDBY OTHERS Complete sectionBfortypical processing oruse operations involving the new chemical substanceatsites you do notcontrol. Importers do nothaveto completethis section foroperations outside the U.S.; however, you mustreport any processingoruse activities afterimport. See the InstructionsManual. Complete aseparate sectionB foreach type of processing, oruse operation involvingthe newchemicalsubstance. Ifthe same operationis performed atmore than one sitedescribethe typical operation commonto these sites. Identify additional sites on acontinuation sheet. 1. OperationDescription

-- Toclaim information in this section as confidential, circleorbracketthe specificinformation thatyou claim as confidential. (1)

-- Diagramthemajorunitoperationsteps and chemical conversions, includinginterim storageandtransportcontainers (specify

- e.g. 5 gallonpails, 55 gallon drums, rail cars, tanktrucks, etc). Onthe diagram, identifyby letterand briefly describeeach workeractivity. (2)

- Providethe identity, the approximate weight (by kg/day orkglbatch, on an 100%new chemical substancebasis), and entrypointofall feedstocks (includingreactants, solvents and catalysts, etc) and all products, recycle streams, and wastes. Include cleaning chemicals (notefrequency ifnotused daily orperbatch). (3) --IdentifY by numberthe points ofrelease, including small or intermittentreleases, to the environment ofthe new chemical substance. (4) Pleaseenterthe#ofsites (remember to identifY the locations of these sites on acontinuation sheet): 7 # ofsites 

# [X] 

# eBI

D Mark (X)this boxifyou attach acontinuation sheet. 2. WorkerExposnrelEnvironmentalRelease (I)

­ Fromthe diagram above, providethe letterfor eachworker activity. Complete 2-8 foreachworkeractivitydescribed. (2)

--Estimate the numberofworkers exposed for all sites combined. (4)

- Estimatethe typical duration ofexposureperworker in (a) hours perday and (b) days peryear. (6)

- Describe physical form ofexposure and %new chemical substance (ifinmixture), and any protective equipmentand engineering controls, ifany, usedto protectworkers. (7)

--Estimatethe percentofthe new substanceas formulated when packaged orused as afinal product. (9)

--From the process diagram above, enterthe numberofeach release point. Complete 9-13 for each release pointidentified. (10)

-- Estimatethe amountofthenew substance released (a) directly tothe environmentor (b) intocontrol technology to theenvironment(in kg/day orkglbatch). i (12)

-- Describe mediaofrelease i.e. stack air, fugitive air(optional-see InstructionsManual), surface water, on-site oroff-siteland orincineration, POTW, orother 

## I (specifY) and controltechnology, ifany, thatwill beused to limitthe release ofthe new substanceto the environment. (14)

-- IdentifYbyproductswhich mayresultfrom theoperation.

(3), (5), (8), (II), (131and (15)

­ Mark (X)this column ifany ofthe proceediDlilentries are confidential business information (CBI). iLetter #of CBI Duration CBI Protective Equip. I %in CBI Release Amounlof CBI MediaofRelease CBI

of Workers of Engineering Controlsl Form- Number New & Control i Act- EXPQSed Exposure Physical Form and % ulation Substance Technology

ivily new substance Released (I) (2) (3) (4a) (4b) (5) (6) (7) (8) (9) (lOa) (lOb) (II) (12) (13) X X X X X i

14) Byproducts: (15)



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

### OPTIONALPOLLUTIONPREVENTIONINFORMATION Toclaim information in this section as confidential circle orbracketthe specific information that youclaim as confidential. 

### Inthis sectionyoumayprovide informationnotreported elsewherein this form regarding your effortsto reduce orminimizepotential risks associated with activities surrounding manufacturing, processing, use and disposal ofthePMN substance. Please includenewinformationpertinenttopollutionprevention, including source reduction, recycling activities and saferprocesses orproducts availabledue tothe newchemical substance. Source reduction includesthereduction inthe amount ortoxicityofchemical wastes bytechnological modification,process andproceduremodification, productreformulation, rawmaterials substitution, and/or inventory controL Recyclingrefersto thereclamationofuseful chemical componentsfrom wastesthatwould otherwisebetreated orreleased as airemissions orwater discharges, orland disposal. Descriptionsofpollution prevention, sourcereduction andrecycling should emphasizepotentialriskreduction subsequentto compliance withexistingregulatoryrequirements and canbe eitherquantitative or qualitative. TheEPA is interested inthe informationto assess overallnetreductions intoxicityorenvironmental releases and exposures,notthe shifting ofrisksto other environmental mediaor non-environmental areas (e.g., occupational or consumerexposure). In addition, informationontherelative cost orperformance characteristics ofthePMN substancetopotential alternatives maybeprovided. 

### All informationprovided inthis sectionwill betaken into consideration duringthereview ofthis substance. SeeInstructionsManualandPollutionPrevention Guidance manualforguidance and examples. Describethe expectednet benefits, such as (1) an overallreduction inriskto human health orthe environment; (2) a reduction in the volumemanufactured; (3) a reduction in the generation ofwaste materialsthrough recycling, source reduction orothermeans; (4) a reduction in potential toxicity orhumanexposure and/orenvironmental release; (5) an increase inproductperformance, a decrease in the costofproduction and/orimproved operation efficiency ofthenewchemical substance in comparisonto existingchemical substancesused in similarapplication; or(6) the extentto which the newchemical substance maybe a substitute for anexisting substancethatposes a greater overallrisktohumanhealth orthe environment.



# o Mark (X) this box ifyou attach a continuation sheet. EPA FORM 7710-25 (Rev. 5-95) Page 11

### Part III··LIST OFATTACHMENTS Attach continuation sheets for sectionsofthe form and test data and otherdata (includingphysical/chemicalproperties and structure/activityinformation), andoptionalinformation afterthispage. Clearly identify the attachment and the sectionofthe form to which itrelates, ifappropriate. Number consecutively thepages ofthe attachments. In the columnbelow, enterthe inclusive page numbers ofeach attachment. Mark (X) the "Confidential"boxnext to any attachmentname you claimas confidential. Read the Instructions Manual for guidance on how to claimany information in an attachment as confidential. Youmust include withthe sanitized copy ofthe notice form a sanitized version ofany attachment in whichyou claiminformation as confidential. Attachmentname Attachment pagenumber(s)

134-141 MaterialSafetyDataSheet(MSDS) Inventory Expert Service Report 133 Confidential
Magnusson & Klingman Maximisation Study in the Guinea Pig 14-64 Primary Skin Irritation - Rabbits 65-76 Acute Oral Toxicity, LD50 - Rats 77-105 Acute Toxicityto Photobacteriumphosphoreum (Screen test) 106-113 Acute Toxicityto Sheepshead Minnow 114-121 Acute Toxicityto Fathead Minnow 122-129 Raw Ready Biodegradability Closed BottleTestdata 130 131-132 X



# N Mark (X) this boxifyou attacha continuation sheet. Enter the attachmentname andnumber.
EPA FORM 7710-25 (Rev. 5-95) Page 12· PHYSICALAND CHEMICAL PROPERTIES WORKSHEET ToassistEPA'sreviewofphysical andchemical properties data, please complete the following worksheetfor data you provideand includeitin the notice. Identifythe propertymeasured, the page ofthe notice on whichthe property appears, the valueoftheproperty, the unitsin which the propertyis measured (as necessary), and whetherornot thepropertyis claimedas confidential. Thephysical stateofthe neat substanceshouldbeprovided. These measuredproperties shouldbe for the neat (100% pure) chemicalsubstance. Properties that are measuredfor mixtures orformulations shouldbe sonoted (% PMN substancein_). You arenotrequired to submit this worksheet; however, EPA stronglyrecommends that you do so, as it willsimplifyreviewandensure thatconfidential information is properly protected. You should submitthis worksheet as a supplementto your submissionoftestdata. This worksheet is not a substitutefor submissionoftestdata. Property Mark (X)if (a) provided X Physical stateofneatsubstance Vaporpressure @ Temperature 

### °C X Density/relativedensity Solubility @ Temperature 

### °C

Solvent Solubilityin water @Temperature 

### °C Meltingtemperature Boiling I sublimationtemperature@ torrpressure Spectra Dissociationconstant Particle size distribution Octanollwaterpartition coefficient Henry'sLawconstant Volitalizationfrom water Volitalization from soil X I'H@concentration 5%in water Flammability Explodability Adsorption I coefficient Other

- Specify Page Value number

(b) (c) (s) X(l)

18 0.863

18 11.5

- 11.9 (g)
Torr g/cm3

gIL gIL 

### °C °C jM.easured or ConliEstimate dential MIIIi«X) (MorE) (d)

E

M

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ATTACHMENT PAGE 13

# ]



# 1 

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# - CONFIDENTIAL



## STUDY SPONSOR: ENV93

## p1201 : MAGNUSSON 

## &KLIGMAN MAXIMISATION STUDY IN THE GUINEA PIG

PROJECT NUMBER: 654/1

AUTHOR: ' 

### R. Driscoll



## ISSUED BY: Safepharm Laboratories Limited 

## P.O. Box No. DERBY DEI 2BT U.K. Telephone: Facsimile: 

### Telex: 

## 45

DERBY (0332) 792896 (0332) 799018 377079 SAFPHM G PAGE 1 OF 51 PAGES

# III 

# • -2­ 

### PROJECT NUMBER: 654/1 

# la QUALITY ASSURANCE REPORT 

# .It 

# •**~i'** '

..... '" •. 

### The routine inspection of short term studies at Safepharm Laboratories is**~':.i\** e 

# ··· carried out as a continuous process designed to encompass all major phases of 

# I 

# I each study type once per**month~** 

### Dates of the most recently completed series of monthly inspections relevant to the study type(s) in this report are given below. 

# I Date{s) of Inspection and Reporting: 

# I 01, 09, 15, 16, 21, 24, 28 February 1994 

# '1 This report has been audited by Safepharm Laboratories Quality Assurance Unit . 

# .. It is considered to be an accurate account of the data generated and of the 

# I procedures followed. 

# I '3 .•. Date of Report Audit: 

# I**~** 

### 28 March 1994

# ~ I 

# 1 I J.R. 

### Paternan C. 

### Biol., M.L 

### Biol. 

### FOR SAFEPHARM QUALITY ASSURANCE UNIT 

# 1
DATE: 

# ........................·······13D·O·O 1 5

### PROJECT NUMBER: 654/1

GlP COMPLIANCE STATEMENT

I, the undersigned, hereby declare that the objectives laid down in the protocol were achieved and as nothing occurred to adversely affect the quality or integrity of the study, I consider the data generated to be valid. This report fully and accurately reflects the procedures used and data generated. The work described was performed in compliance with the UK Principles of Good Laboratory Practice (The United Kingdom Compliance Programme, Department of Health 1989). These Principles are in accordance with GlP standards published by DECO in 1982; and are in conformity with, and implement, the requirements of Directives 87/18/EEC and 88/320/EEC. ,These international standards are acceptable to the United States

Environmental Protection Agency and Food and Drug Administration, and fulfil the requirements of 40 CFR Part 160, 40 CFR Part 792 and 21 CFR Part 58 (as amended)
. 

## 2,-Li--Q l,


### . ........

# ~~~~.~ .............. DATE: 

## .............. \

# ....**\~** 

## ....... ,. ..... 

### R. Driscoll B.Tech. (Hons.) Study Director for Safepharm Laboratories



# 000016

## PROJECT NUMBER: 654/1 CON TEN T S PAGE
7 

## SUMMARY 9 

### 1. 

## INTRODUCTION 2. TEST AND POSITIVE CONTROL MATERIALS 10 2.1 Test Material 10 2.2 Positive Control Material 10 

## 3. TEST SYSTEM 11 

### 3.1 Specification 11 

### 3.2 Husbandry 11 

## 4. PROCEDURE 11 

### 4.1 Selection of Concentrations for Main Study (Sighting Tests) 11 4.2 Main Study 13 

## 5. EVALUATION OF DATA 18 

## 6. ARCHIVES 19 

## 7. RESULTS 20 

### 7.1 Intradermal and Topical Sighting Tests 20 

### 7.2 Main Study 20 

### 7.3 Bodyweight 23 

## 8. CONClUSION 24 

## TABLES TABLE 1 Individual Skin Reactions in Test Animals at Challenge: 

## KEY TO TABLES 25
26 

### ENV 93 1201 Study 

# ·····11 

# '. '--, TABLE 2 Individual Skin Reactions in Control Animals at Challenge: 27 

### ENV 93 1201 Study



# · .... TABLE 3 Individual Skin Reactions in Test Animals at Cha11enge: 

# ......·.l 28



# ....! 

### ONCS Study TABLE 4 Individual Skin Reactions in 

### Control Animais at Challenge: 29

.- .•....•....

# ~**.~,,'** ONCS Study 

# O 

# 'J ,"'I r"'I ," " -" , I r I I, , ,)
....) J ; I-;)­ 

### PROJECT NUMBER: 654/1 CON TEN T S (continued)

- PAGE

30 

### APPENDICES 31 

### KEY TO APPENDICES APPENDIX I Intradermal Sighting Test
- Summary of Results: 32 ENV 93 

### 1201 Study APPENDIX II Intradermal Sighting Test
- Summary of Results: 33 

### DNCS Study APPENDIX III Topical Sighting Test For Induction Application

- 34 

### (48-hour Exposure)

- Individual Skin Reactions: ENV 93 

### 1201 Study APPENDIX IV Topical Sighting Test for Induction Application

- 35 

### (48-hour Exposure)

- Individual Skin Reactions: DNCS Study APPENDIX V Topical Sighting Test For Challenge Application

- 36 

### (24-hour Exposure)

- Individual Skin Reactions: ENV 93 

### 1201 Study APPENDIX VI Topical Sighting Test For Challenge Application

- 37 

### (24-hour Exposure)

- Individual Skin Reactions; ONCS Study APPENDIX VII Intradermal Induction· Individual Si<in Reactions 39 

### in Test Animals: ENV 93 1201 

### Study APPENDIX VIII Intradermal Induction

- Individual Si<in 

### Reactions 40 

### in Control Animals: ENV 93 1201 

### Study APPENDIX IX Intradermal Induction

- Individual**S~in** 

### Reactions 41 in Test Animals: 

### ONeS Study 

# 000018-6­ 

### PROJECT NUMBER: 654/1 CON TEN T S (continued)
PAGE
APPENDICES (contd) APPENDIX X Intradermal Induction

- Individual Skin Reactions 42 

### in Control Animals: ONCS Study APPENDIX XI Topical Induction

- Individual Skin Reactions 43 

### in Test Animals: ENV 93 1201 Study APPENDIX XII Topical Induction

- Individual Skin Reactions 44 in Control Animals: ENV 93 1201 Study APPENDIX XIII Topical Induction

- Individual Skin Reactions 45 in Test Animals: ONCS Study APPENDIX XIV Topical Induction

- Individual Skin Reactions 46 in Control Animals: ONCS Study APPENDIX XV Individual Sodyweights and Sodyweight Gains 47 

### of Test Animals: ENV 93 1201 Study APPENDIX XVI Individual Bodyweights and Bodyweight Gains 48 

### of Control Animals: ENV 93 1201 Study APPENDIX XVII Individual Bodyweights and Bodyweight Gains 49 

### of Test Animals: DNCS Study APPENDIX XVIII Individual Bodyweights and Bodyweight Gains 50 

### of Control Animals: ONeS Study

"'1 

### APPENDIX XIX GLP Statement of Compliance -,,,, 

# c( ,-.r-,49 I**~J** 

## UU 

## l
...

# i 

# i -7­ 

### PROJECT NUMBER: 654/1 SUMMARY 

# - STUDY SPONSOR PETROLITE CORPORATION

, 

### PROJECT NUMBER 654/1
TEST MATERIAL ENV 93 1201

1. Astudy was performed to assess the skin contact sensitisation potential of the test material in the albino guinea pig. The method used followed that described in the OECD Guidelines for Testing of Chemicals No. 4Q6 "Skin Sensitisation" (adopted 17 July 1992) and Method B6 of Commission Directive 92/69/EEC (which constitutes Annex Vof Council Directive 67/548/EEC)
. The results may be used as a basis for classification and labelling under Annex VI of Council Directive 67/548/EEC (as adapted to technical progress by Commission Directive 91/325/EEC)
.. 2. Twenty test and ten control animals were used for the main study. Based on the results of sighting tests, the concentrations of test material for the induction and challenge phases were selected as fa11ows: Intradermal Induction 0.1% w/v in distilled water Topical Induction 10% v/v in distilled water Topical Challenge 5% and 1% v/v in distilled water 3. The test material produced a 0% (0/20) sensitisation rate and was classified as a non-sensitiser to guinea pig skin. The test materiai was also classified as a non-sensitiser according to EC labelling regulations. No risk phrase is required. 

# 000J20-0­ 

### PROJECT NUMBER: 654/1
SUM MAR Y {continued

4. 

## Aconcurrent study using ten test and five control animals was performed with the known positive contact sensitiser, 2,4-Dinitrochlorobenzene (ONCS). Based on the results of sighting tests the concentrations of positive control material for the induction and challenge phases were selected as follows: Intradermal Induction 0.1% w/v in arachis oil B.P. Topical Induction 0.75% w/v in absolute ethanol Topical Challenge 0.75% and 0.5% w/v in absolute ethanol The positive control material, ONCS, produced a 100% (10/10) sensitisation rate. The positive reactions seen with ONCS were consistent with the known contact**~ensitisation** 

### potential of the material.



# 000021-9­ 

### PROJECT NUMBER: 654/1
ENV 93 1201: 

### MAGNUSSON &

### KLIGMAN MAXIMISATION STUDY IN 

## THE GUINEA PIG

1. 

### INTRODUCTION This study was performed to assess the skin contact sensitisation potential of the test material (Safepharm Definitive Protocol Number 654/1). The method used followed the recommendations of the OECD Guidelines for the Testing of Chemicals No. 406 "Skin Sensitisation" (adopted 17 July 1992) and Method 86 of Commission Directive 92/69/EEC (which constitutes Annex Vof Council Directive 67/548/EEC). The results may be used as a basis for classification and labelling under Annex 

### VI of Council Directive 67/548/EEC (as adapted to technical progress by Commission Directive 91/325/EEC). The test system was chosen because the guinea pig has been shown to be a suitable species for this type of study and is recommended in the test method. The strain used in these laboratories has been shown to produce a satisfactory sensitisation response using known positive sensitisers. The results of the study are believed to be of value in predicting the likely contact sensitisation potential of the test material to man. The study was conducted in accordance with the internationally accepted general principles of Good Laboratory Practice and Safepharm Standard Operating Procedures. The study was performed between 7 February 1994 and 12 March 1994. 

# 000022-.I.V

### PROJECT NUMBER:**~54/1**


### 2. TEST AND POSITIVE CONTROL MATERIALS 2.1 

### Test Material a} Description. Identification and Storage Conditions The test material was supplied by the study sponsor as follows: Sponsor's identification ENV 93 1201 

### Batch number 93-73731 

### and 93-73732 

### Activity 70% 

### Date received 26 January 1994 

### Description brown slight1y viscous liquid Container c1ear glass bottle Storage conditions room temperature Data relating to the identity. purity and stability of the test material are the responsibility of the sponsor. b) Method of Preoaration For the purpose of this study the test material was freshly pre­ pared as follows: Intradermal Induction 0.1% 

### wlv in distilled water 0.1% 

### w/v in a mixture of Freund's Complete Adjuvant plus distilled water (l: 1) 

### Topical Induction 10% 

### v/v in distilled water Topical Challenge 5% and 1% 

### v/v in distilled water The concentration,**homogeneit~** 

### and stability of the formulations were not determined by analysis. 2.2 Positive Control Material The positive control material 2,4-dinitrochlorobenzene (ONCS) was supplied by Sigma, Poole, Dorset, U.K. For the purpose of the study the 

### positive controi material was freshly prepared as follows: 

# 000023-11­ 

## PROJECT NUMBER: 654/1 2.2 Positive Control Material (contd) Intraderma1 Induction 0.1% 

### w/v in arachis oil 0.1% 

### w/v in a mixture of Freund's Complete Adjuvant plus distilled water (1: 1) 

### Topical Induction 0.75% 

### w/v in absolute ethanol Topical Challenge 0.75% 

### and 0.5% 

### w/v in absolute ethanol 3. TEST SYSTEM 3.1 Specification Sixty-four female, albino Dunkin-Hartley guinea pigs were supplied by David Hall limited, Burton-an-Trent, Staffordshire, U.K. At the start of the main study the animals weighed 337

- 444g, and were approximately eight to twelve weeks old. After a minimum acclimatisation period of five days, each animal was selected at random and given a number unique within the study which was written on a small area of clipped rump using a black indelible marker-pen. 3.2 Husbandry The animals were housed singly or in pairs in solid-floor polypropylene cages furnished with woodf1akes. Free access to mains tap water and food (Guinea Pig FDI Diet, Special Diets Services limited, Witham, Essex, U.K.) was allowed throughout the study. The animal room was maintained at a temperature of 20

-**23~C** 

### and relative humidity of 46

- 67%. 

### The rate of air exchange was approximately 15 changes per hour and the lighting was controlled by a time switch to give 12 hours light and 12 hours darkness. 4. 

## PROCEDURE The method used for assessing the sensitising properties of the test material was based on the Guinea Pig Maximisation test of Magnusson B. 

## & Kligman A.M., J. 

### Invest. Dermatol. (1969) 52: 268

- 276. 

### 4.1 Selection of Concentrations for Main Study 

### (Siahtino Tests) The concentrations of test material and ONeS to be used at each stage of the main study were determined by 'sighting tests' in which groups of guinea pigs were treated with various concentrations of test material. The procedures were as follows: 

## O**I~l** 

### n 

## 0'
. I**~** I I**~.J ~.J**  .

### -12­ PROJECT NUMBER: 654/1

4.1 Selection of Concentrations for Main Study (Sighting Tests) 4.1.1 Test Material a) Selection of Concentration for Intradermal Induction Four animals were intradermally injected with preparations of test material (O.l%, 0.5%, 1% or 5% w/v in distilled water). The degree of erythema at the injection sites was assessed approximately 24, 48 and" 72 hours and 7 days after injection according to the Draize" scale shown in Section 4.2.1a. The degree of oedema was not evaluated. Any evidence of systemic toxicity was also recorded. The highest concentration that did not cause local necrosis, ulceration, severe irritation or systemic toxicity, was selected for the intradermal induction stage of the main study. b) Selection of Concentration for Topical Induction Two guinea pigs (intradermally injected with Freund's Complete Adjuvant 9 days earlier) were treated with four preparations of the test material (10%, 5%, 2% and 1% v/v in distilled water). Two more guinea pigs (intradermally injected with Freund's Complete Adjuvant twelve days earlier) were treated with two concentrations of the test material (50% and 25% w/v in distilled water). These animals were killed for humane reasons immediately after dressing removal due to severe dermal necrosis. The h.ighest concentration producing only mild to moderate dermal irritation after a 48-hour occlusive exposure, was selected for the topical induction stage of the main study. c) Selection of Concentration for Topical Challenae Four preparations of the test material (10%, 5%,**2~'** 

### and 1% v/v in distilled water) were applied occlus;vely to the flanks of two guinea pigs for a period of 24 hours. These guinea pigs did not form part of the main study but had been treated identically to the control animals of the main study, up to Day 14. The highest non-irritant concentration of the test material and one lower concentration were selected for the topical challenge stage of the main study. 

# 000025

### -13­ 

### PROJECT 'NUMBER: 654/1

Induction 4.1 Selection of Concentrations for Main Stud 4.1.2 

### Positive Control Material a) Selection of Concentration for Intradermal Three animals were intradermally injected with preparations of test material (1%, 0.5% or 0.1% w/v in arachis oil B.P.). The injection sites were evaluated as in Section 4.1.1a. The highest concentration that did not cause 10ca1necrosis,**ulceration~1 ~evere**  

### irritation or systemic toxicity, was selected for the intradermal induction stage of the main study. b) 

### Selection of Concentration for Topical Induction Two guinea pigs (intraderma1ly injected with Freund's Complete Adjuvant 9 

### days earlier) were treated with four preparations of the test material (3%, 1.5%, 0.75% and 0.3% w/v in absolute ethanol). The highest concentration producing only mild to moderate dermal irritation after a 48-hour occlusive exposure, was selected for the topical induction stage of the main study. c} Selection of Concentration for Topical Challenge Four preparations of the test material (0.75%, 0.5%, 0.25% and 0.1% w/v in absolute ethanol) were applied occ1usively to the flanks of two guinea pigs for a 

### period of 24 hours. TheSe guinea pigs did not form part of the main study but had been treated identically to the control animals of the main study, up to Day 14. Two more guinea pigs, which had been intradermally injected with Freund's Complete Adjuvant 11 days.,earlier, were treated with four concentrations of the test material (2%, 1.5%, 1% and 0.75% 

### w/v in absolute ethanol). The highest non-irritant concentration of the test material and one lower concentration were selected for the topical challenge stage of the main study.**~** 

# I , 

# ···'·' 4.2 

### Main Study A total of forty-five guinea pigs were used for the study: twenty test and ten control animals were used to determine the skin sensitisation potential of the test material and a further ten test 

# I

and five control animals comprised the concurrent positive contro' 

# B

..".·.·. 

# r study. 

# n n n';)**f~**

### -14­ 

### PROJECT NUMBER: 654/1 4.2 Main Study (contd) 4.2.1 Test Material a) Induction of the ENV 93 1201 Test Animals Shortly before treatment on Day 

# a the hair was removed from an area approximately 40 mm x 60 mm on the shoulder region of each animal with veterinary clippers. Arow of three injections (0.1 ml each) was made on each side of the mid-line. The injections were:

. i) 

### 'Freund's Complete Adjuvant'plus distilled**wah~~in'~fhe** 

### ratio 1:1 ii) a 0.1% w/v dilution of test material in distilled water iii) a 0.1% 

### w/v dilution of test material in a 1:1 

### mixture of Freund's**Complet~** 

### Adjuvant plus distilled water Approximately 24 

### and 48 

### hours after intradermal injection the degree of erythema at the test material injection sites (i.e. injection site ii) was evaluated according to the following scale from Draize J.H., (1977) 

### "Dermal and Eye Toxicity Tests ll 

### In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington 

### D.C. p.31: 

# I EVALUATION OF 

## SKIN REACTIONS Erythema and Eschar Formation Value No erythema
................................................... 0 Very slight erythema (barely perceptible)
..................... 1 We11 -defined erythema
......................................... 2 Moderate to severe erythema
................................... 3 Severe erythema (beet redness) to slight eschar formation

(injuries in depth)
.......................................... 4 

# I Oedema Formation 

# I No oedema
..................................................... 0 Very slight oedema (barely perceptible)
....................... 1 

# , 

# , Slight oedema (edges of area well-defined by definite ra1 slng)
..................................................... 2 Moderate oedema (raised approximately 1 millimetre)
........... 3 

# I Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure)
............., ................_ .. 4 

# 000027

### (contd) -15­ 

### PROJECT, NUMBER: 654/1

4.2.1 Test Material (contd) a) Induction of the ENV 93 1201 Test Animals One week later (Day 7), the same area on the shoulder region used previously for intradermal injections was clipped again and treated with a topical application of the test material formulation (10% ,;i,v/v'in distilled water). The test material formulation was applied to saturation on filter paper (WHATMAN No.4: approximate size 40 mm x 20 mm) which was held in place by a strip of surgical adhesive tape (BLENOERM: approximate size 50 mm x 30 mm) and covered with an overlapping length of aluminium foil. The patch and foil were further secured by a strip of elastic adhesive bandage (ELASTOPLAST: approximate size 250 mm x 35 mm) wound in a double layer around the torso of each animal. This occlusive dressing was kept in place for 48 hours. The degree of erythema and oedema was quantified one and**twenty~** 

### four hours following removal of the patches using the Draize scale shown previously. Any other reactions were also recorded. b) Induction of the ENV Intradermal injections 93 1201 Control Animals were administered using an identical procedure to that used for the test animals, except that the injections were: ;) Freund's Complete Adjuvant plus distilled water in the ratio 1:1. ii) 

### distilled water. iii) 50% w/v distilled water in a 1:1 mixture of Freund's Complete Adjuvant plus distilled water The topical applications followed the same proceaure as for the test animals except that the vehicle alone was applied to the filter paper. Skin reactions were quantified as for the test animals. rU' ('; Ii

# I,.--:, 0 '.) 'J i.,.; 

### L. 

### u

### -16­ PROJECT**~UMBER:** 

### 654/1

4.2.1 Test Material (contd) c) Challenge of ENV 93 1201 Test and Control Animals Shortly before treatment on Day 21, an area, approximately 50 mm x 70 mm on both flanks of each animal, was clipped free of hair with veterinary clippers. Aquantity of 0.1

- 0.2 ml of the test material formulation (5% v/v in distilled water) was applied to the shorn right flank of each animal on a square of filter paper (WHATMAN No.4: approximate size 20 mm x 20 mm) which was held in place by a strip of surgical adhesive 'tape (BLENDERM: approximate size 40 mm x 50mm). To ensure that the maximum non-irritant concentration was used at challenge, the test material at a concentration of 1% v/v in distilled water was also similarly applied to a skin site on the left shorn flank. The patches were occluded with an overlapping length of aluminium foil and secured by a strip of elastic adhesive bandage (ELASTOPLAST: approximate size 250 mm x 75 mm) wound in a double layer around the torso of each animal. After 24 hours, the dressing was carefully cut using blunt-tipped scissors, removed and discarded. The challenge sites were swabbed with cotton wool soaked in diethyl ether to remove residual material. The position of the treatment sites was identified by using a black indelible marker-pen. Prior to the 24-hour**observ~tion** 

### the flanks were clipped using veterinary cl ;ppers to remove regrown hair. d) Evaluation of Skin Reactions The degree of erythema and oedema was quantified approximately 24 and 48 hours after challenge dressing removal using the Draize scale shown in Section 4.2.1a. Any other reactions were also recorded. 

# 000029

# I I 7 -17­ 

### PROJECT NUMBER: 654/1

4.2.2 Positive Control Material a) 

### Induction of ONCS Test Animals Shortly before treatment on Day 0 

### the hair was removed from an area approximately 40 mm x 

### 60 mm on the shoulder region of each animal with veterinary clippers. Arow of three injections (0.1 ml each) was made on each side of the mid-line. The injections were: i) 

### Freund's Complete Adjuvant plus distilled water in the ratio 1:1 ii) 

### a 0.1% w/v dilution of ONCS in arachis oil 

## B.P. iii) 

### a 0.1% w/v dilution of ONCS in a 1:1 mixture of Freund's Complete Adjuvant plus distilled water Approximately 24 and 48 hours after intradermal injection the degree of erythema at the test material induction site (i.e. injection site ii) was evaluated using the Oraize scale shown in Section 4.2.1a. The topical applications for induction of the ONC8 test animals followed the same procedure as for the ENV 93 

### 1201 animals except that ONCS at a concentration of 0.75% 

### w/v in absolute ethanol was applied. b) 

### Induction of ONCS Control Animals Intradermal injections were administered using an identical procedure to that used for the ONCS test animals, except that the injections were: i) Freund's Complete Adjuvant plus distilled water ii) 

### arachis oil B.P. 

### iii) 50% w/v arachis o"il B.P. 

### in a 1:1 

### mixture of Freund's Complete Adjuvant plus distilled water The topical application for the ONCS control animals followed the same procedure as the 

### ONCS test animals except that 

### absolute ethanol was applied.

### -18­ 

### PROJECT ,NUMBER: 654/1
4.2.2 Positive Control Material (contd) c) Challenge of DNCS Test and Control Animals The challenge applications for the ONCS animals fallowed the same procedure as the ENV 93 1201 animals except that 0.1

- 0.2 ml of DNCS at concentrations of 0.75% and 0.5% w/v in absolute ethanol was applied to the skin. d) Evaluation of Skin Reactions The degree of erythema and oedema was quantified approximately 24 and 48 hours after challenge dressing removal using the Draize scale shown in Section 4.2.1a. Any other reactions were also recorded. 5. EVALUATION OF DATA The percentage of test animals that showed a more severe reaction at the test material challenge site than the most severe reaction seen in the control animals, was compared with the following scale:**~** 

### of animils Classification of sensitiseg sensitisation Qotential 

# a non-sensitiser > 

## a 8 weak sensitiser > 

### 8 

# - 28 mild sensitiser > 

### 28 

## - 64 moderate sensitiser > 

### 64 

# - 80 

### strong sensitiser > 

### 80 

# - 100 extreme sensitiser The data obtained may be used to classify the test material under the Council Directive 67/548/EEC, as amended by Commission according to Directive 91/325/EEC, on the classification, packaging and labelling of dangerous substances. These classification criteria are also in the UK Approved Code of Practice "Classification and Labell ing of Substances Dangerous for**Supply~.** 

### Aresponse in at least 30% of the animals is considered positive and the risk phrase 

### R43 "MAY CAUSE SENSITISATION 

### BY SKIN CONTACT" is required. 

# 000031

### -19­ 

## PROJECT NUMBER: 654/1 I: 

# o \...I/!;, -..t L,1',{!lJ 

### 6. 

## ARCHIVES 1./

## (.!:!J Unless instructed otherwise by the sponsor, all original data

## and61J~pp' ' 4...J,"**~** l 	 . 

### of the final report will be retained in the archives of**Safepharm(J~** 

### Laboratories for a period of ten years. After this period, the sponsor's instructions will be sought.

### -20­ 654/1


### 7. 

### RESULTS 7.1 Intradermal and Topical Sightino Tests A summary of the results of the intradermal sighting test**and~e** 

### individual skin reactions observed in the topical sighting tests , are given in Appendices I to VI. Based on these results the following concentrations were selected for the main study. Test Material 

### [NV 93 1201 Intradermal Induction 0.1% w/v in distilled water Topical Induction 10% v/v in distilled water Topical Challenge 5% and 1% v/v in distilled water Positive Control Material ONCS Intradermal Induction 0.1% w/v in arachis oil B.P. Topical Induction
. 0.75% w/v in absolute ethanol Topical Challenge 0.75% and 0.5% w/v in absolute ethanol 7.2 Main Study 7.2.1 Test Material a) Skin Reactions Observed After Intradermal Induction Individual reactions observed at the intradermal induction sites of test and control group animals are given in Appendix VII and VIII. Very slight to well-defined erythema was noted at the intradermal induction sites of all test group animals at the 24-hour observation and in nineteen test group animals at the 48-hour observation. No skin reactions were noted at the intradermal induction sites of control group animals at the 24 and 48-hour observations. b) Skin Reactions Observed After Topical Induction Individual reactions observed at the topical induction sites of the test and control group animals are given in Appendices XI and XII. 

# 000033

### -21­ 

### PROJECT 

## NUMBER: 654/1

7.2.1 Test Material (contd)

",", . , 

### b) Skin Reactions Observed After Topical Induction (contd) ."1....:, 

### Very slight to moderate to severe erythema with or without very slight to slight oedema was noted at the induction sites of all test group animals at the l-hour observation. Very slight to well­ "defined erythema with or without very slight oedema was noted at

the induction sites of seventeen test group animals at the 24-hour observation. Other skin reactions noted were bleeding caused by animal scratching intradermal induction site, dried blood, desquamation, hardened dark coloured scab and small scattered scabs. Skin reactions prevented accurate evaluation of erythema at the induction sites of three test group animals at the 24-hour observation. j. •**"~** (



### Very slight erythema with or without very slight oedema was noted at the treatment sites of seven control group animals at the l-hour observation. No skin reactions were noted at the treatment sites of control group animals at the 24-hour observation. c) Skin Reactions Observed After Topical Challenae Individual reactions at the challenge sites of test and control group animals are given in Tables land 2. No skin reactions were noted at the challenge sites of the test or control group animals at the 24 and 48-hour observations. 7.2.2 Positive Control Material 

## (DNCB) a) Skin Reactions Observed After Intradermal Induction Individual reactions observed at the intradermal induction sites of test and control group animals are given in Appendices 

## rx and X. Very slight to well-defined erythema was noted at the intradermal induction sites of all 

### ONe8 test group animals at the 24 and 48­ hour observations. 

# 000034

### 654/1

Induction (contd) -22­



### Positive Control Material (ONCSl (contd) a) Skin Reactions Observed After Intradermal Very slight erythema was noted at the intradermal induction sites of all control group animals at the 24-hour observation and in four control group animals at the 48-hour observation. b) Skin Reactions Observed After TODical Induction Individual reactions observed at the topical induction sites of test and control group animals are given in Appendices XIII and XIV. Yellow-coloured staining caused by the ONCS was noted at the induction sites of all ONCS test group animals at the 1 and 24-hour observations. The staining did not affect evaluation of skin responses. Well-defined to moderate to severe erythema and slight oedema were noted at the induction sites of all ONCS test group animals at the I-hour observations with well-defined erythema and slight oedema at the 24-hour observation. Very slight erythema was noted at the treatment site of one control group animal at the I-hour observation. No skin reactions were noted at the treatment sites of control group animals at the 24­ hour observation. c) Skin Reactions Observed After Topical Challenge Individual reactions at the challenge sites of test and control group animals are given in Tables 3 and 4. Pale yellow-coloured staining caused 

### by the 

### ONeS was noted at the challenge sites of all test and control group animals at the 24 and 48-hour observations. The staining did not affect evaluation of skin responses. 

## n 

# '0':; r.: I '...;' '-"-..) 

# O O

### -23­ 

### PROJECT NUMBER: 654/1
7.2.2 Positive Control Material (ONCS} (contd) c) Skin Reactions Observed After Topical Challenoe (contdJ 0.75% w/v in Absolute Ethanol Positive skin responses, well-defined erythema (grade 2) with or without very slight to slight oedema were noted at the challenge sites of nine test group animals at the 24 and 48-hour observations. Other incidents of skin reactions noted were dark brown-coloured scab, small· superficial scabs and well-defined erythema surrounding other skin reactions. Skin reactions prevented accurate evaluation of erythema and oedema at the challenge site of one test group animal at the 24 and 48-hour observations. Very slight erythema was noted at the challenge site of one control group animal at the 24 and 48-hour observations.**O.5~** 

### w/v in Absolute Ethanol Positive skin responses, well-defined erythema (grade 2) with or without very slight to slight oedema were noted at the challenge sites of all test group animals at the 24-hour observation and at the challenge sites of six test group animals at the 48-hour observation. Very slight erythema with or without very slight oedema was noted at the challenge sites of four test group animals at the 48-hour observation. Isolated incidents of small superfical scabs were also noted. Very slight erythema was noted at the challenge sites of**t'l~O** 

### control group animals at the 24 and 48-hour observations. 7.3 Bodvweight Individual bodyweights and bodyweight gains of test and control group animals of the ENV 93 1201 and 

## ONeS studies are given in Appendices XV to XVIII. Bodyweight gains of guinea pigs in the test groups, between Day 0 and Day 24, were comparable with those observed in the concurrent control group animals over the same period. 

# 000036

### -24­ 

### 654/1


### 8. CONCLUSION The test material, ENV 93 1201, produced a 0% (0/20) and was classified as a NON-SENSITISER to guinea pig skin. The test material was also classified as a non-sensitiser according to EC labelling regulations. No risk phrase is therefore required. The positive control material, ONCS, produced a 100% 

### (l0/10) sensitisation rate. The positive reactions seen with ONCS were consistent with the known contact sensitisation potential of the material.



# 000037

### -25­

KEY TOT A 8 L E S 

### PROJECT NUMBER: 654/1

Er 

## = erythema Oe 

### = oedema



### - = no other skin reactions Ss 

## =small superficial scabs St 

## =dark brown-coloured scab STA 

### =yellow-coloured staining at challenge site not affecting evaluation of skin responses We 

## =well-defined erythema surrounding other skin reactions ?e 

# =evaluation of erythema precluded by other skin reactions ?Od 

## = evaluation of oedema precluded by other skin reactions



# 000038

## [NV 93 1201 : MAGNUSSON 

## &KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG TAO t 

## E I 

## JNOIVIOlJAL SKIN REACTIONS IN TEST ANIMALS AT CHALLENGE: ENV 93 1201 STUDY CIlAU[NGE CONCENTRATIONS: 5% AND 1% v/v VEtIICLE: DISTILLED WATER Skin Reactions (Hours After Removal of Dressing) 24 hours 48 hours Animal Number 1% 5% 1% 5% 

## Er Oe Other Er Oe Other Er Oe Other Er Oe Other 1 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 

### 3 0 0 0 0 0 0 0 0 N I 0'1 4 

## o . 0 0 0 0 0 0 0 I 

## o . 5 0 0 0 0 0 0 0 . 0 6 0 0 0 0 0 0 0 7 0 0 0 0 0 0 0 0 8 0 0 0 0 0 0 0 0 

### 9 0 0 0 0 0 0 0 0 10 0 0 0 0 0 0 0 0 11 0 0 0 0 0 0 0 0 12 0 0 0 0 0 0 0 0 '"1;l ;;c 13 0 0 0 0 0 0 0 0**~** 14 0 0 0 0 0 0 0 0 r'T1**-~** 15 0 0 0 0 0 0 0 0 16 0 0 0 0 0 0 0 0 :z c: 

# o b-. 17 0 0 0 0 0 0 0 :I: -f'i t= 

### 0 18 0 0 0 0 0 0 0 

# o**.~** r'T1 

### 0 19 0 0 0 0 0 0 0 

# o

# .~ 

# .. :;0 

### 0 0 20 0 0 0 0 0 0 0 

# o**c-~]** 01 0'1 (J) -'"'



### ...... 

# 11<


# ­ ill

### ENV 93 1201 : MAGNUSSON 

### &KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG T A 8 

### L. [ 2 INDIVIDUAL. SKIN REACTIONS IN CONTROL ANIMALS AT CIIALLENGE: ENV 93 1201 STUDY CIIALL.ENGE CONCftHRATIONS: 5% AND 1% v/v VEIIICLE: DISTILLED WATER Skin Reactions (ltours After Removal of Dressing) 24 hours 48 hours Animal Number 

### 1% 5% . 1% 5% Er Oe Other Er Oe Other Er Oe Other Er Oe Other 

### 21 0 0 

### 0 0 0 0 0 0 N I "-J 

### 22 0 0 0 0 0 0 

### 0 0 I 23 0 0 0 0 0 

### 0 0 0 

### 24 0 0 0 0 0 0 0 0 

### 25 0 0 0 0 0 0 0 0 26 0 0 0 0 0 0 0 27 0 0 0 0 0 0 0 -0 28 0 0 0 0 0 0 0 

# 2**~** 29 0 0 0 0 0 0 0 ...,.j :iii!: 30 0 0 0 0 0 0 0 c::: :J: .=:;' CD -'**~** 

### 0 

### C) 0\ C) tn .,D

....... I-' 

### Cl•• 

## ENV 93 1201 : MAGNUSSON 

## &KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG TAD l E 3 

## INDIVIDUAL SKIN REACTIONS IN TEST ANIMALS AT CllALLENGE: DNCO STUDY CIiAlLENGE CONCENTRATIONS: 0.75% AND 0.5% w/v VEIlICLE: ADSOLUTE ETIlANOl Skin Reactions (Hours After Removal of Dressing) 24 hours 48 hours Animal Number 0.5% 0.75% 0.5% 0.75% Er Oe Other Er Oe Other Er Oe Other Er Oe Other

N 31 

### 2 

## 0 STA 2 

## 0 STA 2 

## 0 STA 2 

## 0 STA I (Xl 32 

### 2 1 

## STA 2 1 

## STA 1 1 

## STA 2 1 

## STA I 33 

### 2 1 

## STA 2 2 

## STA 2 1 

## STA 2 1 

## STA 34 2 

### 1 

## STA ?e ?Od StWeSTA 1 

## 0 STA ?e ?Od StWeSTA 35 2 1 

## STA 2 

### 2 

## STA 2 1 

## STA 2 

### 1 

## STA 36 

### 2 1 

## STA 2 1 

## STA 2 1 

## STA 2 

### 1 

## STA 37 

### 2 2 

## STA 2 2 

## STA 2 

### 1 

## STA 2 

### 1 

## STA ;;.a 

# " 30 2 

### 2 

## STASs 2 2 

## STASs 2 I 

## STASs 2 

## 0 STASs 

## E ,." 39 

### 2 2 

## STA 2 2 

## STASs 1 1 

## STA 2 1 

## STASs n -I

­ ;z: c:: 

### 40 2 1 

## STA 2 2 

## STA 1 

## 0 STA 2 2 :I: OJ ,." :;a 0 0 0'\ 

## STA·6 

# .g lT1 

### CJ**~**
......... jooo& 

## .r:::.**~**

### ENV 93 1201 : HAGNUSSON 

## &KLIGMAN MAXIHISATION STUDY IN TilE GUINEA PIG T A [} L E 4 

### INDIVIDUAL SKIN REACTIONS IN CONTROL ANIMALS AT CIIALlENGE: DNCD STUDY CIIAttl:NG[ CONC[NTIU\TIONS: 0.75% ANn 0.5% w/v 

### VEIlICLE.: ADSOlUTE. ETHANOL Skin Reactions (Hours After Removal of Dressing) 24 hours 

# . 48 hours Animal Number 0.5% 0.75% 0.5% 0.75% 

### Er Oe Other Er Oe Other Er Oe Other Er Oe Other I N 1.0 I 

### 41 0 0 

### STA 0 0 

### STA 0 0 

### STA 0 0 

### STA 42 0 0 

### STA 0 0 

### STA 0 0 

### STA 0 0 

### STA 43 0 

### STA 0 0 

### STA 1 0 

### STA 0 0 

### STA 44 0 

### STA 1 0 

### STA 1 0 

### STA 1 0 

### STA 

# " 45 0 0 

### STA 0 0 

### STA 0 0 

### STA 0 0 

### STA 

# .... ' ;0**~** m n -1 z c:: 

### 0 co**~.'** , m 

### 0 

# 1'1 3: /'" ... :;0 0 en 

### C:J ,t-, 01**!~** t:," 

## f (,,- ""­ 

## r,) 

# #'

### -30­ 

### PROJECT 

### NUMBER: 654/1



## A P .p 

## END ICE S



# 000043

### -31­ 

## PROJECT NUMBER: 654/1

KEY T 0



### Er :: erythema Oe :: oedema

- :: no other skin reactions Ss 

## =small scattered scabs St :: hardened dark-coloured scab STA :: yellow-coloured staining not affecting evaluation of skin responses ?e 

## = evaluation of erythema precluded by other skin reactions ?s :: yellow-coloured staining prevented accurate evaluation of erythema Cr = 

### superficial cracking of skin Bs :: bleeding caused by animal scratching intradermal induction site 

### Bd :: dried blood



## D= 

### desquamation


# 000044

### [NV 93 1201 : HAnmSSON 

### &KLIGMAN MAXIMISATION STUDY IN TIlE GUINEA PIG 

## A P P f 

## N 0 I X I INTlMOfRHAl SIGHTING TEST - SUMMARY 

### OF RESULTS: ENV 

### 93 1201 STUDY 

### VEIIICl£: nISTILlED WATER Concentration of Anima1 Time of Grade of Eviden'ce Evidence of Test Material Erythema at . of local Systemic Identification Observation 

## (% w/v) Injection SHe Necrosis Toxicity 24 

### hours 2 - 4 

### Necrosis None 48 

### hours 3 - 4 

### Eschar None A 

### 72 

### hours 3 - 4 

### Eschar None 7 

### days 2 - 4 

### Eschar None I w N I 

### 24 

### hours 4 

### Eschar None 48 

### hours 4 

### Eschar None B 5 

### 72 hours 4 

### Eschar None

7 days 4 

### Eschar None 24 

### hours 2 - 3 

### None None E 

### 48 

### hours 0-1 

### None None O. I '"C 

### 72 

### hours 0 

### None None ;;0



### 7 

### days 0 

### None None

# ~ E 

# I !'1"\ n -1 

### 24 

### hours 2 - 4 

### Necrosis :z c::: 

### None 0 

### x 

### 48 

### hours 2 - 4 

### Necrosis None F 

### 0.5 c:l !'1"\ 0 

### 72 

### hours 2 - 4 

### Necrosis None' J :;0 

### C) 7 

### days 4 

### Eschar None 

# .. CO) 0'1 U1 

# .r:.. l**~** ........ 

### G"lThe concentration of the test material selected for the intradermal induction stage of the main study waif::­

....... 

### 0.1% w/v in distilled water [r

## fNV 93 1201 : MAGNUSSON &

## KLIGMAN MAXIMISATION STUDY IN TUE GUINEA PIG A P P f 

## N D I X I I INTRADERMAL SIGIITING TEST

- SUMMARY OF RESULTS: ONes STUDY VftlIClf: ARACIIIS 

## OIL D.P. Concentration of Anima1 Time of Grade of Evidence Evidence of Test Material Erythema at of Local Systemic Identification Observation Injecti

### on Site Necrosis Toxicity (% 

### w/v) 24 hours 2 

### None None 40 hours 2 

### None None C 

## 0.1 72 hours 2 

### None None

.... I 

### . 7 days - 2 

### None None w w I 

### 24 hours 2 - 3 

### None None 48 hours 3 

### None None 0 

## 0.5 72 hours 3 - 

### 4 Eschar/focal eschar None

7 days 2 - 

### 4 Eschar/focal eschar None 

# I -c 

### 24 hours 2 - 

### 4 Necrosis None ;0 

### G 

### 48 hours 1 2 - 

### 4 Eschar None 

## E rrt 

### 72 hours 2 - 

### 4 Eschar None n



### 7 days 1 - 4 Eschar None --4 :z c:: 

### :::r. 

### The cOllcentralion of the test material selected for the intradermal induction stage of the main study was**~~_** rrt C) ;0 

# .. 0 

### 0.1% w/v ill 

### arachis ojl 

### B.P. C) 01 01 

# :,1 OJ C)**~-.** j .:.. r, . ""­ 

# .r::a ..... 

### C0

## [NV 93 1201 : MAGNUSSON &

## KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG A P PEN 0 J 

## X I I I TOPICAL SIGIITING TEST FOR INDUCTION APPLICATION (48-1I0UR EXPOSURE)

- INDIVIDUAL SKIN REACTIONS: ENV 93 1201 

## STUDY V[lIIClE: 

## IHSTILLED UATEn Skin Reactions Concentration of Anilllal (liours After Removal of Patches) Test Material ldentificatlon 1 

### 24 

## 48 (% v/v) Er Oe Other Er Oe Other Er Oe Other 10 II 5 

### 2 1

;' 

### t I W .;.. I

'"'0 

# E ,." n :z c: :J: D:I ,." ;;0 

# ..**~** U'1 .;..



### ....... 

# - 10

5 

### 2 1 CJ 

## c=) c=) 

# o P 50 25 1 0 0 0 0 0 0 0 

### 1 0 

### 1 0 

### 1 0 0 0 

### Animal 2 1 

### 2 1 0 0 0 0 0 0 0 0 0 0 0 0 

### 2 1 2 

### 1 0 0 0 0 0 0 0 0 0 0 0 0 

### killed immediately after dressing removal due to severe necrosis -J () 50 

### Animal killed immediately after dressing removal 25 

### due to severe necrosis The 

### concentration of the testlnaterial selected for the main study topical induction was 10% 

### v/v 1n distilled water -i

### [NV 93 1201 : 

### MAGNUSSON 

## &KLIGMAN MAXIMISATION STUDY IN TItE GUINEA PIG

A P PEN 0 I X I V TOPICAL SIGIITING TEST FOR INDUCTION APPLICATION (48-1I0UR EXPOSURE)

- INDIVIDUAL SKIN REACTIONS: ONCB STUDY VEIHCLE: ABSOLUTE ETlIANOl Skin Reactions Concentration of Animal

. (Hours After Removal of Patches) Test Mater;a

### 1 Identification ('Yo 

## v/v) 1 24 48 

### Er Oe Other Er Oe Other Er Oe Other

I 

### 3 2 2 

### STA 2 2 

### STA 1e 2 

### STA W l) 

### 1.5 2 1 

### STA 2 

### 1 

### STA 1 1 

### STA en 

### 0.75 2 

### 0 STA 1 

### 0 STA 0 0 STA I 

### 0.3 1 

### 0 STA 0 0 STA 0 0 STA 3 

### 2 2 

### STA 1e 2 

### STACr 1e 2 

### STASt STA STA
in absolute K 

### 1.5 2 

### 1 

### STA 2 2 

### STA 1 1 

### STA 0.75 2 

### 1 

### STA 1 

### 0 STA 0 0 ", 

### 0.3 2 

### 0 STA 0 0 STA 0 0 

# ,"**~ ~**  

### The concentral1on of the test material selected for the main study topical induction was 0.75% w/v**~** z 

### ethanol**~** CD

### ~ 

### o 

# o 01 01 

## o .j:Io

....... 

## o I-' ..t::lI (X)

### ENV 93 1201 : MAGNUSSON &

### KLIGMAN MAXIHISATION STUDY IN 

### TilE GUINEA PIG API}ENOIX V TOPICAL SIGIITING TEST FOR CHALLENGE APPLICATION (24-IIOUR EXPOSURE)

- INDIVIDUAL SKIN REACTIONS: ENV 93 1201 STUDY V[JtICL(: OISTlll.EO WATER Skin Reactions Concentratlon of Anlmal**(H~urs** 

### After Removal of Patches) Test Material Identification (% v/v) 1 

### 24 48 

### Er De Other Er De Other Er De Other
L 10 5 0 0 0 0 1 0 0 0 1 0 0 0 I W 0'1 I 

### 2 0 0 0 0 1 0 1 0 0 0 0 0 0 10 

### 1 0 1 0 0 0**~1** 5 0 0 0 0 0 0



### 2 0 0 0 0 0 0 1 0 0 0 0 0 0 ""0 

# E m

### ~ Tile 

### concentrations of the test material selected for the main study topical challenge were**5~** 

### and 1% v/v in :z 

## i distilled water a:'l m :;c;l 

# .. o 0'1 

## o**.~!** 01**~** 

# o I-' 

# - a .r.::. ill

### fNV 93 1201 MAGNUSSON & 

### KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG A P PEN 0 I X V I TOPICAL SIGIITING TEST FOR CIiAllENGE APPLICATION (24-1I0UR EXPOSURE)

- INDIVIDUAL SKIN REACTIONS: DNCO STUDY VEIIICLE: 

### ABSOLUTE ETIIANOL Skin Reactions Concentration of Animal (Hours After Removal of Patches) Test Material Identification w I 

### 1 24 

### 48 (% 

### v/v) 

### Er Oe Other Er Oe Other Er Oe Other I 

### 0.15 0 0 

### STA 0 0 

### STA 0 N 

### 0.50 0 0 

### STA 0 0 0 0.25 0 0 

### STA 0 0 

### STA 0 0.10 0 0 

### STA 0 0 0

'"'C ;::0 

### 0.15 0 0 

### STA 0 

### 0 

### STA 0 0 0 c..... 

## o 0.50 0 0 

### STA 0 0 0 0 f"T1 n 

### 0.25 0 0 

### STA 0 0 0 0 -4 

### 0.10 0 0 

### STA 0 0 

### 0 0 z: c::: 3: co f"T1 :;a 

# .. o o 01 

# '" .$00 

## o
........ 

## .....**C~)** 

## Ul CJ

### [NV 93 1201 : MAGNUSSON & 

### KLIGMAN MAXIMISATION STUDY IN TIlE GUINEA PIG 

### A P P [ N D I X V I TOPICAL SIGIITING TEST fOR CttAllENGE APPLICATION 

### (contd) .'\ 

### (24-IIOUn EXPOSURE)

- INDIVIDUAL SKIN REACTIONS: ONCS STUDY V[IIICLE: ABSOLUTE ETIIANOL. 

### Skin Reactions Concentration of Anima1



### .(llours After Removal of Patches) Test Material Identi 

### fication (% 

### v/v) 1 

### 24 

### 48 

### Er Oe Other Er Oe Other Er Oe Other I W I 

### 2 

### 1s 1 2 1 

### STA 2 1 

### STA 00 

### p 

### l.5 

### 1s 1 

### 2 1 

### STA 1 1 

### STA 1 

### 15 0 

### 2 

### 1 

### STA 1 1 

### STA 0.75 

### 1s 0 0 0 

### STA 0 0 

### STA 2 

### 1s 1 

### 2 1 

### STA 1 1 Q 

### 1.5 

### 1s 1 

### 2 

### 1 

### STA 1 0 

### 1 

### 1s 0 

### 2 0 

### STA 1 1 0.75 

### 1s 0 0 0 

### STA 0 0


### The 

### STASs STA STA .-., , 

### STA C i-. """

.

, -,.........**:~~>**

## ...

t. ..**~ ~**  -l 2: c:: X OJ

### ~ 

# .. Ol 01 .+:0-

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

### concentrations of the 

### test material selected for the main study topical challenge were 0.75% 

### and 0.5% 

### w/v**(i'~** 

### absolute ethanol



## o 

# o C:> 

## o 01 ........:L

# 'I

fNV 93 1201 : MAGNUSSON 

## &KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG A P 

## PEN 0 I X V I I INTRADERMAL INDUCTION

- INDIVIDUAL SKIN REACTIONS IN TEST ANIMALS: ENV 93 120) STUDY INTRADERMAL INDUCTION CONCENTRATION: 0.1% w/v 

## VEIIIelE: D ISTIllED WATER Skin Reactions at Observation Time: Animal Number 24 hours 48 hours left Side Right Side left Side Right Side
1 

### 2 

### 2 

### 1 1 

### 2 2 2 

### 1 1 3 2 

### 2 1 1 W I 1.0 

### 4 1 1 1 1 I 

### 5 2 

### 2 1 1 6 

### 2 2 

### 1 1 

### 7 1 

### 2 

### 1 1 8 2 

### 2 1 2 9 1 

### 1 

### 1 1 10 

### 2 

### 2 1 0 

### 11 1 1 

### 1 0 12 

### 2 2 1 1 } -0

. ;;0 

### 13 1 1 1 1 

## E 14 1 1 

### 1 2 J'TI 15 

### 2 

### 2 0 1 -i 

### n

, 16 

### 2 2 0 0 :z c:: 

### 17 2 2 2 

### 1 :I: 

### 0 18 

### 2 

### 1 0 

### 1 J'TI :;;d 

### 0 

### 19 2 2 2 

### 2 •• C) 

### 20 2 2 0 

### 1 

# .. CD <:) 0"1 U1 {:ft-
....... 

## ..... N

# .,

[NV 93 1201 : MAGNUSSON 

## &KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG A P P [ N 0 I X V I I I 

## INTRADERMAL INDUCTION

- INDIVIDUAL SKIN REACTIONS

. IN CONTROL ANIMALS: [NV 93 1201 STUDY VEIIICLE: lHSTIlL[1l WATEn Skin Reactions at Observation Time: Animal Number 24 hours 48 hours Left Side Right Side. Left Side Right Side 21 0 

### 0 0 0 I 

### 22 0 

### 0 0 0 0 I 

### 23 0 

### 0 0 0 

### 24 0 

### 0 0 0 25 0 

### 0 0 0 

### 26 0 

### 0 0 

### 27 0 

### 0 0 "'l:l 

### 28 0 

### 0 0 0 {' 'J ;;: 

# E 

# Ii 29 0 

### 0 0 0 m n 

## ( -f 

### 30 0 

### 0 0 0 '""
_ :z: c: 

### ::r: CD 

### m 

# .. C) 0\ 01 

### U1**~** 

### ....... 

### CD
......

## fNV 93 1201 : MAGNUSSON 

## &KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG A P PEN 0 I X I X INTRADERMAL INDUCTION

- INDIVIDUAL SKIN REACTIONS IN TEST ANIMALS: DNCB STUDY INTllAOERHAl INDUCTION CONCENTRATION: 0.1% w/v 

## VEtIlClE: ARACIHS OIL B.P. Animal Number Skin Reactions 24 hours at Observation Time: 48 hours Left Side Right Side left Side Right Side 31 2 2 1 0 I .Po


### ..... 32 2 

### 2 2 2 I 

### 33 1 1 1 1 

### 34 1 1 

### 1 35 

### 2 2 2 

### 2 36 

### 1 1 1 2 

### 37 2 2 2 2 

# , -c 

### 38 2 2 2 

### 2 l:::: ' ;;c 

## E "., 

### 39 1 1 2 

### 2 n {r:.::. .' /' -; 

### 40 2 2 1 1 

### r'
. z c: 3: CD "., 0**~~,** ;;c 

# .. 0 

### CJ 0'1 U1 

### 01 .Po 

# (;1

........ 

### .l:::!i 

## .....

## fNV 93 1201 MAGNUSSON 

## &KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG A P P f 

## N D I X X INTRADERMAL INDUCTION

- INDIVIDUAL SKIN REACTIONS IN CONTROL ANIMALS: DNCO STUDY VEHICLE: ARACIIIS OIL D.P. Skin Reactions at Observation Time: Animal Number 24 hours· 46 hours Left Side Right Side Left Side Right Side

I .l;Ia I 

### 41 1 

### 1 1 N 

### 42 1 1 

### 1 1 

### 43 0 1 0 

### 1 

### 44 1 1 

### 1 

### 45 1 0 U**~J** '"C

## ~ ,." n -I ::z: c: 3: rr1**~** 

### 0 

# fIJ to



# .. 0**C~** m 01 .l;Ia 

### 0-1 

# I -)
........ 

## <.n .......

### fNV 93 1201 : MAGNUSSON 

## &KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG 

### A P P f 

### N 0 I X X I TOPICAL INDUCTION

- INDIVIDUAL SKIN REACTIONS IN TEST ANIMALS: ENV 93 1201 STUDY INDUCTION CONCENTRATION: 10% v/v VEIHCLE: DISTILLED WATER Skin Reactions (Hours After Removal of Dressing) Animal Number 1 hour 24 hours Er Oe Other Er Oe Other 1 

### 2 1 

### Bs 2 

### 1 BdSt 2 

### 2 1 

### 1 1 

### OSt 3 

### 2 

### 1 1 

### 1 5tO 4 2 1 

### 1 1 StSsO I -Po 

### 5 2 

### 1 Bs ?e 1 StOBd w I 6 3 

### 2 

### ?e 1 StOBd 7 

### 2 1 

### 1 0 8 

### 2 1 Bs 1 0 9 

### 2 1 

### Bs 2 

### 1 5tOBd 10 

### 2 

### 1 

### Bs 2 

### 1 StDlJd II 3 

### 2 

### Bs ?e 1 StO 12 

### 2 1 1 

### 1 13 3 

### 2 2 

### 1 14 

### 2 1 

### 1 1 15 

### 2 1 1 0 16 

### 2 1 1 0 

### 17 

### 2 2 1 0 18 

### 2 1 

### Bs 1 

### I 

### 0 19 1 0 1 0 

### CJ 20 

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### ENV 93 1201 : MAGNUSSON &

### KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG A P 

### PEN D I X X I I TOPICAL INDUCTION

- INDIVIDUAL SKIN REACTIONS IN CONTROL ANIMALS: [NV 93 1201 STUDY VEIIIClE: DISTILLED WATER Skin Reactions (Uours After Removal of Dressing) Animal Number 1 hour 24 hours Er Oe Other 

# . Er Oe Other 21 1 1 0 0 

## t I I 

### 22 0 0 0 0 23 1 0 0 0 

### 24 0 0 0 0 

### 25 1 0 0 0 

### 26 1 0 0 0 

### 27 

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### [NV 93 1201 : MAGNUSSON 

## &KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG A P 

### PEN 0 I X X I I I TOPICAl. INDUCTION

- INDIVIDUAL SKIN REACTIONS IN TEST ANIMALS: DNCD STUDY INnUCTION CONCENTHATION: 

## o.**75~** 

### w/v VEIIICLE: ADSOLUTE ETIIANOL Skin Reactions (Hours After Removal of Dressing) Animal Number 1 hour 24 

### hours Er Oe Other Er Oe Other

..&k 31 

### 2 2 

### STA 2 2 

### STA I lit 32 

### 2 2 

### STA 2 2 

### STA I 33 3 

### 2 

### STA 2 2 

### STA 34 2 

### 2 

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### STA 35 3 

### 2 

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### STA 36 3 

### 2 

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### STA 37 3 

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### STA 38 2 

### 2 

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### STA 39 3 

### 2 

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# ,;'

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## KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG A P P [ N 0 I X X I V TOPICAL INDUCTION

- INDIVIDUAL SKIN REACTIONS IN CONTROL ANIMALS: DNC8 STUDY VEIlICLE: ABSOLUTE HIIANOL Skin Reactions (liours After Removal of Dressing) 

### Animal Number 1 

### hour 24 hours 

## Er Oe Other 

## Er Oe Other



# .,. I Q Q Q 0'1 

### 41 0 I 

### 42 

## o 0 0 Q 

### 43 

## o 0 0 Q 

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### [NV 93 1201 : MAGNUSSON & 

### KLIGMAN MAXIMISATION STUDY IN TilE GUINEA PIG A P P [ ND 

### I X X V INDIVIDUAL 80DYWEIGIITS AND 80DYWElGIlT GAINS OF TEST ANIMALS: ENV 93 1201 STUDY Bodyweight (g) Bodyweight (g) 

### Animal Number Day 0 Day 24 Increase

0 0 

### 1 2 3 

### 4 5 6 1 8 9 

### 10 11 12 13 14 15 16 

### 17 18 

### 19 20 368 402 358 371 381 398 386 404 391 364 361 318 312 409 427 371 399 407 384 367 551' 601 492 555 565 594 622 608 613 539 558 578 565 537 575 551 606 574 567 506 183 199 134 184 184 196 236 204 222 175 

### 191 200 193 128 148 180 207 167 183 139 I .$0­

........ I



# " 

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### ENV 93 1201 : MAGNUSSON & 

### KLIGMAN MAXIMISATION STUDY IN TIlE GUINEA PIG



## A P PEN D I X X V I INDIVIDUAL BODYWEIGIITS AND BOOYWEIGIIT GAINS OF CONTROL ANIMALS: ENV 93 1201 STUDY 

## Bodyweight (9) Bodywelght (g) Animal Number Day 0 Day 24 

### Increase
21 396 553 155 22 366 532 164 I 

### 23 375 581 206 I 

### 24**,~** 

### 414 596 182 25 387 555 168 26 

### 375 530 155 27 427 575 148 26 430 606 176 29 

### 415 576 161**f~** '"'C c'

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### ENV 93 

### 1201 : MAGNUSSON 

## &KLIGHAN MAXIMISATION STUDY IN TilE GUINEA PIG A P PEN D I X X V I I INDIVIOlJAl OODYWEIGIITS AND BODYWEIGIiT GAINS OF TEST ANIMALS: DNCO STUDY 

### Bodyweight (g) Oodyweight (g) Animal Number Day 0 Day 24 

### Increase 31 366 581 215 I 

### 32 385 579 194**~** 1.0 

### 33 381 569 188 I 

### 34 352 526 174 35 363 550 187 36 411 572 161 37 

### 341 509 168 38 

### 444 545 101 -0 

### 39 

### 337 

### 471 134 

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### £NV 93 1201 : MAGNUSSON 

## &KLIGMAN MAXIMISATION STUDY IN TII£ GUIN£A PIG 

## A P PEN 0 I X X V I I I INIHVIOUAL 800YW£IGlITS AND BODYWEIGIIT GAINS OF CONTROL ANIMALS: ONCB STUDY 

## Bodyweight (g) Bodyweight (g) 

## Animal Number Day 0 Day 24 

## Increase I U1 CI I 

### 41 420 

### 576 156 42 380 

### 594 214 43 

### 380 

### 549 169 

### 44 398 580 

### 182 ""C 

# .'­ 45 386 

### 570 184 

## ::.:; 

# 2 /"1"1 n --I :z c: :J: CD 

# o /"1"1 

# o 

# .. o 0'\ 

### c-:::>**~** ()) I-' 0-) ,',

# - 

# - 01

.,

### -51­ 

## PROJECT NUMBER: 654/1 

## A P PEN D I X X I X

TIIE DEPARTMENT OFHEALTH OF

### TIm GOVERNMENT OF

### TIm UNITED KINGDOM GOOD IABORATORY PRACI1CE**STATE~NTOFCOMP~CE** IN ACCORDANCE WITHDIRECTIVE 88/320 EEC LABORATORY 

### SafePharm Laboratories Limited POBox No 45 Derby DEl 2ET

DATE OF INSPECI10N 31 January 1994 A general inspection for compHance with [he Principles of Good Laboratory PraC'"oice was carried out at [he above laboratory 

### as part ofthe UK GLP Compliance Programme. At the time of the inspection no deviations were found of sufficient magnitude to affect the vaiidicy ofstudies performed at these facilities.



# _ .. n00064 D. 1:". Moori!' Direaor UK GLP Monitoring Unit

### ~- 

# .

. ,
, "1·' 

# BIOSEARCH INCORPORATED P.o. Box 8598, Philadelphia, Pennsylvania 19101
' 

### ; 

### Telephone: (215) 739-4499

It·····
..

# ··~.··'.'·:.···.····'.··. Fax: (215) 423-9210 

# :1

. . :1'·.··.···

.



# .··1



# l'**'~** 

### Project Number: 

### 93-7958A 

### SUM:MARY OF RESULTS OF PRIMARY SKIN IRRITATION STUDY Conducted for: 

### Test Article: 

### ENV931201 Primary Skin 

### Irritation: The test article, when administered as supplied, is corrosive to the skin ofalbino rabbits.


# 1 

# 1 1 t i 

# r , 

## ; 

# 000065 

# 1

### StudyTitle PrimarySkin Irritation**~** Rabbits Test Article ENV931201

Sponsor

Author George 

### E. Moore 

### Study.CompletionDate 03/21/94 PerformingLaboratory Biosearch ll1corporated 

### 3408-50 "B" Street Philadelphia, Pennsylvania 19134 BiQsearch ProjectNumber 93-7958A

Page 1 of 11 

# 000066

### Page 2 of 11 

# ,­

....,i

PAGE RESERVED FOR CONFIDENTIALITY STATEMENT



### - "



### . .J

j 

# J J 

# -J



# J .

.-r 

# J '1 

## .J 

# ] 

# 'J 000067 

# "] BIOSEARCH, L"lCORPORATED· P.O. BOX 8598

- PHIUDELPHIA, PE'J:-JSYLVANlA 19101 (215) 739-4499

### , Page 3 of 11 

# BIOSEARCH INCORPORATED P.o. Box 8598, Philadelphia, Pennsylvania 19101 Telephone: (215) 739-4499 Fax: 

### (215) 423-9210 

### ..,..." .J:;) _, 

### GOOD LABORATORY PRACTICE COMPLIANCE

### .~T~~f~NT**,,~ }~/!**  , "'4,**I';~** .', r....... rv .. 

### Project Number: 93-7958A ''<", ..!,**J.,...,.t;~** 

## . "', 

### ,""
. Sponsor:

- ....

; i 

### TestArticle: ENV931201 i .", j ., j 

### Test: Primary Skin Irritation

- Rabbits (fSCA)

...., ProtocolNumber: PSI-B06 (Rev. 02193) 

# .J

. 

# ...,**~J** 

### This study was carried out in 

### compliance with the 40 CFR 792 and OECD Regulations for "Good LaboratoryPractice". 

# ] ] ]
3-;;2. J-

## 9'-1

# ~reC:;.s.~ Date StudyDirectorlLaboratory Director

01110/94 93 

### 040 7958 116 

### Form QA-700A 

# 000068

### Project Number: Sponsor: TestArticle: Test:

, 

### Protocol Number:



# ... StudyInitiated: StudyAudit:


# ]

''''

I i -wi

-,! J



# .!**1jlU~bAl,(L.t1 ll~LUl'(YUl{Al**  

## hV P.O. Box 8598, Philadelphia, Pennsylvania 19101 Telephone: (215) 739-4499 Fax: (215) 423-9210 QUALITY ASSURANCE STATEMENT



### ENV931201 Primary Skin Irritation

- Rabbits (TSCA) Sponsor PSI-806 (Rev. 02193) 

### SignedProtocol: 12122193 01102194 

### Study Completed: 03/21194 

### Critical phases for this studyor this type ofstudywereinspected on: Individual StudyAudits: Shaving TestArticle Admin. Final Report 01/03/94 01104194 03121/94 

### StudyTypeAudits: Test Article Preparation 01125194 

### Any findings protocol were of deviation from standard operating procedure immediately reported to the Study Director and or to management. The QAU Study Inspection Report to the Study Director and management was submittedby 02111/94
.. 

### The results shown in 

### this 

### report accurately reflectthe rawdata records.

02128/94930407958 116 

### Form**QA~700B** 

# 000069

### Project Number:**93~7958A** 

### P,age 5 of 11 TABLE OF CONTENTS
Title Page
................................................................................................... l' 

### ConfidentialityStatement
.............................................................................. 2 Good Laboratory Practice Compliance Statement
.............................................. 3 "I



### , i 

### QualityAssurance Statement
......................................................................... 4 

### Table ofContents
.......................................................................................... 5
......., 

# ,J Objective ofTest
.....................................................,.......................... :......... 6 Reference
.................................................................................................... 6

# ~] Justification for Test System
.......................................................................... 6 

# J Animals
...................................................................................................... 

### 6 Husbandry Conditions
................................................................................. 7 

# J Procedure
......................................................:............................................ 7 

### Data Retention
............................................................................................. 7 

# ] 

# ] TestArticle Characterization
.......................................................................... 7 

### Results
....................................................................................................... 8 Conclusion
.................................................................................................. 8 

# J Signatures
................................................................................................... 8 Table 1**~** 

### Primary Skin Irritation Scores
........................................................... 9 

# J ] AppendixA - Draize Scale for Scoring Skin Reactions
..................................... 10 Appendix B**~** 

### Protocol Deviation
................................................................... 11 

# ] ] 

# J J ] 

# 000070

BIOSEARCH, L"ICORPORATED· P.O. BOX 8598· PHILADELPHIA, PEJ'<NSYLVANLA 19101 (215) 739-1499u 

# BIOSEARCH INCORPORATED

Project Number: frotocol Number: TestingFacility: Sponsor:
TestArticle: Test Article Description: Test Article Received: In-Life Study Initiated: Objective ofTest: Reference: Justification for Test System:
Animals: P.O. Box 8598, Philadelphia, Pennsylvania 19101 Telephone: (215) 739-4499 Fax: (215) 423-9210 93-7958A PSI-806 (Rev. 02/93) Biosearch Incorporated 

### 3408-50 liB" Street Philadelphia, PA 19134

ENV931201 Amberliquid 12123/93 

### In-Life 01/04194 

### StudyCompleted: 01106/94 

### PrimarySkin Irritation

- Rabbits To determine the degree ofirritation, if

### any, which the test article mayproduce when appliedto the skin ofalbino rabbits. The methods employed in 

### this studywere similar to those described in 40 CPR Part798, EPA Health Effects Testing Guidelines

- Subpart E - Specific OrganffissueToxicity, September 1985. The laboratory albino rabbit was the system of choice since it is required in the federal guideline for this type of study and has a longhistory ofuse in dermal irritation studies. The New Zealand White rabbit was used for this study. The animals were obtained from Davidson Mill Farm, Jamesburg, 

### NT 08831 (U.S.D.A. License #22-A-7). The animals were individually housed and maintained in accordance with standards set forth in the Guide for the Care and Use ofLaboratory Animals (NIH Publication No. 86-23). The rabbits were acclimated to the laboratory for at least 5 days prior to the initiation of the study. The animals were individually identified by an ear tag and each cage was identified with a cage card, displaying the project number, animal number, sex, date initiated and responsible technician's initials. 

# 000071
BfOSEA.RCH. u'ICORPORATED
. P.O. BOX 8598·PHILADELPHJA. PE"INSYLVANIA 19101 (215) 7394499

### Project Number: 93-7958A Page 7 of 11

Wayne 15% Rabbit Ration was provided as a maintenance diet Based on our current . ENV931201 Primary Skin Irritation

- Rabbits Hllsbandry


## .. Conditions: Temperature: 21.2°C

- 21.8°C Relative Humidity,%: 

### 41

- 47 Light: 12-hour light/dark cycle Diet: and tap water was provided ad 

### libitum. knowledge, no contaminants are known to be in this 

### diet or water which might be expected to interfere with the objectives 

## i ofthe study. 1 

### Caging: Stainless steel elevated wiremesh flooring, 

### 1rabbit/cage. Procedure: A group of six rabbits was clipped over a wide area on their backs approximately 24 hours prior to application. Animals

### withhealthyintact skin were used. The testarticle was applied dermally as supplied. A 

### 0.5 

### m1 portion of the test article was applied dermally to the test site (approximately 

### 6 crn2 ). An 

### adjacent area of untreated skin served as a control for the test. Gauze patches on non-irritating tape were placed over the treated and control areas. A suitable semi-occlusive dressing (gauze) was wrapped around the trunk 

### of the animal and secured with elastic tape. The wrapping was removed at the end of the four-hour skin contact period and the test sites were wipedwith deionized water. The treated areas were then examined for signs of erythema and edema, using the Draize method of scoring (Appendix A), within 30 

### to 60 

### minutes after patchremoval. Readings were also made after 24 and

### 48 hours. Due to the necrosis produced by the test article, the study was terminated after the 48 hour readings and the animals were euthanized for humanereasons. . 'Data Retention: The ra:w, da!a,aIlda 

### copyof this report are filed in the archives of Biosearch, Incorporated, Philadelphia, Pennsylvania. Test Article Characterization: The Sponsor was responsible for test article characterization and stability determination. Details regarding the identity, strength, stability, purity, and composition of the test article are in 

### the possession of the Sponsor. Information supplied to Biosearch, Incorporated regarding characterization of the test article is on file in the archives of Biosearch, Incorporated, Philadelphia, Pennsylvania. 

# 000072
BIOSEARCH. lNCORPORATED· po, BOX 8598· PHILADELPHIA. PEN'NSYLVANIA 19101 (215) 739-4499

### Project Number: 93-7958A Page 8 of 11 ENV931201



### .... Primary Skin Irritation

- Rabbits



### ....

l 

### Results: See Table 1 . CQnclusion: The test article, when administered as supplied, is corrosive to fhe

### ,~idn ofalbino rabbits.
..**'''~,** 

# ] Signatures: Submitted by Biosearch, Incorporated: 

# ] ]

# ~kC?/~ 3h.)**J9'~** G;orgeRoore, B.S.

## ~ Laboratory Director/StudyDirector 

# ] Reviewedf Auditedby: 

# J ] 

# J

# pa~ Manager, QualityAssurance Approved by: 

# ]

# ?#;If~ es ong, Sc.D. Date 

### .I ].' 

### Vice Pr sident, Laboratory Operations

... & 

### RegulatoryAffairs



# 000073
B[OSEARCH, INCORPORATED

- P.O. BOX 8598·PHlL>\DELPHlA, PENNSYLVANIA 19101 (215)739-4499.

### ..., Project Number: 93-7958A Page 9 of 11

- ENV931201, applied dermally as supplied.


## , 

# - Primary Skin Irritation

- Rabbits TABLE 1 

### Primary Skin Irritation Scores 

## -,

i 

### Rabbit Number 

### 1 

### 2 3 .4 

### 5 

### .6 

## 1 .J 

### 5..ex:t M 

## M M E E 

### E Initial 

# ] BodyWeight (kg): 2.71 2.49 2.38
. 2.63 2.57 2.85 

### EI)!thema and. Reading 

## 1 J 

### Eschar Formation (Hours) IntactSkin "'1 N N N N N N 

# ] IntactSkin 24 

### N N N N N N IntactSkin 48 

### N N N N N N 

# ] IntactSkin ""'72 

# 1
Edema Formation Intact Skin "'1 

## o 2 

### 3 

### 3 2 

### 3 IntactSkin 24 

### 3 3 3 

### 3 4 

### 4 IntactSkin 48 3 

### 3 3 3 4 4 

### Intact Skin
Primary Irritation Index: Corrosive 

### '" Readings were made 30 to 60 ininutes afterpatch removal, after a 4-hour exposure. Due to the necrosis of the test sites, the animals were euthanized for humane reasons after the 48-hour readings. 

### t M 

## = Male F

## = Female 

# 000074
BIOSEARCH. INCORPORATED

- P.O. BOX 8598

- PHILADELPP.IA
. PENNSYLVA,"llA 19101 (215) 739-1499

### Project Number: 93-7958A Appendix A - Page 10 of 11 ( - ENV931201 Primary Skin Irritation

- Rabbits



### ,.... 

# l Draize Scale for Scoring Skin Reactions» 

# l

..... Erythema and Eschar Formation Value No erythema 

## o Very slighterythema (barely perceptible) 1 

### Well defined erythema 2 

### Moderate to severe erythema 

### 3 Severe erythema (beetredness) to slight eschar formation 

# J (injuryin depth) 4
Edema Formation Value No edema 0 Very slight edema (barely perceptible) 1 Slight edema (edges ofarea well defined

### by definite raising) 2 Moderate edema (raised approximately 1 mm) 3 

### ;:;1,', Severe edema (raised more than 1 mm and extendingbeyond the 

### .,J area ofexposure) 4 

# :J**+"'~** 

# '-J:

i, , 

# J ]

,.. Association of Food and Drug Officials of the United States, APPRAISAL OF THE 

# J SAFETY OF o-rEMICALS IN FOODS, DRUGS AND COSNIETICS, p. 48. Bureau ofFoods and Drugs, Texas State Department ofHealth, Austin, Texas, 1959. 

# ] 

# J 00007b B[OSEARCH. J.NCORPORATED· P,O, BOX 3598 PHILADELPHIA. PENNSYLVANIA 19101 (215) 739-4499Appendix B- Page 11 of 11

ProjectNumber: Protocol Number: Sponsor Test Article: Test: GLP Compliance: Deviation:

Submittedby:

Approvedby:

Acknowledgedby: 

# BIOSEARCH INCORPORATE.D P.O. Box 8598, Philadelphia, Pennsylvania 19101 Telephone: (215) 739-4499 Fax: (215) 423-9210 DEVIATION FROM PROTOCOL #1 93-7958A PSI-806 (Rev. 02/93) ENV931201 Primary SkinIrritation

- Rabbits (TSCA) 40 CFR 792 OECD Due to the necrosis produced by the test article, the study was terminated ·after the 48-hour readings and the' animals were euthanized for humane reasons.
This deviation is notthoughtto affect the integrityofthis study.

## ;11:1.t/q~ Date LaboratoryDirector/StudyDirector 

# J/I/r Ct () Date Manager, QualityAssurance 

# Jle5**E~g:s;D.** Vice President, Laboratory Operatios & Regulatory Affairs
Sponsor RepresentL
. Feb 2, 1994 93 040 7958 116 Form QA-500A 

# 000076:

# ... Studv , 

### Title Acute Oral Toxicity, LD
- Rats 50


### TestArticle ENV931201



### Sponsor 

# J 

# 1 ;] Author David Gabriel 

# j Study Completion Date 

# 1] 05112194 

# J I:erforrningLaboratory 

# J Biosearch Incorporated 3408-50 "B" 

### Street Philadelphia, Pennsylvania 19134 

# -1

.:J -Jl!III!I



## J ! 

### Biosearch Project Number 93-7958A



### Page 1 of28 

# 000078

### Page 2 of 28

PAGE RESERVED FOR CONFIDENTIALn-V STATEMENT



# 000079

### Page 3 of 28 

# BIOSEARCH INCORPORATED P.o. Box 8598, Philadelphia, Pennsylvania 19101 

### Telephone:(215) 739-4499 Fax: (215) 423-9210
GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT Project 

### Number: 93-7958A

,

# . Sponsor: Test Article: 

### ENV931201
Test: 

### Acute Oral Toxicity, LD
- Rats (TSCA) so 

### Protocol Number: AOT-800 (Rev. 02193)
This study was carried out in 

### compliance with the 40 CFR 792 and OECO Regulations for "Good Laboratory Practice".

i.-_,4) 

# D-=: < 

# bC
..**~ ~~**  5'-12-

# '1Lf David Gabriel, B. 

### A. Date StudyDirector

. ....1
-, 

## J

; 

# 000080 03/28/94 93 

### 040 7958 1485 Form 

### QA-700A

### 12122193 05112194

### .t)lU~hAl{LH 

# lNCORPORATED P.O. Box 8598, Philadelphia, Pennsylvania 19101 Telephone: (215) 739-4499 Fax: (215) 423-9210 QUALITY ASSURANCE STATEMENT Project Number: 93-7958A Sponsor: Test Article: ENv931201 Acute Oral Toxicity, LD50

- Rats (TSCA) Test: Sponsor Protocol Number: AOT-800 (Rev. 02193) 

### Signed Protocol: Study Initiated: 12129/93 

### Study Completed: Study Audit: Critical phases for this studywere inspected on: Individual Study Audits: Dose Calculation 02110/94 

### Test Article Admin. 02110/94 

### Initial Weights 02110194 

### Euthanasia 03/03/94 

### Necropsy/Gross Path. 03/03/94 Necropsy/Gross Path. 03/08/94 

### Observations 03121/94 

### Final Report 05113/94 

### Any findings of deviation from standard operating procedure or protocol were immediately reported to the Study Director and to management. The QAU Study Inspection Report to the Study Director and management was submitted by04108/94. The results shown in this report accurately reflect the raw data records.

05/13/94 

### 93 0407958 1485 Form QA-700B 

# 000081



# ..

### . Project Number: 93-7958A rage 5 of28 TABLE OF CONTENTS

... .,"''' Title Page
......................................................................................**~.:.:**:'
..

## j7)';';;., 1 Confidentiality Statement
....................................................................... :.:::::

### ·2:, 

# .'
..**-/./:.>~** Good LaboratoryPractice Compliance Statement
.............................................. 

### 3 • Quality Assurance Statement
.......................................................................... 4 Table of Contents
......................................................................................... 5 Objective ofTest
.......................................................................................... 

### 6



## .... Reference
..................................................................................................... 6

, , Justification for Test System
.......................................................................... 6 Animals
...................................................................................................... 

### 6 Husbandry Conditions
................................................................................. 7 

# .,**~** Procedure
................................................................................................... 7 Data Retention
..............................:............................................................. 8 Test Article Characterization
.......................................................................... 8 Results
....................................................................................................... 8 Conclusion
.......................................................................:.......................... 8 Signatures
................................................................................................... 9 Table 1 - Mortality & Average Body Weights

- Males
....................................... 10 Table 2 - Observations

- Males
...................................................................... 11

.. j



# - Table 3 - Gross Pathology

- Males
................................................................. 13**~** Table 4 - Mortality &Average Body Weights

- Females
.................................... 14 Table 5 - Observations

- Females
................................................................. 15 Table 6 - Gross Pathology

- Females'
.............................................................. 17 Table 7 - Individual Body Weights

- Males
..................................................... 19 Table 8 Individual Body Weights

- Females
.................................................. 23 Appendix A - Deviation from Protocol
.......................................................... 28

# 000082

...., BlOSEARCH. INCORPORATED· PO. sox 8598

- PHIL'\DElPHIA. P!::''-.'NSYLVAMA 19101 • (215) 739--499

### Project Number: Protocol Number: Jesting Facility Sponsor:

.-,

# .• Test Article: Jest Article Description: Jest Article Received: In-Life StudyInitiated: 

### Ie.s.:t: 

# J Objective ofTest: Reference:
Justification for 

# J Test System: 

# J Animals: 

# ] 

# J ] BIOSEARCH INCORPORATED P.O. Box 8598, Philadelphia, Pennsylvania 19101 Telephone: (215) 739-4499 Fax: (215) 423-9210



### 93-7958A AOT-800 (Rev. 02193) 

### Biosearch Incorporated 3408-50 

### "B" Street Philadeluhia, PA 19134 J.



### ENV931201 Amber liquid 12123/93 In-Life 01128/94 

### Study Completed: 03/23/94 

### Acute Oral Toxicity, LD50

- 

### Rats To determine the acute oral LD ofthe test article in rats. 50 

### The methods employed in the testing of this test article are similar to those described in 40 

### CFR Part 798, EPA Health Effects Testing Guidelines

- Subpart B - General Toxicity Testing, September 1985. The laboratory rat was the system of choice since it is required in the federal guideline for this type of study and has a long history of use in acute oral toxicity studies. The outbred Sprague-Dawley rat, weighing 200 

### to 300 

### grams, was used for this study. The animals were obtained from Buckshire Corp., Perkasie, PA 18944 (U.S.D.A. License #23-BL).


# 000083 I3[OSEARCH. [NCORPORATED· ?o. GOX 3593· PH[L-'.OELPH['\, PENNSYLVANfA 19101
. (215) 739-+199

### t:NV931201 Acute OralToxicity, LD50 

### - Rats
'j L i**,~,** 

### Animals: The animals were housed and maintained in accordance with standards' ,".,;­ (continued) set forth in the Guide for the Care and Use of Laboratory Animals (NIH 

### Publication No. 86-23). The rats were acclimated to the laboratory for at least 5 days prior to the initiation of the study. Animals were individually identified by ear punch. Each cage was identified with a cage card, displaying the number of animals, sex, project number, date administered and responsible technician's initials. Husbandry Temperature: 20.5°C

- 24.0°C Conditions: Relative Humidity, %: 

### 32

- 60 Light: 12-hour light/dark cycle Diet: Teklad Rodent Diet (W) 

### 8604 and tap water were provided 

### ad libitum. Based on our current knowledge, no contaminants are known to be in this diet or water which might be expected to interfere with the objectives ofthe study. 

## J! 

### Caging: Stainless steel elevated wire mesh flooring,S rats/cage by sex. _..J '" 

### Procedure: The test article was administered as supplied. 

# J A group of 85 (35 male & 

### 50 female) albino rats was used in this study. The animals were deprived of food but not water overnight prior to 

# ] dosing. Each animal was weighed and dosed by direct administration of the test article into the stomach by gavage. 

# J The following dose levels were administered: Males: Females: 

# ] 2.00 g/kg 2.00 glkg LOO 

### glkg* 1.00 glkg 0.944 glkg 0.794 glkg* 

# ] 0.891 glkg* 0.631 glkg* 0.794 glkg* 0.562 g/kg 0.631 glkg 0.50 glkg* 

# J 0.50 

### glkg 0.473 glkg 

### 0.447 glkg 0.398 g/kg* 

# J 0.355 glkg * 

### Levels used to calculate LD. and 95% confidence limits employing the Thompson moving averages and interpolation**-~ethod.** 

### The fourth level of 1.12 g/kg used to calculate the LDso 

### for the males was inferred. 

# 000084
3ICSEARCH. INCORPORATED. P,O. [lOX 8598· PH!L'.DELPI-'JA. PE.NNSYLVANIA \9101 (215) 739-4499


# .........**--------------------~**l ­ 

### ENV931201 Acute Oral Toxicity, LDso
- Rats Procedure: (continued) Following administration, the animals were allowed food and water 

### ilii libitum for the 14-day observation period during which time, the rats were observed for signs of toxicity and mortality. Animals were observed frequently on the day of dosing, and twice per day (morning and afternoon) on weekdays. On weekends and holidays, the animals were observed once per day. Individual weights were recorded on the day ofdosing, weekly thereafter, and prior to sacrifice. The animals were euthanized with carbon dioxide at the conclusion of the observation period. Gross necropsies were performed on all animals. The LD and its 95% confidence interval were calculated employing the Thompi8n, W.R., Use of moving averages and interpolation, to estimate median effective dose, Bacteriological Rev. 11: 115-145, 1947. Data Retention: The raw data and a copy of this report are filed in the archives of Biosearch, Incorporated, Philadelphia, Pennsylvania. Tes", Article Characterization: The Sponsor was responsible for·test article characterization and stability determination. Details regarding the identity, strength, stability, purityand composition of the test article are in the possession of the sponsor.. Information supplied to Biosearch, Incorporated regarding characterization of the test article is on file in the archives of Biosearch, Incorporated, Philadelphia, Pennsylvania. Results: See Tables 1 through 8. Conclusion: The test article, when administered as supplied to male albino rats, has an acute oral LD of 0.901 gfkg with 95% confidence limits from 0.852 to 0.953 g/kg. 50 

### The test article, when administered as supplied to female albino rats, has an acute oral LD of 0.562 glkg with 95% confidence limits from 0.494 to 50 

### 0.640 glkg.



# 000085
S[OSE.-\RCC1, [NCORPORATED

- P.O. [lOX 8598

- PHILADELPH[A. ?ENNSYL'/A,'1IA t9101

- (215) 739-4499- ENV931201 

### Acute OralToxicity, LD
- Rats 50 

### Signatures: Submitted by Biosearch, Incorporated:

# ~-**hc:::c-~** $'·\2-1'\ David Gabriel, B.A. Date Study Director



### 1 f I 

### Reviewed!Audited by:**~** 

## i .1
___f

# ~ Manager, QualityAssurance Approved by:

# "~**<"~/?k_**

# P.!i?s~~D.**~.** Vice President, Laboratory Operations 

# 1 .J & 

### RegulatoryAffairs 

# l

...... 

# J

J 

# 000086 BlOSEARCH, fNCORPORATED

- P.o. BOX 8598

- PHrL
....OElPHlA, ?ENNSYL,,'ANIA 19101

- (215) 739-1499.I. ""DC:: .l.V Vi ':"'U 7,J-j 7")U1-'" 

### !JroJect l'lUmuer:



### _ENV93120l, administered as supplied. Acute Oral Toxicity, LDso
- Rats TABLE 1 Mortality & 

### Average Body Weights

- Males Density: 0.93 glml Dose Levels, glkg: 2.00 1.00 0.944 0.891 0.794 0.631 0.50 Animals:· Number 5 5 5 5 5 5 

### 5

1 i


## .­**~** 

## 1 J

.., 

# J Mortality:

14-DayTotals: Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Dead Live 1 4

5 0 0 

### 0 2 ,2

1

5 

### 0 0 0 0 0 2 0 0 

### 0 0 

### 0 0 0 0 0 0 2 3 0 0 0 1 1 0 0 0 0 0 

### 0 0 0 0 0 2 3 0 0 0 0 0 0 0 0 

### 0 

### 0 0 0 0 

### 0 0 0 5 0 0 0 

### 0 0 

### 0 0 0 0 

### 0 0 0 0 0 0 0 5 

### 0 0 0 0 

### 0 

### 0 0 

## a 0 0 0 0 

### 0 0 0 

### 0 5 "1 i j

,

, 

### Average Body Weights, g: Initial 7th Day Final 290 NA NA 295 NA NA 249 259 331 262 265 304 214 247 297 207 260 316 262 311 356

i



## ... NA = 

### Not Applicable

..., 

# 000087

1lIOSEARCH, !NCORPORAfED

- p,O gOX 3598

- PHILADELPHIA.**PE.'1NSYLVN~:,\** 19101 - (215) 739-4499

### Page 11 of28

. - ENV931201, administered as supplied. 

### -..,

I 

### Acute Oral Toxicity, LD50 - 

### Rats TABLE 2 Obseryations

- Males 

## l

. .J 

### 2.00 glkg: Immediate

- All animals appeared to be nonna!. 1 

### 1 Hour All animals appeared to be lethargic and exhibited

j 

### ruffled fur. 4 Hours 5/5 

### animals appeared to be lethargic, and exhibited yellow anogenital staining and ruffled fur. 5 Hours 115 

### animals found dead. Day 1 415 animals found dead. 

# 1 1.00 glkg: Immediate

- All animals appeared to be nonnaL 4 Hours 2/5 animals appeared to be lethargicr 

### 2/5 animals j 

### appeared to be lethargic and exhibited gasping, and 115 

### animals appeared to be lethargiC and exhibited 

# .--

I gasping andbrown oral staining. Day 1 2/5 animals appeared to be lethargic and exhibited brown oral and anogenital staining and ruffled fur. 3/5 

### animals appeared to be lethargic and exhibited brown oral and anogenital staining, ruffled fur and gasping. Day 2 2/5 animals found dead. 3/5 

### animals exhibited brown body and anogenital staining and piloerection. Day 3 2/5 animals found dead. 115 

### animals exhibited brown


# - body and anogenital staining andpiloerection. Day4 115

### animals found dead. 0.944 glkg: Immediate

- All animals appeared to be normal. 4 Hours All animals appeared to be normal. Day 1 All animals exhibited dark anogenital stainingand ruffled fur. Day 2 All animals exhibited dark anogenital staining. Day 3 All animals appeared to be normal. Day4 2/5 animals found dead. 3/5 

### animals exhibited ruffled fur. Days 5&6 

# - 3/5 

### animals exhibited ruffled fur. Days 7-14 

## - 3/5 

### animals appeared to be normal.



# 000088 B[OSE.
...RCH. t."1CORPORATED P.o. BOX 8598· PHILADELPHIA. PENNSYLVAJ"1A 19101 • (215) 739-4499

### - ENV931201, administered as supplied. Acute Oral Toxicity, LD50



### 0.891 glkg: Immediate

­ 4 Hours Day 1 Day 2**~.,** " 

### Day 3 Day 4 DayS Day 6 Day 7

, 

## - Days 8-10

­ Days 11-14

­



### ......

, 

### 0.794 glkg: Immediate4 Hours Day 1 Days 2&3 Day4 Days 5-14 - Rats TABLE 2 (continued) Qbservations

- Males (continued)
All animals appeared to be normal. All animals appeared to be normal. All animals appeared to be normal. All animals exhibited dark body and anogenital staining. 1/5 animals found dead. 4/5 animals exhibited dark body and anogenital staining. 115 animals found dead. 3/5 animals exhibited dark body and anogenital staining. 3/5 animals exhibited dark body and anogenital staining. 3/5 animals exhibited dark anogenital staining. 

### 215 animals exhibited dark anogenital staining. 

### 115 animals exhibited dark anogenital stainingand appeared thin. 

### 215 animals appeared to be normaL 

### 1/5 animals exhibited dark anogenital stainingand appeared thin. 3/5 

### animals appeared to be normaL A loss ofbodyweight (60 grams) was exhibited by 115 animals (#75) between Days 0 and 7. This animal did not regain to its initial weightby 14 days. All animals appeared to be normal. All 

### animals appeared to be normal. All animals appeared to be normal. All animals exhibited ruffled fur and anogenital staining. All animals exhibited ruffled fur. All animals appeared to be normal. 0.631 glkg: All animals appeared to be normal throughoutthe 14-day observation period. 0.50 g/kg: All animals appeared to be normal throughout the 14-day observation period.

, 

# -

000089

afOSEi\RCH, fNCORPORATED· P.O. flOX 5595· PH!LAOELPHrA, PENNSYLVANrA 19101
. (215) 739-4G9



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

### - ENV931201, administered as supplied. Acute Oral Toxicity, LDso
- Rats TABLE 3

2.00 glkg:

__

I 

### 1.00 glkg:

0.944 glkg:
0.891 glkg:



### ....,
. ; .J 

### 0.794 g/kg:**~ ~** 
-: 

### 0.631 glkg: -' 

### 0.50 glkg: .-J 

### Gross Pathology

- Males
Yellow anogenital staining, ocular opacity, ruffled fur, amber liquid in the stomach and intestines, and an irritated stomach lining (dark red) were observed in the one animal found dead at five hours. Yellow anogenital staining, amber liquid in the stomach and intestines, and an irritated stomach (50% dark red or red spots) were observed in the first two animals necropsied on Day 1. 

### Yellow anogenital staining, amber liquid in the stomach and red liquid in the intestines, and an irritated stomach (50% red spots) were observed in the third animal found dead on Day 1. Yellow anogenital staining, amber liquid inthe stomach and intestines were observed in the fourth animal found dead on Day 1. Red-brown muzzle and yellow-brown anogenital staining, and dark liquid in the stomach and lower gastrointestinal tract were observed in the first animal found dead on Day 2. Red-brown muzzle and yellow-brown anogenital staining, and dark material and gas in the stomach,'and gas and yellow liquid in the lower gastrointestina! tract were observed in the second animal found dead on Day 2. Red-brown muzzle and genital staining, and dark liquid in the stomach and lower gastrointestinal tract were observed in the 2J5 

### animals found dead on Day 3. Red-brown muzzle and genital staining, and dark liqUid in the stomach and lower gastrointestinal tract were observed in the one animal found dead on Day4. Tan, dry anogenital staining was observed in the two animals found dead on Day 4. No gross abnormalities were observed in any of the three animals necropsied at the conclusion ofthe 14-day observation period. Dark muzzle and body staining, dark liquid in the stomach and lower gastrointestinal tract were observed in the one animal found dead on Day 3. Dark muzzle, body and anogenital staining, and clear liquid in the stomach and lower gastrointestinal tract were observed in the one animal found dead on Day 4. No gross abnormalities were observed in any of the three animals necropsied at the conclusion ofthe 14-day observation period. No gross abnormalities were observed in any of the five animals necropsied at the conclusion ofthe 14-day observation period. No gross abnormalities were observed in 

### any ofthe five animals necropSied at the conclusion ofthe 14-dayobservation period. No gross abnormalities were observed in any of the five animals necropsied at the conclusion ofthe 14-day observation period
. - ' ••::1



# 000090
I3[OSEARCH, INCORPORATED P.O. cox 8598
.**PHIL~DELPHlA.** PE."fNSYlVAN!A :9101
. (215i 739-1499E!'-JV931201, administered as supplied. Acute Oral Toxicity, LD50**~** Rats TABLE 4 Mortality & Average BodyWeights**~** Females Density: 0.93 glmi Dose Levels, glkg: 2.00 l.00 0.794 0.631 0.562 0.50 0.473 0.447 0.398 0.355 Animals: Number 5 5 5 5 5 5 5 5 5 5 Mortality: Day 0 1 1 0 0 0 

## a a a a 0 Day 1 

### 3 2 2 0 

## a a a a 0 

## a Day 2 1 1 2 

## a c' 

### 3 0 0 0 0 Day 3 1 1 1 0 1 

## a 0 0 0 Day 4 0 0 0 0 

## a a a Day 5 

## a a 0 0 

## a a a Day 6 0 

## a 0 0 

## a 0 0 Day 7 

## a a 0 0 0 

## a 0 Day 8 0 

## a 0 0 0 

## a a Day 9 0 

## a a a a a a Day 10 0 

## a a a 0 0 

## a Day 11 

## a a a 0 0 0 

## a Day 12 0 0 0 

## a 0 0 0 Day 13 

## a 0 

## a a 0 

## a a Day 14 0 

## a a a 0 

## a 0 14-Day Totals: Dead 5 5 5 1 0 4 0 

## a a a Live 0 

## a a 4 5 1 5 5 5 5



## ... Average Body Weights, g: Initial 223 235 206 223 218 221 213 237 218 216 7th Day NA NA NA 240 233 235 236 254 238 242 Final NA NA NA 255 251 238 247 266 246 256 NA = Not Applicable



# -

.. 

# 000091
[lIOSEARCH. lNCORl'ORATED· P.o. !lOX 5598· PHlw\DELPHfA. PENNSYLVANIA 19101 (215) 739-4499. ENV931201, 

### administered as supplied. Acute Oral Toxicity, 

### LDso
- Rats 

### TABLE 5 Qbservations

- Females
2.00 

### glkg: Immediate

­ 1 Hour 4 

### Hours 5 Hours Day 1 

### Day 2 LOa 

### glkg: Immediate

­**~** 4 

### Hours**.~**

.

### ," Day 1 

# ,-

I


### Day 2 

### Day 3 0.794 

### glkg: Immediate2 Hours 4 Hours Day 1 

# J Day 2 Day 3 ! ;

1 

### ..... 

### J 1 

## J All animals appeared to be normaL All animals appeared to be lethargic and exhibited ruffled fur. All animals appeared to be lethargic and exhibited ruffled fur and yellow anogenital staining. 115 

### animals found dead. 3/5 

### animals found dead. 115 

### animals appeared to be lethargic and exhibited ruffled fur and yellow anogenital staining. 115 

### animals found dead. 115 

### animals exhibited audible breathing. 415 

### animals appeared to be normal. 215 

### animals appeared to be lethargic. 115 

### animals exhibited gasping and red-brown nasal staining. 115 

### animals appeared lethargic and exhibited gasping and red-bro;wn nasal staining. 115 

### animals appeared to be lethargic and exhibited gasping and red-brown nasal staining, and was found dead at5 1/2 

### hours. 115 

### animals found dead. 115 

### animals appeared to be lethargic and exhibited ruffled fur and brown oral and anogenital staining in the morning and was found dead in 

### the afternoon. 215 

### animals appeared to be lethargic and exhibited gasping, ruffled fur, brown oral and anogenital staining, andred-brown nasal staining. 1/5 

### animals found dead. 115 

### animals exhibited piloerection and brown body and anogenital staining. 1/5 

### animals found dead. All animals appeared to be normal. All animals appeared to be lethargic. All animals appeared to be lethargic, and exhibited ruffled fur. 215 

### animals found dead. 3/5 

### animals appeared to be**lethargic~** 

### and exhibited ruffled fur 

### and dark body staining. 2/

5 animals found dead. 1/5 

### animals appeared to be lethargic and exhibited ruffled fur and dark body staining. 115 

### animals found dead.

8[OSEARCH, [NCORPORATED· P.O. BOX8598

- PHILADELPHIA, PENNSYLVANIA 1910J· (21S) 739-4499 

# 000092

### PrQject Number: 93-7'35(5A yage Ib 

### or L.1:5



### . ENV931201, administered as supplied. Acute Oral Toxicity, LD
- Rats 50 

### IABLE 5 (continued) Observations

- Females (continued) 0.631 g/kg: Immediate- All animals appeared to be normaL 4Hours All animals appeared to be normal. Days 1&2

- 415 animals exhibited ruffled fur, and brown body staining. 1/5 animals exhibited ruffled fur, and brown body and anogenital staining. Day 3 115 

### animals found dead. 415 animals exhibited ruffled fur and brown body staining. Day4 415 animals exhibited ruffled fur and brown body staining.
. Days 5&6 415 animals exhibited ruffled fur. Days 7-14

- 415 animals appeared to be normaL



### "'".\ 

### 0.562 glkg: Immediate- All animals appeared to be normal. 4 Hours All animals appeared to be normal. 

### --1

, 

### Days 1-4 415 animals exhibited brownbody staining and ruffled fur. 115 

### animals exhibited brown body and anogenital stainingand ruffled fur. Days 5&6

- 415 animals exhibited ruffled 

### fur. 

# .-

.

# , Days 7-14

- 415 animals appeared to be normaL

1 

### 0.50 glkg: Immediate

- All animals appeared to be normal. 

# ,­

, 

### 4 Hours All animals appeared to be normal. 1 

### Day 1 All animals appeared to be lethargic and exhibited



# - yellow anogenital staining. Day 2 3/5 animals found dead. 215 animals appeared to be


## ..... 

# J lethargic and exhibited yellow anogenital staining. Day3 115 

### animals found dead. 115 

### animals appeared lethargic and exhibited yellow anogenital staining. Day 4 115 

### animals appeared to be lethargic and exhibited yellow anogenital staining. Days 5-14

- 115 

### animals appeared to be normal. 0.473 glkg: Immediate- All animals appeared to be normal. 4 Hours All animals appeared to be normal. 

# -

; 

### Day 1 All animals exhibited ruffled fur. 

# J Days 2-14

- All animals appeared to be normal.



# -­

, 

### 0.447 glkg: All animals appeared to be normal throughout the 14-day observation 

## j period. 0.398 g/kg: 

### All animals appeared to be normal throughout the 14-day observation 

# - period. 

# J 0.355 glkg: All animals appeared to be normal throughout the 14-day observation period. 

# 000093
OIOSEARCH, [NCORPORATED

- P.o. BOX 8598 PHILADELPHIA, PENNSYLVAN!A 19101 (215) 739-4499 

# J- ENV931201, administered as supplied. Acute Oral Toxicity, LD
- Rats 50

2.00 g/kg:

, .

1.00 glkg:

0.794 glkg:

0.631 glkg:

0.562 g/kg: 

### TABLE 6 GrQSS PatholQgy

- Females

Ruffled fur, yellow anogenital staining, ocular opacity, brown liquid in the stomach and intestines, and an irritated stomach lining (dark red) were

.observed in the one animal found dead at five hours. Yell<?w anogenital

staIning, amber liquid in the stomach and intestines were observed in the first animal found dead on Day 1. Yellow anogenital staining, corneal opacity and amber liquid in the stomach and intestines were observed in the second animal found dead on Day 1. Yellow anogenital staining, brown liquid in the stomach and intestines, and an irritated stomach (50% dark red spots) were observed in the third animal found dead on Day 1. Yellowfbrown anogenital staining, yellowfbrown liquid in the stomach and intestines, and an irritated stomach (50% dark red spots) were observed in the one animal found dead on Day 2. Red-brown nasal staining, 25% of the stomach lining colored with dark red spots, and yellow liquid in the stomach and intestines were observed in the one animal found deag at 5 1/2 hours. Nasal and body staining, gas in the stomach and yellow liquid in the lower gastrointestinal tract were observed in the first animal found dead on Day 1. Nasal, body and genital staining, gas in the stomach and yellow liquid in the lower gastrointestinal tract were observed in the second animal found dead on Day 1. Red-brown muzzle and yellow-brown anogenital staining, dark liquid in the stomach and pink liquid in the lower gastrointestinal tract and no formed feces were observed in the one animal found dead on Day 2. Red-brown muzzle and genital staining, dark liquid in the stomach and lower gastrointestinal tract were observed in the one animal found dead on Day 3. Dark muzzle and genital staining and dark liquid in the lower gastrointestinal tract were observed in the two animals found dead on Day 1. Dark muzzle and genital staining and dark liquid in the lower gastrointestinal tract were observed in the two animals found dead on Day 2. Dark muzzle and genital staining, dark liquid in the lower gastrointestinal tract were observed in the one animal found dead on Day 3. Brownish anogenital staining and dark liquid in the lower gastrointestinal tract were observed in the one animal found dead on Day 3. No gross abnormalities were observed in any of the four animals necropsied at the conclusion of the 14-day observation period. No gross abnormalities were observed in any of the five animals necropsied at the conclusion of the 14-day observation period. • 

# 000094 B[OSEARCH, INCORPORATED· P.O. BOX 8598 • PHILJ\DELPHlA. PENNSYLVANIA 19101 • (215) 739-+499ENV931201, administered as supplied. Acute Oral Toxicity, LD
- Rats so TABLE 6 (continued) Gross Pathology

- Females (continued) O.SO g/kg: Dark muzzle, body and genital staining and dark liquid in the stomach and lower gastrointestinal tract were observed in the three animals found dead on Day 2. Dark muzzle and genital staining, and dark liquid in the stomach and lower gastrointestinal tract were observed in the one animal found dead on Day 3. No gross abnormalities were observed in the one animal necropsied at the conclusion of the 14-day observation period. 0.473 g/kg: No gross abnormalities were observed in any of the five animals necropsied at the conclusion of the 14-day obserration period. 0.447 glkg: No gross abnormalities were observed in any of the five animals necropsied at the conclusion of the 14-day observation period. 0.398 

### glkg: No gross abnormalities were observed in any of the five animals necropsied at the conclusion of the 14-day observation period. 0.355 

### glkg: No gross abnormalities were observed in any of the five animals necropsied at the conclusion ofthe**l~day** observation period.



# 000095 UlOSEARCH, INCORPORATED· PO. BOX 3598· PHILADELPHIA, PENNSYLVANIA 19101
. (215) 739--\499

### l:'age 19 of 28 1. lY)C,\",L 1'fUlllJJC1.



### - ENV93120l, administered as supplied.

.. Acute Oral Toxicity, LDso
- Rats TABLE 7 Individual BodyWeights MALES Dose Level: 2.00 glkg Individual Body Weights (g) Individual Body Weight Change (g) 

### I2.ay: Intery:al: Animal#···0 7 14 Animal# (0-7) (7-14) (0-14)

16 278 FD FD 

### 16 FD FD 

### FD 1.7 290 FD FD 

### 17 FD FD 

### FD 18 293 FD 

### FD 18 FD FD 

### FD 

# ;;1 19 294 FD FD 

### 19 FD FD 

### FD 20 296 FD FD 

### 20 FD FD FD Mean 290 NA NA Mean NA NA NA S.D. 7 NA NA S.D. NA NA NA N 5 

### NA NA N NA NA NA Dose Level: 1.00 glkg Individual BodyWeights (g) Individual BodyWeight Change (g)**~:** 

### InterYal: i (ur. ! 

### Animal# 0 7 14 Anirnal# (0-7) (7-14) (0-14) 

## ..J 1 288 FD FD 

### 1 FD FD 

### FD 2 

### 291 

### FD FD 

### 2 FD FD FD 

### 3 299 FD 

### FD 3 FD 

### FD FD 4 

### 299 FD FD 4 FD 

### FD FD 5 

### 300 FD FD 

### 5 FD FD 

### FD Mean 295 NA NA Mean NA NA NA S.D. 6 NA NA S.D. NA NA 

### NA N 5 NA NA N NA NA NA FD = 

### Animal found dead. 

# - NA Not applicable.



# - 000096 [JIOSEARCH, INCORPORATED· P.O. BOX 8598· pHIL-\DELPH!A, PENNSYl.'/
. .1.,"'_-\ 19101
. (llS) 739-4499ENV931201, administered as supplied. Acute Oral Toxicity, LDso
- Rats TABLE 7{continued) Individual 

### BQd¥Weights "**.~**....... , . MALES Dose Level: 0.944 glkg Individual BodyWeights (g) Individual BQdy Weight Change (g)**~:** Interval:

.AniriuH#: 0 7 14 Animal#: (0-7) (7-14) (0-14)

31 240 251 321 31 11 70 81 32 254 FD 

### FD 32 FD 

### ill FD 33 242 FD 

### FD 33 FD FD FD

}-

## .• 34 234 237. 306 34 3 69 72 

# j 35 275 288 366 35 13 78 91 Mean 249 259 331 .Mean 9 72 81 S.D. 16 26 31 S.D. S 5 10 N S 3 3 N 3 3 3 

# J Dose Level: 0.891 glkg 

# ] Individual Body Weights (g) Individual BQdy Weight Change (g) 

# J I&y: Interval: Animal#: 0 7 14 Animal# (0-7) (7-14) (0-14)

71 274 

### FD FD 71 FD FD 

### FD 72 266 FD 

### FD 72 FD FD FD 

# J J 73 265 305 351 73 40 46 86 74 218 264 316 74 46 52 98 75 285 225 244 75 -60 19 41 Mean 262 265 304 Mean 9 39 

### 48 S.D. 26 40 55 S.D. 60 18 77 ., ,.., 

# .] N 5 3 3 N 3 ..) 

# " 

# J, 

# - FD 

## = Animal found dead. 

# 000097
srOSEARCH. INCORPORATED

- P.O. BOX 8598· PH!LADELPHIA. PENNSYLVANIA 19101 (115) 739-499..L J.V)>'>J. J.**'f'rl..j,L~y** ..... ,.L. ..,...., . "" ......

## -...... - -'0

--.. -- 

### -­

- ENV931201, administered as supplied. Acute Oral Toxicity, LD50 - Rats IABLE 7(continued) Individual Body Weights MALES Dose Level: 0.794 glkg Individual BodyWeights 

### (g) Individual Body Weight Change (g) 

## 1J..B¥: InteIYial: " Animal# 0 7 - 14 Animal# (0-7) (7-14) , (0-14) 21 235 269 322 21 34 53 87 22 205 216 279 22. 11 63 74 " 23 219 265 311 23 46 46 92 :" 24 200 220 275 24 20 55 75 25 209 263 297 25 54 34 88 Mean 214 247 297 Mean 33 50 83 S.D. 14 26 20 S.D. 18 11 8 ."0 N 5 5 5 N 5 5 

### 5 

# .• Dose Level: 0.631 glkg Individual BodyWeights (g) Individual BodyWeight Change (g) -' r:la}!: InteIYal: 

# -

i Animal# 0 7 14 Animal# (0-7) (7-14) (0-14)**~** 1 207 253 311 1 46 58 104


# - 2 218 264 316 2 46 52 98 3 211 283 3"'''' 3 72 50 122 .;X) 4 200 260 315 4 60 55 115 5 200 239 306 5 39 67 106 Mean 207 260 316 Mean 53' 

### 56 109 S.D. 8 16 10 S.D. 13 7 9 r- N 5 5 5 N :J 5 5


# 000098
BIOSEARCH, INCORPORATED

- P,O. BOX 8598

- PHILADELPHIA, PENNSYLVANIA 19101 - (2!5) 739,4499

### Project Number: 93-7958A Page 22 of 28

- E:Nv931201, administered as supplied. Acute Oral Toxicity, LD50

- Rats TABLE 7(continued) Individual Body Weights MALES Dose Level: 0.50 glkg Individual Body Weights 

### (g) Individual Body Weight Change (g) 

### D.e.y: InteIval:
. Animal# 

# a 7 14 Animal# (0-7) (7-14) (0-14)

51 255 302 346 51 47 44 

### 91 52 273 325 378 52 52 53 105 53 262 318 362 53 56 44 

### 100 54 272 

### 322 360 54 50 38 88 55 247 287 334 55 40 47 87 Mean 262 311 356 Mean 49 45 94 S.D. 11 

### 16 17 S.D. 6 5 

### 8 N 

### 5 5 5 N 

### 5 5 

### 5



# 000099
1l10SEARC:l:, INCORJ'ORATED· po. nox 3598 PHILADELPHIA, PENNSYLVANIA 19101· (215) 739.4499l-'TOJect 

### NumQer: 'j;j-/'j:JOA



### - ENV931201, administered as supplied. Acute Oral Toxicity, LDso
- Rats TABLE 8 Individual BodyWeights FEMALES Dose Level: 2.00 glkg Individual Body Weights (g) Individual Body Weight Cbange (g)



## .p.g.y: Interval: Animaltf 0 

### 7 14 Ariimaltf (0-7) (7-14) (0-14)

11 

### 216 FD 

### FD 11 FD FD FD 12 234 FD FD 

### 12 FD 

### FD ill 

### 13 218 

### FD FD 13 

### FD FD FD 14 225 

### FD FD 14 

### FD FD FD 15 222 FD FD 15 

### FD FD FD Mean 223 NA NA Mean NA NA NA S.D. 7 NA NA S.D. NA NA NA N 5 NA NA N 

### NA NA NA Dose Level: 1.00 glkg Individual Body Weights (g) Iudividual BodyWeight Change 

### (g:) J::2gy: Intex:vgl: Animaltf 0 7 14 Animal# (0-7) (7-14) (0-14) 

### 6 218 FD FD 

### 6 

### FD FD 

### FD 7 234 

### FD FD 7 

### FD FD 

### FD 8 

### 243 FD 

### FD 8 FD FD 

### FD 9 247 FD FD 9 FD 

### FD FD 10 

### 234 FD 

### FD 10 FD FD 

### FD Mean 235 NA NA Mean NA NA NA S.D. 11 

### NA NA S.D. NA NA NA N 5 

### NA NA N NA NA NA FD = 

### Animal found dead. NA
- Not 

### applicable.



## ... 

# 000100
13rOSEARCH. INCORPORATED· P.O. BOX 8598· PHILADELPHIA. PENNSYL'/AN[A 1910t
. (215) 739.4499

### PrQject Numbe.r: 93-7958A Page 24 of28

- ENV931201, administered as supplied. Acute Oral Toxicity, LD
- Rats 50 

### TABLE 8(cQntinued) Individual BQdy Weights FEMALES Dose Level: 0.794 glkg Individual BQdy Weights 

### (g) Individual BQdy Weight Change (g) Day: 

### InterYs:!l: Animal# 0 7 14 Animal# (0-7) (7-14) (0-14)

71 207 FD FD 71 FD 

### FD FD 72 

### 200 FD 

### FD 72 FD FD FD 

### 73 201 FD FD 73 

### FD FD FD 74 208 FD FD 74 

### FD FD FD 75 215 FD FD 7S FD FD FD Mean 206 NA NA Mean NA NA NA S.D. 6 

### NA NA S.D. NA NA NA N 5 NA NA N NA NA NA Dose Level: 0.631 glkg Individual BQdy Weights (g) Individual BQdy Weight Change 

### (g) ........



## I


# -**129~D** 

### Interval: Animal# 0 7 14 Animal# (0-7) (7-14) (0-14)

16 226 FD 

### FD 16 

### FD FD FD 17 217 236 242 17 19 6 

### 2S 18 218 240 243 18 22 3 2S 19 228 232 252 19 4 20 24

,....., 

### 20 228 2S0 281 20 22 31 -J.:J**?~** 

### Mean 223 240 _:J:J 

### Mean 17 15 32 S.D. 5 8 18 S.D. 9 13 14



# - N 5 4 4 

### N 4 4 4 FD = 

### Animal found dead. NA = 

### Not applicable. 

# - 000101 llrOSC:ARCH, INCORPORATED· P.O. BOX 8598
. PHilADELPHIA, ?S'\INSYLVANfA 19101
. (215) 739.<1499

# -- A 1 VJ el. L 1'1 UH (Vel.



### - ENV931201, administered as supplied. Acute Oral Toxicity, LDso
- Rats TABLE 8(continued) Individual BodyWeights FEMALES Dose Level: 0.562 glkg Indiyidual BodyWeights 

### (g) Individual Body Weight Change (g)**~:** 

### InteIYsl:

.. 

### -Anim"al#' 0 7 14 Animal# (0-7) (7-14) (0-14)

11 225 226 242 11 1 16 17 12 205 230 238 12 25 8 33 13 214 232 250 13 18 18 36 14 221 233 258 14 12 25 37 15 223 246 266 15 23 20 43 Mean 218 233 251 Mean 16 17 33 S.D. 8 8 11 

### S.D. 10 6 10 

### N 5 5 5 N 5 5 5 Dose Level: 0.50 glkg Individual 

### Bod:y Weights (g) Individual BodyWeight Change (g) Day: 

### InteIYal: Animal# 0 7 14 Animal# (0-7) (7-14) (0-14)

56 235 FD FD 56 FD FD FD 

### 57 214 235 238 57 21 3 

### 24 58 212 FD FD 

### 58 FD 

### FD 

### FD 59 214 FD FD 

### 59 FD FD FD 

### 60 232 FD FD 

### 60 FD 

### FD 

### FD Mean 221 NA NA Mean NA NA NA S.D. 11 NA NA S.D. NA NA NA N 

### 5 NA NA N NA NA NA FD = 

### Animal found dead. NA 

## = Not applicable. 

# 000102 mOSEARCH. INCORPORATED

- PO. flOX 8598· PHILADELPHIA. PENNSYLVANIA 19101
. (215) 739-+;99

### Project Number: 93-7958A Page 26 of 28

. E"N'V931201, administered as supplied. Acute Oral Toxicity, LD50

- Rats TABLE 8(continued) Individual Body Weights FEMALES Dose Level: 0.473 glkg Individual Body Weights (g) 

### Individual BodyWeight Change (g) 

### D..gy: InteIYal: Animal# 0 7 14 Animal# (0-7) (7-14) (0-14)

36 204 229 235 36 25 6 31 37 215 235 239 37 20 4 24 38 215 239
. 259 38 24 20 44 39 229 260 277 39 31 17 48 40 200 216 225 40 16 9 25 Mean 213 236 247 Mean 23 11 

### 34 S.D. 11 

### 16 21 S.D. 6 7 11 N 

### 5 5 5 N 

### 5 5 5 Dose Level: 0.447 glkg Individual BodyWeights (g) 

### Individual BodyWeight Change (g)**~:** 

### InteIYal: Animal# 0 7 14 Animal# (0-7) (7-14) (0-14)

76 224 239 253 76 15 14 29 77 237 248 262 77 11 

### 14 25 78 238 257 277 78 19 20 39 79 239 254 260 79 15 6 21 80 245 274 279 80 29 5 34 Mean 237 254 266 Mean 18 12 30 '7 I 

### S.D. S 

### 13 11 

### S.D. 6 7 N 5 

### 5 5 N 

### 5 5 5


# 000103

B£OSEARCH. INCORPORATED

- P.O. BOX 8598

- PHIL·\DELPHIA. PENNSYLVANIA 19101

- (215) 739-4499Project Number:**~J-'/~.JtiA** t'age L.I or**L.~**

- ENV931201, administered as supplied. Acute Oral Toxicity, LD50 - Rats TABLE 8(continued) Individual Body Weights FEMALES Dose Level: 0.398 glkg Individual Body Weights 

### (g) Individual Body Weight Change 

### (g) I2ay: Intl:!nml: Animal# 0 7 14 Animal# (0-7) (7-14) (0-14)

26 207 225 238 26 18 13 31 27 220 242 251 27 22 9 31 28 228 245 254 28 17 9 26 29 212 238 239 29 26 1 27 30 223 240 249 30 17 9 26



# - Mean 218 238 246 Mean 20 8 28 S.D. 

# a 8 7 S.D. 4 4 3 N 5 5 5 N 5 5 5**~** Dose Level: 0.355 glkg IndividualBodyWeights 

### (g) Individual BodyWeight Change (g) Day: InterY!ill: j
,

'. Animal# 0 7 14 Animal:t (0-7) (7-14) (0-14)
, 6 230 236 257 6 6 21 27


# - 7 207 236 249 7 29 13 42 8 214 238 252 8 24 14 38 9 211 257 272 9 46 15 61 10 217 241 248 10 24 7 31 Mean 216 242 256 Mean. 26 14 40**~**



### S.D. 9 9 10 S.D. 14 5 13 N 5 5 5 N 5 5 j


# 000104
8!OSEARCH. INCORPORATED P.O. 80X 3598· PH!LADELPHIA. PENNSYLVANIA 19101
. (215) 739-4499Project Number. Protocol Number. Sponsor Test Article: Test: GLP Compliance: Deviation:

•...i SubmiHedby:
'-t



# -I

'''''"l 

# J Approvedby: 

# -

-i



### ....

, Acknowledged by:



### .... Appendix A - Page 28 of28 

# BIOSEARCH INCORPORATED P.o. Box 8598, Philadelphia, Pennsylvania 19101

### ..-" Telephone: (215) 739-4499 I Fax: (215) 423-9210 DEVIATION FROM PROTOCOL #1 93-79S8A AOT-800 (Rev. 02/93) ENV931201 Acute OralToxicity, LDSO

- Rats (TSCA) 40 CFR792 OECO Due to a technician error, afternoon observations for males in the 6.31 glkg dose level were made but not recorded on Day 9. In addition, due to technician error, morning observations on Day 10 for these animals were made butagainnotrecorded
.

This deviation"is notthoughtto affectthe integrityofthis study. 

# C;::;/";\ David Gabriel, B. A. 

# , . S

# ~

I alityAssurance

Vice President, Laboratory Operations & Regulatory Affairs

# Jf~~'**o;::~**

# ~13-4~ Date



# t;-;:;

# -ii Date 

# '!f
te

# J 

# I / 

# 1Y

# -f/?~ Date


### 5----

## Ie!

- c: )

, /;, Dare lvfay 12, 1994 93 040 7958 1485 Form QA-500A 

# 000105

=1,...; 

### Acute Toxicity of

to Photobacteriumphosphoreum 

### (Screen Test)

,

### .....

July 1995 by

, 

# -

000106Title: Acute Toxicityof to Photobacterium phosphoreum

-. Issue Date: July 1995 Report Preparation: 

# Skl,\).,1 l. 

# srt$'u..·-rrod Sheryl L. Spotswood Study Director Environmental Services Group 

## l i Report Approval: 

# ] 

# J Report and Study Inspection: Date: ['2- - 

# 5 u.b..

- 

## qIS

Date:

## Il-1~-/47S

I Date: 

# a.:::r**c.,~** 

# f{' I

Date oftesting: Protocol: Data/Archive Reference: Date Protocol Approved:

# ;JI22~·)r~ WIlliam J. Kuc Study Inspector Environmental Services Group

December 2, 1993 Microtox Basic Test Procedure (Microbics Corp., 1991) SX-2081 May 30, 1991 (Microbics Corp., 1991)


# 000107i :.. .' ! ill,

',,­**'-~·l;.,** Test Material: Batch/Lot: 93-72733 Test Date: December 2, 1993 Summary: The 15 minute ECSO ofSX-2081 to Photobacterium phosphoreum is estimated to be 1 - 10 mg/L. Species: Photobacterium phosphoreum Source: Petrolfte Corporation 15 Minute EC50: 1 - 10 mg/L (estimation)
Phenol Standard Test: 5 minute EC50

# = 23 mg/L (Acceptable range 

## =13 - 26 mg/L)

.Control 10: 

### 89 Temperature: 15°C



# 000108Introduction This Microtox bioassay was performed by the The Microtox test system determines toxicity by measuring the decrease in light production ofthe luminescent marine bacterium, Photobacterium phosphoreum, following a timed exposure to a test material. During these organisms' normal cellular respiration, about ten percent oftheir metabolic energy is converted into visible light. Changes in this metabolic process cause a change in light output ofthis species. A reduction in cell metabolism, caused by exposure to a test sample, is directly proportional to the measured reduction in bioluminescence (Bulich and Isenberg. 1981). The concentration oftest sample which produces a 50% reduction in bioluminescence, following a tiined exposure, is considered the effective concentration (EC50). Microtox bioassay EC50s are usually calculated following 5 and 15 minute exposures, however 30 minute exposure times are sometimes required. This study followed the Basic Test protocol which was published by Microbics Corporation (Microbics, 1991). All original raw data is retained in the' archives. Methods and Materials Test Organism A Microtox reagent comprised ofa lyophilized (freeze-dried) preparation of Photobacterium phosphoreum was rehydrated with Microtox Reconstitution Solution. This bacterial suspension was maintained in a separate well ofthe analyzer at 5°C and allowed to equilibrate a minimum of 30 minutes priorto testing. Test Substance and Stock Preparation A sample of.
. batch 93-72733, was obtained from I , facility. is a brown, water soluble liquid. A stock of was prepared by weight to a preCision ofone milligram and diluted to volume in volumetric glassware with deionized water
. 

### • Test Conditions This toxicity bioassay was performed using a Microtox Model 500 analyzerand software manufactured by Microbics Corporation (Carlsbad, CAl. The software was loaded onto an IBM-compatible computer which interfaced with the analyzer. All supplies used were those obtained from or recommended by



## ... Microbics Corporation.
. Unused cuvettes were added to rows A and B ofthe analyzer. Micratox Osmotic Adjustment Solution and Diluent (a 2% saline solution) were added to the cuvettes as required by protocol for a test with a dilution factor of 10. All were maintained at a constant temperature of 15°C for 15 minutes. Subsequently, 10 pi of the bacterial suspension were pipetted into row B cuvettes containing 500 pi of diluent and allowed to equilibrate for 15 minutes. 

# 000109

7Four dilutions of the stock were made in row A with Microtox Diluent while the bacteria were I equilibrating. Following the equilibration time, light emissions ofthe bacteria in row B were recorded as initial values (10)' Immediately, 500 pi from each cuvette in row A were transferred into the



## ... corresponding row B cuvettes. At 5 and 15 minutes following the sample dilution, light emissions were recorded (15 and 115)'
. Statistical Analvsis The ratio of light lost to the light remaining in each of the cuvettes in row B is used to calculate the EC50. A ratio for each concentration is expressed as a gamma value. That concentration of sample which was calculated to produce 50% reduction in light ora gamma of 1.0, following a given timed "expos'ure,"Was designated as the EC50 value. EC50 values are expressed in mg/L, with lower EC50 values corresponding to higher toxicity levels. Ninety-five percent confidence limits were calculated for each EC50. Results and Discussion Upon addition ofthe product to the test cuvette itwas observed that appeared soluble in the

.

# ,-

! test media
. I The 15 minute EC50 of to Photobacterium phosphoreum is estimated to be 1 - 10 mg/L. The phenol standard test which was performed simultaneously with the " Basic Test resulted in an acceptable EC50 value for the 5 minute exposure. 

# , This assay was a range-finding orscreen'test. A definitive test was not performed. 

## :. .J

References 

# J BUlich, A. A. and Isenberg, D.L, 1981, "Use ofthe Luminescent Bacterial system for the Rapid Assessment ofAquatic Toxicity." ISA Transactions, Vol.20, NO.1. Microbics Corporation, 1991, "Microtox Model 500 Toxicity Test System Manua'" (Preliminary 

# -

i Release), Microbics Corp., Carlsbad, CA.**~**



## .J .J 

# 000110

# --

# --- ------------ ---------- --------- TEST DATE: TEST TIME:---------­ MICROTOX DATA REPORT FILE NAME: .K15 Investigator: 

## C:;;.

# see**~u** ..rr:-c(J 1

# 0**~** 

# ";-'**-~~** Approved by:

_______ 

### Sample Descriptidn: Test date;\.",12/2/93, i 

### (93-72733)
. Procedure: BASIC '"
.. __/ Osmotic Adjustment: MOAS Initial Concentration: 9.876 MG/L Dilution Factor: 10 Test Time: 15 minutes Concentration Units: MG/L


### .... NUMBER IO/IT CONC. GAMMA 1 82.53/ 74.66 0.0099 0.05284# 2 81.11/ 76.08 0.0988 0.01541# 3 

### 85.30/ 69.30 0.9876 0.,17234#

. ".**:,,,~,** 4 " 

### 89.55/ 0.06 

## ., 9:8760 > 

### 999 

## *

, 

### CONTROL IT/IO 

## = 84.31/ 88.52 ......1 

### CORRECTION FACTOR 

# = 0.9524 .IC25 0.5602 (95% CONFIDENCE RANGE EXCEEDS LIMITS)**~jECOMMEND:** 

### RERUN ASSAY AT HIGHER CONCENTRATIONS**~** 

### # Used for calculations 

### * Invalid data or controls



# ...

I 

## ..J

-.lIIIj

, ,

2**.~**



# - 000111



# -

### • STATISTICAL REPORT FOR DATA FILE
- _.K15



## .. ESTIMATING EQUATION: LOG C 

## = 0.9338 

### x LOG +0.1939

'.f! 

### NUMBER OF DATA PAIRS USED: 3 RESIDUAL VARIANCE: 1.52054 95% CONFIDENCE FACTOR: %9999.00000 FOR IC25 CORRELATION COEFFICIENT: R =: 

### 0.48962 

# .,. COEFFICIENT OF DETERMINATION: RU2 =: 

### 0.23973 INHIBITING CONCENTRATIONS (95% CONFIDENCE RANGE) : ICSO =: 

### 1.5627 MG/L ( 0.0000 TO 

### % 628.6954E+26)*** IC20 =: 

### 0.4282 MG/L ( 0.0000 TO 

### % 446.7338E+14) ICI0 = 

### 0.2008 MG/L ( 0.0000 TO % S30.4764E+08) ICOl =: 

### 0.0214 MG/L ( 0.0000 TO % 746.4709E+13) *** EXTRAPOLATED RESULTS

'""'I 

# JI



### i 

## .J



# - - 000112

## ------------ TEST TIME:**-----...:.~~**



### -----­ Osmotic Adjustment: MOAS MICROTOX DATA REPORTTEST DATE:

..K5 

### FILE NAME:**-"** 

### ­ Investigator: 

## <;-:**s-r±tw'1~J**

# ~**~Approved** by: 

### date,,- '-1.2/2/93 I 

### (93-72733). Sample Descriptio: Test

.Procedure: BASIC**~.** 

### : 9.876 MG/L Dilution 

### Factor: 10 Initial Concentration Test Time: 5 minutes NUMBER IO/IT 1 82.53/ 74.15 2 81.11/ 73.91 3 85.30/ 75.74 

### 4 89.55/ "0.44 

### CONTROL IT/IO 

## = 81.03/ 88.52 

### CORRECTION FACTOR 

# = 0.9154 i

. \ 

### Concentration Units: MG/L CONC. 0.0099 0.0988 0.9876 

### "9.8760 GAMMA 0.01884 0.00456* 0.03093 185.30194*



### ........... NO TOXICITY AT CONCENTRATIONS TESTED 

## * Invalid data or controls

.. "",

i

'

# ..

.

# ­

, 

## ......
.­

,



# -

000113ACUTE TOXICITY OF

TO THE SHEEPSHEAD MINNOWk Cyprinodon variegatus 

# '.

July 1995 by


# - 000114''''I


# 1 ....!

.

# '-

1 -' 

# J!

,

# I

..... -, 

## JI



## , 

# - .J TITLE: Acute Toxicity of variegatus.

ISSUE DATE: July 1995

REPORT PREPARED BY: Date: 

# 11::) IF;.)

# 1I1£~~1~v .-/ Scientific Investigator

REPORT APPROVED BY: Date:

### 17:S~**/7i~**

DATES OF TESTING: January 17 - 21, 1994.
PROTOCOL NO: FM-01-5A 

### Quality Assurance Statement This study was not conducted according to Good Laboratory Practices (40CFR792). The test data was reviewed on January 21, 1994 by the Quality Assurance Officer to assure that Standard Operating Procedures and the protocol used forthis study were adhered to. This report is an accurate reflection of the raw data.

Date: Sheryl 

### L. Spotswood Designated Quality Assurance Officer 2 

# 000115. i 

# - MATERIAL: BATCH/LOT No.: (93-72731 & 93-72732)



# -

i TEST DATE: January 17 - 21, 1994. 

# -, ...l SUMMARY: 

# l 

# .­ SPECIE: SOURCE: The 96 hour static LCso of , in synthetic seawater to the sheepshead minnow is 2.2 mg/l. Sheepshead minnow, Cyprinodon variegatus. Aquatic Research Organisms, Hampton, New Hampshire 96 HOUR LCso AND 95% CONFIDENCE LIMITS: 2.2 (1.0

- .3.2) mg/l 

# -J 96 HOUR NO OBSERVED EFFECT CONCENTRATION (NOEC):



# ] '""l 

## .J WATER TYPE:

TEMPERATURE:



# - pH:



# - SALINITY:



# -



# - 1.0 mg/I Synthetic seawater

15.5 -17.5 0 C

8.3··

31 parts-per-thousand

3 

# 000116INTRODUCTION

.)."-, This static acute bioassay was performed by the - ,. '.\'­ during the period of January 17 - 21, 1994.

# 'J.""~"".'"**;~... "'~**  Bioassay procedures described below are patterned after those described by the " . American Public Health Association (1), American Society of Testing and Materials (2), Office of Pesticides and Toxic Substances (3), the US EPA (4) and the 

# ., Organization for Economic Cooperation and Development (5). Test procedures 

## J follow I. Standard Operating • 

# ] Procedures which were compiled following the intent of Good Laboratory Practices (6). All original raw data is retained in the archives. METHODS AND MATERIALS 

# .] The sheepshead minnows, 

### Cyprinodon variegatus, used for this study were obtained from Aquatic Research Organisms, Hampton, New Hampshire. Test fish were cultured in an activated carbon filtered, recirculating synthetic seawater 

# ] system for at least ten days prior to testing. All test fish were maintained according to the procedures outlined by Brauhn 

### eta/(7) on a 16 hour daylight photoperiod. Fish were fed dry commercial fish food or live 

### Artemia nauplius larva on a daily basis (Monday thru Friday) during holding. Fish appeared in good health and mortalities during the 48 hours immediately preceding the test were less than 3%. Fish used in this test were randomly selected from the stock culture. Twenty-four hours before initiating the test, fish were taken off feed. Ten fish were introduced at random into the control and each exposure vessel. The fish had a mean (10 organism) total length of 1.9 cm and a mean weight of 0.09 grams. Biological loading was 0.06 g/1. Artificial seawater used during this study was prepared by dissolving Hawaiian 

# ] Marinemix sea salts in deionized water; deionized watertest was prepared by passing St. Louis municipal water through multiple deionization beds. Test water had a salinity of 31 parts-per-thousand and a pH of 8.3. Salinity was determined with a YSI Model 33 SCT meter, pH with a CORNING pHI/on Meter Model 135 and dissolved oxygen with a YSI Model 54 oxygen meter. Temperature was determined with a handheld glass thermometer. Environmental control was achieved through the use of a Heinicke HC 982M walkin environmental chamber. Thermostatic controls were adjusted to achieve a nominal environmental chamber test temperature of 17

### .±. 1°C. Temperature range in the test tanks was 15.5 - 17.5 °C while the photoperiod consisted of a 16 hour daylight regime. The study employed two separate experiments, a preliminary or rangefinding study and a definitive study. The rangefinding study delineated a narrower range of test concentrations which were utilized for the definitive study.
4 

# 000117

### L'\,,::..,

,

.**~** . A sample of fa 

### 50/50 composite sample of batches 93-72731

## ~~3 72732) was obtained from Technology Department, <'. '>b was received in a clear glass bottle and stored at**roo~** temperature until use. The test material, , , is a brown, water soluble liquid:-'"" Seven concentrations and a control were tested to determine the toxicity of**~** to the sheepshead minnow. A 10 mgtml stock solution of was prepared by weight and diluted to volume with deionized water. Test tank position was randomized in the test chamber using random number generating program (RAND). The study was initiated by adding the toxicant to test vessels containing 15 liters of dilution water, thoroughly mixing, and then randomly introducing the fish. Dissolved oxygen, pH and temperature were determined initially and every 24 hours thereafterfor the control and all concentrations tested. Salinity in the control tank was determined initially and at 96 hours. Dead organisms were removed from test tanks every 24 hours; behavioral observations of surviving fish were performed at this time. Mortality data were used to calculate the median lethal concentration (LC ) and so their respective 95% confidence limits for the appropriate exposure periods. Calculations were performed using a computer program developed by Stephan stal (8). RESULTS AND DISCUSSION Test solutions of - were completely soluble in all concentrations tested.

......, The 96 hour LCso with 95% confidence limits for to the sheepshead



# i minnow is 2.2 (1.0 - 3.2) mg/l. The 96 hour No Observed Effect Concentration {NOECI is 1.0 mgt!. Mortalities and LCso values with asso.ciated 95% confidence limits are presented in Tab/es 1 and 2. Behavioral observations conducted during the study are presented in Table 3. Chemical and physical parameters monitored during the test are presented in Tables 4, 5, and 6. DEVIATIONS FROM PROTOCOL The following temperature deviations from the specified 17.0 ± 1.0°C occurred: 1.0 mgtl at 0 hours (15.5°C) and 0.56 mgtl at 24 hours (15.8°C). It is not felt that these'deviations'should effectthe outcome of the study. "'­ ""'':v'-.

5 

# 000118

# ...

...

.

# -**.~** ,

;

j 

# "' ...:



### ..,.

I

.,J

, .

# .,

,J 

# .] "J
" .

. " 

### , I ,0IIi 

# ",

J

i

.......

J j 

## .....

.

### ...,


# ..1

!



## ...

... REFERENCES 1. American Public Health Association. 1992. Standard Methods for the Examination of Water and Wastewater, 18th ed. Washington, D.C. 2. American Society of Testing and Materials. 1980. Standard Practice for Conducting Acute Toxicity Tests with Fishes, Macroinvertebrates and Amphibians. E-729, Philadelphia, PA
. 3. U.S. E.P.A. 1982 - Environmental Effects Tests Guidelines (EG-9), EPA 560/6-82-002. 4. Office of Pesticides and Toxic Substances. 1980. Fish Acute Toxicity Test Standard Toxic Substances Control Act. U.S. E.P.A., Washington, D.C; 5. OECD Guidelines for Testing of Chemicals. 1984. Organization for Economic Co-operation and Development, Paris
. 6. U.S. E.P.A. July 1, 1986. Good Laboratory Practice Standards. Code of Federal Regulations. 40 CFR Part 792. Washington, D.C. 7. Brauhn, J. L., R. A. Schoettger, and L. H. Mueller. 1975. Acquisition and Culture of Research Fish. Rainbow Trout, Fathead Minnows, Channel Catfish and Bluegills. EPA-660/3-75-001.
. 8. U.S. E.P.A. 1978 - Stephan, C.E. and R.W. Andrews. A Computer Program for Calculating an LCso, Duluth, MN.

6 

# 000119

....
m**~.**

: ....

.' ,:

. :' i
... ... ,.... 

### Table(s) 1, 2, 3: Toxicity to the Sheepshead Minnow,". Cyprinodon variegatus .,,:'
..'" 

### Mortality Observations, LC50 Calculations and Behavioral Observations / j

A# /'! ,f " ./ Table 1: Mortality Observations (#Dead out of 10)

'-'



### , ,

",
........;.;.:..<;.::;:; :::;. 

### o o o 2 10 

### o 10 10

### !i:4~:i)fm::::m .;.; .::::=;::::: :;:;:;:::;:::.;. 

### o 

### o o 2 10 

### o 10 10 

### :::::::i??Wflf::::}:: 

# "] o o 2 

### o 10 

### o 10 10 Table 2: LCso Calculations 

# ,]

1.8

- 3.2 Binomial 1.0

- 3.2 Binomial 1.0

- 3.2 Binomial 1.0

- 3.2 .Binomial Table 3: Behavioral Observations

1. GENERAL a. quiescent b. hyperexcitable c. irritated d. surfacing e. sounding f. twitching g. tetanus h. flaccid i. normal 2. SWIMMING 3. PIGMENTATION a. ceased a. light discolored b. erratic b. dark discolored c. gyrating c. varidiscolored d. skittering e. inverted f. on side 4. INTEGUMENT a. mucus shedding b. mucus coagulation c. hemorrhagic

.- Data not collected after 100% mortality occurs. 5. RESPIRATION a. rapid b. slow c. irregular d. ceased e. gulping air f. labored

7 

# 000120

### Table(s) 4,5,6: Toxicity to the Sheepshead Minnow, Cyprinodon variegatus 

### Chemical and Physical Parameters

. pH



# ..,


# J -,

j? ---" 

# J I -" 8.3 8.3 8.2 8.2 8.2 8.2 8.2 8.2 8.2 8.2 8.2 8.2 8.2 8.2 8.2 8.2 8.2 8.2 8.2 8.2 8.2 8.1 Dissolved Oxygen, ppm

7.9 7.9 • 7.9 8.0 8.0 7.7 7.S 7.8 7.7 7.9 7.7 7.8 8.0 7.9 8.0 7.7 7.9 8.0 7.9 7.8 Temperature °C

16.0 16.0 15.8 16.0 16.1 16.3 16.2 16.3 16.3 16.2 16.3 16.4 16.2 16.4 16.5 16.3 1'6:5 16.3 16.5 16.3 Control Tank Salinity,**parts-per~thousand** 30.5 31.0

- Data not collected after 100% mortality. 

# 000121 8

# ]

ACUTE TOXICITY OF 

# ] TO THE FATHEAD MINNOW, Pimephalespromelas 

# oJ 

# ,-

l 

# .J



# - '-'

! 

### ..J



# - - - July 1995 by 

# - 000122 

# -

# , TITLE: Acute Toxicity of co the Fathead Minnow I Pimephales promelas.

ISSUE DATE: July 1995

REPORT PREPARED BY: 

# 1

# ?y~~~.**I;::~**
- . .-J William 

### J. Kuc 

# t:1 t Scientific Investigator 

# ] REPORT APPROVED BY:



# ..J

] DATES OF TESTING: January 31 - February 4, 1994. 

# ] PROTOCOL NO: FF-01-3A ",- . /"" Date: 

## 17.Jv2r**l1Y'~**

Date:

### '9:.r~ 177

# S

Quality Assurance Statement This study was not conducted according to Good laboratory Practices (40CFR792). The test data was reviewed on February 

### 2, 1994 by the Quality Assurance Officer to assure that Standard Operating Procedures and the protocol used for this study

, were adhered to. This report is an accurate reflection of the raw data.

. ...-;
Date: Sheryl L. Spotswood Designated Quality Assurance Officer 2 

# 000123MATERIAL:



### ...... BATCH/LOT No.: (93-72731 & 93-72732)



### ...... TEST DATE: January 31 - February 4, 1994.

"SUMMARY':' The 96 hour static LCso of in reconstituted freshwater to the fathead minnow is 0.80 mg/l. SPECIE: Fathead minnow, Pimephalespromelas SOURCE: Aquatic Research Organisms, Hampton, New Hampshire 96 HOUR LCso AND 95% CONFIDENCE LIMITS: 0.80 (0.56 : 1.00) mg/l 

# J 96 HOUR NO OBSERVED EFFECT CONCENTRATION (NOEC): 0.32 mg/l
WATER TYPE: Reconstituted freshwater TEMPERATURE: 15.5 - 17.4 0 C pH: 

### 8.3 HARDNESS: 160 mg/I as CaCOa " .r ,,., , ALKALINITY: 11 3 mg/l as CaCOa

3


# -INTRODUCTION This static acute bioassay was performed by the during the period of January 31 - February 4, 1994. Bioassay procedures described below are patterned after those described by the American Public Health Association (1), American Society of Testing and Materials (2), Office of Pesticides and Toxic Substances (3), the US EPA (4) and the Organization for Economic Cooperation and Development (5). Test procedures**~.:':** . i,
.."< r'
. f()lIo.w.
. . .... . . Standard Operating 'ProCecll.ireswhich were compiled following the intent of Good Laboratory Practices (6). All original raw data is retained in the archives. 

## 1
,....

I METHODS AND MATERIALS The fathead minnows, 

### Pimephalespromelas, used for this study were obtained from Aquatic Research Organisms, Hampton, New Hampshire. Test fish were held in recirculating, 29 gallon glass aquariums for five days prior to testing. Holding water was reconstituted hard water prepared from deionized St. Louis municipal water. All test fish were maintained according to the procedures outlined by Brauhn 

### etal (7) on a .16 hour daylight photoperiod. Fish were fed dry commercial fish food or live

### Artemia nauplius larvae on a daily basis (Monday thru Friday) during holding. Fish appeared in good health and mortalities during the 48 hours immediately preceding the test were less than 3%. Fish used in this test were 

# ] randomly selected from the stock culture. Twenty-four hours before initiating the test, fish were taken off feed. Ten fish were introduced at random into each control and exposure vessel. The fish had a mean (10 organism) total length of 2.2 

# ] cm and a mean weight of 

### 0.08 grams. Biological loading was 

### 0.05 gil. 

# J Dilution water for the test was prepared by the carbon filtration and deionization of municipal water. Treated water was then reconstituted according to procedures outlined by Marking and Dawson (8). The static fish bioassays were conducted in 21 liter glass-silicone aquaria containing 15 liters of reconstituted water. , -1 Reconstituted water was characterized as having a pH of 8.3, total hardness of 1 .­ 160 mg/l as CaC03, total alkalinity of 113 mg/l as CaC03, and a specific conductance of 

### 410 pmhos/cm. Hardness and alkalinity were determined according to standard analytical procedures APHA (1), pH with a CORNING pHllon Meter Model 135, conductivity with a YSI Model 33 conductivity meter, dissolved oxygen with a YSI Model 54 oxygen meter, and temperature with a handheld thermometer.



# -- Environmental control was achieved through the use of a Heinicke HC 982M walk­ in environmental chamber. Thermostatic controls were adjusted to achieve a nominal environmental chamber test temperature of 17 ± 1

# ac. Temperature range in the test tanks was 15.5
. 17.4

# ac while the photoperiod consisted of a 16 hour daylight regime.

4 

# 000125

# -
The study employed two separate experiments, a preliminary or rangefinding study and a definitive study. The rangefinding study delineated a narrower range of test concentrations which were utilized for the definitive study. A sample of (a 50/50 composite sample of batches 93-72731 & 93-72732) was obtainedfrom'
. was received in aclear glass bottle and stored at room temperature until use. The test material, I, is a brown, water soluble liquid. Eight concentrations and a control were tested to determine the toxicity of to the fathead minnow. A 10 mg/ml stock solution of was prepared by weight and diluted to volume**,~,i~h,d.eionized wat~r.**  Testtank position was randomized in the test chamber using random number generating program (RAND). The study was initiated by adding the toxicant to test vessels containing 15 liters of dilution water, thoroughly mixing, and then randomly introducing the fish. Dissolved oxygen, pH and temperature were determined initially and every 24 hours thereafterforthecontrol and all concentrationstested. Dead organisms were removed from test tanks every 24 hours; behavioral observations of surviving fish were performed at this time. Mortality data were used to calculatethe median lethal concentration (LC ) and their so respective 95% confidence limits for the appropriate exposure periods. Calculations were performed using a computer program developed by Stephan 

### et

### at (9). RESULTS AND DISCUSSION Test solutions of were completely soluble in all concentrations tested. The 96 hour LCso with 95% confidence limits for , to the fathead minnow is 0.80 (0.56 - 1.00) mg/1. The 96 hour No Observed Effect Concentration (NOEC) is 0.32 mg/I. Mortalities and LCso values with associated 95% confidence limits are presented in Tables 1 and 2. Behavioral observations conducted during the study are presented in Table 3. Chemical and physical parameters monitored during the test are presented in Tables 4, 5, and 6. DEVIATIONS FROM PROTOCOL Test organisms were held for 5 days ratherthan 10 days as specified in the protocol. The following temperature deviation from the specified 17.0 

### ± 1.0°C occurred: 0.56 mg/l at 0 hours (15.5°C). ' It is not felt that these deviations should effect the outcome of the study, since the fish appeared in excellent health during holding and no mortalities or aberrant behavior was evident in the control tank during testing.

5 

# 000126

### ..., REFERENCES 1. American Public Health Association. 1992. Standard Methods for the Examination



## .... of Water andWastewater, 18th ed. Washington, D.C
. 2.American SocietyofTesting and Materials. 1980. Standard Practicefor Conducting



### .....

! Acute Toxicity Tests with Fishes, Macroinvertebrates and Amphibians. E-729, Philadelphia, PA. 3. U.S. E.P.A. 1982 - Environmental Effects Tests Guidelines IEG-9), EPA 560/6-82-002. 4. Office of Pesticides and Toxic Substances. 1980. Fish AcuteToxicityTestStandard 

### ! 

### J Toxic Substances Control Act. U.S. E.P.A., Washington, D.C. 

### 5. OECD Guidelines for Testing of Chemicals. 1984. Organization for Economic Co-operation and Development, Paris. 

# .-

, 6. U.S. E.P.A. July 1, 1986. Good Laboratory Practice Standards. Code of Federal 

# J Regulations. 40 CFR Part 792.**W~shington,** D.C.



## ..... 7. Brauhn, J. L., R. A. Schoettger, and L. H. Mueller. 1975. Acquisition and Culture 

# .J of Research Fish. Rainbow Trout, Fathead Minnows, Channel Catfish and Bluegills. EPA-660/3-75-001. 8. Marking, L.L. and V. K. Dawson. 1973.

### Toxicity of Quinaldine Sulfate to Fish. Fish Control No. 48. U. S. Fish Wildlife Service, Washington, D. C. 9. U.S. E.P.A. 1978 - Stephan, C.E. and R.W. Andrews. A Computer Program for Calculating an LCso, Duluth, MN.

6 

# 000127**~< «/~** 

. ... /.**I~J-:-:!**r":"" .".' 

### Table{s} 1, 2, 3: Toxicity to the Fathead Minnow, 

### Pimephalespromefas Mortality Observations, LC50 Calculations and Behavioral**Observatforis"~i<;** < Table1: Mortality Observations I#Dead out of 10) .•,



# -

! ..J

• 

# J ]

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# ««J



# ., J

< -r !



# -**~ .~**  

# - o o o o 

### o 6 10 

### o 0< 8 10 

### o 

### o o 9 10 Table 2: LCso Calculations 10 10 10 10 10 10 10 10 10

Table 3: Behavioral Observations 0.93 0.83 0.80 1.00

­ 1.80 0.56

­ 1.80 0.56

- 1.80 0.56

­ 1.00 Binomial Binomial Binomial Binomial

li li 1ad.2b 3b.5f 1i 1a.3b 1ad,3b Sf 1i 1ad,3b 1ad,3b, 5f 1i 1a,3b 1ad,3b Sf 1. GENERAL 2. SWIMMING 3. PIGMENTATION a. quiescent a. ceased a. light discolored b. hyperexcitable b. erratic b. dark discolored c. irritated c. gyrating c. varidiscolored d. surfacing d. skittering e. sounding e. inverted f. twitching f. on side 4. INTEGUMENT g. tetanus a. mucus shedding h. flaccid b. mucus coagulation i. normal c. hemorrhagic

-- Data not collected after 100% mortality occurs. 7 5. RESPIRATION a. rapid b. slow c. irregular d. ceased e. gulping air f. labored



# 000128 i

### Table(s) 4,5,6: Toxicity to the Fathead Minnow, Pimephales promelas 

### Chemical and Physical Parameters pH

.

# .., ! -1



# -, 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3

, ,**~-.j** Dissolved Oxygen (ppm) .-,



## I -,

f •. 1 9.8 9.2 9.3 9.2 9.8 9.8 9.8 9.8 9.9 9.5 9.4 9.6 9.6 9.7 9.6 9.6 9.7 9.4 9.3 9.3 9.4 Temperature. 

## °e**~**



# - ,7.1 17.4 17.4 17.3 16.0

, 6.7 16.5 16.2 16.7 16.5 16.8 16.8 16.5 16.5 16.8 16.3 16.6 16.5 16.8 16.8 16.8

- Data not collected after 100% mortality.


# 00012'3 8Protocol'90-0'.-01 !'est**Sn~l~** 

## I Batch/tot' IDate Started IInvestigator RfAOY BIOVfGRADAB'lr,Y CLOSED BOrTLE TfST Innoculum Data Buffers Added •...,1. C

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# I I"VI._.fAl'fI."" 

# I - Medium High Form' 005a Medium High lOM
. A Issue Date: July 1990

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Reference Standard S..,le Oflotfon Ueter Date seed Collected: lyp!! Seed Collt'Cted:**~$tlel** Seed location: "SO-Sissell Poi Seed concentration: 

### O.**o·~'** trial 1 mill 6.53% Potassfum Phosphate 1 mIll 2.25% Magnesium**~ulfate** 0.6 mg/l Ma "<:etate Source: lab trade delonfzed Filtrat(on: 0.22 _(eron

Data Botth!



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## POG)E? \30

### AQUATIC TOXICOLOGY Static Acute Freshwater Toxicity PARAMETERS Specie: Fathead minnow Temperature: 17 degrees C Duration: 96 hours Static Acute Saltwater Toxicity PARAMETERS Specie: Sheepshead minnow Temperature: 17 degrees C Duration: 96 hours Specie: Photobacterium phosphoreum Test Type: Microtox Duration: 15 minutes 

### ENVIRONMENTAL FATE Oxygen Demand PARAMETERS Test Concentration: 100 ppm Ready Biodegradability PARAMETERS Media: Freshwater Inoculum Acclimation Per: 0 days Test Concentration: 2 ppm RESULTS Lethal Concentration 50% No Effect Concentration ­

RESULTS Lethal Concentration 50% No Effect Concentration ­ - 96 hours 96 hours

- 96 hours 96 hours
Effective Concentration 50% - 15 minutes

RESULTS Chemical Oxygen Demand

RESULTS Biodegradability, 5 days Biodegradability, 14 days Biodegradability, 20 days Biodegradability, 28 days VALUES 0.8 mg/l 0.32 mgtl

VALUES 2.2 mgtl 1.0 mg/l
1 - 10 mgtl

VALUES 192 mg

VALUES

26% 39% 57% 78 % NOTE: Percent biodegradability is based on a chemical oxygen demand (COD) value of 192 mg per 100 ppm product.

Page 1of 2 April 13, 2004
Disclaimer of Liability: , warranls to purchaser, but nO third parties 0( others Ihe specifICations fO( the product shall fa" within a generally recognized range for typiCal physical properties eslablished by_when the product departs -. point oforigin and thai any services shall be performed in accordance with applicable written documenls,**~AKES** NOOTHERWARRENTYOR GUARANTEE OF ANY KIND, EXPRESS OR IMPUED, INCLUDING NO IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, REGARDING ANY SERVICES PERFORMED OR PRODUCT SUPPUED. _will give purchaser Ihe benellt of_bestjudgemenl in making interpretationsofdata, butdoes notguaranteethe accuracyofcorrectness ofsuch interpretations,

_ recO(Omendations contained herein areadviSO(y onlyand without representafions as to Ihe resufts,
.. shall not be liablefor any**ind~ect,** special, punitive, exemplary0(consequential damagesorlosses frO(O any causewhatsoever including but not limited to ils negligence,MAMMALIAN TOXICOLOGY Acute Oral Toxicity PARAMETERS Specie: Rat-female Observation Period: 14 days Specie: Rat-male Observation Period: 14 days Primary Skin Irritation PARAMETERS Specie: Rabbit Scoring Method: Draize Dermal Sensitization PARAMETERS Specie: Guinea Pig Method: Magnussen-Kligman Maximization RESULTS Lethal Dose 50% Lethal Dose 50%

RESULTS Skin Irritation Index

RESULTS Sensitization VALUES 0.56 g/kg
0.90 g/kg

VALUES Corrosive

VALUES Non-sensitizer

Page20f 2 April 13, 2004 Disclaimer of Uability: ,warrantsto purchaser, but no Ihird parties or others the specifICations for the product shall fall within a generally recognized range for Iypical physical properties established by" when Ihe product departs _poinl of origin and that any services shall be performed in accordance with applicable written documents,

___MAKES NOOTHERWARRENTYOR GUARANTEE OF ANY KIND. EXPRESS OR IMPLIED. INCLUDING NO IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARllCULAR PURPOSE. REGARDING ANY SERVlCES PERFORMED OR PRODUCT SUPPLIED.
_ will give purchaser the benefil of
_ bestjudgement in making interprelalionsofdala. butdoes notguaranteetheaccuracyof correctnessofsuch inlerprelatiOIlS, --"ecommendationsconlained herein are advisory only and without representations as to the**resu~s,** _ shall not be liable for any indirect, special, punitive, exemplaryorconsequential damagesor losses from any causewhatsoever including but not limited 10 its negligence,A division oftheAmerican Chemical Society Phone: 800-631-1884. 614-447-3870 Fax: 614-447-3747 E-mail: answers@cas.org Internet http://www.cas.org/Supportlclienthtml

INVENTORY EXPERT SERVICE REPORT

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

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

# Material Safety Data Sheet

Product Name Supplier

Material Uses 24 Hour Emergency Numbers CANUTEC 613-996-6666 (Canada24 hours) CHEMTREC Int'l 01-703-527-3887 (International 24 hour) National Fire Protection Association (U.S.A.)

# ~**amme.bllity** HeaD. 

### 3

# ~ 0 Reactivi" Speclnc Hazard Code Version 1.0

Effective Date 

### 411312004 Print Date 4114/2004

Quaternary ammonium hydroxide Trade secret. 30-60 Fattyamine Trade secret 10-30 Isopropanol 67-63-0 30-60

Ethylene Glycol 107-21-1 1-5 Not available. Not available. ACGIH (UnitedStates).

TWA: 490 mg/m3 STEL: 980 mg/m3 TWA: 200 ppm STEL: 400 ppm OSHA (United States).

TWA: 400 ppm STEL: 500 ppm TWA: 980 mg/m3 STEL: 1225 mg/m3 ACGIH (United States).

CEIL: 100 mg/m3 Form: Aerosol only. OSHA (United States).

CEIL: 50 ppm CEIL: 125 mg/m3Not applicable.

skin and tract. CERCLA Reportable Qua Hazard Summary

Routes of Potential Acute Health Effects

Medical Conditions aggravated by Exposure SeeToxicol Additional Hazard Identification Remarks DANGER. May cause chronic effects. Flammable liquid. Vapors can form an ignitable or explosive mixture with air. Can form explosive mixtures at temperatures at or above the flash point. Vapors can flow along surfaces to a distant ignition source and flash back. Static discharges can cause ignition or explosion when container is not bonded. May be corrosive to cause central nervous effects if inhaled.

Eyes May be corrosive to the eyes. May cause eye burns and permanent eye injury. SkinMay be corrosive. Skin contact may produce burns. Inhalation May cause central nervous system (CNS) effects if inhaled. May be severely irritating to the lungs. Ingestion Notconsidered a likely route of exposure, however, may be corrosive ifswallowed. Exposure to this product may aggravate medical conditions involving the following: kidneys, nervous system, respiratorytract, skin/epithelium, eyes. May be harmful if ingested. This product may be aspirated into the lungs during swallowing or vomiting of swallowed material. Aspiration into the lungs may produce chemical pneumonitis, pulmonary edema, and hemorrhaging.
Eye Contact Skin Contact
Inhalation Ingestion

Additional FirstAid Remarks Immediately flush eyes with plenty of water for at least 15 minutes, occaSionally lifting the and lower Get medical attention imm Remove contaminated clothing and shoes immediately. Wash affected area with soap and mild detergent and large amounts of water until no evidence of chemical remains (apprOXimately 15-20 mi Get medical attention if irritation occurs. Remove to fresh air. Oxygen may be administered if breathing is difficult. If not breathing, administer artificial respiration and seek medical attention. Get medical attention if symptoms Get medical attention immediately. If swallowed, do not induce vomiting unless directed to do so by medical personnel. Wash out mouth with water if person is conscious. Never induce vom or mouth to avictim who is unconscious or havi convulSions. Not Not available.Flammability ofthe Product OSHA Flammability Class Autoignition ra Flash Points Flammable Limits Products of Combustion Fire Hazards in Presence ofVarious Substances Fire Fighting Media and Instructions
ProtectiveClothing Special Remarks on Fire Hazards Flammable liquid. Vapors can form an ignitable or explosive mixture with air. Can form explosive mixtures attemperatures at or above the flash point. Vapors can flow along surfaces to a distant ignition source and flash back. Static discharges can cause ignition or explosion when container is not bonded. IC Not available. Closed L.E.L. Not available. U.E.L. Not available. These products are carbon oxides (CO, C02) nitrogen oxides (NO, N02
...). Open Flames/Sparks/Static. Heat.
In case of fire, use foam, drychemicals, or C02fire extinguishers. Evacuate area andfightfire from a safe distance. Water spray may be used to keep fire-exposed containers cool. Keep water run off out of sewers and public waterways. Note that flammable vapors mayform an**;~~,;+..,Ikl"** mixture with air. V travel considerable distances andflash back if innitari Do not enterfire area without proper personal protective equipment, including NIOSH approved self-contained breath Notavailable.

Spill

OtherStatements Put on appropriate personal protective equipment. Keep personnel removed and upwind of spill. Shut off all ignition sources; no flares, smoking, or flames in hazard area. Approach release from upwind. Shut off leak if it can be done safely. Contain spilled material. Keep out of waterways. DIke large spills and use a non-sparking or explosion-proof means to transfer material to an appropriate container for disposal. For small spills add absorbent (soil may be used in the absence of other suitable materials) scoop up material and place in a sealed, liquid-proof container. Note that flammable vapors may form an ignitable mixture with air. Vapors may travel considerable distances from spill and flash back, if ignited. Waste must be of in with federal state and local environmental control ulations. If RQ (Reportable Quantity) is exceeded, report to National Spill Response- Office at 1-800-424-8802. Additional Accidental Not available. Release Measures RemarksHandlingandStorage Put on appropriate personal protective equipment. Avoid contact with eyes, skin, and clothing. Avoid breathing vapors or spray mists. Use only with adequate ventilation. Store in a dry, cool and well ventilated area. Keep away from heat, sparks and flame. KE!ep away from incompatibles. Keep container tightly closed and dry. To avoid fire or explosion. ground container mentand before handl Additional Handlingand Notavailable. Storage Remarks

or to ns vapors or particles below their respective threshold limit value. Ensure that eyewash stations and showers are to the work-station location. Personal Protection Personal Protective Equipment recommendations are based on anticipated known manufacturing and use conditions. Theseconditions are expected to result in only inCidental exposure. A thorough review of thejob tasks and conditions bya safety professional is recommended to determinethe level of personal protective equipmentappropriatefor thesejob tasks and conditions.Eyes Chemical safetygoggles. Usefull face shield ifsplashes could occur. BodyWear long sleeves and chemical resistant apron to prevent repeated or prolonged skincontact. Respiratory Respirator use is not expected to be necessary under normal conditions of use. In poorly ventilated areas, emergencysituations or if exposure levels areexceeded, use NIOSH approved full face respirator. Hands Chemical resistant gloves.

FeetChemical resistant boots or overshoes. Otherinformation Nitrile

### or neoprene gloves. Protective Clothing (Pictograms)

Additiofuil Exposure Control Remarks Notavailable.

Color Li htAmber.Pour Point Solu Boilin Point Physical Chemical Notavailable. Notavailable. Soluble Notavailable. Notavailable. ments Stability and Reactivity**..:.T.:..:.he:::....t:.p:....:ro:..:::d~uc::..:t;..:;is:::...;;;..;st::::;;a;;;.;bl.=..e:....**_________________________ Conditions of Instabil Notavailable. Incompatibilitywith VariousSubstances Oxidizing material. Acid. Hazardous Decomposition Products Notapplicable.
Hazardous Hazardous polymerization is notexpected to occur. Special Stability & Not available. Reactivity Remarks
ComponentToxicological Information AcuteAnimalToxicity Quaternaryammonium hydroxide Fattyamine Isopropanol Ethylene Glycol

ChronicToxicity Data 1) Quaternaryammonium hydroxide Not available. 2) Fattyamine Not available. 3) Isopropanol Not available. ORAL (LD50): Acute: 3000 mg/kg [Mouse]. ORAL (LD50): Acute: 5045 mg/kg [Rat]. 3600 mg/kg [Mouse]. DERMAL (LD50): Acute: 12800 mg/kg [Rabbit]. VAPOR (LC50): Acute: 16000 ppm 8 hour(s) [Rat]. ORAL (LD50): Acute: 5500 mg/kg [Mouse]. 4700 mg/kg [Rat]. 4000 mg/kg [Female rat]. DERMAL (LD50): Acute: 10600 mg/kg [Rabbit].Isopropanol, a component of this product. Ingestion has produced hyperglycemia (high blood sugar) in humans (Lacouture, P, et ai, 1983, "American Journal of Medicine" and Chan K-M, et ai, 1993, "Clinical Chemistry"). In a four month study, inhalation of isopropanol vapors for 20 hours per week by laboratory animals produced bronchitis, pneumonia, and blood effects (International Program of Chemical Safety, 1990, Environmental Health Criteria 103: 2-propanol, World Health Organization). Ataxia (a jerky or shaky movement that occurs during voluntary muscle movement) and microscopic hyaline droplets (fungal or branched structures) in the kidneys were seen in rats exposed tc isopropanol at concentrations up to 5000 ppm for 6 hours per day, 5 days per week, for 13 weeks (Burleighflayer et ai, 1994). Inhalation of high levels of isopropanol (4,000 and 8,000 ppm for 8 hours) has produced congestion in the liver, lungs, and spleen of laboratory animals (Laham S, et al. 1980, "Drug and Chemical Toxicology). In addition, isopropanol has shown to be a fetotoxin in tests on laboratory animals [Nelson, BK et al (1988), Food and Chemical Toxicology, 26(3), pps 247-254]. 4) Ethylene Glycol Ethylene glycol (EG) is a component of this product. Chronic ingestion has shown to cause adverse kidney, liver, bladder, and blood effects in laboratory animals (NTP Technical Report, 1993; Fund. Appl. Toxicol. 7:547-65; FD Cosmet Toxico!. Vol. 3:229-34; Drug and Chern ToxicoI13(1):43-70). EG is an animal teratogen at doses which produced mild toxicity to the mother. EG given at doses up to 5,000 mg/kg/day to pregnant rats or up to 3,000 mg/kg/day to mice induced a wide variety of fetal malformations, including those of the musculoskeletal, bone marrow, and spleen (RTECS. 1996). Ethylene glycol is used to cryopreserve embryos of many mammalian species, including pigs, goats, cows and horses (Otoi et ai, 1995; Fieni et ai, 1995; Hochi et ai, 1994). This makes it unlikely that ethylene glycol itself is the activeteratogen in whole animal studies. The EG metabolite, glycolic acid, was active in contrast to EG itself for inducing developmental defects in whole rat embryos in culture (Carney et ai, 1996). EG inhibited metabolic cooperation of Chinese hamster cells in vitro, afinding which may have implications for its mechanism of teratogeniCity (Loch-Caruso et ai, 1984).
ProductToxicological Information Not nervous ..,,,,,,,rern OtherAdverse Effects This product results from a reaction involving ethylene oxide (EtO) (CAS No 75-21-8). It may, therefore contain residual EtO (less than 0.1%) which can accumulate in the container headspace and be released to the ambient environment when opened. This release of EtO would tend to increase when the product is agitated during unloading or blending operations. The Occupational Safety and Health Administration (OSHA) has set the worker exposure level for EtO at 1 ppm (8-hour time weighted average). The STEL is 5 ppm (15 minutes). This standard regulates occupational exposure to EtO from sources including products containing residual EtO. It is the responsibility of the employer to comply with OSHA EtO Standard (29 CFR 1910.1047). The OSHA action level TWAfor EtO is 0.5 ppm. The OSHA excursion level is 5 ppm as averaged over a sampling period of 15 minutes. The International Agency for Research on Cancer (IARC) has identified EtO as a "human carcinogen (Group 1); human evidence is limited; animal evidence is sufficient". EtO has been listed as an "anticipated carCinogen" by the National Toxicology Program (NTP). Eto has also been designated as a carcinogen and reproductive toxin under the State of California's Safe Drinking Water and Toxic Enforcement Act (Proposition 65). A no significant risk level of 2 meg/day (an acceptable intake level is 20 meg/day) was established under Proposition 65.8005 and COD Not available. Biodegradable/OECD Not available.

Not available. Special Remarks Toxicity ofthe Products Not available. ofB

Responsibility for proper waste disposal rests with the generator of the waste. Dispose of any waste material in accordance with all applicable federal, state and local regulations. Note that these regulations may also apply to empty containers, liners and rinsate. Processing, use, dilution or contamination of this product may cause its physical and chemical erties to Additional Waste Notavailable. Remarks

DOT Reportable Marine Pollutant Additional DOT information Emergency Response Guide Page Number

U.S. Federal Regulations Environmental Regulations CORROSIVE LIQUID, FLAMMABLE, N.O.S. (Contains: Quaternary ammonium hydroxide, Isopropanol), 8(3), UN2920, "

Not applicable. Not icable. Not available. 132

Extremely Hazardous Substances: Not applicable to any components in this product. SARA 313Toxic Chemical Notification and Release Reporting: Ethylene Glycol; SARA 3021304 Emergency Planning and Notification substances: Not applicable to any components in this product. Hazardous Substances (CERCLA 302): Not applicableto any components in this product. SARA 311/312 MSDS distribution - chemical inventory - hazard identification: fire; immediate health hazard;Clean Water Act (CWA) 307 Priority Pollutants: Notapplicableto any components in this product. Clean Water Act (CWA) 311 Hazardous Substances: Notapplicable to anycomponents in this product. Clean Air Act (CAA) 112(r) Accidental Release Prevention Substances: Notapplicableto any components in this product. Threshold Notapplicable. Planning Quantity (TPQ) TSCA Inventory This product or its components, ifa mixture, are not listed on theTSCA inventory. Status This product is manufactured and distributed in compliancewith the United States Toxic Substance Control Act (TSCA) Research and Development (R&D) Exemption Regulation, 40 CFR 720.36. It is intended for R&D use only and maycontain one or more ingredientsthat are not listed on theTSCA Inventory. This product must be handled bya technicallyqualified (trained) individual or under the supervision of atrained individual. This productdoes not contain anycomponents that are subject to the reporting requirements of TSCA Section 12(b) if exported from the United States. States...,.....,·+O" information is available u on from State International Regulations**",n,,...n,,.n~C!** are included on the Canadian Domestic Substances List. Canada WHMIS(Canada)**~~~~~** __________________________________________________**~** European Union Not all components are included on the European Inventoryof Existing Commercial Chemical orthe Euro Listof Notified Substances. International inventorystatus information is available upon requestfrom for the i fol countries: Austral Korea Phil HarmonizedTariff Code Notavailable. Other Regulatory Nofurther regulatory information is available. Information

OtherSpecial Notavailable. ConsiderationsDisclaimer NOTE: The.'informatioifdrfthis MSDS is basedon data which is consideredto beaccurate:_. . however, makes noguarantees orwarranty, eitherexpressedorimpliedofthe accuracyorcompleteness ofthis information. The conditions ormethods ofhandling, storage, use anddisposal oftheproductare beyondourcontrolandmaybe beyondourknow/edge. Forthis andotherreasons, we do notassume responsibilityandexpresslydisclaim liabilityfor loss, damage orexpensearising out oforin any wayconnectedwith the handling, storage, use ordisposal ofthis product. This MSDS waspreparedandis to be usedforthis product. Iftheproductis usedas a componentin anotherproduct, this MSDS information maynotbe applicable.