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## UNITEDSTATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON D

.C., 20460



# VIA FACSIMILE: 201-437-4801 April 19,2012 Attention: GlennMerritt(201-437-4000) OFFICE OF PREVENTION, PESTICIDESAND TOXIC SUBSTANCES



# Subject: Follow-upto fax transmissionof4/13/12 (ePMA

# removalofCBIclaimsregarding identified studies) 

# DearMr. Merritt: Thisis sentto you pursuantourconversationofseveralminutesago. In LarryRobinson'sfax of April 13th 

# whilethereisawaiverofconfidentialbusinessinformation (CBI)claimsinthe identified studiesthereis aanassertionofcopyrightprotectionsand someadditionaltextthat providesin part
... "Reproduction,reference,summarization,retransmissionorotheruse ofthis copyrighteddocument.

..includingelectronictransmission,usingtheinternetoremail

....is strictlyprohibited

...." Whileweunderstandthebasisforthecopyrightprotections, we have a concernthatthelanguage inthe fax mightbeinterpretedto meanthatePMAisassertingthatthestudies submittedperthe TSCAactivityarenottobemadeavailableinan Agencye-docket(electronicdocket)orbe includedin EPAwebsitesamongthecollectionofTSCAhealthand safety studies. Wearethinkingthattheidentifiedlimitations noted inthefax arenotintendedto applyto EPA's normal proceduresforthehandlingofnon-CBI materialssubmittedby entities subjectto TSCA regulations(includingthepostingofsuchmaterialsongovernmentwebsites) and submittedby suchentitiesinorderto complywithaTSCArequirement,but areratherintendedto limit entitiesoutsidethe government from the unauthorizeduseorappropriationofthe studies. Canyoupleaseconfirm CPMA'sintent? Manythanks.

ScottM. Sherlock,AttorneyAdvisor Environmental AssistanceDivision OfficeofPollutionPreventionandToxics

# CONFIDENTIAL BUSINESS INFORMATION



# APPROVAL STATEMENT PRELIMINARY FINAL REPORT

Study Title Acombined 28-day repeated dose toxicity studywith the reproduction/developmental toxicityscreening testofClI. Pigment Blue 61 inrats by oral gavage NOTOX Substance 183591/A NOTOX Project 488231

STUDY DIRECTOR STATEMENT The undersigned declares thatthis preliminaryfinal report constitutes atrue accountofthe procedures adopted and the results obtained inthe performance ofthis study. This report has not yet been inspected bythe NOTOX QualityAssurance Unitto confirm that the methods and results accurately and completely reflectthe raw data. The audit isplanned for Weeks 47 and 48 2008.Any comments will be incorporated inthe final report.

Study director

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

## MWEW. Beekhuijzen, M.Sc. date:

APPROVAL Management

D~rs. A.W.M. van Rozendaal d ate:

# CONFIDENTIAL BUSINESS INFORMATION



# PRELIMINARY FINAL REPORT

StudyTitle 

# A COMBINED 28-DAY REPEATED DOSE TOXICITY STUDY WITH THE REPRODUCTIONIDEVELOPMENTAL TOXICITY SCREENING TEST OF C.I. PIGMENT BLUE 61 IN RATS BY ORAL GAVAGE

Author KE.W Beekhuijzen, M.Sc. Studycompletion date

Test Facility NOTOX B. Hambakenwetering 7 5231 DD's-Hertogenbosch The Netherlands Laboratory Study Identification NOTOX Project 488231 (488232) 

## NOTOX Substance 183591 -Page1of249-

## C.I. PIGMENT BLUE61 NOTOX Project 488231 1. CONTENTS

I CONTENTS...........................2 2. STATEMENT OF GLP COMPLIANCE................................................ 4 3. LEAD QUALITY ASSURANCE STATEMENT................................................... 5 4. SUMMARY
....................................................................................... 6 5. INTRODUCTION............................................................ ...................... 7 5.1. Preface
...................................................................................... 7 5.2. Studyschedule............................................................................... 8 5.3. Project numbers .............................................................................. 8 5.4. Aims of study................................................................................. 8 5.5. Guidelines................................................................................... 9 5.6. Storage and retention of recordsand materials............................................ 9 6. MATERIALS ANDMETHODS................................................................... 10 6.1. Testsubstance.............................- ............................................... 10 6.1.1. Testsubstance information (170676/A)............................................ 10 6.1.2. Studyspecific test substance information.......................................... 10 6.1.3. Testsubstance formulation......................................................... 10 6.1.4. Chemical analysis ofdose preparations ........................................... 11 6.2. Testsystem................................................................................. 11 6.3. Allocation................................................................................... 12 6.4. Animal husbandry........................................................................... 12 6.5. Treatment.................................................................................. 13 6.5.1. Parental animals..................................................................... 13 6.5.2. Offspring.............................................................................. 13 6.6. Observations................................................................................ 14 6.6.1. Parental animals ..................................................................... 14 6.6.2. Offspring .................................................................... ......... 15 6.7. Clinical laboratory investigations........................................................... 16 6.8. Pathology Parental Animals................................................................ 17 6.8.1
. Termination........................................................................... 17 6.8.2. Macroscopic examination ........................................................... 18 6.8.3. Organ weights........................................................................ 19 6.8.4. Histotechnology...................................................................... 19 6.85. Histopathology....................................................................... 19 6.9. Pathology Offspring......................................................................... 20 6.9.1. Termination........................................................................... 20 6.9.2. Macroscopic examination ........................................................... 20 6.10. Calculations .......................................................................... 20 6.11. Electronic data capture .......................................................... ... 21 6.12. Interpretation......................................................................... 21 6.13. Listofdeviations ..................................................................... 21 6.13.1. List of protocol deviations........................................................ 21 6.13.2. List ofstandard operating procedure deviations............................... 21 7. RESULTS ......................................................................................... 22 7.1. Analysis of Dose Preparations (see alsoAppendix 3) ................................... 22 7.2. Observations ................................................................................ 22 7.2.1, Mortality .............................................................................. 22 7.2.2. Clinical Signs......................................................................... 22 7.2.3. Body Weights......................................................................... 22 7.2.4. Food Consumption................................................................... 22 7.2.5. Functional observations ............................................................. 23 7.3. Clinical Laboratory Investigations.......................................................... 23 7.3.1. Haematology......................................................................... 23 7.3.2. Clinical Biochemistry................................................................. 23 7.4, Pathology
................................................................................... 23 7.4.1. Macroscopic Examination........................................................... 23 7.4.2. OrganWeights ....................................................................... 24
Page 2

## C.I. PIGMENT BLUE 61 NOTOX Project488231 7.4.3. Microscopic Examination (see alsoAppendix 4).................................. 24 7.5. Reproduction................................................................................ 24 7.6. Breeding Data............................................................................... 25 7.7. Pup Development........................................................................... 25 8. DISCUSSION AND CONCLUSION............................................................. 26 9. REFERENCES
.................................................................................. 27 APPENDICES APPENDIX 1 SUMMARY TABLES ............................................................... 1-27 Clinical Signs ...................................................................... 2 Body Weight.............I........................................................... 5 Body Weight Gain................................................................. 7 Food Consumption
........ ... .................. 9 Relative Food Consumption.................................................. 11 FunctionalObservations......................................................... 13 Motor Activitytest................................................................ 14 Haemnatology..................................................................... 15 Clinical Biochemistry............................................................. 17 Macroscopic Findings............................................................ 19 Organ weights!/Organ to BodyWeight Ratios................................ 20 Reproduction Processes........................................................ 24 Mating Performance ............................................................. 25 Breeding Data per Group ....................................................... 26 Mean Body Weights of Pups.................................................... 27 APPENDIX 2 INDIVIDUAL TABLES............................................................... 1-85 Mortalitydata...................................................................... 2 Clinical Signs ...................................................................... 4 Body Weight...................................................................... 12 BodyWeight Gain................................................................ 15 Food Consumption

........................ 18 Relative Food Consumption..................................................... 20 Functional Observations......................................................... 22 MotorActivity test................................................................ 24 Haemnatology..................................................................... 26 Clinical Biochemistry............................................................. 31 Macroscopic Findings............................................................ 35 Organ weights/Organ to Body Weight Ratios................................ 38 Breeding Data per Litter......................................................... 46 Mean Body Weights of Pups per Litter......................................... 50 BodyWeights of Pups........................................................... 53 Individual Pup Data.............................................................. 61 Corpora Lutea.................................................................... 83 Implantation Sites................................................................ 84 Keyto Missing Values/Remarks Clinical Laboratory Investigations.........85 APPENDIX 3 FORMULATION ANALYSIS........................................................ 1-11 APPENDIX 4 PHASE REPORT HISTOPATHOLOGY ........................................... 1-97 APPENDIX 5SUMMARY DOSE RANGE FINDING............................................... 1-2
TOTALNUMBEROF PAGES ............ .......................................................... 249

Page 3

## C.I. PIGMENT BLUE 61 NOTOX Project 488231
2. STATEMENT OF GLP COMPLIANCE NOTOX B.V., 's-Hertogenbosch, The Netherlands The study described inthis report has been correctly reported andwas conducted incompliance with: The Organization for Economic Cooperation and Development (OECD) Good Laboratory PracticeGuidelines (1997). Which essentially conform to: The United States Food and DrugAdministration Good Laboratory Practice Regulations. The United States Environmental Protection Agency Good Laboratory Practice Regulations Thesponsor isresponsible for Good Laboratory Practice (GLP) compliance forall test substance information unless determined by NOTOX
TheGLP statement forthe delegated phase histopathology isincluded inAppendix4ofthis report.
NOTOX B.V.
M.E.W. Beekhuijzen, M.Sc. Drs. A.W.M. van Rozendaal Study Director Section Head Toxicology

Date:........................................ Date:........................................

Page 4

## C.I PIGMENT BLUE 61 NOTOX Project488231 3. LEAD QUALITY ASSURANCE STATEMENT NOTOX B. 's-Hertogenbosch, The Netherlands This reportwas inspected bythe NOTOX QualityAssurance Unitto confirm thatthe methods and resultsaccurately and completely reflect the raw data. The dates ofQualityAssurance inspections aregiven below. During the on-site process inspections procedures applicable tothistype ofstudy were inspected. The reporting date isthedate of reporting to theStudy Director. The QAU report was then forwarded to theTest Facility Management. Type of Phase ISection Start End Reporting date inspections Inspection Inspection date(s) date(s) Protocol (Study) DD\MMM\JJ DD\MMM\JJ DD\MMM\JJ On-site (Process) DD\MMM\JJ DD\MMM\JJ DD\MMM\JJ On-site (Study) DD\MMM\JJ DD\MMM\JJ DD\MMM\JJ Report (Study) DD\MM M\JJ DD\MMM\JJ DD\MMM\JJ TheQualityAssurance programme forthe delegated phase histopathologywas performed by theQualityAssurance appointed bythetestsite management and aQuality Assurance statement isincluded inAppendix 4ofthis report.

Head ofQualityAssurance C.J Mitchell B.Sc.

Date: ...................

Page 5

## C.I. PIGMENT BLUE 61 NOTOX 

## Project488231 4. SUMMARY Combined repeated dose toxicitystudywith reproduction/developmental toxicity screeningtest with ClI. Pigment Blue61 administered by oralgavage inWistar rats. The purpose ofthisstudywas toevaluatethe potentialtoxic effects of0.1. Pigment Blue 61 when administeredto rats foraminimum of28 days and toevaluatethe potential ofthetest substance toaffect male andfemale reproductive performance such asgonadal function, mating behaviour, conception, parturition and early postnatal development. Thestudy was based on the followingguideline: Organisation of Economic Co-operation and Development Guidelines (OECD) for testing of Chemicals Guideline 422, Combined Repeated DoseToxicity Studywith the Reproduction/Developmental ToxicityScreening Test, March 1996. After acclimatisation, four groups often male and ten femaleWistar Han rats were exposed by oral gavage to thetest substance at 0,50, 250 and 1000 mg/kg/day. Males were exposed for30 days, i.e.2weeks priorto mating, during mating, and upto termination. Females wereexposed for 41 to45 days, i.e. during 2weeks prior to mating, during mating, during post-coitum, and during at least4days of lactation. The following parameters wereevaluated: clinicalsigns, functionalobservations, bodyweights, food consumption, reproduction processes, observations offspring, clinical pathology, macroscopy, organ weights, and histopathology. Chemical analyses offormulationswere conducted once during the study toassess accuracy, homogeneity and stability. RESULTS 

## Accuracy, homogeneity and stabilityofformulations weredemonstrated byanalyses. Notreatment-related toxicologicallysignificant changes were noted inany ofthe parameters investigated inthis study (i.e. mortality, clinical appearance, functional observations, body weight,food consumption, clinical laboratory investigations, macroscopic examination, organ weights, microscopic examination, reproduction, breeding and pup development). CONCLUSION 

## From the results presented inthis report, theparental, reproduction, breeding and developmental No ObservedAdverse Effect Level (NOAEL) for 0.1. PIGMENT BLUE 61 ofat least 1000 mg/kg/daywas established.

Page 6

## C.I. PIGMENT BLUE 61 NOTOX Project488231 5. INTRODUCTION 5.1. Preface This isamulti-site study.The studywas performed according to theOECD Consensus document Number 13 TheApplication ofthe OECD Principles ofGLP tothe Organization and ManagementofMulti-Site studies. Information concerning the delegated phase isgiven intheapplicable appendix. Phase: Histopathology Principal Investigator: Dr. A.Pilling, ToxPath Consultancy Ltd, UK (see Appendix4) Sponsor Color Pigments Manufacturers Association, Inc. 300 N.Washington St. Ste. 102, ALEXANDRIA, VA 22314 USA Study Monitor Mr. L.Robinson jlr@cpma.com Study Director M.EW. Beekhuijzen, M.Sc. NOTOX B.V. Hambakenwetering 7 Is-Hertogenbosch The Netherlands Tel: +31.73 640 6700 Fax: +31.73 640 6799 Email: manon.beekhuiizen~cnotox.nI Coordinating Biotechnician M.M.A. Rijkers (NOTOX ByV.) Clinical Pathology J.E. van Kesteren (NOTOX B.V.) Analytical Chemistry Ir.M.J.C. Brekelmans (Principal Scientist, NOTOX B.V.) Histotechnology Ing T.A. Mijnders (NOTOX ByV.) Necropsy J.H. van den Brink, DVM (NOTOX BYV) Lead QA C.J. Mitchell, B.Sc. (NOTOX B.V.): christine.mitchell(ftotox.nl Testfacility Management H.H. Emmen, M.Sc. (NOTOX ByV.): harrv.emmena-notox. ni Test Facility NOTOX B.V. Hambakenwetering 7 5231 DD s-Hertogenbosch The Netherlands

Page 7

## C.I. PIGMENT BLUE 61 NOTOX Project488231 5.2. Study schedule Deliveryofanimals 11 June 2008 Experimental start date 12 June 2008 (allocation) Start treatment 17 June 2008 Start mating 01 July 2008 Necropsy of males 17July 2008 Delivery of litters 23 to 27July2008 Necropsy offemales and pups 28Julyto 01 August 2008 Experimental completion date 01 August2008 (end in-life) 5.3. Project numbers Two projectnumbers were used to collectonlinedata. Eventually, alldata were reported under NOTOX Project 488231 Project number Online data 488236 Parental animals: Clinical signs Functional observations Body weights andfood consumption pre-mating Body weights mating period Clinical laboratory investigations Macroscopic findings and organ weights 488231 Allother data 5.4. Alms ofstudy Thepurpose ofthisstudy wasto evaluatethe potentialtoxic effectsofthetest substance when administered to rats for aminimum of28days and to evaluate thepotential ofthetest k substanceto affect male and female reproductive performance such as gonadalfunction, mating behaviour, conception, parturition andearly postnatal development. ANo Observed Adverse Effect Level (NOAEL) was evaluated. This studyshould provide part ofarational basis fortoxicologicalrisk assessment inman.

Page8

## C.l. PIGMENT BLUE61 NOTOX Project 488231 5.5. Guidelines The protocol was reviewed and agreed bythe Laboratory Animal Welfare Officer and the Ethical Committee ofNOTOX (DEC NOTOX 07-13) as required bythe Dutch Acton Animal Experimentation (February 1997). The study procedures described inthis reportwere based onthe following guideline: 1)Organisation of Economic Co-operation and Development Guidelines (OECD) fortesting of Chemicals Guideline 422, Combined Repeated Dose Toxicity Studywith the Reproduction/Developmental Toxicity Screening Test, March 1996. 5.6. Storage and retention ofrecords and materials Records and materials pertaining tothe study, includingprotocol, raw data, specimens (except specimens requiring refrigeration orfreezing) and thefinal report, will be retained inthe NOTOX archives foraperiod ofat least 2years afterfinalization ofthe report. After this period, the sponsorwill be contacted todetermine whether raw dataand specimens should be returned to them, retained or destroyed on their behalf. Those specimens requiring refrigeration orfreezing will be retained by NOTOX for as long as the quality ofthe specimens permits evaluation but no longerthan three months afterfinalization ofthe report. NOTOX will retain atest substance sample untilthe expiry date, butno longerthan 10 years after finalization ofthe report. Afterthis period the samplewill be destroyed. Storage and retention of records and materials bythe testsitesforthe delegated phasesare described intheapplicable appendices.

Page 9

## ClI. PIGMENT BLUE 61 NOTOX Project488231 6. MATERIALS AND METHODS 6.1. Test substance 6.1.1. Test substance information (170676/A) Identification C.l. Pigment Blue 61 Structure

N0

S03H 

# Molecularformula C3 7

## H2 9

## N3

## 03 S 

## Molecular weight 596 GAS Number 1324-76-1 Description Dark blue powder with granules Batch 2084701 Composition 96.830% Test substancestorage At room temperature inthedark Stability under storage conditions Stable Expirydate 02January 2018 6.1.2. Study specifictestsubstance information pH 4.3 at concentration of>1% Solubility invehicle: 0Propylene glycol Yes, <1% 6.1.3. Test substanceformulation Vehicle Propylene glycol, specificgravity 1.036 (Merck, Darmstadt, Germany). Rationalefor vehicle Based on trialformulations performed at NOTOX Stabilityoftestsubstance invehicle Formulations at 10 and 200 mg/mI were stablewhen stored at room temperature forat least6hours (determined during this project; seeAppendix 3). Method offormulation Formulations (wlw)were prepared dailywithin 6hours priorto dosing and were homogenised to avisually acceptable level. Adjustment was made for specific gravityofthevehicle (1.036). Storage conditions At ambient temperature.

Page 10

## C.I. PIGMENT BLUE 61 

## NOTOX Project488231 6.1.4. Chemical analysis ofdose preparations Analyses were performed on asingle occasion during the treatment phase (30 June 2008) according to avalidated method (NOTOX Project 488235) as specified below: Group Analysis (type ofsample)
1 acc(M) 2 acc +

## hom +

## stabt.0 

## (TMB), stabt-6, RT

## (S) 

## 3 acc(M) 

# 4 acc+

# homn +

# stabt0 

## (TMB), stabt-6.RT

## (S) Duplicate sampleswere analysed acc=accuracy, hom=homogeneity, stab=stability (hours), T=top, MWmiddle, B=bottom position ofcontainer S=stability sample taken at middle position of container RT=room temperature 

## The

## accuracy ofpreparation was considered acceptable ifthemean measured concentrations were 90-110% ofthe target concentration forsolutions, or85-1 15% 

## forsuspensions. Homogeneity was demonstrated ifthecoefficient ofvariation was :5 10%. Formulations were considered stable ifthe relative difference before and after storage was maximally 10%. 6.2. Test system Test System Rat: Cr1 WI(Han) (outbred, SPE-Quality). Untreated animals and virginfemaleswere used atinitiation ofthe study. Rationale This speciesand strain of rat has been recognized as appropriate forgeneral and reproductivetoxicitystudies. NOTOX BV has general and reproductive historical data inthis speciesfrom the same strainand source. Thisanimal model has been proven to be susceptibleto theeffects of reproductive toxicants. Source Charles RiverDeutschland, Sulzfeld, Germany. Age atstart Fo-treatment Approximately 10 weeks. Number of F 0

## -animals 40females and 40 males. Acclimatisation FO At least 5days priorto start oftreatment. Health check F 0 

## Ahealth inspection was performed priorto commencement of treatment toensure thattheanimals were inagood stateof health. Randomisation F 0 

## Priorto commencement oftreatment, bycomputer-generated random algorithm accordingto body weight,with all animalswithin ±

## 20% ofthe sex mean. Identification F 0 

## Earmark andtattoo.

Page 11

## ClI. PIGMENT BLUE 61 NOTOX Project 488231 Breeding procedures Following aminimum of 14 daysof exposure forthe males and females, one female was cohabitatedwith one male ofthesame treatment group, avoiding sibling mating (Charles River supplied non-litter mates). Detection of matingwas confirmed byevidence ofsperm inthevaginal lavage or bythe appearance ofan intravaginal copulatory plug. Thisday wasdesignated Day 0post- coitum. Once mating had occurred, the males and femaleswere separated. Amaximum of 14dayswas allowed for mating. Parturition F 0 

## The femaleswere allowed to litter normally. Day 1of lactationwas defined as thedaywhen alitterwasfound completed (i.e. membranes, placentas cleaned up, nest build up and/or feeding of pups started). Females that were littering were left undisturbed. Lactation F 0 

## Deficiencies inmaternal care, such as inadequate construction or cleaning ofthe nest, pups left scattered and cold, physical abuse of pups orapparently inadequate lactation orfeeding, were recorded. Number of pups Approximately 480 pups (40 litters x12 pups). Identification offspring On Day 1oflactation, all pups were individually identified by means of intracutaneous injection of Indian ink. 6.3. Allocation

Group Dose level Number ofanimals Animals numbers mg/kg b.w./day F 0

## males 

# IF 0

## females males females 1 0 10 10 01-10 41-50 

# 2 

# I 50 

# I 10 

# I 10 11-20 51-60 3 250 10 10 21-30 61-70 4 1000 1 10 10 1 31-40 71-80

*Dose 

## levels followingwere based on the dose range finding study(NOTOX Project 488232; seeAppendix5). 6.4. Animal husbandry Animals were housed inroom 16. Conditions Animals were housed inacontrolled environment, inwhich optimal conditions were considered to be approximately 15 airchanges per hour, atemperature of21 ±

## 3 0

## C(actual range: 19.8

- 

## 22.40

## C), arelative humidity of30

- 

## 70% (actual range: 41

-

## 87%) and 12 hours artificial lightand 12 hours darkness perday. Temporary fluctuationsfrom the light/dark cycle (with amaximum of 1hour) occurred dueto performance offunctionalobservations inthe room. Cleaning procedures inthe room might have causedthetemporary fluctuations abovethe optimal level of 70%for relative humidity. Based on laboratory historical data these conditionswere considered notto haveaffectedthe study integrity.

Page 12

## C.I. PIGMENT BLUE 61 NOTOX Project488231

Accommodation Pre-mating Animals were housed ingroups of5animals/sex/cage inMacrolon cages (MIV type, height 18cm). Mating Femaleswere cagedtogetherwith maleson aone-to-one-basis inMacrolon cages (Milltype, height 18 cm). Post-mating Maleswere housed intheir homecage (Macrolon cages, MIV type, height 18 cm) with amaximum of5animals/sex/cage. Females were individually housed inMacrolon cages(Milltype, height 18 cm). Lactation Offspring was keptwith the dam until termination. General Sterilised sawdustas bedding material (Litalabo, 

# S.P.P.S., Argenteuil, France) and paper ascage-enrichment (Enviro-dri, Win. Lillico &Son (Wonham Mill Ltd), Surrey, United Kingdom) were supplied. Certificates ofanalysis were examined and then retained inthe NOTOX archives. During overnightactivity monitoring, animalswere housed individually inMacrolon cages (MiIl type; height 15 cm)with sterilised sawdustas bedding material. No cage-enrichment was provided during ovemnight activitymonitoring. Diet Free accessto pelleted rodentdiet (SM R/M-Z from SSNIFF® Spezialdiaten GmbH, Soest, Germany). Resultsofanalyses for nutrients and contaminants ofeach batch were examined and archived. Water Free access totap-water. Certificates ofanalysis (performed quarterly) wereexamined and archived. Analysisofbedding, paper, diet and waterdid not reveal any findingsthatwere considered to have affected study integrity. 6.5. Treatment 6.5.1. Parental animals Method Oral gavage, using aplasticfeeding tube. Formulations were placed on amagnetic stirrer during dosing. Frequency Once dailyfor

# 7days perweek, approximately the same time each daywith amaximum of6hours difference betweenthe earliest and latestdose.Animals weredosed upto the day priorto scheduled necropsy. Exposure period Males were exposed for30 days, i.e.2weeks priorto mating, during mating, and up totermination. Females were exposedfor 41-45 days, i.e. during 2weeks prior to mating, during mating, during post-coitum, and during at least

# 4days of lactation. Female 78was not dosed forone occasionas thisfemale was littering atthe time ofdosing. Dose volume 

# 5mL/kg bodyweight.Actual dosevolumes were calculated according tothe latest bodyweight. 6.5.2. Offspring Offspring was nottreated.

Page 13

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 6.6. Observations The following animalswere selected (first5mated males andfirst 5femaleswith live offspring) forfunctional observations, clinical laboratory investigations, macroscopic examination and determination oforgan weights. GroupAnimals numbers

SMales 

## (n=5) Females (n=5) 1 3 7 10 42 1;43 4 48 50 2 11 12 114 1 15~ 16 52 155 56157-1-58 3 1 2 31 41 5 1 63 64 65 67 4 132 34 135 136 37 73 174 75 76 77* 6.6.1. Parental animals Mortality/Viability At least twice daily. Clinical signs At least oncedaily, detailed clinicalobservationswere made inall animals. Once priorto startoftreatment and at weeklyintervals thiswasalso performed outside the home cage inastandard arena. Arena observationswere notperformed when the animals were mating, or housed individually.The time ofonset, degreeand duration was recorded. Allsymptoms were recorded and graded according to fixedscales: Maximum grade 1: grade 0=absent, grade 1= 

## present Maximum grade 3or4: grade 1=

## slight, grade 2 =

## moderate, grade3 =

## severe, grade 4= 

## very severe Cage debris of pregnant femaleswere examined to detectabortion or premature birth, ifapplicable. Signsofdifficult or prolonged parturition were recorded, ifapplicable. Functional Observations The following tests were performed inthefirst 5mated malesand thefirst 5females with liveoffspring, from each group:

- hearing ability, pupillary reflex, static righting reflex and grip strength (Score 0 =

## normal/present, score 1 abnormal/absent).

- 

## motor activitytest (recording period: 12 hours during overnight forindividual animals, using acomputerised monitoring system, Pearson Technical Services, Debenham, Stowmarket, England). During the motoractivity test, maleswere caged individuallyand females were caged with theiroffspring. The assigned maleswere tested during week 4oftreatment and the assigned femaleswere tested during lactation (all before blood sampling). Inorderto avoid hypothermia ofpups, damswere removedfrom the pupsfor not more than 30-40 minutes. Bodyweights Males and femaleswere weighed onthe first day ofexposure and weekly thereafter. Mated femaleswere weighed on Days 0,4,7, 11, 14,17and 20 ofgestation, and during lactation on Days 1and 4.

Page 14

## C.I. PIGMENT BLUE 61 NOTOX Project 488231

Food consumption Weekly, 

# formales and females. Food consumption was not recorded duringthe breeding period. Food consumption ofmated females was measured on Days 0,4,7,11, 14,17 and 20 postcoitum and afterdelivery on Days 1and 4post-partum. Water consumption Subjective appraisal was maintained during the study, but no quantitative investigation introduced as no effectwas suspected. Reproduction processes Male number pairedwith, matingdate, confirmation of pregnancy, and deliveryday was recorded. 6.6.2. Offspring Each litterwas examined to determine the following, if practically possible: Mortality/Viability The numbersof live and dead pups at the First LitterCheck ( 

## checkat Day 1of lactation) and dailythereafter were determined. If possible, defects orcause ofdeath were evaluated. Clinicalsigns At least oncedaily, detailed clinicalobservations were made inall animals. Bodyweights Live pupswereweighed during lactation on Days 1and4. Sex Was determined forall pups on Days 1and4of lactation (by assessment ofthe ano-genitaldistance).

Page 15

## C.I. PIGMENT BLUE 61 NOTOX 

## Project488231 6.7. Clinical laboratory investigations Blood samples were collected from thefirst 5mated males and thefirst 5females with live offspringfrom each group under iso-flurane (Abbott Laboratories Ltd., Zwolle, The Netherlands) anaesthesia immediately prior to scheduledpostmortem examination, between 7.00and 10.30 am. Theanimals werefasted overnight (with amaximum of20 hours) before blood sampling, butwaterwas provided. Blood sampleswere drawn fromthe retro-orbital sinus and collected into tubes (Greiner Bio-One, Bad Hailer, Austria) prepared with EDTAfor haematological parameters (0.5 ml), with citratefor clotting tests (0.9 ml) and Li-heparin treated tubesforclinical biochemistry parameters (0.5 ml). The following parameters were determined: Parameter Abbreviation Unit Haematologya 

## White blood cells WBC 10 9

## /LDifferential leucocyte count %WBC neutroph is, lymphocytes, monocytes, eosinophils, basophils 

## Red blood cells 1 0' 2



### /L 

## Reticulocytes %RBC Red blood cell distribution width RDW% Haemoglobin mmol/L Haematocrit L/L Mean corpuscularvolume MCV flMean corpuscular haemnoglobin MCH find Mean corpuscular haemoglobin MCHC mmol/L concentration Platelets 10 9 

## '/L ClottingPoeta Prothrombin time PT s Activated Partial thromboplastin time APTT s ClinicalBiochemistryc 

## Alanine aminotransferase ALAT U/L Aspartate aminotransferase ASAT U/L Alkaline phosphatase ALP UIL Total Protein gIL Albumin g/L Total Bilirubin pinolIL Urea minol/L Creatinine pinol/L Glucose minolL Cholesterol minoilL Sodium inmolIL Potassium minol/L Chloride inmol/L Calcium mmol/L Inorganic Phosphate lnorg. Phos inmol/L

'Instrumentation: AOVIA 120 (Bayer Diagnostics) 'Instrumentation: STA Compact (Roche Diagnostics). 'Instrumentation: Olympus AU400 (Goffin Meyvis)



## Page 16

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 6.8. Pathology Parental Animals 6.8.1. Termination All animalswere fasted overnight (with amaximum of20 hours) priorto planned necropsy, but waterwas provided. Females Which Deliver On lactation Day 5,the Fo-females were anaesthetised using iso-flurane (Abbott Laboratories Ltd., Zwolle, The Netherlands) and subsequently exsanguinated. Organ weightswere collected and tissueswere preserved forpossible future histopathological examination asdescribed in thefollowing paragraphs. The numberofformer implantation siteswere counted and recorded. Corpora lutea were alsocounted and recorded. Gross lesionswere saved for possiblefuture histopathological examination. Females Which Failed to Deliver On post-mating Day25-27 (females with evidence of mating), the F 0 

## -femaleswhich had not deliveredwere anaesthetised using iso-flurane (Abbott Laboratories Ltd., Zwolle, The Netherlands) andsubsequently exsanguinated. No organ weights were collected. Tissueswere preserved for possiblefuture histopathological examination asdescribed inthe following paragraphs, with the following exceptions: Nongravid uteriwere stained using the Salewski technique (Salewski, 1964) inorderto determine any former implantation sites (Salewski staining prepared at NOTOX using Ammoniumsulflde-solution 20% (Merck, Darmstadt, Germany) and Milli-Rowater (Millipore Corporation, Bedford, USA)). Ifevidence of macroscopic implantations was present, the number of implantation sitesand corpora lutea was recorded. Gross lesionswere saved for possiblefuture histopathological examination. Termination Procedures for Fo 0

## -Males Following completion ofthe mating period (29days of dose administration), the F 0

## -males were anaesthetised using iso-flurane (Abbott Laboratories Ltd., Zwolle, The Netherlands) and subsequently exsanguinated. Organ weightswere collected and tissueswere preserved for possiblefuture histopathological examination as described inthefollowing paragraphs.

Page 17

## C.I. PIGMENT BLUE 61 NOTOX Project488231 6.8.2. Macroscopic examination After sacrifice all parental animals weresubjected to macroscopic examination ofthecranial, thoracic and abdominal tissuesand organs, with specialattention being paidto the reproductive organs. Descriptions ofall macroscopic abnormalities was recorded. Samples ofthefollowing tissuesand organswere collected and fixed inneutral phosphate buffered 4%formaldehyde solution (Klinipath, Duiven, The Netherlands): From thefirst 5males pergroup and thefirst5femaleswith live offspring per group:

Identification marks: not processed Ovaries Adrenal glands Pancreas Aorta Peyer's patches (jejunum, ileum) ifdetectable Brain (cerebellum, mid-brain, cortex) Pituitary gland Caecum Preputial gland Cervix Prostate gland Clitoral gland Rectum Colon (Salivary glands

- 

## mandibular, sublingual) Coagulation gland Sciatic nerve Duodenum Seminal vesicles Epididymides* (Skeletal muscle) (Eyes* with optic nerve (if

## detectable) and (Skin) Harderian gland) Spinal cord -cervical, midthoracic, lumbar (Female mammary gland area) Spleen (Femur includingjoint) Sternum with bone marrow Heart Stomach Ileum Testes* Jejunum Thymus Kidneys Thyroid including parathyroid (if detectable) (Larynx) (Tongue) (Lacrimal gland, exorbital) Trachea Liver Urinary bladder Lung, infusedwith formalin Uterus Lymph nodes

- 

## mandibular, mesenteric Vagina (Nasopharynx) Allgross lesions Oesophagus From all remaining animals: Cervix Prostate gland Clitoral gland Seminal vesicles Coagulation gland Testes* Epididymides* Uterus Ovaries Vagina Preputial gland Allgross lesions Identification marks: not processed

*Fixed 

### inmodified Davidson's solution (prepared at NOTOX using Formaldehyde 37.40%, Ethanol, Aceticacid (glecial)(al Merck, Darmstadt, Germany) and Milli-Ro water(Millipore Corporation, Bedford, USA)) and transferred to formalln after fixation forat least24 hours. Tissues/organs mentioned inparentheseswere notexamined bythe pathologist, since no signsoftoxicity were noted at macroscopicexamination.



## Page 18

## C.I. PIGMENT BLUE 61 NOTOX Project488231 6.8.3. Organweights The following organ weights (and terminal bodyweight) was recorded: From thefirst 5males per group andthefirst 5femaleswith liveoffspring per group: Adrenal glands Liver Brain Spleen Epididymides Testes Heart Thymus Kidneys From all remaining males: Epididymides Testes 6.8.4. Histatechnology Allorgan and tissuesamples. asdefined under Histopathology (following),were processed, embedded and cutat athickness of2-4 micrometers and stained with haemnatoxylin and eosin (Klinipath, Duiven, The Netherlands). Oftheselected 5males/group ofthe control and highdose group, additional slides ofthetestes were prepared to examine staging of spermatogenesis. Thetestes was processed, sectioned at 3-4 microns, and stainedwith PAS/haematoxylin (Klinipath, Duiven, The Netherlands). 6.8.5. Histopathology The delegated phase was performed bythe Principal Investigatorfor Histopathology. See Appendix 4. Thefollowing slideswere examined byapathologist:

-The 

## preserved organs and tissues oftheselected animals ofGroups 1and 4.

-The 

## additional slidesofthe testesoftheselected 5males/group of Groups 1and 4to

examine staging ofspermatogenesis

-All 

## gross lesions ofall animals (all dose groups)

-The 

## reproductive organs* ofall animals thatfailed to mate, conceive, sire or deliverhealthy

offspring. These animals included:

- 

## Group 1 Male 1and Female 41 (non-pregnant).

- 

## Group 2 Male 14and Female 54 (non-pregnant).

- 

## Group 3 Male 26 and Female 66 (non-pregnant).

- 

## Group 4 Male 32 and Female 72 (implantation sitesonly). Histological examination was extendedto the mesenteric lymph nodes oftheselected males and females of Group 2based on possible treatment-related changes noted for mid and high dose animals. All abnormalitieswere described and included inthe report.An attempt was made tocorrelate gross observations with microscopic findings.
* 

## Reproductive organs included cervix, clitoralgland, coagulation gland, epididymides, ovaries, preputial gland, prostate gland, seminal vesicles, testis, uterus, and vagina.

Page 19

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 6.9. Pathology Offspring 6.9.1. Termination Pups were killed bydecapitation on Day 5of lactation orshortly thereafter. 6.9.2. Macroscopic examination Alloffspring wassexed and externally examined ifpractically possible. The stomach was examined forthe presenceof milk. Descriptions ofall macroscopicabnormalities were recorded. Ifpossible, defects orcause ofdeathwas evaluated. Anyabnormal pup, organ ortissuewas preserved in10% buffered formalin, for possible further examination. 6.10. Calculations Foreach dosegroup reproduction parameters were expressed intwo ways:

-

## As amean (with standard deviation) ofthe numberobserved foreach litter

- 

## Relative to asecond parameter and calculated on atotal group basis Foreach group the following calculationswere performed: Percentage mating Number offemales mated Ix 100 Number offemales paired Fertility index Number ofpregnantfemales x 100 Number offemales paired Conception rate Number oforeanantfemales x 100 Numberoffemales mated Gestation index Number offemales bearing~livePups x 100 Number ofpregnantfemales Duration ofgestation Number ofdays between confirmation ofmating and the beginning ofparturition Percentage live males Number of live male pups at First Litter Checkx 100 at First LitterCheck Number of live pupsat First Litter Check Percentage live females Number of livefemale pups at First Litter Checkx 100 at First LitterCheck Number of live pups at First Litter Check Percentage ofpostnatal Number ofdead ouPs on Day 4ipostpartumx 100 loss Days 0-4postpartum Number of live pups at First LitterCheck Viability index Number of livepups on Day 4Postpcartumx 100 Number of pups born alive

Page20

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 6.11. Electronic data capture Observations/measurements inthe studywere recorded electronically using the following programme(s):

- 

## REES version 1.5 (REES Scientific, Trenton, NJ, USA): Environmental monitoring.

-

## TOXOATA version 8.0 (NOTOX B.V., 's-Hertogenbosch, The Netherlands): Mortality/Clinical signs/ Body weights/Food consumption / Reproduction processes/Observations offspringI Organ weights.

-Advia 120 Version V.3.1.8.0.MS (Siemens Medical Solution Diagnostics, Mijdrecht, The Netherlands): Haemnatology.

-

## Sta Compact version 1.06.06 (Stago Instruments, Gennevilliers, France): Clotting parameters

-

## Olympus AU 400 version 8.2A (Goffin-Meyvis, Etten-Leur, The Netherlands): Clinical laboratory investigations.

- 

## MAMS version6.5 (Pearson Technical Services, Suffolk, Great Britain): Motoractivity measurement. -Empower 

## version 5.00 (Waters, Milford, MA, USA): System control, data acquisition and data processing for analytical chemistry. 6.12. Interpretation Thefollowing statistical methods will be usedto analysethe data:

- 

## Ifthevariables could beassumed to follow anormal distribution, the Dunnett-test (Dunnett, 1955) (many-to-one t-test) based on apooled varianceestimate was applied forthe comparison ofthe treated groups and the control groups for eachsex.

- 

## The Steel-test (Miller, 1981) (many-to-one ranktest)was applied ifthe datacould not be assumedto follow anormal distribution.

- 

## The Fisher Exact-test (Fisher, 1950) was appliedtofrequency data. All testswere two-sided and inall cases p< 

## 0.05 was accepted asthe lowest levelof significance. Nostatistical analysiswas performed on histopathologyfindings. 6.13. List ofdeviations 6.13.1. List of protocol deviations 1 Noorgan weight was determined ofthetestes and epididymides ofanimal 30. Evaluation: Sufficient datawere available forevaluation. 2 Afewtissueswere notavailable forhistopathology. Reasons forthis included that these tissueswere notdiscernableat necropsy ortrimming, orwere erroneously not collected at necropsy.Tissues are listed inraw dataand pathology report. Evaluation: Sufficientdata was availablefor evaluation. 3 Tissuesfor histopathology were preserved and organ weights were determined for another subsetof animals (out of20 selected males, 5weredifferent: Group 1males 4 and 5instead of7and 10, Group2male 13 instead of16, and Group 4males31 and 33 instead of36 and 37) than testedfor functional observations and clinical laboratory investigations. Evaluation: No toxicitywas noted upto the high dose levelfor all parameters; therefore itwas notessential to have results ofthese parameters on the same subset of animals. The study integritywas notadversely affected bythe deviations. 6.13.2. Listofstandard operating procedure deviations Therewere nodeviations from standard operating procedures.

Page21

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 7. RESULTS 7.1. Analysis of Dose Preparations (see also Appendix3) The concentrations analysed intheformulations of Group 2,Group 3and Group 4were in agreement withtarget concentrations (i.e. mean accuracies between 85% and 115%). No test substancewas detected intheGroup 1formulations. Theformulations ofGroup 2and Group4 were homogeneous (i.e. coefficientofvariation :5 10%). Formulations of Group 2and Group 4 were stable when stored atroom temperature for at least6hours. 7.2. Observations 7.2.1. Mortality No mortality occurred during the study period. 7.2.2. Clinical Signs Therewere no clinicalsigns oftoxicitynoted duringtheobservation period. Blue discolouration ofthefaeces was noted inall animals ofboth sexesat 50, 250,and 1000 mg/kg. Blue staining of several body parts (tail, back, mouth, snout, abdomen, forelegs, flank) was also observedat these dose levels but atalower incidence.These findings were considereddue to the staining properties ofthe test substance and notto betoxicologically relevant. Slight salivation was noted for one lowdosemale (1day),five mid dosemales (1 -3days), six high dose males (2-18 days) and two high dosefemales (1day). Salivationwas considered to be aphysiological responseto thetaste of theformulation rather than asign ofsystemictoxicity, consideringthe nature, minor severity oftheeffect and its time ofoccurrence (i.e. afterdosing). Therefore, thiswas considered nottoxicologically relevant. Incidental findingsthatwere noted included alopecia and scabbing on several body parts, rales, piloerection and abroken tailapex. These findings are commonly noted inrats ofthis ageand strain which are housed and treated underthe conditions inthis study. At the incidence observed, these were considered signs of no toxicologicalsignificance. 7.2.3. BodyWeights Body weightsand bodyweight gain oftreated animals remained inthe same range as controls overthestudy period. 7.2.4. Food Consumption Food consumption (absolute and relative) oftreated animals inallgroups remained inthesame range ascontrols overthe study period. Two female rats (numbers 55 and 60) ofthe lowdose groupshowed very low foodconsumption on Days 11-14 post-coitum. As bodyweight gainsforthese damswere normal, these low valueswere probably caused byaweighing error ratherthan being true valuesand were therefore considered nottoxicologicallyrelevant.

Page 22

## C.I. PIGMENT BLUE 61 NOTOX Project488231 7.2.5. Functional observations No changes wereobserved inhearing ability, pupillary reflex, static righting reflex and grip strength inthe animals treated with 0.1. Pigment Blue 61, when compared tocontrol animals. Thevariation inmotor activitydid not indicate arelation with treatment. 7.3. Clinical Laboratory Investigations 7.3.1. Haematology Haemnatological parameters oftreated rats were not affected bytreatment upto 1000 mg/kg. Femalestreated with 1000 mg/kg showed statisticallysignificantly lower prothrombin time than controls. Thisfinding isconsidered to be ofno toxicological significancesincethe valuewas within normal limits and no corroborative findingswere noted. Inaddition,the statistically significantlyincreased haemnatocrit valuefor lowdose females was notconsidered treatment related as no dose response relationship was noted. 7.3.2. Clinical Biochemistry Notoxicologically relevant changes occurred inclinical biochemistry parameters oftreated rats. Males treated with 1000 mg/kg had statisticallysignificantly highercholesterol measures than controls. This was considered to be ofnotoxicological significance as the valuewaswithin normal limits and no corroborativefindings were noted. 7.4. Pathology 7.4.1. Macroscopic Examination Notoxicologically relevant macroscopic findingswere noted upto 1000 mg/kg bodyweight/day. Greenish or bluish discolouration ofthe mesenteric lymph nodes was noted forthree lowdose animals, seventeen mid dose animalsand all highdose animals. Seven high dosefemales had bluish contents inthe gastro intestinaltract. Bluishdiscolouration ofthe tail was observed for twomales. These findingswere considered due to thestaining properties ofthetestsubstance andnot tobe toxicologically relevant. Incidental findings included benttail, hard yellowish noduleon the tail ofthe epididymidis, alopecia atseveral body parts, reduced seminal vesiclesize, dark redor reddish discolouration ofthe mandibular lymph nodes, reddish discolouration ofthe pancreas, and red-brown discolouration ofthe adrenal glands.These findingsare occasionally seen among rats used in thesetypes ofstudies and inthe absence ofcorrelated microscopicfindings theywere not considered changesoftoxicologicalsignificance.

Page23

## C.l. PIGMENT BLUE 61 NOTOX Project 488231 7.4.2. Organ Weights No toxicologically relevant organ weight changeswere noted foranimals treated up to 1000 mg/kg bodyweight/day. Thestatistically significantly increased relative spleen weight notedfor highdose males was considered notto be asign oftoxicity intheabsence ofcorroborative (microscopic) findings. 7.4.3. Microscopic Examination (see alsoAppendix 4) No treatment related histopathological changes were observed inthe tissues ofanyanimal receiving 0.1. Pigment Blue61. No abnormalities were noted on evaluation of PAS stained sections oftestes of F 0

## males.All seminiferous tubules were normal with respectto their stage inthe spermatogenic cycleand the integrity ofthe various celltypes present withinthe differentstages. No pathologicalfindings were present inthe reproductive organs ofmale andfemale animals thatfailed to produce offspring. Small amounts of blue material wereobserved inthe lumen oftheduodenum, ileum,jejunum, colon or rectum inafew group4animals. Blue material was also noted inthe superficialskin (keratin layer)ofthe tail inonegroup 4male.These findings are considered to represent the presence ofthe test substance and insomecases correlatedwiththe macroscopic observations. No microscopicfindings wereobserved inthe mesenteric lymph nodesto account forthe discolouration ofthesetissues recorded atnecropsy. Itisconsidered thatthe discolouration is due tothe presence ofthe testsubstance and that the pigment was lostfrom thetissues inthe processing orstaining procedures. Allother microscopic findings were considered to be incidental and unrelated to treatment with thetest substance. 7.5. Reproduction Reproduction parameters are described intables Iand 11: Table 1.Reproduction Data. Number offemales Group 1 Group2 Group 3 Group 4 Control 50 mg/kg 250 mg/kg 1000 mg/kg Paired 10 10 10 10 Mated 10 10 10 10 Non-pregnant 1 1 1 0 Pregnant 9 9 9 10 Implantations sitesonly 0 0 0 1(F72) Number offemaleswith 9 9 9 9 living pups atfirst litter check

Page24

## CAI. PIGMENT BLUE 61 NOTOX Project488231 Table i1. Fertility FO-generation. Group 1 Group2 Group 3 Group4 Control 50 mg/kg 250 mg/kg 1000 mg/kg Percentage mating 100 100 100 100 (Females mated /Females paired) *10 

## Fertility index 

## 90 90 90 100 (Females achieving apregnancy/Females paired) *100 

## Conception rate 90 90 90 100 (Females achieving apregnancy/Females mated) *100 

## Gestation index 100 100 100 90 (Number offemales with living pups at first litter check/ Number offemalespregnant) * 

### 100 

## Reproduction parameters were unaffected bytreatment up to 1000 mg/kg bodyweight/day. Notreatment related findings were observed for mating performance, fertility parameters, duration ofgestation, number ofdead and living pups atfirst littercheck, number of implantation sites and number ofcorpora lutea. 7.6. Breeding Data Breeding parameters were unaffected by treatment up to 1000 mg/kg bodyweight/day. Postnatal loss and viabilityindexwere similarfor thecontrol and treated groups. 7.7. Pup Development Development of pups was unaffected by treatment up to 1000 mg/kg bodyweight/day. (Mean) bodyweights were similarfor the control and treated groups. Incidental clinicalsymptoms consisted ofsmall size ofthe pup, red spot on the nose orhead, and pale appearance. At macroscopic examination, autolysis and absence of milk inthe stomach was notedforafew dead pups. No relationshipwith treatment wasestablished for these observations and theywere considered to be within the normal biologicalvariation for rats ofthis age and strain.

Page25

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 8. DISCUSSION AND CONCLUSION C.I. PIGMENT BLUE 61 was administered by dailyoral gavage to male and femaleWistar Han rats at dose levelsof50, 250 and 1000 mg/kg/day. The males were exposed for 2weeks prior to mating, during mating, and upto termination (for 30 days). Thefemales were exposedfor2 weeks priorto mating, during mating, during post-coitum, and at least 4days of lactation (for41- 45days). Formulation analysisshowedthat the formulations were prepared accurately, were homogeneous and were stableforat least 6hours at room temperature. Notreatment-related toxicologicallysignificantchanges were noted inanyofthe parameters investigated inthisstudy (i.e. mortality, clinical appearance, functional observations, body weight,food consumption, clinical laboratory investigations, macroscopicexamination, organ weights, microscopic examination, reproduction, breeding and pup development). From the results presented inthis report, the parental, reproduction, breeding and developmental NoObservedAdverse Effect Level (NOAEL) forClI. PIGMENT BLUE 61 of at least 1000 mg/kg/day was established.

Page 26

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 9. REFERENCES Dunnett C.W., AMultiple Comparison Procedurefor Comparing SeveralTreatments with a Control, J.Amer. Stat. Assoc. 50, 1096-1121 (1955). Fisher R.A., Statistical Methods for Research Workers, Oliverand Boyd, Edinburgh (1950). Miller R.G., Simultaneous Statistical Inference, SpringerVerlag, NewYork (1981). Salewski, E.Farbemethode zummakroskopischen Nachweis von lmplantationsstellen am Uterusder Ratte. [Staining method foramacroscopic

# testfor implantation sites intheuterus of the rat]. Naunyn

- 

## Schmiedebergs Archlyfor Experimentelle Pathologie und Pharmakologie, 247, 367 (1964).

Page27

## C.I. PIGMENT BLUE 61 NTXPoet483 APPENDIX 1IOO rjet483

APPENDIX 1 SUMMARY TABLES

-Page 1

-

## C.!. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1 CLINICAL SIGNS SUMMARY,WEEKLY MALES PRE MATING REPRO PHASE SIGN (MAX. GRADE) WEEK: 1. 1.. 4. (LOCATION)
GROUP II(CONTROL) Various

Broken (1) G:

. 1

(Tail apex) %,1 GROUP 2(50 MG/KG) Skin/fur

Scabs(3) G:

. 1

(Neck) %:

. 1 Bluestaining(1) G:11 Blue discolouration (1) G: 11 ill

(Faeces) %: PAA Secretion/excretion

Salivation(3) 0:

... 1 %:. 1 GROUP 3(250 MG/KG) Skin/fur

Alopecla (3) 0:...

(Forelegs) %/:

.... 1 Scabs (3) G:

. 1

(Neck) %:

.. 1. Blue staining (1) G: .1.

(Back) %:

.. 1 Blue staining (1) G:

....

(Tail) %:

.. Blue discolouration (1) G: II11

(Faeces) %: PAA Secretion /excretion

Salivation (3) G:

.. 1 %: .24 GROUP 4(1000 MG/KG) Skin/fur

Alopecia (3) G:
. .11

(Foreleg right) %:

. .11 Alopecia (3) G:

.. 121

(Forelegs) %:

.. 11 Scabs (3) G:

... 1

(Foreleg right) %:

... 1 Bluestaining (1) G: 1
. .. (Back) %: 1'. .. Blue staining (1) G:

.. 11 Bluediscolouration (1) G: 11 ill

(Faeces) %: PA Secretion/excretion

Salivation (3) G: I 1 %: .6 653

G: Median value ofthe highest Individual weeklygrades %:Percent ofaffected animals (OlIess than 5%, 1=between 5%and 15%,.,A=more than 95%)

Observation performed, sign not present

- Page 2 - 

## fr24Oct08 15h45

## C.I. PIGMENT BLUE 61 NOTOX 

## Project488231 APPENDIX 1 CLINICAL SIGNS SUMMARY,WEEKLY FEMALES PRE MATING REPRO PHASE SIGN (MAX. GRADE) WEEK:1. 1-.. (LOCATION) GROUP I(CONTROL) Skin/fur

Piloerection (1) G:

... 1 Alopecia (3) G: 11 1..

(Cheek left) %: 11 11
... Alopecia (3) G: .1 1..

(Cheek right) %: .1 1.. Alopecia (3) G: 11 11.

(Back) %: 23 32
... (Neck) %: 43 4441. .Alopecia(3) G:

. . . 22

(Abdomen) %

. ."..22 Alopecia (3) G:

..... 11

(Flank left) %

. ... 12 Alopecia (3) G:

..... 11

(Thigh hind left) %

. ... 12 Alopecia (3) G:

.. .

(Foreleg left) %

. .11. Alopecia(3) G:

.. ... 33

(Forelegs) %: 12 Scabs (3) G:

. . 1

(Mouth) %.

... 1. Scabs (3) G:

. 1.

(Snout) %

... GROUP 2(50 MG/KG) Skin/fur

Piioerection (1) G.

. %.1 Alopecia (3) G: 111..

(Neck) %: 12 1... Alopecia (3) G:

... 1

(Abdomen) %:

... 1. Blue staining (1) G: .1

(Tail) %

. 1. . . Blue discolouration (1) G: 11 11111

(Faeces) %: A AAAAA GROUP 3(250 MG/KG) Breathing

Rales (3) G:

. .12. %11 Skin/fur

Piloerectlon (1) G:

... .. %1. Alopecia (3) G, 111..

(Cheekrfight) %: 121.. Bluestaining(1) G:

... 1

(Mouth) %:

..... 1 Blue staining (1) G: 1....

(Snout) %: 1.... Bluestaining(1) G: 111..

(Back) %: 111.. Blue staining (1) G: 1

(Abdomen) %:

.... 1 Blue staining (1) G:

. .. 1

(Forelegs) %:...1 Bluediscolourstion(1) G: 11111

(Faeces) %: AA AAAAM

G-Medianvalue of the highest individual weekiygrades %:Percent ofaffected animais (O=iess than 5%, 1=between 5%and 15%..Amore than 95%)

Observation performed, sign not present

- Page 3 24Oct08 1.5h45

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1 CLINICAL SIGNS SUMMARY,WEEKLY FEMALES PRE MATING REPRO PHASE SIGN (MAX. GRADE) WEEK:1.1.4 (LOCATION)
GROUP 4(1000 MG/KG) Breathing

Rales (3) G: I. %: 1. Skin/fur

Alopecia (3) G:

... 1.

(Back) %

. . 1. Alopecia (3) G: 1.

(Neck) %: 1. Alopecia (3) G: .*.1

(Abdomen) 1. Alopecia (3) G:

.. .. 333

(Forelegs) %

. . .112 Alopecia (3) G:

.. .. 22

(Hindlegs) %..12 Scabs (3) G: I .1 (Mouth) %

.. 1. Blue staining (1) G: .1 1111.

(Tail) % .1 1111. Bluestaining (1) G:

... 1
...

(Flank ight) %

. 1... Blue d~scolouration (1) G: 11 11111

(Faeces) %: AA AAA Secretion /excretion

Salivation (3) G: 1 1. %:

. 1.

G: Median value ofthe highest individual weekly grades %:Percent ofeffected animals (O=Iess than 5%, I between5% and 15%..Amore than 95%)

Observation performed, sign not present -Page 4

- 100- 24Oct08 15h45

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX I BODY WEIGHTS (GRAM) SUMMARY MALES
GROUIP1 GROUP 2 GROUP 3 GROUP4 CONTROL 50 MG/KG 250MG/KG 1000 MG/KG
PRE MATING DAY 1 MEAN 302 301 303 303 WEEK I ST.DEV 15.1 13.9 8.4 9.2 N 10 10 10 10 DAY 8 MEAN 325 322 328 321 WEEK 2 ST.DEV 18.5 19.6 12.0 10.9 N 10 10 10 10 MATING PERIOD DAY I MEAN 346 345 352 340 WEEK 1 ST.DEV 21.0 23.1 14.6 10.6 N 10 10 10 10 DAY 8 MEAN 360 349 366 352 WEEK 2 ST.DEV 25.2 18.7 13.2 9.5 N 10 10 10 10 DAY 15 MEAN 378 372 383 369 WEEK 3 ST.DEV 26.9 30.3 16.8 9.3 N 10 10 10 10 

## FEMALES
GROUIP1 GROUP2 GROUP3 GROUP4 CONTROL 50MG/KG 250 MG/KG 1000 MG/KG
PRE MATING DAYl MEAN 193 194 193 192 WEEK 1 ST.DEV 8.1 7.5 6.9 6.2 N 10 10 10 10 DAY8 MEAN 198 203 199 198 WEEK 2 ST.DEV 8.5 9.1 6.4 5.6 N 10 10 10 10 MATING PERIOD DAYl MEAN 208 212 211 208 WEEK 1 ST.DEV 7.6 6.5 7.1 10.1 N 10 10 10 10

*r Dunnett-test based on pooledvariance significant at 5% ()or 1%

### (-) level

- Pages5

- JV~ 29Sep08 09h35

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1 BODY WEIGHTS (GRAM) SUMMARY FEMALES FO-GENERATION
GROUP 1 GROUP2 GROUP3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
POST COITUM DAY 0 MEAN 209 214 212 210 ST.DEV. 9.7 7.3 4.4 6.9 N 9 9 9 10 DAY 4 MEAN 226 230 230 228 ST.DEV. 11.9 10.2 7.0 5.8 N 9 9 9 10 DAY 7 MEAN 233 238 238 236 ST.DEV. 10.7 13.3 7.8 67 N 9 9 9 10 DAY 11 MEAN 246 251 253 249 ST.DEV 13.1 14.1 8.5 7.0 N 9 9 9 10 DAY 14 MEAN 258 263 265 259 ST.DEV. 18.5 15.8 10.1 9.1 N 9 9 9 10 DAY 17 MEAN 281 285 287 280 ST.DEV. 17.8 18.4 12.7 15.5 N 9 9 9 10 DAY 20 MEAN 314 317 319 312 ST.DEV. 26.3 21.7 17.8 28.9 N 9 9 9 10 LACTATION DAY I MEAN 241 245 248 238 ST.DEV. 14.5 14.7 8.0 7 1 N 9 9 9 9 DAY 4 MEAN 252 250 256 251 ST.DEV. 15.3 17.6 6.8 10.6 N 9 9 9 9

/D unnet-test based on pooled variance significantat 5%

## ()or 1%

### (**) level Explanations for excluded dataare listed inthe tables ofthe individual values

- Page 6

- ON 23Oct05 16h07

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 BODY WEIGHT GAIN 

## (%) SUMMARY MALES
GROUPi GROUP2 GROUP3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG

PRE MATING DAY1 MEAN 0 0 0 0 WEEK 1 ST.OEV 0.0 0.0 0.0 0.0 N 10 10 10 10 DAY 8 MEAN 8 7 8 6 WEEK 2 ST.DEV 2.5 2.3 1.7 2.0 N 10 10 10 10
MATING PERIOD DAYl MEAN 15 14 16 12 WEEK 1 ST.DEV 4.3 2.9 24 3.2 N 10 10 10 10 DAYB8 MEAN 19 16 21 16 WEEK 2 ST.OEV 5.9 4.6 2.1 4.1 N 10 10 10 10 DAY 15 MEAN 25 24 26 22 WEEK 3 ST.DEV 6.4 4.9 3.2 3.1 N 10 10 10 10 

### FEMALES
GROUP 1 GROUP2 GROUP 3 GROUP4 CONTROL 60 MG/KG 250 MG/KG 1000 MG/KG

PRE MATING DAYl MEAN 0 0 0 0 WEEK 1 ST.DEV 0.0 0.0 0.0 0.0 N 10 10 10 10 DAY8 MEAN 3 4 4 3 WEEK2 ST.DEV 1.8 1.8 30 1.9 N 10 10 10 10 MATING PERIOD DAYl MEAN 7 9 10 8 WEEK 1 ST.DEV 22 3.0 5.0 4.4 N 10 10 10 10

*1'Dunnett-test based on pooled variance significant at 5% 

### ()or 1%

### ()level

- Page 7

- 29Sep08 091136

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1
BODY WEIGHT GAIN

## (%) SUMMARY FEMALES FO-GENERZATION
GROUP 1 GROUP2 GROUP 3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 

### MG/KG
POST COITUM DAYO0 MEAN 0 0 0 0 ST.DEV. 0.0 0.0 0.0 0.0 N 9 9 9 10 DAY4 MEAN 8 7 8 9 ST.DEV. 2.0 1.9 2-3 1.8 N 9 9 9 10 DAY7 MEAN 11 11 12 13 ST.DEV. 2.8 3.0 2.6 1.8 N 9 9 9 10 DAY11 MEAN 18 17 19 19 ST.DEV. 3.2 34 2.9 2.8 N 9 9 9 10 DAY14 MEAN 23 23 25 23 ST 0EV. 5.9 4.2 4.2 4.9 N 9 9 9 10 DAY17 MEAN 34 33 35 34 ST.DEV. 5.8 5.1 64 8.2 N 9 9 9 10 DAY20 MEAN 50 48 50 49 ST.DEV. 10.4 6.5 9.1 14.6 N 9 9 9 10 LACTATION DAYl MEAN 0 0 0 0 ST.DEV. 0.0 0.0 00 0.0 N 9 9 9 9 DAY4 MEAN 5 2 3 5 ST.DEV. 3.0 2.5 2.8 2.8 N 9 9 9 9

IDunnett-test based on pooled vaniance significantat5% 

### ()or 1 %

### (**) level Explanations for excludeddata are listed in the tables ofthe individual values

- Page 8

- 23Oct08 16h07

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1 FOOD CONSUMPTION (GIANIMAL/DAY) SUMMARY MALES
GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
PRE MATING DAYS 1-8 MEAN 21 21 22 21 WEEKS 1-2 ST.DEV 2.2 0.0 0.1 0.4 N(CAGE) 2 2 2 2 DAYS 8-15 MEAN 22 22 22 22 WEEKS 2-3 ST.DEV 2.7 0.5 0.2 0.2 N(CAGE) 2 2 2 2 MEAN OF MEANS OVER PRE MATING MEAN 22 21 22 22 

## FEMALES
GROUPI GROUP2 GROUP3 GROUP4 CONTROL 50MG/KG 250 MG/KG 1000 MG/KG

PRE MATING DAYS 1-83 MEAN 14 14 14 14 WEEKS 1-2 ST.DEV 0.6 0.4 0.5 0.3 N(CAGE) 2 2 2 2 DAYS 8-15 MEAN 15 14 15 15 WEEKS 2-3 ST.DEV 0.5 0.2 0.5 0.4 N (CAGE) 2 2 2 2 MEAN OF MEANS OVERPRE MATING MEAN 14 14 14 15

/~Dunnett-test based on pooled variance significant at5% 

## ()or 1%()level

- Page 9

- '29Sep08 

### 09h35

## C.I. PIGMENT BLUE 61 NOTOX 

## Project488231 APPENDIX 1
FOOD CONSUMPTION (G/ANIMALIDAY) SUMMARY FEMALES FO-GENERATION

GROUP 1 GROUP2 GROUP 3 GROUIP4 

### CONTROL 50MG/KG 250 MG/KG 1000MG/KG
DAYS 0-4 MEAN 17 17 17 17 ST DEV. 1.4 2.5 1.9 1.3 N 9 9 9 10 DAYS 4-7 MEAN 18 18 19 19 ST.DEV. 1.9 3.0 2.1 0.8 N 9 9 9 10 DAYS 7-1l MEAN 19 18 20 19 ST.DEV. 14 1.7 2.0 1.7 N 9 9 9 10 DAYS 11-14 MEAN 20 17 23 20 ST.DEV. 3.2 7.7 5.1 1.8 N 9 9 9 10 DAYS 14-17 MEAN 20 21 22 20 ST.DEV. 2.9 3.6 18 2.6 N 9 9 9 10 DAYS 17-20 MEAN 22 22 22 21 ST.DEV. 3.3 3.3 17 2.8 N 9 9 9 10 MEAN OF MEANS 20 19 20 19 

### LACTATION DAYS 1-4 MEAN 31 27 26 31 ST DEV. 7.1 8.5 5.6 10.7 N 9 9 9 8

/Dunnett-test based on pooled variance significantat 5%

### ()or 1 %

### ()level Explanationsforexcluded data are listed inthe tables ofthe individual values -Page 10 -h 23Oct08 16h07

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1
RELATIVE FOOD CONSUMPTION (GIKG 

## BODYWEIGHTIDAY) SUMMARY MALES
GROUP 1 GROUP 2 GROUP3 GROUP4 CONTROL 50MG/KG 250 MG/KG 1000 MG/KG

PRE MATING DAYS 1-8 MEAN 66 65 66 66 WEEKS 1-2 ST.DEV 3.5 1.7 1.3 0.9 N(CAGE) 2 2 2 2 DAYS 8-15 MEAN 69 68 68 68 WEEKS 2-3 ST.DEV 5.0 0.2 1.5 0.3 N(CAGE) 2 2 2 2 MEAN OF MEANS OVER PRE MATING MEAN 67 66 67 67 

## FEMALES
GROUP 1 GROUP 2 GROUP 3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG

PRE MATING DAYS 1-8 MEAN 71 71 71 72 WEEKS 1-2 ST.DEV 4.3 1.9 3.1 0.3 N(CAGE) 2 2 2 2 DAYS 8-15 MEAN 74 71 73 75 WEEKS 2-3 ST.DEV 4.0 1.1 3.3 1.1 N(CAGE) 2 2 2 2 MEAN OF MEANS OVER PRE MATING MEAN 73 71 72 73

/~Dunnett-test based on pooled variancesignificant at5% ()or I% ()level -Page 11- 29Sep08 09h35

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 RELATIVE FOOD CONSUMPTION (GIKG BODYWEIGHT/DAY) SUMMARY FEMALES FO-GENERATION
GROUP 1 GROUP2 GROUP3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
POST COITUM DAYS 0-4 MEAN 75 74 75 74 ST.DEV. 5.1 8.3 6.4 5.5 N 9 9 9 10 DAYS 4-7 MEAN 79 76 78 79 ST.DEV. 7.7 8.5 7.1 3.2 N 9 9 9 10 DAYS 7-11 MEAN 78 73 77 77 ST.DEV. 4.6 7.6 59 57 N 9 9 9 10 DAYS 11-14 MEAN 79 64 85 78 ST.DEV 9.3 27.5 17.9 6.7 N 9 9 9 10 DAYS 14-17 MEAN 72 74 75 72 ST DEV. 9.6 7.9 5.1 6.7 N 9 9 9 10 DAYS 17-20 MEAN 70 69 69 67 ST.DEV. 7.5 5.8 2.6 5.1 N 9 9 9 10 MEAN OF MEANS 76 72 77 74 LACTATION DAYS 1-4 MEAN 122 105 102 124 ST.DEV. 24.8 263 21.7 39.1 N 9 9 98

SDunnett-test based on pooled variance significant at5%

### ()or 1%

### ()level Explanations forexcluded data are listed inthe tablesof the individual values

- Page 12

- 23Oct08 16h07

## 0.1. PIGMENT BLUE61 NOTOX Project 488231 APPENDIX 1 FUNCTIONAL OBSERVATIONS SUMMARY MALES
GROUP 1 GROUP 2 GROUP3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
AT WEEK 4 HEARING MEDIAN 0 0 0 0 SCORE 0/1 N 5 5 5 5 PUPIL L MEDIAN 0 0 0 0 SCOREO/1 N 5 5 5 5 PUPILR MEDIAN 0 0 0 0 SCOREO/1 N 5 5 5 STATIC R MEDIAN 0 0 0 0 SCOREO011 N 5 5 5 5 GRIP MEDIAN 0 0 0 0 SCOREO/1 N 5 5 5 5 

## FEMALES
GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
AT WEEK 4 HEARING MEDIAN 0 0 0 0 SCOREO/1 N 5 5 5 5 PUPILL MEDIAN 0 0 0 0 SCORE 0/1 N 5 5 5 5 PUPILR MEDIAN 0 0 0 0 SCOREO/1 N 5 5 5 5 STATIC R MEDIAN 0 0 0 0 SCOREO/1 N 5 5 5 5 GRIP MEDIAN 0 0 0 0 SCOREO/1 N 5 55

+/++ Steel-test significant 

### at 5% (4)or1%(++) 

### level

- Page 13

- 08Oct08 12h2l

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 MOTOR ACTIVITY MEASUREMENTS SUMMARY MALES WEEK 4 GROUP 1 GROUP 2 GROUP 3 GROUP4 CONTROL 50MG/KG 250 MG/KG 1000 MG/KG 

## total high MEAN 3328 2361 2081 1777 

## sensorcount ST.DEV 1736 578 573 581 N 5 5 5 5 total low MEAN 4533 4690 4204 4759 

## sensorcount ST.DEV 1009 929 266 1030 N 5 5 5 5



## FEMALES LACTATION GROUP 1 GROUP 2 GROUP 3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
total high MEAN 2238 1917 2124 1422 sensor count ST.DEV 617 1072 1239 644 N 5 5 55 total low MEAN 16563 6827 18471 18503 sensor count ST.DEV 3736 4455 6655 6789 N 5 5 55

I~Dunnett-test basedon pooled variance significantat5%()or 1 %

## ()level

- 

### Page 14-

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX I HAEMATOLOGY SUMMARY MALES
GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
END OF TREATMENT WBC MEAN 7.1 8.5 8.3 7.6 10E9/L ST.DEV 1.4 1.1 1.6 1.4 N 5 5 5 5 Neutrophils MEAN 15.4 13.6 15.9 12.1 %WBC ST.DEV 2.7 2.3 3.4 3.8 N 5 5 5 5 Lymphocytes MEAN 80.7 83.1 80.5 84 8 %WVBC ST.DEV 3.2 2.5 4.2 3.8 

### N 5 5 5 5 Monocytes MEAN 2.2 2.2 2.3 1.9 %WBC ST.DEV 0.3 0.5 0.7 0.6 N 5 5 5 5 Eosinophils MEAN 1.3 0.8 0.9 0.8 %WBC ST.DEV 0.6 0.3 0.2 0.4 N 5 5 5 5 Basophils MEAN 0.4 0.4 0.4 0.4 %WBC ST.DEV 0.1 0.1 0.1 0.2 N 5 5 5 5 Red blood cells MEAN 8.11 8.67 8.76 8.69 10E12/L ST.DEV 0.44 0.53 0.51 0.31 N 5 5 5 5 Reticulocytes MEAN 2.8 2.7 2.6 2.4 %RBC ST.DEV 0.3 0.7 0.3 0.3 N 5 5 5 5 ROW MEAN 12.7 13.6 12.8 15.0 %ST.DEV 0.9 2.1 0.4 5.0 N 5 5 5 5 Haemoglobin MEAN 9.7 9.9 10.1 9.9 mmolIL ST.DEV 0.4 0.6 0.4 0.2 N 5 5 5 5 Haematocrit MEAN 0.424 0.444 0455 0.448 L/L ST.DEV 0.025 0029 0.020 0.010 N 5 5 5 5 MCV MEAN 52.4 51.2 51 9 51.6 f'L ST.DEV 0.9 1.7 0.9 1.0 N 5 5 5 5 MCH MEAN 1.20 1.14 1.15 1.14 fmol ST.DEV 0.04 007 0.03 0.034 N 5 5 5 5 MCHC MEAN 22.91 22.27 22.12 22.09 mmol/L ST.DEV 0.68 066 0.30 0.48 N 5 5 5 5 Platelets MEAN 805 862 873 981 10E9/L ST.DEV 179 93 90 71 IN 5 5 5 5 PT MEAN 17.9 18.1 17.4 17.6 s ST.DEV 1.4 0.9 0.5 0.6 N 4 5 5 5 APTT MEAN 18.9 20.7 19.2 19.4 s ST.DEV 1.1 1.7 2.8 2.4 IN 4 5 5 5

+/++ Steel-test significantat 5% (+)or 1 %(++) level /~Dunnett-test based on pooled variance signlificanlt at 5% ()or 1 %

## ()level

- Page 15

- 29Sep08 09h39

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1 HAEMATOLOGY SUMMARY FEMALES
GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50MG/KG 250 MG/KG 1000 MG/KG

END OF TREATMENT WBC MEAN 4.0 4.0 4.5 5.7 10E9/L ST.DEV 0.5 1.4 1.7 1.1 N 5 4 5 5 Neutrophils MEAN 21.9 26.1 28.3 24.8 %WBC ST.DEV 6.3 15.6 30 9.8 N 5 5 5 5 Lymphocytes MEAN 74.6 71.4 68.6 73.2 %WBC ST.DEV 5.9 15.5 3.1 8.7 N 5 5 5 5 Monocytes MEAN 2.6 2.0 2.1 1.4 %WVBC ST.DEV 0.9 0.65 0.6 1.0 N 5 5 5 5 Eosinophils MEAN 0.7 0.3 0.6 0.5 %WGC ST.DEV 0.6 0.4 0.5 0.5 N 5 5 55 Basophils MEAN 0.2 0.1 0.4 0.1 %WBC ST.DEV 0.2 0.2 0.4 0.1 N 6 5 5 5 Red blood cells MEAN 7.29 7,59 7.32 7.62 10E12/L ST.DEV 0.17 0.47 0.28 0.23 N 5 5 5 5 Reticulocytes MEAN 4.8 4.8 5.7 5.3 %RBC ST.DEV 1.0 1.9 1.9 0.9 N 5 5 5 5 ROW MEAN 14.4 15.6 158 14.0 %ST.DEV 1.8 3.7 1.5 1.2 N 5 5 5 5 Haemoglobin MEAN 689 9.3 8.8 9.2 mmol/L ST.OEV 0.3 0.1 04 0.2 N 5 5 5 5 Haemnatocrit MEAN 0.386 0.405 0.386 0.405 LUL ST.DEV 0.008 0.009 0.014 0.014 N 5 5 5 5 MCV MEAN 52.9 535 52.7 53.1 fL_ ST.DEV 1.2 2,2 04 1.1 N 5 555 MCII MEAN 122 1.23 120 1.20 fmol ST.DEV 0.04 0.08 0.03 0.05 

### N 5 5 

### 5 5 MOHO MEAN 2309 22.94 2276 22.64 mrnol/L ST.OEV 0.56 0.55 0.64 0.58 N 5 5 5 5 Platelets MEAN 1117 1320 1385 1195 10E9/L ST.DEV 315 95 274 322 N 5 5 5 5 PT MEAN 17.1 16.5 16.3 15.6 sST.DEV 0.5 06 0.7 0.5 N 4 5 5 5 APTT MEAN 17.7 18.4 16.4 15.9 s ST.DEV 23 3.4 1.5 3.4 N 4 5 55

+/++ Steel-test significant at 5%(+) or 1%(++) level

SDunnett-test based on pooled variance significant at 5%

## ()or 1%

### (**) level
-Page 16 - N10- 29Sep08 09h39

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 CLINICAL BIOCHEMISTRY SUMMARY MALES
GROUPI1 GROUP2 GROUP3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
END OF TREATMENT ALAT MEAN 44.7 52.0 46.8 47.8 UIL ST.OEV 7.5 14.1 6.7 6.2 N 5 5 5 5 ASAT MEAN 81.9 82.9 79.7 

### 82.6 UIL ST.DEV 17.0 10.1 5.3 5.5 N 5 5 5 5 ALP MEAN 154 173 150 143 U/L ST.DEV 40 28 23 33 N 5 5 5 5 Total protein MEAN 63.3 65.2 65.4 66.2 gIL ST.DEV 2.8 2.3 2.1 1.3 N 5 5 5 5 Albumin MEAN 33.4 33.7 

### 34.0 34.3 gIL ST.DEV 1.3 0.8 0.5 0.4 N 5 5 5 5 Total bilirubin MEAN 2.2 2.4 2.2 2.0 umol/L ST.DEV 0.3 0.3 0.3 0.2 N 5 5 5 5 Urea MEAN 8.8 9.5 8.6 8.4 mmol/L ST.DEV 2.3 1.3 0.6 1.1 N 5 5 5 5 Creatinine MEAN 42.8 46.0 46.3 43.0 umol/L ST.DEV 1.7 6.8 4.7 1.5 N 5 5 5 5 Glucose MEAN 8.07 7.49 7.32 7.20 mmol/L ST.OEV 1.49 1.70 0.98 1.56 N 5 5 5 5 Cholesterol MEAN 1.65 1.97 1.88 

### 2.13 mmol/L ST.DEV 0.31 0.20 0.27 0.33 N 5 5 5 5 Sodium MEAN 142.3 142.0 1426 143.4 mmolIL ST.DEV 0.4 0.7 0.8 1.7 N 5 5 5 5 Potassium MEAN 3.77 4.03 3.81 4.03 mmolIL ST.DEV 0.23 0.16 0.07 0.26 N 5 5 5 5 

### Chloride MEAN 104 104 105 105 mmol/L. ST.IJEV 1 0 1 1 N 5 5 5 5 Calcium MEAN 2.78 2.71 2.73 2.73 mmolIL ST.DEV 0.04 0.05 0.08 0.04 N 5 5 5 5 Inorg.Phos MEAN 2.07 2.05 2.18 2.04 rnmol/L ST.OEV 0.21 0.25 0.13 0.13 N 5 5 5 5

Dunnet-test based on pooled variancesignificant at5% ()or I% 

### (**) level

- Page 17 - * 

### 29Sep08 09h39

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 CLINICAL BIOCHEMISTRY SUMMARY FEMALES
GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50MG/KG 250 MG/KG 1000 MG/KG
END OF TREATMENT ALAT MEAN 58.1 67.8 666 46.2 U/L ST.DEV 13.0 19.6 8.2 6.3 

### N 5 5 5 5 ASAT MEAN 80.9 75.1 80.1 70.2 U/L ST.OEV 11.7 12.0 10.2 5.9 N 5 5 5 5 ALP MEAN 65 72 70 69 U/L STODEV 22 19 20 44 N 5 5 5 5 Total protein MEAN 65.8 65.3 67.7 65.6 gIL ST.OEV 1.1 2.9 2.2 2.2 N 5 5 5 5 Albumin MEAN 32.6 32.1 33.8 32.7 g/L ST.DEV 0.9 2.2 1.2 1.0 N 5 5 5 5 Total bilirubin MEAN 2.4 2.0 2.5 1.8 umol/L ST.OEV 0.4 0.4 0.6 0.3 N 5 5 5 5 Urea MEAN 9.4 10.1 10.0 9.7 mmoll ST.DEV 2.0 1.2 0.9 1.2 N 5 5 5 5 Creatinine MEAN 46.7 47.2 46.6 48.0 umol/L ST.DEV 3.5 1.6 5.0 3.0 N 5 5 5 5 Glucose MEAN 6.48 5.92 5.47 5.81 mmol/L ST.DEV 1.28 1.23 0.82 0.76 N 5 5 5 5 Cholesterol MEAN 1.45 1.69 1.69 1.75 mmol/L ST.DEV 0.21 0.32 0.15 0.33 N 5 5 5 5 Sodium MEAN 141.5 141.0 141.2 141.2 mmol/L ST.DEV 1.9 0.9 1.2 0.5 N 5 5 5 5 Potassium MEAN 3.58 3.79 3.67 3.61 mmol/L ST.DEV 0.24 0.16 0.19 0.46 N 5 5 5 5 Chloride MEAN 99 99 97 97 mmol/L ST.DEV 2 2 1 2 N 5 5 5 5 Calcium MEAN 2.78 2.75 2.81 2.74 mmol/L ST.DEV 0.03 0.06 0.09 0.04 N 5 8 5 5 Inorg.Phos MEAN 2.27 2.23 2.59 2.31 mmol/L ST.DEV 0.34 0.32 0.26 0.19 N 5 5 5 5



## *r.Ounnetl-test based on pooled variance significant at 5%()or 1%

## ()level -Page 18- 29Sep0809h39

## ClI. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1 MACROSCOPIC FINDINGS SUMMARY MALES GROUP 1 GROUP 2 GROUP 3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG NECROPSY Animals examined 10 10 10 10 Animals without findings 9 7 1 0 Animals affected 1 3 9 10 Epididymides

Nodule(s) 0 0 2 0 Seminal vesicies

Reduced insize 0 0 0 1 MesentericInode

Discolouration 0 2 9## 10 ## Skin

Alopecia 0 0 0 1 Discolouration 0 1 0 1 Bone

Tailbent 1 0 0 0 

## FEMALES GROUP 1 GROUP 2 GROUP 3 GROUP4 CONTROL 50MG/KG 250 MG/KG 1000 MG/KG
NECROPSY Animals examined 10 10 10 10 Animals without findings 9 8 2 0 Animals affected 1 2 8 10 General observations

Contents: 0 0 07# Pancreas

Discolouration 0 1 0 0 Adrenal glands

Discolouration 0 0 0 1 Mesenteric Lnode

Discolouration 0 1 8## 9 # Mandibular Lnode

Discolouration 1 2 0 1 Skin

Alopecia 1 0 0 1

## Fisher's Exacttest significant at5%#)or1 %(#level

- 

### Page 19 00- 

### 29Sep08 09h39

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 ORGAN WEIGHTS (GRAM) SUMMARY MALES
GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50MG/KG 250 MG/KG 1000 MG/KG

NECROPSY BODY W, MEAN 355 349 358 348 (GRAM) ST.DEV 26 27 17 7 N 10 10 10 10 BRAIN MEAN 2.04 2.02 2.02 2.08 (GRAM) ST.DEV 0.06 0.04 0,07 0.04 N 5 5 5 5 HEART MEAN 0.909 0.943 0.991 1.005 (GRAM) ST.DEV 0.035 0.053 0083 0.083 N 5 5 5 5 LIVER MEAN 8.44 8.22 9.03 9.23 (GRAM) ST.DEV 0.92 1.03 1.04 0.85 N 5 5 5 5 THYMUS MEAN 0.355 0.369 0.394 0.341 (GRAM) ST 0EV 0.062 0.091 0.038 0.043 N 5 5 5 5 KIDNEYS MEAN 2.43 2.40 2.35 2.41 (GRAM) ST DEV 0.05 0.12 0.16 0.20 N 5 5 5 5 ADRENALS MEAN 0.069 0.058 0.085 0.075 (GRAM) ST.DEV 0.012 0.010 0.005 0.013 N 5 5 5 5 SPLEEN MEAN 0.559 0.590 0.627 0.672 (GRAM) ST DEV 0.114 0.080 0,077 0.032 N 5 5 5 5 TESTES MEAN 3.36 3.28 3.30 3.39 (GRAM) ST.DEV 0.15 0.42 0.38 0.34 N 10 10 9 10 EPIDIDYMIDES MEAN 1.139 1.138 1.207 1182 (GRAM) ST.DEV 0.086 0.114 0.136 0.124 N 10 10 9 10

/~Dunnett-test based on pooled variance significantat 5% ()or 1 %

## ()level

- Page 20

- 29Sep08 09h39

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 ORGANIBODY WEIGHT RATIOS I(%) SUMMARY MALES
GROUP 1 GROUP2 GROUP3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
NECROPSY BODY W. MEAN 355 349 358 348 (GRAM) ST.DEV 26 27 1 7 7 N 10 10 10 10 BRAIN MEAN 0.60 0.60 0.58 0.60 

## ()ST.DEV 0.03 0.03 0.03 0.01 N 5 5 5 5 HEART MEAN 0.268 0.278 0.283 0.288 

## ()ST.DEV 0.014 0.017 0.012 0.020 N 5 5 5 5 LIVER MEAN 2.48 2.42 2.57 2.64 

## ()ST.DEV 0.17 0.20 0.16 0.20 IN 5 5 5 5 THYMUS MEAN 0.104 0.108 0.112 0.098 

## ()ST.DEV 0.016 0.024 0.006 0.012 N 5 5 5 5 KIDNEYS MEAN 0.72 0.71 0.67 0.69 

## %)ST.OEV 0.03 0.03 002 0.04 N 5 5 5 5 ADRENALS MEAN 0.020 0.020 0019 0.021 %)ST.DEV 0.003 0.003 0.002 0.003 N 5 5 5 5 SPLEEN MEAN 0.164 0.174 0.178 0.193 

### (%) ST 0EV 0026 0.016 0015 0.007 N 5 5 5 5 TESTES MEAN 0.95 094 0.92 0.98 

## ()ST.OEV 0.09 0.14 0.08 0.10 N 10 10 9 10 EPIDIDYMIDES MEAN 0.323 0.327 0.339 0.340 

## ()ST.DEV 0.040 0.036 0.035 0.033 IN 10 10 9 10

/ unnett-test based on pooled variancesignificant at 5%()or 1 %

## ()level

- Page 21

- NIP 29Sep0809h39

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1
ORGAN WEIGHTS (GRAM) SUMMARY FEMALES
GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50MG/KG 250MG/KG 1000 MG/KG
NECROPSY BODY W. MEAN 225 220 228 221 (GRAM) ST.DEV 17 18 2 15 N 5 5 5 5 BRAIN MEAN 1.84 1.79 1.83 1.86 (GRAM) ST.DEV 0.05 0.04 0.06 0.01 N 5 5 5 5 HEART MEAN 0733 0.725 0.785 0.716 (GRAM) ST.DEV 0.073 0055 0.041 0043 N 5 5 5 5 LIVER MEAN 7,23 7.07 781 7.11 (GRAM) ST.DEV 0.52 0.78 0.32 0.63 

### N 5 5 5 5 THYMUS MEAN 0.160 0.170 0.171 0.179 (GRAM) ST.DEV 0.055 0.022 0.022 0.026 

### N 5 5 

### 5 5 KIDNEYS MEAN 1.64 1.62 1.74 1.63 (GRAM) ST 0EV 0.21 008 0.08 0.16 N 5 5 55 ADRENALS MEAN 0.079 0.084 0.082 0.088 (GRAM) ST.DEV 0.005 0.009 0.008 0.014 N 5 6 5 5 SPLEEN MEAN 0.457 0.462 0.538 0.501 (GRAM) ST.DEV 0.088 0.057 0.091 0.061 N 5 5 5 5



### IDunnett-test based on

## pooledvariancesignificantat 5% ()or 

## 1 %()level

- Page 22 - *I" 29Sep08 09h39

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 ORGANIBODY WEIGHT RATIOS 

## (%)SUMMARY FEMALES
GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
NECROPSY BODY W MEAN 225 220 228 221 (GRAM) ST.DEV 17 18 2 15 N 55 5 5 BRAIN MEAN 0.82 0.82 0.80 0.84 

## ()ST.DEV 008 0.07 0.03 0.06 N 5 5 5 5 HEART MEAN 0.327 0.331 0.344 0.325 ()ST.DEV 0.035 0,026 0016 0.021 N 5 5 5 5 LIVER MEAN 3.22 3.21 3.42 3.23 

## ()ST.DEV 0.27 013 0.15 0.28 N 5 5 5 5 THYMUS MEAN 0071 0,077 0.075 0.081 ()ST.DEV 0023 0.010 0.010 0.014 N 5 5 5 5 KIDNEYS MEAN 0.73 0.74 0.76 0.74 

## ()ST.DEV 0.11 005 0.03 0.10 N 5 5 5 5 ADRENALS MEAN 0035 0.038 0.036 0040 

## ()ST.DEV 0.003 0.003 0.003 0,005 N 5 5 5 5 SPLEEN MEAN 0.202 0.211 0.236 0.227 ( 0)ST.DEV 0.035 0.031 0.042 0.029 N 5 5 5 5

*r* Dunnett-test based on pooled variance significant at5% ()or 1 %

### (**) level -Page 23 - 00- 29Sep08 09h39

## C.I. PIGMENT BLUE61 NOTOX Project488231 APPENDIX 2 MOTOR ACTIVITY TEST MALES WEEK 4 Counts per sample period (hour) Animal Location of number sensor 1 2 3 4 5 6 7 8 9 10 11 12 Total
GROUP I(CONTROL) 1 High 172 330 112 410 368 110 202 35 182 130 148 54 2253 1 Low 410 465 224 702 625 440 292 283 445 430 549 238 5103 2 High 1919 189 1117 556 768 63 354 276 186 405 213 363 6407 2 Low 457 336 441 505 446 156 245 468 325 245 348 120 4092 3 High 224 138 409 565 330 263 67 247 198 142 179 120 2882 3 Low 425 328 616 1125 488 645 176 372 301 625 590 335 6026 7 Hiugh 85 166 278 499 339 217 169 55 112 143 216 222 2501 7 Low 346 248 483 591 226 199 343 86 198 330 425 313 3788 10 High 305 209 226 320 189 259 207 53 250 160 305 115 2598 10 Low 139 370 513 537 236 310 355 66 300 120 338 373 3657 GROUP 2(50 MG/KG) 11 High 94 225 369 186 319 240 191 237 2 70 268 172 2373 11 Low 339 455 503 339 765 805 251 690 29 95 481 391 5143 12 High 140 134 98 246 216 ill 0 47 242 108 132 242 1716 12 Low 137 277 169 471 795 236 26 120 398 248 329 634 3840 14 High 109 183 309 424 227 33 101 123 175 90 110 167 2051 14 Low 359 559 404 587 115 258 159 176 422 308 177 246 3770 15 High 107 513 313 690 463 445 8 37 190 194 200 109 3269 15 Low 161 742 557 596 561 812 131 261 340 554 589 680 5984 16 High 65 126 315 332 284 179 148 44 149 243 316 194 2395 16 Low 436 385 330 673 564 130 536 202 291 640 425 103 4715 GROUP 3(150 MG/KG) 21 High 225 119 70 159 146 330 196 7 19 227 245 73 1816 21 Low 418 195 158 466 314 468 273 129 295 943 562 306 4527 22 High 74 110 284 383 333 0 42 454 108 0 17 6 1811 22 Low 217 295 681 574 570 2 345 870 146 9 138 35 3882 23 High 16 208 125 184 132 50 96 219 96 100 248 100 1574 23 Low 217 361 430 447 331 84 661 243 432 386 503 310 4405 24 High 308 235 286 252 397 197 287 23 297 120 433 202 3037 24 Low 365 283 185 386 490 482 415 102 186 423 396 305 4018 25 High 161 138 287 313 258 182 221 104 93 139 110 81 2067 25 LOW 341 439 402 513 397 223 512 251 211 326 354 218 4187 GROUP 4(500MG/KG) 32 High 89 11 5 66 137 29 1 237 1 79 119 112 886 32 Low 331 119 116 461 436 140 73 699 43 280 3715 345 3418 34 High 258 297 297 252 106 204 322 9 90 100 90 147 2172 34 Low 582 857 655 404 484 790 601 172 132 213 285 323 5498 35 High 300 23 216 449 721 28 283 1 22 15 126 171 2355 35 -Low 632 313 350 750 881 223 672 154 381 339 741 611 6047 36 High 68 130 102 170 205 340 168 65 218 0 50 44 1560 36 Low 470 494 131 417 430 687 439 200 609 85 229 269 4460 37 High 216 25 226 185 243 140 384 73 113 38 148 123 1914 37 Low 578 159 455 422 456 249 718 293 245 86 386 327 4374

-Page

### 24-

## 0.1. PIGMENT BLUE61 NOTOX Project488231 APPENDIX 1 REPRODUCTION PROCESSES FO-GENERATION
- 

## POST COITUM
FEMALE MALE MATING PREGNANT SCHEDULE DELIVERY NECROPSY NUMBER NUMBER DATE RECORDED DATE
GROUP 1(CONTROL) 41 <NP> 1 02JUL2008 NO ---- 29JUL2008 42 2 02JUL2008 YES BREEDING 23JUL2008 28JUL2008 43 3 02JUL2008 YES BREEDING 23JUL2008 28JUL2008 44 4 05JUL2008 YES BREEDING 27JUL2008 OIAUG2008 45 5 OSJUL2008 YES BREEDING 27JUL2008 CiAUG2008 46 6 05JUL2008 YES BREEDING 27JUL2008 OIAUG2008 47 7 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 48 8 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 49 9 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 50 10 02JUL2008 YES BREEDING 23JUL2008 28JUL2008 GROUP 2(50 MG/KG) 51 11 04JUL2008 YES BREEDING 26JUL2008 31JUL2008 52 12 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 53 13 05JUL2008 YES BREEDING 27JUL2008 01AUG2008 54 <NP> 14 02JUL2008 NO ---- 29JUL2008 55 15 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 56 16 02JUL2008 YES BREEDING 23JUL2008 28.JUL2008 57 17 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 56 18 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 59 19 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 60 20 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 GROUP 3(250 MG/KG) 61 21 02JUL2008 YES BREEDING 23JUL2008 28JUL2008 62 22 04JUL2008 YES BREEDING 26JUL2008 31JUL2008 63 23 02JUL2008 YES BREEDING 23JUL2008 28JUL2008 64 24 02JUL2008 YES BREEDING 23JUL2008 28JUL2008 65 25 02JUL2008 YES BREEDING 23JUL2008 28JUL2008 66 <NP> 26 04JUL2008 NO ---- 29JUL2008 67 27 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 68 28 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 69 29 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 70 30 05JUL2008 YES BREEDING 26JUL2008 31JUL2008 GROUP 4(1000MGKG) 71 31 05.JUL2008 YES BREEDING 27JUL2008 01AUG2008 72 <10> 32 02JUL2008 YES -- -- 29JUL2008 73 33 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 74 34 02JUL2008 YES BREEDING 23JUL2008 28JUL2008 75 35 02JUL2008 YES BREEDING 23JUL2008 28JUL2008 76 36 03JUL2008 YES BREEDING 24JUL2008 29JUL2008 77 37 02JUL200B YES BREEDING 23JUL2008 28JUL2008 78 38 05JUL2008 YES BREEDING 27JUL2008 01AUG2008 79 39 05JUL2008 YES BREEDING 26JUL2008 31JUL2008 80 40 05JUL2008 YES BREEDING 26JUL2008 31JUL2008

<10> implantation sitesonly <NP > Non -pregnan t P g 42lc O 6 O

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1 MATING PERFORMANCE FO-GENERATION
- 

## POST COITUM
DAY OFTHE GROUP 1 GROUP2 GROUP3 GROUP4 PAIRING PERIOD CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
NUMBER OF FEMALES MATED DURING THE FIRST PAIRING PERIOD 1 4 2 4 4 2 1 3 3 7 5 1 4 3 1 1 4 MEDIAN PRECOITAL TIME 3 3 3 3 MEAN PRECOITAL TIME 2.5 2.7 2.3 2.5 N 10 10 10 10

+/++ Steel-test significantat 5% ()or 1%(++) level

- Page 25

- 23Oct08 10h08

## C.L PIGMENT BLUE61 NOTOX Project488231 APPENDIX 1 BREEDING DATA PER GROUP FO-GENERATION
- 

## LACTATION
GROUPi GROUP2 GROUP3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
LITTERS

TOTAL 9 9 9 9 DURATION OF GESTATION

MEAN (+) 21.3 21.2 21.1 21.2 ST.DEV. 0.5 0.4 0.3 0.4 N 9 9 9 9 DEAD PUPS AT FIRST LITTER CHECK

LITTERS AFFECTED 

## ()0 0 1 2 TOTAL 0 0 1 2 MEAN (+) 0.0 0.0 0.1 0.2 ST DEV. 0.0 0.0 0.3 0.4 N 9 9 9 9 LIVING PUPS AT FIRST LITTER CHECK

%OF MALES/FEMALES(# 44/56 47/53 50/50 50/50 TOTAL 104 103 101 114 MEAN (+) 11.6 11.4 11.2 12.7 ST.DEV. 3.3 1.6 3.9 1.7 N 9 9 9 9 POSTNATAL LOSS

%OFLIVINGPUPS 1.0 1.0 0.0 1.8 LITTERS AFFECTED 

### (#) I 1 0 2 TOTAL(#) 1 1 0 2 MEAN (+) 0.1 0.1 0.0 0.2 ST DEV. 0.3 0.3 0.0 0.4 N 9 9 9 9 VIABILITY INDEX 

### (#) 99.0 99.0 100.0 98.2



### Viabilityindex =(Number 

### ofalive pups before planned necropsy/Number ofpups born alive) *100 -+/++Steel-test significant at5%

## (+)or 1 %(++) 

### level #1 Fisher's Exact testsignificantat 5% 

## (#)or 1 %(##)level

- Page 26 -23Oct08 

### 16h08

## CI. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX I MEAN BODY WEIGHTS OF PUPS PER GROUP (GRAM) FO-GENERATION
- 

## LACTATION
DAY SEX GROUP 1 GROUP 2 GROUP 3 GROUP4 CONTROL 50MG/KG 250 MG/KG 1000 MG/KG 1 M MEAN 6.0 6.1 6.1 5.9 ST.DEV. 0.6 0.7 0.4 0.4 N 9 9 9 9 F MEAN 5.8 5.7 5.6 5.6 ST.DEV. 0.5 0.7 0.4 0.4 N 9 9 8 9 M+F MEAN 5.9 5.9 5.9 5.8 ST.DEV. 0.5 0.7 04 0.4 N 9 9 9 9 4 M MEAN 8.6 9.1 9.0 8.5 ST.DEV. 0.9 1.1 0.9 0.6 N 9 9 9 9 F MEAN 8.4 8.6 8.3 8.2 ST.DEV. 1.0 1.0 0.9 0.8 N 9 9 8 9 M+F MEAN 8.5 8.8 87 8.3 ST.DEV 09 1.1 1.0 0.7 N 9 9 9 9

Dunnett-test based on pooled variance significantat 5%()or 1%()level

- 

### Page 27 - 23Oct08 16h08

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2

APPENDIX 2 INDIVIDUAL TABLES

-Page 1

-

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 MORTALITY DATA MALES
ANIMAL SCHEDULED TREATMENT SACRIFICE FROM TO
GROUP 1 (CONTROL) 1 17JUL08 17JUNOB 16JUL08 2 17JUL08 17JUN08 16JUL08 3 17JUL08 17JUN08 16JUL08 4 17JUL08 17JUN08 16JUL08 5 17JUL08 17JUN08 16JUL08 6 17JUL08 17JUN08 16JUL08 7 17JUL08 17JUN08 16JUL08 8 17JUL08 17JUN08 16JUL08 9 17JUL08 17JUN08 16JUL08 10 17JUL08 17JUN08 I6JUL08 

### GROUP 2

## (50 MGIKG) 11 17JUL08 17JUN08 16JUL08 12 17JUL08 17JUN08 16JUL08 1 3 17JUL08 17JUN08 16JUL08 k14 17JUL08 17JUN08 16JUL08 15 17JUL08 17JUN08 16JUL08 16 17JUL08 17JUN08 16JUL08 17 17JUL08 17JUN08 16JUL08 18 17JUL08 17JUN08 16JUL08 19 17JUL08 17JUN08 16JUL08 20 17JUL08 17JUN08 16JUL08 GROUP 3(260 MG/KG) 21 17JUL08 17JUN08 16JUL08 22 17JUL08 17JUN08 16JUL08 23 17JUL08 17JUN08 16JUL08 24 17JUL08 17JUN08 16JUL06 25 17JUL08 17JUN08 16JUL08 26 17JUL08 17JUN08 16JUL08 27 17JUL08 17JUN08 16JUL08 28 17JUL08 17JUN08 16JUL08 29 17JUL08 17JUN08 16JUL08 30 17JUL08 17JUN08 16JUL08 GROUP 4(1000 MG/KG) 31 17JUL08 17JUN08 16JUL08 32 17JUL08 17JUN08 16JUL08 33 17JUL08 17JUN08 16JUL08 34 17JUL08 17JUN08 16JUL08 36 17JUL08 17JUN08 16JUL08 36 17JUL08 17JUN08 16JUL08 37 17JUL08 17JUN08 16JUL08 38 17JUL08 17JUN08 16JUL08 39 17JUL08 17JUN08 16JUL08 40 17JUL08 17JUN08 16JUL08

- Page2

- 29SepO8lOh2l

## ClI. PIGMENT BLUE 61 NOTOX 

## Project488231 APPENDIX 2
MORTALITY DATA FEMALES
ANIMAL SCHEDULED TREATMENT SACRIFICE FROM TO
GROUP I (CONTROL) 41 29JUL08 17JUN08 28JUL08 42 28JUL08 17JUN08 27JUL08 43 28JUL08 17JUN08 27JUL08 44 01AUG08 17JUN08 31JUL08 46 01AUG08 17JUN08 31JUL08 46 01AUG08 17JUN08 31JUL08 47 30JUL08 17JUN08 29JUL08 48 30JUL08 17JUN08 29JUL08 49 30JUL08 17JUN08 29JUL08 50 28JUL08 17JUN08 27JUL08 GROUP 

### 2(50 MG/KG) 51 31JUL08 17JUN08 30JUL08 52 30JUL08 17JUN08 29JUL08 53 01AUG08 17JUN08 31JUL08 54 29JUL08 17JUN08 28JUL08 55 30JUL08 17JUN08 29JUL08 56 28JUL08 17JUN08 27JUL08 57 30JUL08 17JUN08 29JUL08 58 30JUL08 17JUN08 29JUL08 59 30JUL08 17JUN08 29JUL08 60 30JUL08 17JUN08 29JUL08 GROUP 3 (250 MG/KG) 61 28JUL08 17JUN08 27JUL08 62 31JUL08 17JUN08 30JUL08 63 28JUL08 17JUN08 27JUL08 64 28JUL08 17JUN08 27JUL08 65 28JUL08 17JUN08 27JUL08 66 29JUL08 17JUN08 28JUL08 67 30JUL08 17JUN08 29JUL08 68 30JUL08 17JUN08 29JUL08 69 30JUL08 17JUN08 29JUL08 70 31JUL08 17JUN08 30JUL08 GROUP 4(1000MG/KG) 71 01AUG08 17JUN08 31JUL08 72 29JUL08 17JUN08 28JUL08 73 30JUL08 17JUN08 29JUL08 74 28JUL08 17JUN08 27JUL08 75 28JUL08 17JUN08 27JUL08 76 29JUL08 17JUN08 28JUL08 77 28JUL08 17JUN08 27JUL08 78 01AUG08 17JUN08 31JUL08 79 31JUL08 17JUN08 30JUL08 80 31JUL08 17JUN08 30JUL08

-Page 3

- 

### 29Sep08 10h21

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 CLINICAL SIGNS MALES PRE MATING REPRO PHASE SIGN (MAX. GRADE) WEEK: 1............................ ... 4.... (LOCATION) DAY: 123456712345671234567123456712345671234567123

GROUP I (CONTROL) ANIMAL I 

### No clinicalsigns noted ANIMAL 2 No clinicalsigns noted ANIMAL 3 

### Various

Broken(1) G:.................. ....... 111

(Tell apex) ANIMAL4 No clinical signsnoted ANIMAL5 

### No clinicalsigns noted ANIMAL 6 No clinicalsigns noted ANIMAL 7 No clinicalsigns noted ANIMAL 8 No clinicalsigns noted ANIMAL 9 No clinicalsigns noted ANIMAL 10 No clinicalsigns noted GROUP 2(50 MG/KG) ANIMAL 11 Skin/fur

Bluediscolouration(1) G: .... 111111111111111111111111

(Faeces) Secretion/excretion

Salivation (3) G:............. I........... 1... ANIMAL 12 

### Skin/fur

Bluediscolouration (1) G: .... 111111111111111111i111111

(Faeces) ANIMAL 13 Skin /fur

Bluediscolouration (1) G: .... 1111111111111111111111111

(Faeces) ANIMAL 14 Skin/fur

Bluediscolouration(1) G: .... 111111111111111111111111

(Faeces) ANIMAL 15 Skin/fur

Bluestaining(1) G:................1111111111111111

(Tail) Bluediscolouration (1) G:

. 111111111111111111111111

(Faeces) ANIMAL 16 SkinI fur

Bluediscolouration(1) G: 111111111111111111111111

(Faeces) ANIMAL 17 Skin /fur

Blue discolouration (1) G: .... 1111111111111111111111111

(Faeces)

G0Highest dailygrades

Observation performed, sign not present

- 

### Page 4 -fr24Oct08 15h46

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 CLINICAL SIGNS MALES PRE MATING REPRO PHASE SIGN (MAX. GRADE) WEEK:I.............I .................... ... (LOCATION) DAY: 123456712345671234667123456712345671234567123

GROUP 2(50 MG/KG) ANIMAL 16 Skin /

### fur

Scabs(3) G:..........I.............. 11111111

(Neck) Blue discolouration (1) G: 111. II111111111111111111111 (Faeces) ANIMAL 19 Skin/fur

Bluediscolouration (1) G: .... 111111111111111111111111

(Faeces) ANIMAL 20 Skin/fur

Bluediscolouration (1) G: .... iiiiiiiiiiiiiiiiiiiiiii

(Faeces) GROUP 3(250 MG/KG) ANIMAL 21 Skin/fur

Bluediscolouration (1) G: ... .11111111111111111111111111 (Faeces) Secretion/ excretion

Salivation (3) G...................... ANIMAL 22 Skin/fur

Bluediscolouration (1) G: ... 111111111111111111111111111l (Faeces) ANIMAL23 Skin/fur

Alopecia (3) G:..........
.......... I.........11

(Forelegs) Bluediscolouration (1) G: . .. iiiiiiiiiiiiiiiiiii'liiii

(Faeces) Secretion /excretion

Salivation (3) G:......I.............. ........ 1 ANIMAL24 Skin/fur

Bluediscolouration (1) G: ... .1111111111111111111111111111 (Faeces) ANIMAL 25 Skin/fur

Bluedscolouration (1) G: ... 111111111111111111111111111

(Faeces) 

### Secretion/excretion

Salivation (3) G:.............. I.............. 1. ANIMAL 26 Skin / fur Bluediscolouration (1) G: ... .111111111111111111111111111 (Faeces) ANIMAL27 Skin/fur

Bluediscolouration (1) G:

... 1111111111111

(Faeces) Secretion/excretion

Salivation (3) G:..............................1

G: Highest daily grades

Observation performed, sign not present

-PageS 5 24Oct08 15h46

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 CLINICAL SIGNS MALES PRE MATING REPRO PHASE SIGN (MAX. GRADE) WEEK: 1
............ 1...................4..... (LOCATION) DAY: 123456712345671234567123456712345671234567123

GROUP 3(250 MG/KG) ANIMAL28 Skin/Ifur

Scabs(3) G:..........................1

(Neck) Blue discolouratIon (1) G:

. 111111111111111111111111111

(Faeces) Secretion/excretion

Salivation (3) G:.......I................
.... 1. ANIMAL 29

Skn

/fur Blue discolouration (1) G:

. . .111111111111111111111111.

(Faeces) ANIMAL30 Skin/Ifur

Blue staining (1) G: . . . . . . . . . . .

(Back) Blue staining (1) G..........................1I

(Tail) (Faeces) GROUP 4(1000 MG/KG) ANIMAL 31 Skin/fur

Alopecia(3) G:...............
... .. I.......11

(Foreleg right) Scabs(3) 0:.......................111....

(Foreleg ight) Bluediscolouratlon (1) G: 1111111111111111111111111111ii

(Faeces) ANIMAL32 Skin/Ifur

Bluediscolouration (1) G:

.. 111111111111111111111111111

(Faeces) ANIMAL 33 Skin /fur

Blue discolouration (1) G
.. 111111111111111

(Faeces) Secretion/excretion

Salivation (3) G:..........11111111111111111.... I1 ANIMAL34 Skin/Ifur

Alopecia(3) G:.................... 111112111111

(Forelegs) Bluediscolouration (1) G:

.. 1111111111 1111111111111111111

(Faeces) Secretion/excretion

Salivation (3) G:...............1I...11... ANIMAL 35 Skin/fur

Bluediscolouration (1) G:

.. iiiiiiiiiiiiii

(Faeces) ANIMAL36 Skin/fur

Bluediscolouration (1) G

.. 11111111111111

(Faeces) Secretion/excretion

Salivation(3) G:............111l,... 1.. 11.
. 1..

G: Highest dailygrades

Observation performed, sign not present

- Page 6 -fr24Oct08 15h46

## C.I. PIGMENT BLUE61 NOTOX Project488231 APPENDIX 2 CLINICAL SIGNS MALES PRE MATING REPRO PHASE SIGN (MAX. GRADE) WEEK: 1.............1 .... ............ ....... (LOCATION) DAY: 123456712345671234567123456712345671234567123

GROUP 4(1000 MG/KG) ANIMAL 37 Skin /fur

Bluediscolouration (1) G, .1111111111111111111111111111

(Faeces) ANIMAL 38 Skin /fur

Bluediscolouration (1) G:

. . 11111111111111111111111111111

(Faeces) Secretion/excretion

Salivation (3) G:............... 1 . ... ANIMAL39 Skin/fur

Bluestaining(1) G:....................111111111111

(Tail) Blue discolouration (1) G:

. . 11111111111111111111111111111

(Faeces) Secretion/ excretion

Salivation (3) G- ..... 1.111...11. 11..1 ANIMAL 40 Skin /fur

Blue staining (1) G: .. III1......................

(Back) Bluediscolouration (1) G,

.. 11111111111111

(Faeces) Secretion/excretion

Salivation (3) G:.......... 1.... 1
... 11111 .1.1 

## FEMALES PRE MATING REPRO PHASE SIGN (MAX. GRADE) WEEK: 1I.............I ................. .. ... (LOCATION) DAY: 123456712345671234567123456712345671234567123

GROUP I(CONTROL) ANIMAL41 Skin/fur

Alopecia (3) G:.......... I.......I.......................1

(Abdomen) ANIMAL 42 Skin/fur

Scabs(3) G:.............................
...... 11111i..

(Mouth) Scabs(3) G:........................1ill...............

(Snout) ANIMAL43 No clinical signs noted ANIMAL44 No clinicalsigns noted ANIMAL45 No clinicalsigns noted ANIMAL 46 Skin/fur

Alopecia(3) G:............... 11111111
.. .....................

(Back) (Neck)

G: Highestdaily grades

Observation performed, sign not present

- Page 7

- 

# ~24Oct08 l5h46

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 CLINICAL SIGNS FEMALES PRE MATING REPRO PHASE SIGN (MAX. GRADE) WEEK: I.............I ................... 4.... (LOCATION) DAY: 123456712345671234567123456712345671234567123

GROUP 1(CONTROL) ANIMAL 47 Skin/fur

Piloerection (1) G:....
.......... I............. ..... 1.. Alopecia(3) G: .11111
............ .......................

(Back) Alopecia (3) G:......................... 12222222

(Abdomen) Alopecia (3) G:............................... 1111111

(Flank left) Alopecia(3) G:....................
................ 111

(Thigh hind left) Alopecia(3) G:... ............................ 1111..

(Foreleg left) Alopecie (3) G:........................
........... .2222233

(Forelegs) ANIMAL48 Skin/fur

Alopecia(3) G: 1111111111111111111111111111111
............

(Neck) ANIMAL 49 Skin/fur

Alopecia(3) G, .1111111111111..............

(Back) (Neck) ANIMAL 50 Skin/fur (Cheek left) Alopecia(3) G:...........11111111......................

(Cheek right) (Back) Alopecia(3) G: 111111
.... 11111111111111111111111111.

(Neck) GROUP 2(50 MG/KG) ANIMAL 51 Skin/fur

Alopecia(3) G: ... .111111...........
.......... (Neck) Blue discolouration (1) G.........11111111111111111111111111111111111111i

(Faeces) ANIMAL 52 Skin/fur

Blue discolouration (1) G:

...... 1111111111111111

(Faeces) ANIMAL53 Skin/fur

Piloerection (1) G: .. . . .I. . .. . . . . . . Blue discolouration (1) G: ..... iiiiiiiliiiii iiiiiii iiilii

(Faeces) ANIMAL 54 Skin/fur

Alopecia (3) G:..........
....... ....... .... 11111111111111

(Abdomen) (Faeces)

G: Highest daily grades

Observation performed, sign not present

- Page 8

- 

# ~24Oct08 15h46

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 CLINICAL SIGNS FEMALES PRE MATING REPRO PHASE SIGN (MAX.GRADE) WEEK: 1............ 1...................4.... (LOCATION) DAY: 123456712345671234567123456712345671234567123

GROUP 2(50 MG/KG) ANIMAL 55 Skin/fur

Bluediscolouration (1) G: 1111111111111111111111

(Faeces) ANIMAL 56 Skin/fur

Alopecia(3) G:..........1111111.
... ...................

(Neck) Blue discolouration (1) G: ...... 111111111111111111111111111111lil

(Faeces) ANIMAL 57 Skin/fur

Bluestaining(1) G:.........111111
......... I................ ...

(Tail) Bluediscolouration (1) G- .. 111111111111111111111111111111111111 il (Faeces) ANIMAL 58 SkiI/fur

Blue discolouration (1) G:

...... 1111111111111111

(Faeces) ANIMAL 59 Skin/fur (Faeces) ANIMAL 60 Skin/fur (Faeces) GROUP 3(250 MG/KG) ANIMAL 61 Skin/fur

Blue discolouration (1) G:
. .. 11111111111111111111111111111111111111

(Faeces) ANIMAL 62 Breathing

Rales (3) G:.......................
............ 12... Skin/fur

Bluestaining(1) G:...............1111l..... .....................

(Back) Blue discolouration (1) G: ... .11111111111111111111111111111111111111111 (Faeces) ANIMAL 63 Skin/fur

Alopecia (3) G:...........1111111111111i. .................

(Cheek right) Blue staining (1) G:.....................................1....

(Mouth) Blue staining (1) G:....
............................. 1.

(Abdomen) Bluestaining(1) G: .. . . . . . . . . . . . . . . .. .1 . .

(Forelegs) (Faeces) ANIMAL 64 Skin/fur

Alopecia(3) G:........111.................
...

(Cheek right) (Faeces)

G: Highestdaily grades

Observation performed, sign not present

- Page 9

- 24Oct08 15h46

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 CLINICAL SIGNS FEMALES PRE MATING REPRO PHASE SIGN (MAX. GRADE) WEEK:1.................. ............. 4 ... (LOCATION) DAY, 123456712345671234567123456712345671234567123

GROUP 3(250 MG/KG) ANIMAL 65 Skin 

### /fur

Bluediscolouration (1) G: .11111111111111111111111111111111111111

(Faeces) ANIMAL 66 Skin /fur

Piloerection (1) G:......... .. .......................... 1.

(Faeces) ANIMAL67 Skin/fur

Blue staining (1) G:
.. I...................................

(Back) (Faeces) ANIMAL68 Skin

### /fur

Bluestaining(1) G: ... .111......
...................... . (Snout) (Faeces) ANIMAL 69 Skin 

### /fur (Faeces) ANIMAL 70 Skin/fur (Faeces) GROUP 4(1000 MG/KG) ANIMAL71 Skin/fur

Alopecia(3) G: 111111......................................

(Neck) Alopecia (3) G:........................................11l...

(Abdomen) Alopecia (3) G:.................._............ 2222333333333333

(Forelegs) 

### Alopecia (3) G: .................................. 11222222 

## ( (H-indlegs) (Faeces) ANIMAL 72 Skin Ifur

Blue discolouration (1) G: .11111111111111111111

(Faeces) ANIMAL73 Skin/Ifur (Tail) (Faeces) ANIMAL 74 Skin/fur (Faeces) Secretion/excretion



### Salivation(3) G:..............1...... ....... .........

G: Highest dailygrades

Observation performed, sign not present

- 

### Page 10
- 00 24Oct08 15h46

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 CLINICAL SIGNS FEMALES PRE MATING REPRO PHASE SIGN (MAX. GRADE) WEEK: 1 ............. 1
................... 4.... (LOCATION) DAY: 123456712345671234567123456712345671234567123

GROUP 4(1000 MG/KG) ANIMAL75 Skin/fur

Blue discolouration (1) G:

. .1111111111i111111111111111111111111iii

(Faeces) 

### ANIMAL76 Skin/fur (Faeces) ANIMAL 77 Skin/Ifur

Alopecia (3) G:........
............................. 11111i

(Back) (Faeces) ANIMAL 78 SkinIfur (Faeces) ANIMAL 79 Breathing

Rales (3) G: 1 ........................................ Skin/fur

Scabs(3) G:....................................1 1..

(Mouth) (Faeces) Secretion/excretion

Salivation (3) G:................. .... 1 .......... ......... ANIMAL80 Skin/Ifur

Blue staining (1) G:.........................1.1..................

(Flank right) (Faeces)

G: Highest dailygrades

Observation performed, sign not present -Page 11

- 24Oct08 15h46

## C.I PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 BODY WEIGHTS (GRAM) MALES PRE MATING MATING PERIOD DAYS 1 8 1 8 15 22 29 36 WEEKS 1 2 1 2 3 4 5 6 ANIMAL GROUP1I(CONTROL) 1 293 322 351 366 386 2 274 288 306 322 339 3 292 320 343 355 373 4 303 313 323 325 342 5 306 325 339 348 366 6 313 339 371 397 421 7 295 323 348 357 368 8 329 350 369 382 398 9 296 315 337 356 372 10 314 350 369 391 411 GROUP 2(50 MG/KG) 11 308 323 347 355 371 12 279 292 317 333 343 13 298 319 336 345 359 14 288 305 323 331 339 15 306 342 362 369 389 16 292 314 335 343 361 17 322 350 385 339 420 18 296 314 330 340 357 19 297 311 333 341 357 20 322 352 378 392 427 GROUP 3 (250 MGIKG) 21 319 344 371 388 409 22 302 335 353 367 378 23 291 309 327 349 363 24 305 328 350 362 372 25 292 309 330 342 355 26 302 334 360 374 392 27 307 334 357 372 390 28 312 340 388 373 395 29 300 321 345 362 375 30 301 329 359 372 397 GROUP 4(1000 MGIKG) 31 314 340 359 368 388 32 290 304 327 348 366 33 312 322 340 351 369 34 304 320 332 346 360 35 291 311 334 346 360 36 301 326 352 368 379 37 294 320 343 354 364 38 307 323 340 351 374 39 314 332 343 344 370 40 298 308 325 340 359

-Page 12

- 29SeP08 101121

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 BODY WEIGHTS (GRAM) FEMALESPRE MATING MATING PERIOD DAYS 1 8 1 8 15 22 29 36 WEEKS 1 2 1 2 3 4 5 6 ANIMAL
GROUP I (CONTROL) 41 206 211 217 42 182 185 201 43 199 196 218 44 204 212 212 45 194 199 204 46 186 193 200 47 185 191 198 48 193 203 210 49 189 195 202 50 196 198 215 GROUP 2(50 MG/KG) 51 195 204 209 52 184 189 210 53 198 210 211 54 183 196 206 55 202 214 220 56 191 193 216 57 197 202 210 58 207 217 225 59 190 197 205 60 196 205 207 GROUP 3 1250 MG/KG) 61 196 200 216 62 203 210 215 63 187 195 214 64 194 198 222 65 191 189 211 66 198 204 205 67 185 204 216 68 196 203 213 69 180 191 202 70 196 200 199 GROUP 4 (1000 MG/KG) 71 198 205 203 72 201 203 221 73 186 196 201 74 196 204 220 75 190 193 214 76 183 195 207 78 195 200 203 79 184 187 190 80 195 201 201

- Page 13

- 

# ~29Sep08 10h21

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 BODY WEIGHTS (GRAM) FEMALES FO-GENEIRATION POST COITUM LACTATION DAYS 0 4 7 11 14 17 20 1 4 ANIMAL
GROUP I(CONTROL) 41 <NP> 222 223 220 225 237 240 245 --- 42 194 216 226 239 250 277 315 235 243 43 220 239 247 267 280 297 332 270 280 44 222 244 243 257 269 296 330 242 261 45 213 228 233 250 266 287 322 239 261 46 209 221 224 238 244 263 272 235 248 47 200 213 219 230 224 252 280 225 227 48 210 225 238 248 272 291 335 246 256 49 199 210 220 230 243 263 292 223 239 50 215 235 243 259 273 301 346 254 256 GROUP 2(50 MGIKG) 51 212 229 234 247 262 279 300 243 247 52 206 218 224 238 247 270 298 225 229 53 223 249 262 277 290 316 352 269 276 54<NP> 207 211 209 209 210 214 222 --- 55 217 232 242 254 264 288 324 239 242 56 208 226 234 251 265 288 325 253 250 57 210 222 227 240 250 271 304 234 230 58 227 243 257 271 287 314 349 265 279 59 207 222 229 239 250 270 299 234 245 60 215 227 232 244 253 271 301 241 253 GROUP 3(250 MGIKG) 61 213 224 229 244 251 275 306 239 247 62 220 235 244 259 265 272 287 257 260 63 211 235 242 257 270 296 339 252 259 64 208 228 234 245 264 287 318 237 252 65 211 232 243 259 270 297 330 254 261 66 <NP> 217 221 225 221 225 222 225 --- 67 217 239 249 262 280 298 334 260 263 68 210 229 236 253 266 299 324 242 259 69 206 216 225 237 247 266 299 248 243 70 214 234 243 257 269 292 332 246 257 GROUP 4(1000 MGIKG) 71 213 232 241 258 271 301 338 246 264 72 216 236 245 243 242 243 237 --- 73 198 218 229 238 251 267 295 230 233 74 217 234 242 255 265 283 309 238 247 75 209 222 230 244 263 286 325 232 248 76 205 223 228 244 250 283 316 232 238 77 214 228 238 253 268 288 332 250 260 78 217 231 241 257 259 286 325 233 256 79 200 224 227 245 256 281 317 241 249 80 209 229 239 253 264 286 324 243 261

<NP> Non-pregnant
-Page 14 -I 23Oct08 16IWO

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 BODY WEIGHT GAIN(% MALES PRE MATING MATING PERIOD DAYS 1 8 1 8 15 222936 WEEKS 1 2 1 2 3 4 5 6 ANIMAL GROUP1I(CONTROL) 1 0 10 2025 32 2 0 5 12 1824 3 0 10 172228 4 0 3 7 7 13 5 D 6 11 14 20 6 0 8 192735 7 0 9 1821 25 8 0 6 12 1621 9 0 6 142026 10 0 11 1825 31 GROUP 2(50 MG/KG) 11 0 5 13 1520 12 0 5 14 1923 13 0 7 13 1620 14 0 6 12 15 18 15 0 12 18 21 27 16 0 8 15 1724 17 0 9 205 30 18 0 6 11 15 21 19 0 5 12 1520 20 0 9 172233 GROUP 3(250 MG/KG) 21 0 8 1622 26 22 0 11 1722 25 23 0 6 122025 24 0 8 15 1922 25 0 6 13 1722 26 0 11 192430 27 0 9 1621 27 28 0 9 182027 29 0 7 15 2125 30 0 9 19 2432 GROUP 4(1000 MG/KG) 31 0 8 14 1724 32 0 5 132026 33 0 3 9 13 16 34 0 5 9 14 18 35 0 7 15 1924 36 0 8 172226 37 0 9 172024 38 0 5 11 14 22 39 0 6 9 10 18 40 0 3 9 14 20



### -Page 15

- 

# ~29Sep08 10h2l

## C.l. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 BODY WEIGHT GAIN I(%) FEMALESPRE MATING MATING PERIOD DAYS 1 8 1 8 152229 36 WEEKS 1 2 1 2 3 4 5 6 ANIMAL
GROUP I (CONTROL) 41 0 2 5 42 0 2 10 43 0 -2 10 44 0 4 4 45 0 3 5 46 0 4 8 47 0 3 7 48 0 5 9 49 0 3 7 50 0 1 10 GROUP 2(50 MG/KG) 51 0 5 7 52 0 3 14 53 0 6 7 54 0 7 13 55 0 6 9 56 0 1 13 57 0 3 7 58 0 5 9 59 0 4 8 60 0 5 6 GROUP 3 (250 MG/KG) 61 0 2 10 62 0 3 6 63 0 4 14 64 0 2 14 65 0 -1 10 66 0 3 4 67 0 10 17 68 0 4 9 69 0 6 12 70 0 2 2 GROUP 4 (1000 MG/KG) 71 0 4 3 72 0 1 10 73 0 5 8 74 0 4 12 75 0 2 13 76 0 7 13 77 0 1 11 78 0 3 4 79 0 2 3 80 0 3 3

-Page 16 -I 29Sep08 10h21

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 BODY WEIGHT GAIN(% FEMALES FO-GENERATION POST COITUM LACTATION DAYS 0 4 7 11 14 17 20 1 4 ANIMAL
GROUP I (CONTROL) 41 -NP> 0 0 -1 1 7 8 10 --- 42 0 11 16 23 29 43 62 0 3 43 0 9 12 21 27 35 51 0 4 44 0 10 9 16 21 33 49 0 8 45 0 7 9 17 25 35 51 0 9 46 0 6 7 1 4 17 26 30 0 6 47 0 7 10 15 12 26 40 0 1 48 0 7 13 18 30 39 60 0 4 49 0 6 11 16 22 32 47 0 7 50 0 9 13 20 27 40 61 0 1 GROUP 2 (50 MG/KG) 51 0 8 10 17 24 32 42 0 2 52 0 6 9 16 20 31 45 0 2 53 0 12 17 24 30 42 58 0 3 54<NP> 0 2 1 1 1 3 7 ---- 55 0 7 12 17 22 33 49 0 1 56 0 9 13 21 27 38 56 0 -1 57 0 6 8 14 19 29 45 0 -2 56 0 7 13 19 26 38 54 0 5 59 0 7 11 15 21 30 44 0 5 60 0 6 8 13 18 26 40 0 5 GROUP 3(250 MG/KG) 61 0 5 8 15 18 29 44 0 3 62 0 7 11 18 20 24 30 0 1 63 0 11 15 22 28 40 61 0 3 64 0 10 13 18 27 38 53 0 6 65 0 10 15 23 26 41 56 0 3 66<NP> 0 2 4 2 4 2 4 ---- 67 0 10 15 21 29 37 54 0 1 68 0 9 12 20 27 42 54 0 7 69 0 5 9 15 20 29 45 0 -2 70 0 9 14 20 26 36 55 0 4 GROUP 4(1000 MGIKG) 71 0 9 13 21 27 41 59 0 7 72 0 9 13 13 12 13 10 --- 73 0 10 16 20 27 35 49 0 1 74 0 8 12 18 22 30 42 0 4 75 0 6 10 17 26 37 56 0 7 76 0 9 11 19 22 38 54 0 3 77 0 7 11 18 25 35 55 0 4 78 0 6 11 18 19 32 50 0 10 79 0 12 14 23 28 41 59 0 3 80 0 10 14 21 26 37 55 0 7

<NP> Non-pregnant -Page 17- 230ct08 16hl4

## C.I. PIGMENT BLUE 61 NOTOX 

## Project488231 APPENDIX 2 FOOD CONSUMPTION (G/ANIMAL/DAY) MALES PRE MATING 

### DAYS 1-8 8-15 WEEKS 1-2 2-3 CAGE
GROUP I(CONTROL) 1 20 20 2 23 24 GROUP 2(50 MG/KG) 3 21 22 4 21 22 GROUP 3(250 MG/KG) 5 22 22 6 22 22 GROUP 4(1000 MG/KG) 7 21 22 8 22 22 

## FEMALES PRE MATING DAYS 1-8 8-15 WEEKS 1-2 2-3 CAGE

GROUP I(CONTROL) 9 14 14 10 15 15 GROUP 2(50 MG/KG) 1 1 15 15 12 14 1 4 GROUP 3(250 MG/KG) 13 14 15 1 4 14 1 4 GROUP 4(1000 MG/KG) 15 15 15 16 14 14

-Page 18- 

## W 29Sep0810h21

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 FOOD CONSUIMPTION (G/ANIMALJDAY) FEMALES FO-GENERATION POST COITUM LACTATION DAYS 0-4 4-7 7-11 11-14 14-17 17-20 1-4 ANIMAL
GROUP I (CONTROL) 41 <NP' 14 16 16 18 15 17 -- 42 1B 20 20 23 21 24 27 43 18 18 21 22 15 26 29 44 18 17 19 18 21 18 41 45 16 17 19 20 21 21 42 46 18 17 19 19 22 19 34 47 15 18 18 15 16 20 20 48 19 22 22 24 22 25 29 49 15 17 17 19 19 19 26 50 18 20 20 24 24 25 29 GROUP 2(50 MG/KG) 51 16 16 17 20 19 20 21 52 17 16 20 17 19 19 21 53 22 24 15 25 26 29 47 54<NP> 17 18 16 18 17 21 -- 55 15 18 19 4 21 22 23 56 17 19 19 20 19 23 25 57 17 16 18 19 19 19 20 58 20 22 22 25 27 26 33 59 15 15 18 18 20 21 24 60 16 18 18 4 19 20 25 GROUP 3(250 MG/KG) 61 16 18 18 20 20 21 27 62 16 18 19 19 19 20 13 63 19 21 20 22 21 24 29 64 16 18 18 20 20 21 30 05 17 21 20 25 22 24 30 66 NP> 18 15 17 15 16 15 -- 67 20 19 22 23 22 24 29 68 17 17 20 21 24 21 29 69 15 15 17 18 22 21 22 70 20 21 23 35 24 23 28 GROUP 4(1000 MG/KG) 71 19 19 22 21 24 23 45 72 17 19 18 20 16 15 -- 73 16 17 17 19 19 20 20 74 15 19 18 21 17 19 77(3) 75 16 20 21 22 23 23 31 76 17 18 20 20 21 20 23 77 15 16 19 21 19 23 29 78 18 19 21 16 21 20 50 79 18 18 20 21 22 25 24 80 17 18 20 20 21 22 28

(3)Tare error <NP> Non-pregnant -Page 19- ' 23Oct08 16h14

## C.l. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2
RELATIVE FOOD CONSUMPTION (GIKG BODY WEIGHT/DAY) MALES PRE MATING DAYS 1-8 8-15 WEEKS 1-2 2-3 CAGE
GROUP I (CONTROL) 1 63 65 2 68 72 GROUP 2 (50 MG/KG) 3 66 68 4 64 68 GROUP 3 (250 MG/KG) 5 67 69 6 65 67 GROUP 4 (1000 MG/KG) 7 66 68 8 67 68 

## FEMALESPRE MATING DAYS 1-8 8-15 WEEKS 1-2 2-3 CAGE

GROUP I(CONTROL) 9 68 71 10 74 77 GROUP 2(50 MG/KG) 11 73 72 12 70 70 GROUP 3(250 MG/KG) 13 73 76 14 68 71 GROUP 4 (1000 MG/KG) 15 73 75 16 72 74

-Page 20 -I'29Sep08 I01121

## CI. PIGMENT BLUE 61 NOTOX 

## Project488231 APPENDIX 2 RELATIVE FOOD CONSUMPTION (GIKG BODYWEIGHT/DAY) FEMALES FO-GENERATION POST COITUM LACTATION DAYS 0-4 4-7 7-11 11-14 14-17 17-20 1-4 ANIMAL
GROUP I (CONTROL) 41 <NP> 63 71 69 75 61 71 -- 42 83 88 85 92 77 76 il1 43 74 73 79 79 52 79 102 44 73 70 74 67 71 56 158 45 70 73 74 75 72 64 160 46 79 74 78 77 85 69 138 47 70 82 77 67 65 70 90 48 82 92 87 89 77 76 112 49 70 76 75 77 72 64 109 50 76 82 75 89 80 73 113 GROUP 2(50 MG/KG) 51 68 67 69 78 67 67 84 52 79 70 82 69 72 65 93 53 89 92 55 86 88 81 170 54 <NP> 79 65 78 86 79 93 -- 55 65 73 74 16 73 67 95 56 73 80 74 74 67 70 99 57 77 72 73 76 69 64 87 58 81 86 79 88 87 75 117 59 65 67 73 72 75 69 98 60 69 76 74 17 70 65 100 GROUP 3(250 MG/KG) 61 70 80 73 81 74 69 109 62 68 74 73 70 69 69 49 63 80 87 78 83 72 72 ill 64 70 77 71 77 69 65 119 65 73 85 75 91 75 72 114 66<NP> 79 65 75 67 71 68 -- 67 85 78 85 81 74 71 109 68 74 72 79 79 79 65 112 69 68 65 73 73 81 70 89 70 83 85 89 130 82 69 109 GROUP 4(1000 MG/KG) 71 82 79 84 79 79 67 169 72 71 78 72 83 64 62

_ 73 75 76 69 76 70 68 86 74 65 79 70 81 60 63 312(3) 75 70 87 84 84 79 70 125 76 75 79 80 81 75 63 98 77 67 77 74 77 67 68 112 78 79 79 82 60 73 61 197 79 79 78 81 82 79 78 95 80 75 75 78 77 75 69 107

(3)Tare error <NP> Non-pregnant -Page 21 -ON- 23Oct08 l6h14

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 FUNCTIONAL OBSERVATIONS MALES ATWEEK 4
ANIMAL HEARING PUPIL L PUPIL R. STATIC R GRIP SCORE 0/1 SCORE 0/1 SCORE 0/1 SCORE 0/1 SCORE 0/1
GROUP I (CONTROL) 1 0 0 0 0 0 2 0 0 0 0 0 3 0 0 0 0 0 7 0 0 0 0 0 10 0 0 0 0 0 GROUP 2 (50 MG/KG) 11 0 0 0 0 0 12 0 0 0 0 0 14 0 0 0 0 0 15 0 0 0 0 0 16 0 0 0 0 0 GROUP 3(250 MG/KG) 21 0 0 0 0 0 22 0 0 0 0 0 23 0 0 0 0 0 24 0 0 0 0 0 25 0 0 0 0 0 GROUP 4 (1000 MG/KG) 32 0 0 0 0 0 34 0 0 0 0 0 35 0 0 0 0 0 36 0 0 0 0 0 37 0 0 0 0 0 

## FEMALES ATWEEK 4
ANIMAL HEARING PUPIL L PUPIL R. STATIC R GRIP SCORE 0/1 SCORE 0/1 SCORE 0/1 SCORE 0/1 SCORE 0/1

GROUP I(CONTROL) 42 0 0 0 0 0 43 0 0 0 0 0 47 0 0 0 0 0 48 0 0 0 0 0 50 0 0 0 0 0 GROUP 2(50 MG/KG) 52 0 0 0 0 0 55 0 0 0 0 0 56 0 0 0 0 0 57 0 0 0 0 0 58 0 0 0 0 0 GROUP 3 (250 MG/KG) 61 0 0 0 0 0 63 0 0 0 0 0 64 0 0 0 0 0 65 0 0 0 0 0 67 0 0 0 0 0

- Page 22 -I 08Oct08 12h22

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 FUNCTIONAL OBSERVATIONS FEMALES AT WEEK 4
ANIMAL HEARING PUPIL L PUPIL R STATIC R GRIP SCORE 011SCORE 011SCORE 01i SCORE 0/1 SCORE (3/1
GROUP 4(1000 MGIKG) 73 0 0 0 0 0 74 0 0 0 0 0 75 0 0 0 0 0 76 0 0 0 0 0 77 0 0 0 0 0

-Page 23 00 

# I O 8Oct08 121122

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2
MOTOR ACTIVITY TEST FEMALES LACTATION
Counts persample period (hour) Animal Location of number sensor 1 2 3 4 5 6 7 8 9 10 11 12 Total GROUPI1(CONTROL) 42 High 141 103 228 462 320 46 184 51 29 0 69 285 1918 42 Low 334 472 1615 2749 2437 327 1930 365 406 21 424 2012 13092 43 High 146 123 174 91 183 115 208 96 268 88 131 112 1733 43 Low 1748 3127 3380 1558 1001 1599 2702 669 2753 1002 470 961 20968 47 High 156 246 130 202 524 479 433 119 244 368 222 151 3274 47 Low 1374 1728 648 2188 3095 2183 2361 134 1088 1392 1158 547 17898 48 High 32 401 180 344 385 254 105 19 101 40 22 60 1943 48 Low 258 2528 1325 2677 2385 2915 1823 444 2128 808 537 767 18595 50 High 407 299 272 96 131 303 378 0 112 218 60 46 2322 50 Low 1573 1790 1316 379 583 1780 2408 6 659 1057 439 276 12266 GROUP 2(50 MG/KG) 52 High 204 210 69 248 285 86 119 56 73 136 11 8 1505 52 Low 970 808 423 1171 1485 1022 627 525 72 421 41 63 7628 55 High 167 132 93 203 331 54 404 24 197 72 136 96 1909 55 Low 1916 1199 776 1136 2139 431 1740 24 1826 767 1469 793 14216 56 High 57 51 1 256 262 532 182 302 61 47 25 1911 3687 56 Low 297 228 10 537 216 1152 72 686 228 235 76 363 4080 57 High 79 218 154 145 54 19 33 56 2 42 0 1 803 57 Low 912 2301 2682 2079 1203 465 866 1216 59 841 0 101 12725 58 High 273 33 228 139 145 232 1 71 104 133 113 211 1683 58 Low 1042 91 694 315 427 981 7 244 118 354 760 552 5485 GROUP3(150 MG/KG) 61 High 134 72 110 136 168 44 145 55 0 58 0 36 958 61 Low 1125 686 905 963 1682 1058 1946 438 0 689 0 416 9908 63 High 375 425 228 292 419 194 140 194 182 10 261 84 2804 63 Low 2660 3379 1624 2570 3178 2539 982 1637 985 0 2088 838 22480 64 High 302 65 483 475 271 317 85 32 72 0 0 48 2150 64 Low 1427 676 2245 2209 2020 2461 2276 295 1123 0 19 182 14933 65 High 185 90 184 18 180 79 54 40 0 30 2 43 905 65 Low 259 615 1961 340 2115 1261 1299 1187 5 636 115 889 10682 67 High 317 161 800 512 458 135 519 251 33 348 121 151 3806 67 LOW 1348 821 4852 3478 3423 926 3940 2300 108 1911 748 496 24351 GROUP 4(500MG/KG) 73 High 50 288 39 295 156 292 117 64 272 161 115 20 1869 73 Low 1046 1975 649 2403 1090 2282 156 573 1742 492 471 115 12994 74 High 424 117 375 217 168 214 153 94 183 57 167 144 2313 74 Low 2226 1285 3293 3601 1168 1103 1204 411 2937 460 1553 426 '19667 75 High 107 97 0 134 57 191 37 54 54 40 0 4 775 75 Low 1580 1472 22 1700 924 2659 599 1163 1561 1140 0 197 13017 77 High 235 61 71 171 129 126 0 77 47 103 0 122 1142 77 Low 2140 383 1240 2531 2841 2629 0 1547 670 1865 1 1480 17327 76 High 263 97 55 119 102 27 64 63 80 102 26 16 1014 76 Low 3683 3707 2398 3739 2836 1415 2978 2844 2778 1245 1505 383 29511

- 

### Page25-

## C.I. PIGMENT BLUE 61 NOTOX 

## Project488231 APPENDIX 2 HAEMATOLOGY MALES END OF TREATMENT
ANIMAL WBC Neutrophils Lymphocytes Monocytes Easinophils 10E9/L %WBC %WBC %WBC %WBC
GROUP I (CONTROL) 1 5.2 19.0 76.1 2.2 2.3 2 69 14.1 82.8 1.9 0.7 3 9.2 13.3 82.6 2.8 1.0 7 6.9 13.2 83.5 2.1 0.9 10 7.4 17.5 78.5 2.1 

### 1.4 GROUP 2(50 MGIKG) 11 7.3 14.0 83.0 1.6 0.8 12 10.3 11.8 85.6 1.7 0.6 14 8.1 16.0 81.0 2.2 0.5 15 7.9 15.7 802 27 1.2 16 6.7 10.7 85.6 2.6 0.7 GROUP3 (250 MG1KG) 21 9.5 20.7 74.7 32 1.1 22 7.7 13.6 83.3 2.1 0.8 23 6.5 18.1 77.7 2.7 1.1 24 7.4 14.9 81.9 17 0.9 25 10.4 12.4 84.8 1.7 0.7 GROUP 4(1000 MGIKG) 32 10.0 8.0 89.6 1.6 0.5 34 7.3 12.2 83.7 28 1.0 35 6.3 12.2 83.6 20 1.5 36 7.8 18.2 79.8 1.3 0.6 37 6.7 9.9 876 1.7 0.6 

## MALES END OF TREATMENT
ANIMAL Basophils Red blood cells Reticulocytes ROW Haemoglobin %WBC 10E12/L %RBC %mmol/L

GROUP I (CONTROL) 1 0.3 7.43 2.8 13.2 9.0 2 0.5 8.42 3.3 136 10.1 3 0.2 8.67 2.6 11.8 9.7 7 0.3 8.06 2.7 13.0 9.7 10 0.6 8.06 2.8 11.7 10.1 GROUP 2(50 MG/KG) 11 0.6 8.09 2.7 13.2 9.6 12 0.4 9.04 2.6 12.7 10.2 14 0.3 9.36 1.9 12.1 10.6 15 0.3 8.24 2.6 12.7 '10.0 16 0.3 8.61 3.8 17.2 9.0 GROUP 3(250 MG/KG) 21 0.3 9.05 2.6 13.2 10.3 22 0.3 7.93 3.0 13.2 9.6 23 0.4 9.04 2.6 12.6 10.4 24 0.5 8.62 2.3 12.6 9.8 25 0.4 9.17 2.3 12.2 10.3

-Page 26 -29Sep08 llh10

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 HAEMATOLOGY MALES END OF TREATMENT
ANIMAL Basophils Red blood cells Reticulocytes ROW Haemoglobin %WBC 10E12/L %/RBC %mmol/L
GROUP 4 (1000 MG/KG) 32 0.4 8.48 2.8 12.4 9.5 34 0.4 8.83 2.5 12,9 10.1 35 0.8 8.89 2.3 12.8 9.9 36 0,3 8.25 2.2 23.9 9.9 37 0.1 8.99 2.0 12.9 9.9 

## MALES END OF TREATMENT
ANIMAL Haematocril MCV MCH MCHC Platelets L/L fL. final mmol/L IO0E9IL

GROUP I(CONTROL) 1 0.382 51.5 1.21 23.44 699 2 0.438 52.0 1.20 23.03 985 3 0447 52.2 1.14 21.75 990 7 0421 52.3 1.20 23.00 770 10 0.434 53.8 1.26 23.33 583 GROUP 2(50 MGIKG) 11 0.418 51.6 1.19 23.10 750 12 0.465 51.4 1.13 21.96 925 14 0.482 51.5 1.13 21.91 906 15 0.437 53.0 1.21 22.84 775 16 0.417 48.4 1.04 21.55 955 GROUP 3 (250 MG/KG) 21 0.468 51.7 1.14 21.95 972 22 0.425 53.5 1.21 22.53 954 23 0.467 51.7 1.15 22.21 792 24 0.442 51.3 1.14 22.17 774 25 0.472 51.5 1.12 21,74 873 GROUP 4(1000 MGJKG) 32 0.441 52.0 1.12 21.61 988 34 0.460 52.1 1.14 21.92 964 35 0.451 50.7 1.12 22.07 1008 36 0.434 52.6 1.20 22 89 875 37 0.453 50.4 1.11 21.96 1069 

## MALES END OF TREATMENT
ANIMAL PT APTT s s 

## GROUP I(CONTROL) 2 17.5 19.9 

### 3 19.5 19.4 7 183 188 10 16.2 17.4



### -Page 27 -M 

### 29SepO8IWhO

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 HAEMATOLOGY MALES END OF TREATMENT
ANIMAL PT AP'Fr $ $
GROUP 2(50 1861KG) 11 1637 18.3 

### 12 1 8.4 21.4 14 19.1 22.6 15 18.4 21.5 16 17.9 19.81 GROUP 3 (250 MG/KG) 21 16.7 16.2 22 17.9 20.7 23 17.6 22.3 24 17.0 16.2 25 17.6 20.7 GROUP 4 (1000 MG/KG) 32 17.4 19.5 34 18.1 17.7 35 18.2 21.8 36 17.0 21.7 37 17.1 16.4 

## FEMALES END OF TREATMENT
ANIMAL WBC Neutrophils Lymphocytes Monocytes Eosinophils 10E9/L %WBC %WBC %WBC %WBC

GROUP 1 (CONTROL) 42 3.3 29.2 68.2 1.8 0.3 43 4.7 12.0 84.0 3.0 1.0 47 4.3 22.8 73.5 2.0 1.6 48 3.7 21.6 75.3 2.4 0.4 50 3.9 24.0 72.0 4.0 0.0 GROUP 2(50 MG/KG) 52 3.4 16.3 81.4 1.3 0.8 55 6.1 15.0 82.9 1.6 0.3 56 --- 53.0 450 2.0 0.0 57 3,5 21.0 77.0 2.0 0.0 58 29 25.4 70,9 2.9 0.6 GROUP 3 (250 MG/KG) 61 5.9 30.2 67.5 1.8 0.2 63 5.9 30.6 66.4 2.3 0.3 64 3.3 27.3 69.8 1.7 0.2 65 5.3 23,5 73.4 1.6 1.4 67 2.2 30,0 66.0 3.0 1.0 GROUP4(1000 MG/KG) 73 5.2 15.5 81.1 2.5 0.7 74 7.5 39.0 61.0 0.0 0.0 75 5.6 27.0 71.0 1.0 1.0 76 4.6 15.5 82.1 1.3 0.8 77 5.6 27.0 71.0 2.0 0.0

-Page28

- 

## Wii 29Sep08 lihG

## 0.1. PIGMENT BLUE61 NOTOX Project488231 APPENDIX 2 HAEMATOLOGY FEMALES END OF TREATMENT
ANIMAL Basophils Red blood cells Reticulocytes RIDW Haemoglobin %WBC IOE12IL %RBC %mmol/L
GROUP I(CONTROL) 42 0.5 7.13 4.2 16.7 9.1 43 0.0 7.34 5.0 14.0 9.1 47 0.2 7.10 4.2 12.3 8.8 48 0,3 7.37 6.4 15.7 8.5 50 0.0 7,50 4.0 13.1 9.0 GROUP 2(50 MG/KG) 52 

### 0.3 7.42 4.1 12.6 93 55 0.1 7.81 3.6 12.3 9.2 56 00 7.06 7.7 17.8 9.2 57 0.0 7.36 5.9 21.0 9.4 58 0.3 8.29 2.9 14.2 9.3 GROUP 3(250 MG/KG) 61 0.3 6.85 7.6 17.2 8.4 63 0.4 7.58 4.3 13.7 8.2 64 1.1 7.32 7.9 16.9 8.4 65 0.1 7.39 4.8 14.8 8.9 67 0.0 7.48 3.9 16.3 9.0 GROUP 4(1000 MGIKG) 73 0.2 7.91 4.3 12.1 9.3 74 0.0 7.66 5.7 14.9 8.8 75 0.0 750 5.2 14.9 9.1 76 0.2 7.30 4.7 13.5 9.3 77 0.0 7.75 6.6 14.8 9.3 

## FEMALES END OF TREATMENT
ANIMAL Haematocrit MGV MCH MCHC Platelets L/L fL fmol mmol/L I10E9/L

GROUP I(CONTROL) 42 0.377 529 1.27 24.01 1102 43 0.396 539 1.24 22.97 1127 47 0.386 54.4 1.24 22.83 652 48 0.380 51.6 1.16 22.45 1540 s0 0.389 51 9 1.21 23.21 1166 GROUP 2(50 MG/KG) 52 0.397 53.6 1.26 23.50 1372 55 0.414 530 1.18 22.28 1328 56 0.395 55.9 1.30 23.30 1443 67 0405 55.1 1.28 23.17 1197 58 0.415 50.1 1.12 22.43 1262 GROUP 3(250 MG/KG) 61 0.361 52.7 1.22 23.18 1660 63 0.399 52.7 1.21 22.93 1312 64 0.389 53.2 1.15 21.64 1670 65 0.388 52.5 1.21 22.94 1273 67 0.391 52.2 1.21 23.12 1022

-Page 29

- .29Sep08 111110

## C.l. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 HAEMATOLOGY FEMALES END OF TREATMENT
ANIMAL Haematocrit MCV MCH MCHC Platelets UJL fl. find 

### mmoIIL 1OE9IL
GROUP 4(1000 MG/KG) 73 0.425 53.7 1.18 21.99 1399 74 0.393 51.3 1.14 22.32 861 75 0.395 52.6 1.21 23.02 1363 76 0.396 54.2 1.27 23.44 1517 77 0.414 53.5 1.20 22.42 835 

## FEMALES END OF TREATMENT
ANIMAL PT APTT S

GROUP I(CONTROL) 42 17.7 19.4 43 16.8 16.0 47---- 48 16.7 19.9 50 17.2 15.4 GROUP 2(50 MVG/KG) 52 16.8 22.0 55 17.4 21.9 56 16.1 14.8 

### 57 15.9 16.9 58 16.4 162 GROUP 3(260 MG/KG) 61 16.6 18.8 63 16.2 16.0 64 17.3 17.0 65 15.8 14.9 67 15.5 15.5 GROUP 4(1000 MG/KG) 73 16.4 21.9 74 15.6 15.5 75 15.6 14.9 76 15.3 13,3 77 15.1 14.0



### -Page 30

- 29Sep08 IlhIO

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 CLINICAL BIOCHEMISTRY MALES END OF TREATMENT
ANIMAL ALAT ASAT ALP Total protein Albumin U/L U/L U/L g/L g/L
GROUP 1 (CONTROL) 1 57.1 71.1 203 65.9 34.4 2 39.8 79.7 140 64.6 34.4 3 45.0 72.8 135 62.2 34.0 7 38.2 74.1 185 64.7 32.8 10 43.3 111.7 105 58.9 31.3 GROUP 2(50 MG/KG) 11 40.9 85.1 197 61.7 32.6 12 56.3 79.3 187 68.0 34.5 14 65.5 77.7 195 65.3 33.6 15 33.6 73.2 142 65.6 33.5 16 63.6 99.4 142 65.3 34.3 GROUP 3(250 MG/KG) 21 40.8 78.3 185 68.3 34.1 22 46.2 72.7 151 65.5 33.9 23 58.2 87.4 126 65.6 34.2 24 43,4 78.8 154 624 33.3 25 45.6 81.4 135 65.4 34.7 GROUP 4(1000 MG/KG) 32 47.1 82.2 120 64.9 33.8 34 43.5 85.7 116 65.8 34.4 35 44.5 84.0 188 65.5 34.0 36 45.5 73.5 120 66.4 34.5 37 58.6 87.8 169 68.2 34.8 

## MALES END OF TREATMENT
ANIMAL Total bitirubin Urea Creatinine Glucose Cholesterol umol/L mmol/L umol/L mmolIL mmol/L

GROUP I(CONTROL) 1 2.6 6.7 42.1 7.05 1.48 2 1.9 6.4 40.6 10.52 1.54 3 2.3 11.9 43.6 8.38 1.44 7 2.1 9.0 45.1 7.50 2.20 10 2.0 9.9 42.8 6.89 1.59 GROUP 2(50 MG/KG) 11 2.0 8.4 47.3 6.23 1.84 12 2.1 10.2 39.8 8.03 2.16 14 2.6 11.2 48.1 10.20 1.83 15 2.4 8.2 39.1 6.93 2.22 16 2.8 9.3 55.6 6.07 1.81 GROUP 3 (250 MG/KG) 21 1.8 8.7 43.6 6.84 2.12 22 2.6 8.1 42.1 7.29 2.21 23 2.4 9.4 46.6 8.31 1.60 24 2.2 7.9 45.1 5.96 1.75 25 2.1 8.7 54.1 8.19 1.73

-Page 31 

# ~ 29SepO81lhIO

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 CLINICAL BIOCHEMISTRY MALES END OF TREATMENT ANIMAL Total bilirubin Urea Creatinine Glucose Cholesterol umol/L mmol/L umol/L mmol/L mmol/L
GROUP 4(11000MG/KG) 32 2.1 7.8 43.6 9.76 2.10 34 2.1 8.3 44.3 6.36 1.72 35 2.0 8.2 41.3 761 1.92 36 1.8 10.3 44.3 6.15 2.33 37 1.8 7.6 41.3 6.11 2.57 

## MALES END OF TREATMENT
ANIMAL Sodium Potassium Chloride Calcium lnorg.Phos mmol/L mmol/L mmol/L mmol/L mmol/L

GROUP 1 (CONTROL) 

### 1 1 42.1 3.94 104 2.80 2.05 2 142.0 3.39 105 2.81 1.75 3 142.0 3.82 104 2.73 2.18 7 142.9 3.74 104 2.83 2.03 10 142.4 3.94 105 2.75 2.32 GROUP 2(50 MG/KG) 11 142.0 388 104 2.64 1.94 12 142.1 4.22 104 2.76 2.08 14 142.5 4.18 104 2.66 1.71 15 142.6 3.88 104 2.73 210 16 140.9 3.97 103 2.74 2.40 GROUP 3(250 MG/KG) 21 143.4 3.91 104 2.82 2.40 22 1420 3.83 103 2.69 216 23 142.1 3.71 105 2.77 2.08 24 1421 3.80 105 2.61 2.10 25 143.5 379 106 2.76 2.15 GROUP 4 (1000 MG/KG) 32 142.6 4.25 104 2.69 1.94 34 143.5 3.88 104 2.77 2.08 35 148.2 3.90 104 2.72 2.21 36 141.9 4.37 108 2.77 2.09 37 142.6 3.77 105 2.72 1.88 

## FEMALES END OFTREATMENT ANIMAL ALAT ASAT ALP Total protein Albumin U/L U/L U/L g1L 

### g/L

GROUP I (CONTROL) 42 62.8 85.7 56 64.6 32.3 43 45.6 85.0 42 65.0 32.2 

### 47 64.8 95.1 102 66.6 32.7 48 43.5 73.9 60 67.1 34.0 

### 50 73.6 64.8 64 657 31.7

- Page 32- 29Sep08 11h10

## C.I PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 CLINICAL BIOCHEMISTRY FEMALES END OF TREATMENT
ANIMAL ALAT ASAT ALP Total protein Albumin U/L U/L U/L g/L gIL
GROUP 2(50 MG/KG) 52 67.2 78.5 47 69.1 33.6 55 45.0 58.4 92 63.7 30.2 56 68.3 76.0 70 64.0 32.6 57 59.7 71.3 61 67.5 34.7 58 98.7 91.5 91 62.1 295 GROUP 3(250 MG/KG) 61 72.4 74.6 86 69.2 35.4 63 587 83.2 61 682 34.1 64 77.5 762 87 67.3 32.5 65 65.1 70.1 76 64.1 32.8 67 59.4 96.3 39 69.5 34.1 GROUP 4(1000 MGKG) 73 36.8 62.5 36 643 31.7 74 42.3 76.9 113 62.6 31.8 75 55.4 72.1 120 665 32.6 76 46.0 73.6 44 68.1 34 0 77 48.5 65.9 31 66.5 33.2 

## FEMALES END OF TREATMENT
ANIMAL Total bilirubin Urea Creatinine Glucose Cholesterol umol/L mmolIL umolIL mmol/l. mmolIL

GROUP I(CONTROL) 42 3.0 7.8 46.5 6.56 1.35 43 2.1 9.3 50.0 495 1.14 47 20 11.3 45.4 8.26 1.58 48 2.3 7.1 41.7 5.63 1.48 50 24 11.4 50.0 7.02 1.69 GROUP 2(60 MG/KG) 52 2.4 9.7 47.6 6.46 2.09 55 15 95 48.3 4.65 1.48 56 2.1 10.1 45.8 5.69 1.40 57 1.9 9.2 45.4 5.06 1.49 58 2.3 12.2 49.0 7.74 1.98 GROUP 3(250 MG/KG) 61 2.8 9.8 45.1 5.24 1.67 63 2.0 9.4 40.2 5.79 1.51 64 2.8 11.5 46.5 5.16 160 65 3.1 9.5 47.2 4.49 1.87 67 1.7 9.7 54.2 6.69 1.59 GROUP 4(1000MG/KG) 73 1.6 11.4 49.8 5.10 1.65 74 1.5 9.9 50.0 5.94 1.33 75 1.7 9.3 50.0 5.23 2.24 76 2.3 9.8 47.4 7.02 1.80 77 2.0 8.0 43.0 5.77 1.71

-Page 33

- 29Sep08 llhlO

## ClI. PIGMENT BLUE61 NOTOX Project 488231 APPENDIX 2 CLINICAL BIOCHEMISTRY 

## FEMALES END OF TREATMENT
ANIMAL Sodium Potassium Chloride Calcium lnorg.Phos mnmol/L mmol/L mmol/L mmol/L mmol/L
GROUP I(CONTROL) 42 142.2 364 96 2.76 264 43 144.3 356 100 2.73 2.35 47 139.2 3.95 102 2.80 1.91 48 140.5 3.42 98 2.78 1.92 50 141.5 3.34 98 2.82 2.54 GROUP 2(50 MG/KG) 52 141.2 363 98 2.70 1.71 55 141.9 400 101 2.82 2.61 56 141.2 368 100 272 2.27 57 141.0 3.72 97 2.82 2.29 58 139.5 3.93 99 2.69 2.26 GROUP 3 (250 MG/KG) 61 140.3 3.49 96 2.88 2.63 63 140.1 3.65 97 291 247 64 142.5 3.57 98 2.81 3.02 65 140.4 3.98 98 2.71 2.49 67 142.5 3.66 97 2.72 2.33 GROUP 4(1000 MG/KG) 

### 73 141.0 3.73 99 2.75 2.61 74 141.2 3.14 98 2.67 2.31 75 141.7 3.38 95 2.76 2.33 76 140.4 4.34 98 2.75 2.10 

### 77 141.7 3.45 95 2.77 2.18

-Page 

### 34

- 

# ~29Sep08 llh10

## ClI. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 MACROSCOPIC FINDINGS MALES ALL NECROPSIES
ANIMALORGAN FINDING DAY OF DEATH
GROUP I(CONTROL) 1 Nofindings noted Scheduled sacrifice, 17Jul2008 2 No findings noted Scheduled sacrifice. 17JuI2008 3 Bone Tail: tailbent. Scheduled sacrifice, 17Jul2008 4 No findings noted Scheduled sacrifice. 17JuI2008 6 Nofindings noted Scheduled sacrifice, M1u2008 6 Nofindings noted Scheduled sacrifice. 17Jul2008 7 Nofindings noted Scheduled sacrifice, 17Jul2008 8 Nofindings noted Scheduled sacrifice, 17Jul2008 g Nofindings noted Scheduled sacrifice, 17,u02008 10 Nofindings noted Scheduled sacrifice, 17Jul2008 GROUP 2(50 MG/KG) I1I Nofindings noted Scheduled sacrifice, 1 7Jul2008 12 Mesenteric Inode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 13 Nofindings noted Scheduled sacrifice, 1 7Jul2008 14 Nofindings noted Scheduled sacrifice, 17Jul2008 15 Skin Tail: discolouration, bluish. Scheduled sacrifice, 17Jul2008 16 Mesenteric Lnode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 17 No findings noted Scheduled sacrifice, 17Jul2008 18 No findings noted Scheduled sacrifice, 17,102008 19 Nofindings noted Scheduled sacrifice, 17Jul2008 20 No findings noted Scheduled sacrifice, 1 7Jul2008 GROUP 3(250 MG/KG) 

### 21 Mesenteric Inode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 22 Epididymides Right side, tall: nodule(s), d=7x5 mm, Scheduled sacrifice, 17Jul2008 Yellowish, hard. Mesentenc Inode Discolouration, greenish. 23 Mesenteric Lnode Discolouralion, greenish. Scheduledsacrifice, 17Jul2008 24 Mesenteric Inode Discolouration, greenish. Scheduledsacrifice, 1 7JuI2008 25 Mesenteric Inode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 26 Nofindings noted Scheduledsacrifice. 17Jul2008 27 Epidldymides Rightside, tall: nodule(s), d=2x2 mm, Scheduledsacrifice, 17Jul2008 Yellowish, hard. Mesenteric Inode Discolouration, greenish. 28 MesentericInode Discolouration, greenish. Scheduledsacrifice, 1 7Jui2008 29 MesentericInode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 30 MesentericInode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 GROUP 4(1000 MG/KG) 

### 31 MesentericLnode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 32 MesentericLnode Discolouration, greenish. Scheduled sacrifice, 17,102008 33 MesentericLnode Discolouration, greenish. Scheduled sacrifice, 1 7Jul2008 34 Mesenteric I.node Discolouration, greenish. Scheduled sacrifice, 17Jul2008 Skin Foreleg, left side: alopecia 35 Mesenteric Inode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 36 Mesenteric Lnode Discolouralion, greenish. Scheduled sacrifice, 1 7Jul2008 37 Mesenteric Inode Discolouration, greenish. Scheduled sacrifice, 17Jut2008 38 MesentericLnode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 39 MesentericInode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 Skin Tall: dIscolouration, bluish. 40 Seminal vesicles Left side: reduced insize. Scheduled sacrifice, 1 7Jul2008 Mesenteric Lnode Discolouralion, greenish.

-Page 35 011- 29Sep08 11110

## 02. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2
MACROSCOPIC FINDINGS FEMALES ALLNECROPSIES ANIMALORGAN FINDING DAYOF DEATH GROUP I (CONTROL) 41 Nofindings noted Scheduled sacrifice,29Jul2008 42 Nofindings noted Scheduledsacrifice,28Jul2008 43 No findings noted Scheduled sacrifice,28Jul2006 44 No findings noted Scheduledsacrifice,OlAug2008 45 Nofindings noted Scheduled sacrifice,OlAug2008 46 Nofindings noted Scheduledsacrifice,OlAug2008 47 Mandibular node Bothsides:discolouration, dark red. Scheduled sacrifice,30Jul2008 Skin Foreleg, left side: alopecia. Abdominal region: alopecia. Foreleg, rightside: alopecia. Flank, right side: alopecia. Flank, left side. alopecia. 48 Nofindings noted Scheduled sacrifice,3OJuI2008 49 Nofindings noted Scheduled sacrifice,30Jul2008 50 Nofindings noted Scheduled sacrifice,28Jul2008 GROUP 2(So MG/KG) 51 Nofindings noted Scheduled sacrifice, 31Jul2008 52 Mandibular inode Both sides:discolouration, reddish. Scheduledsacrifice, 30Jul2008 53 No findings noted Scheduled sacrifice, OlAug2008 54 Nofindings noted Scheduled sacrifice, 29Jut2008 55 No findings noted Scheduledsacrifice, 30JuI2008 56 Pancreas Discolouration, reddish. Scheduled sacrifice, 28Jul2008 MesentericInode Discolouration, bluish MandibularLnode Bothsides: discolouration, dark red. 57 No findings noted Scheduledsacrifice, 30Jul2008 58 Nofindings noted Scheduledsacrifice, 30Jul2008 59 Nofindings noted Scheduledsacrifice, 30Jul2008 60 No findings noted Scheduled sacrifice, 30Jul2008 GROUP 3(250 MG/KG) 61 Mesenteric tLnode Discolouration, bluish. Scheduled sacrifice,28Jul2008 62 Nofindings noted Scheduled sacrifice 31Jul2008 63 Mesenteric Inode Discolouration, bluish. Scheduled sacrifice,28Jut2008 64 Mesenteric Inode Discolouration, bluish. Scheduled sacrifice,28Jul2008 65 Mesenteric Lnode Discolouration, bluish. Scheduled sacrifice,28JuI2008 66 No findings noted Scheduled sacrifice,29Jul2008 67 MesentericInode Discolouration, bluish. Scheduledsacrifice, 30JuI2008 68 MesentericInode Discolouration, bluish. Scheduledsacrifice, 30Jut2008 69 MesentericInode Discolouration, bluish. Scheduledsacrifice, 30Jul2008 70 MesentericInode Discolouration, bluish. Scheduledsacrifice, 31Jul2008 GROUP 4(1000 MG/KG) 71 General observationsG~i-tractus, contents: bluish. Scheduledsacrifice, OlAug2008 Mesenteric Lnode Discolouration, bluish. Skin Foreleg, right side: alopecia. Hindleg, left side: alopecia. Hindleg, right side:alopecla. Foreleg, leftside. alopecia. 72 General observationsGl-tractus, contents: bluish. Scheduled sacrifice, 29Jul2008 73 General observationsGi-tractus,contents: bluish. Scheduled sacrifice, 30Jul2008 MesentericLnode Discolouration, bluish. 74 General observationsGi-tractus,contents: bluish. Scheduled sacrifice, 28Jut2008 Adrenal glands Both sides:discolouration, red-brown. MesentericLnode Discolouration, bluish. Mandibular Lnode Bothsides: discolouration,reddish. 75 Mesenteric Inode Discolouration, bluish. Scheduled sacrifice,28Jut2008 76 General observationsGi-tractus, contents: bluish. Scheduledsacrifice, 29JuI2008 Mesenteric Inode Discolouration, bluish. 77 Mesenteric Inode Discolouration, bluish. Scheduledsacrifice, 28JuI2008 78 General observationsG. i-tractus, contents: bluish. Scheduledsacrifice, 01Aug2008 Mesenteric Inode Discolouration, bluish. 79 Mesenteric Inode Discolouration, bluish. Scheduledsacrifice, 31Jul2008

-Page 36 -I'29Sep08 lihiC

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2
MACROSCOPIC FINDINGS FEMALES ALL NECROPSIES ANIMALORGAN FINDING DAY OF DEATH
GROUP 4(1000 MGIKG) 80 General observatlonsGi-tractus,contents: bluish. Scheduled sacrifice,31Jul2008 Mesenteric Inode Discolouration, bluish.

-Page37 111110 29Sep081lIMO

## 0.1. PIGMENT BLUE61 NOTOX Project 488231 APPENDIX 2 ORGAN WEIGHTS (GRAM) MALES NECROPSY
ANIMAL BODYW. BRAIN HEART LIVER THYMUS (GRAM) (GRAM) (GRAM) (GRAM) (GRAM) GROUPlI(CONTROL) 1 361 2.06 0.918 9.21 0.361 2 316 2.01 0.914 8.02 0.370 3 354 2.07 0,961 9.51 0.435 

### 4 323 

### 1.95 0.872 725 0.262 5 344 2.09 0.881 8.23 0.346 6 401----- - 7 345------- - 8 373------- - 9 346

--------- 10 383

--------- GROUP 2(50 MG/KG) 11 341 1.99 1.019 7.54 0.279 12 332 2.04 0.966 781 0.444 13 337 1.97 0.918 8.47 0.361 14 320 2.03 0.880 7.38 0.283 15 365 2.06 0.930 9.90 0.477 16 335----- - 17 398----- - 18 336----- - 19 335

--------- 20 395 ------ --- GROUP 3(260 MG/KG) 21 379 2.11 1091 10.70 0.425 22 361 1.95 1.055 9.34 0434 23 340 2.08 0889 8.66 0.370 24 347 1.98 0.987 8.33 0.400 25 325 1.97 0.933 8.14 0.343 26 364---- 27 369------- - 28 371 -- - --- 29 351------- - 30 371------- - GROUP 4(1000 MG/KG) 31 363 2.13 1.066 10.23 0.327 32 348 2.08 1.119 9.92 0.407 33 351 2.04 0.962 8.74 C.337 34 340 2.10 0.936 8.16 0.287 35 342 2.03 0.941 9.10 0.347 36 352

--------- 37 343------- - 38 350 --- ------ 39 348------- - 40 341 --- --- ---

-Page 38 -. 

### 29Sep08 IlhIO

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 ORGAN WEIGHTS (GRAM) MALES NECROPSY
ANIMAL KIDNEYS ADRENALS SPLEEN TESTES EPIDIDYMIDES (GRAM) (GRAM) (GRAM) (GRAM) (GRAM)
GROUP I (CONTROL) 1 2.48 0.069 0.647 3.42 1.165 2 2.43 0.061 0.504 3.32 1.254 3 2.46 0.077 0.709 3.43 1.238 4 2.34 0.082 0.434 3.45 1.163 5 2.43 0.066 0.503 3.67 1.153 6 - -- -- -3.30 1.001 7 - -- -- -3.08 1.027 8 -- - -- 3.36 1.187 9 - -- -- -3.29 1.050 10 - -- -- -3.29 1.153 GROUP 2(50 MG/KG) 11 2.54 0.079 0.626 3.88 1.193 12 2.39 0,064 0586 3.47 1.067 13 246 0.071 0.540 2.60 0.969 14 2.21 0.062 0.495 3.40 1.206 15 2.42 0.073 0.703 324 1095 16 --- -- -2.66 0.951 17 - -- -- -3.67 1.177 18 - -- -- -3.21 1.183 19 - -- --- 364 1.281 20 - -- -- -3.06 1.261 GROUP 3 (250 MG/KG) 21 2.54 0.065 0.695 3.71 1.219 22 2.36 0.059 0.663 3.54 1.443 23 2.36 0.070 0.647 3.35 1.232 24 2.39 0.070 0.632 3.24 1.011 25 2.10 0.063 0.496 236 1.079 26 - -- --- 3.33 1.078 27 - -- -- -3.29 1.323 28 - -- -- -3.40 1.195 29 -- 3----.47 1.283 30---------- GROUP 4 (1000 MG/KG) 31 265 0.097 0.710 3.70 1.418 32 2.33 0.067 0.659 3.16 1.061 33 2.60 0.068 0.688 3.70 1.148 34 2.32 0.065 0.677 3.24 1.190 35 2.17 0.077 0.625 3.38 1.165 36 - -- -- -3.47 1.195 37 - -- -- -3.78 1.266 38 - -- -- -2.61 0.959 39 - -- -- -3.46 1.273 40 - -- -- -3.44 1.143

Page39- 29Sep08illI

# o i.PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 ORGANIBODY WEIGHT RATIOS I(%) MALES NECROPSY
ANIMAL BODY W. BRAIN HEART LIVER THYMUS 

### (GRAM) (%) (%)(%()
GROUP I (CONTROL) 1 361 0.57 0.254 2.55 0.100 2 316 0.64 0.289 2.54 0.117 3 354 0.58 0.271 2.69 0.123 4 323 0.60 0.270 2.24 0.081 5 344 0.61 0.256 2.39 0.101 6 401--- - 7 345------- - 8 373------- - 9 346------- - 10 363------- - GROUP 2 (60MG/KG) 11 341 0.58 0.299 2.21 0.082 12 332 0.61 0.291 2.35 0.134 1 3 337 0.58 0.272 251 0.107 14 320 0.63 0.275 2.31 0.088 15 365 0.56 0.255 2.71 0.131 16 335----- - 17 398----- - 18 336------- - 19 335------- - 20 395 - - - - GROUP 3 (250 MG/KG) 21 379 056 0.288 2.82 0.112 22 361 0.54 0.292 2.59 0.120 23 340 0.61 0.261 2.55 0.109 24 347 0.57 0.284 2.40 0.115 25 325 0.60 0.287 2.50 0.106 26 364----- - 27 369------- - 28 371------- - 29 351------- - 30 371------- - GROUP 4 (1000 MG/KG) 31 363 0.59 0294 282 0.090 32 348 060 0.322 2.85 0.117 33 351 0.58 0.274 2.49 0.096 34 340 0.62 0.275 2.40 0.084 35 342 0.59 0.275 2.66 0.101 36 352------- - 37 343------- - 38 350----- - 39 348------- - 40 341----- -

-Page 40

- lI 29Sep08 llhIO

# C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 ORGANIBODY WEIGHT RATIOS(% MALES NECROPSY
ANIMAL KIDNEYS ADRENALS SPLEEN TESTES EPIDIDYMIDES
GROUPI1(CONTROL) 1 0.69 0.019 0.179 0.95 0.323 2 0.77 0.016 0.1659 1.05 0.397 3 0.70 0.022 0.200 0.97 0.350 4 0.72 0.025 0.134 1.07 0.360 5 0.71 0.019 0.146 1.07 0.335 6 - -- -- -0.82 0.250 7 - -- -- -0.89 0.298 8 - -- -- -0.90 0.318 9 ---- 0.95 0.303 10 - -- -- -0.86 0.301 GROUP 2(50 MG/KG) 11 0.74 0.023 0.184 1.14 0.350 12 0.72 0.019 0.177 1.05 0321 13 0.73 0.021 0.160 0.77 0.288 14 0.69 0.016 0 155 1.06 0.377 15 0.66 0.020 0.193 0.89 0300 16 - -- -- -0.79 0.284 17 - ---- -0.92 0.296 18 - -- -- -0.95 0.352 19 - -- -- -1.09 0.382 20 - ----- 0.77 0.319 GROUP 3,(250 MG/KG) 21 0.67 0.017 0.183 0.98 0.322 22 0.65 0.018 0.184 0.98 0.400 23 0.69 0,021 0.190 0.99 0.362 24 0.69 0,020 0.182 093 0.291 25 065 0.019 0.153 0.73 0.332 26 - ----- 0.92 0.296 27 - ----- 0.89 0.359 28 ----- -0.92 0.322 29 ------ 0.99 0.366 30---------- GROUP 4(1000MG/KG) 31 0.73 0.027 0.196 1.02 0.391 32 0.67 0.019 0.189 0.91 0.305 33 0.74 0.019 0.196 1.06 0.327 34 0.68 0.019 0.199 0.95 0.350 35 0.63 0.023 0.183 0.99 0.341 36 ------ 0.98 0.339 37 ------ 1.10 0.369 38 ----- -0.74 0.274 39 ------ 099 0.366 40 - ----- 1.01 0.335

-Page41 295ep08llh10

## C.I. PIGMENT BLUE61 NOTOX Project 488231 

## APPENDIX 2
ORGAN WEIGHTS (GRAM) FEMALES NECROPSY
ANIMAL BODY W. BRAIN HEART LIVER THYMUS (GRAM) (GRAM) (GRAM) (GRAM) (GRAM) GROUP1I(CONTROL) 41 ------ - - 42 243 1.82 0.661 7.00 0.118 43 240 1,81 0.786 7.25 0.237 44-------- 45---------- 46---------- 47 201 1.86 0.650 6.48 0.133 48 220 1.80 0.806 7.66 0,198 49---- 50 222 1.93 0.762 7.77 0.115 GROUP 2(50 MGIKG) 51 ---------- - 52 202 1.76 0,755 6.29 0.151 53

-- 54 ---------- - 55 220 1.79 0.731 673 0161 56 223 1.85 0.727 7.54 0.207 57 207 183 0.634 6.59 0.162 58 248 1,76 0.780 8 18 0.167 59 --- ------- - 60 --- ------- - GROUP 3 (250 MGIKG) 61 225 1.85 0.749 8.17 0.202 62---------- 63 230 1.87 0,752 8.12 0.145 64 227 1.78 0.774 7.46 0.168 65 230 1.76 0.847 7.72 0.180 66---------- 67 230 1.90 0.802 7.59 0.158 68 --- --------- 69---------- 70 --- ---- GROUP 4(1000 MG/KG) 71---------- 72-------- 73 211 1.87 0.667 6.14 0.182 74 247 1.84 0.740 7.32 0.175 75 214 1.86 0.756 7.19 0.152 76 212 1.87 0.672 7.01 0.221 77 220 1.85 0.745 7.89 0.165 78 - -- --- --- -- - 79---------- 80
- -- -- -- --

-Page 42 -29Sep08 IlhlO

## C.I. PIGMENT BLUE 61 NOTOX 

## Project488231 APPENDIX 2
ORGAN WEIGHTS (GRAM) FEMALES NECROPSY
ANIMAL KIDNEYS ADRENALS SPLEEN (GRAM) (GRAM) (GRAM)
GROUP I(CONTROL) 41------ 42 1.58 0.078 0.452 43 1.50 0.077 0.496 44---- - 45---- 46---- - 47 1.49 0.072 0.312 48 2.01 0.084 0.548 49 ----- 50 1.64 0.084 0.477 GROUP 2(50 MG/KG) 51 --- - 52 1.65 0.075 0391 53------ 54---- 55 1.62 0.083 0.475 56 1.62 0.097 0.526 57 1.53 0.076 0.502 58 1.70 0.088 0.415 59 60------ GROUP 3(250 MG/KG) 61 1.69 0.077 0,693 62------ 63 1.88 0.096 0.479 64 170 0.080 0.543 65 1.71 0076 0.497 66 -- - - 67 1.71 0.079 0.478 68------ 69------ 70------ GROUP 4(1000MG/KG) 71---- 72------ 73 1.44 0.065 0400 74 1.48 0.100 0.501 75 1.72 0.088 0535 76 1.68 0.089 0.510 77 1.81 0.098 0.559 78 ---- - 79------ 80------

-Page 43

- 

### 29Sep08 llhlO

## C.!. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 ORGAN/BODY WEIGHT RATIOS I(%) FEMALES NECROPSY
ANIMAL BODY W. BRAIN HEART LIVER THYMUS 

### (GRAM) (%(%(%()
GROUP I (CONTROL) 41-------- 42 243 0.75 0.272 2.88 0.049 43 240 0.75 0.328 3.02 0.099 44 --- ------- - 45---------- 46 --- --------- 47 201 0.93 0.323 3.22 0.066 48 220 0.82 0.366 3.48 0.090 49 --- ----- - 50 222 0.87 0,343 3.50 0.052 GROUP 2(50 MG/KG) 53 ----- 54 --- --------- 55 220 0.81 0.332 3.06 0.073 56 223 0.83 0.326 3.38 0.093 57 207 0.88 0.306 3.18 0.078 58 248 0.71 0.315 3.30 0.067 59 ---------- - 60---------- GROUP 3 (250 MG/KG) 61 225 0.82 0.333 3.63 0.090 62 ---------- - 63 230 0.81 0.327 3.63 0.063 64 227 0.78 0.341 3.29 0.074 65 230 0.77 0.368 3.36 0.078 66-------- 67 230 083 0.349 3.30 0.069 68 ---------- - 69 -------- - 70---------- GROUP 4(1000 MG/KG) 71 ---------- - 72---------- 73 211 0.89 0.316 2Z91 0.086 74 247 0.75 0.300 2.96 0.071 75 214 0.87 0.353 3.36 0.071 76 212 0.88 0.317 3.31 0104 77 220 0.84 0.339 3.59 0075 78 ---------- - 79 ---------- - 80 -------- -

-Page 44 - *I- 29SepOS llhlO

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 ORGAN/BODY WEIGHT RATIOS(% FEMALES NECROPSY
ANIMAL KIDNEYS ADRENALS SPLEEN

GROUP 1 (CONTROL) 41------ 42 0.65 0.032 0.186 43 0.63 0.032 0.207 44------ 45-- 46------ 47 0.74 0.035 0.155 48 0.91 0.038 0.249 49 -- - - 50 0.74 0.038 0.215 GROUP 2(50 MG/KG) 51------ 52 0.82 0.037 0.194 53 -- 54 -- 55 0.74 0.038 0.216 56 0.73 0.043 0236 57 0.74 0037 0.243 58 0.66 0,035 0.167 59------ 60---- GROUP 3 (250 MG/KG) 61 0.75 0.034 0308 62------ 63 0.82 0.042 0.208 64 0.75 0,035 0.239 65 0.74 0.033 0.216 66 --- ---- 67 0.74 0.034 0.208 68------ 69------ 70
------- GROUP 4 (1000 MG/KG) 71 -- 72------ 73 0.68 0.031 0.190 74 0.60 0.040 0.203 75 0.81 0.041 0.250 76 0.79 0.042 0.241 77 0.82 0.045 0.254 78------ 79 --- --- 80
-------

-Page 45

- 29Sep08 llhJ0

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 BREEDING DATA PER LITTER FO-GENERATION
- 

## LACTATION

-- FIRST LITTER CHECKLITTER DURATION OF DEAD PUPS LIVING PUPS P.NATAL LOSS GESTATION M F M F TOT M F
GROUP 1 (CONTROL) 42 21 0 0 2 10 12 0 0 43 21 0 0 4 6 10 0 0 44 22 0 0 10 5 15 0 0 45 22 0 0 4 9 13 0 0 46 22 0 0 1 3 4 0 0 47 21 0 0 3 7 10 0 0 48 21 0 0 6 8 1 4 0 0 49 21 0 0 5 7 12 0 1 50 21 0 0 11 3 14 0 0 TOTAL 0 0 46 58 104 0 1 N 9 9 9 9 9 9 9 9 MEAN 21.3 0.0 0.0 5.1 6.4 11.6 0.0 0.1 ST.DEV. 0.5 0.0 0.0 3.4 2.5 3.3 0.0 0.3

-Page 46

- -23Oct08 10h12

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 BREEDING DATA PER LITTER FO-GENERATION
- 

## LACTATION

-- FIRST LITTER CHECK LITTER DURATION OF DEAD PUPS LIVING PUPS P.NATAL LOSS GESTATION M F M F TOT. M F
GROUP 2(50 MG/KG) 51 22 0 0 5 4 9 0 1 52 21 0 0 4 7 11 0 0 53 22 0 0 5 6 11 0 0 55 21 0 0 7 6 13 0 0 56 21 0 0 8 6 14 0 0 57 21 0 0 4 7 11 0 0 58 21 0 0 7 6 13 0 0 59 21 0 0 4 6 10 0 0 60 21 0 0 4 7 11 0 0 TOTAL 0 0 48 55 103 0 1 N 9 9 9 9 9 9 9 9 MEAN 21.2 0.0 0.0 53 6.1 11.4 0.0 0.1 ST.DEV. 0.4 0.0 0.0 1.6 09 16 0.0 0.3

-Page 47 -'"23Oct08 161112

## C.I. PIGMENT BLUE61 NOTOX Project 488231 APPENDIX 2 BREEDING DATA PER LITTER FO-GENERATION
- 

## LACTATION

-- FIRST LITTER CHECKLITTER DURATION OF DEAD PUPS LIVING PUPS P.NATAL LOSS GESTATION M F M F TOT. M F
GROUP 3 (250 MGIKG) 61 21 0 0 5 6 11 0 0 62 22 0 0 2 0 2 0 0 63 21 0 0 9 6 15 0 0 64 21 0 0 4 9 13 0 0 65 21 0 0 7 7 14 0 0 67 21 0 0 6 5 11 0 0 68 21 0 1 6 6 12 0 0 69 21 0 0 4 5 9 0 0 70 21 0 0 7 7 14 0 0 TOTAL 0 1 50 51 101 0 0 N 9 9 9 9 9 9 9 9 MEAN 21.1 0.0 01 56 5.7 11.2 0.0 0.0 ST.DEV 0.3 00 0.3 2.1 2.4 39 0.0 0.0

-Page 46 8" 23Oct08161112

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 BREEDING DATA PER LITTER FO-GENERATION
- 

## LACTATION

-- FIRST LITTER CHECKLITTER DURATION OF DEAD PUPS LIVING PUPS P.NATAL LOSS GESTATION M F M F TOT. M F
GROUP 4(1000 MG/KG) 71 22 0 0 7 7 14 0 0 73 21 1 0 3 6 9 0 0 74 21 0 0 6 6 12 0 0 75 21 0 0 5 10 15 0 0 76 21 0 0 6 7 13 1 0 77 21 0 0 8 4 12 0 0 78 22 1 0 9 5 1 4 0 0 79 21 0 0 5 8 13 0 1 80 21 0 0 8 4 1 2 0 0 TOTAL 2 0 57 57 114 1 1 N 9 9 9 9 9 9 9 9 MEAN 21.2 0.2 0.0 6.3 6.3 12.7 0.1 0.1 ST.DEV. 0.4 0.4 0.0 1.9 1.9 1.7 0.3 0.3

-Page 49 -23Oct08 10h12

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2
MEAN BODY WEIGHTS OF PUPS PER LITTER (GRAM) FO-GENERATION
- 

## LACTATION
LITTER SEX DAYl1 DAY4
GROUP I(CONTROL) 42 M 6.7 8.6 F 6.0 8.2 M+F 6.1 8.2 43 M 6.3 9.3 F 5.9 8.6 M+F 6.1 89 44 M 5.6 7.9 F 5.7 7.8 M+F 5.6 7.9 45 M 6.1 8.5 F 6.0 8.7 M+F 6.0 8.7 46 M 6.9 10.7 F 6.9 10.6 M+F 6.9 10.7 47 M 5.3 7.5 F 51 7.3 M+F 5.1 7.4 48 M 5.8 8.3 F 5.5 7.9 M+F 5.6 8.0 49 M 5.2 8.1 F 5.2 8.2 M+F 5.2 8.2 50 M 62 8.5 F 5.9 8.0 M+F 6.1 8.4 GROUP 2(50 MG/KG) 51 M 7.0 11.1 F 6.1 10.2 M+F 6.6 10.7 52 M 5.8 8.4 F 5.7 8.3 M+F 5.8 8.3 53 M 7.6 10.8 F 7.3 1 0.3 M+F 7.5 10.5 55 M 5.6 8.5 F 5.5 8.5 M+F 5.6 8.5 56 M 6.3 8.1 F 5.9 7.5 

### M+F 6.1 7.8 57 

### M 5.8 8.6 F 5.4 8.2 M+F 5.5 8.3 58 M 5.8 8.7 F 5.3 8.1 

### M+F 5.6 8.4 59 M 5.8 8.7 F 5.4 82 M+F 5.5 8.4

-Pages50~ 29Sop08l1h3i

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2
MEAN BODY WEIGHTS OF PUPS PER LITTER (GRAM) FO-GENERATION - 

## LACTATION

LITTER SEX DAYlI DAY4
GROUP 2(50 MG/KG) 60 M 56 8.6 F 5.0 7.8 M+F 5.2 8.1 GROUP 3(250 MGIKG) 61 

### M 5.9 8.3 F 5.5 7.6 M+F 5.7 7.9 62 M 6.6 10.3 M+F 8.6 10.3 63 M 5.9 8.3 F 5.4 7.5 M+tF 5.7 8.0 64 M 5.8 7.9 F 5.0 7.2 M+F 5.2 7.4 65 M 5.6 8.0 F 5.6 8.2 M+F 5.6 8.1 67 M 6.8 10.3 F 6.2 9.8 M+F 65 10.1 68 M 6.0 9.4 F 5.5 9.3 M+F 59 9.3 69 M 6.1 9.5 F 5.7 9.0 M+F 5.9 9.2 70 M 6.0 8.8 F 5.4 7.9 M+F 5.7 8.3 GROUP 4(1000 MG/KG) 71 M 6.4 9.1 F 6.2 8.7 M+F 6.3 8.9 73 M 6.0 9.1 fF 5.8 9.1 

### M+F 5.9 9.1 74 

### M 5.4 7.5 F 5.1 7.1 M+F 5.2 7.3 75 M 5.5 7.7 F 5.3 7.4 M+F 5.3 7.5 76 M 5.8 8.1 F 5.4 7.8 M+F 5.6 8.0 77 M 6.1 8.6 F 5.8 8.3 M+F 6.0 8.5 78 M 6.4 9.2 F 64 9.2 M+F 6.4 9.2

-Page 51

- 

# ~29Sep08 11h31

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 MEAN BODY WEIGHTS OF PUPS PER LITTER (GRAM) FO-GENERATION
- 

## LACTATION
LITTER SEX DAYl1 DAY4
GROUP 4(1000 MG/KG) 79 M 5.6 80 F 5.2 7.5 M+F 5.4 7.,7 80 m 5.8 8.9 F 5.8 8.5 M+F 5.7 5.8



### -Page 52 00- 

### 29Sep0811h31

## C.I. PIGMENT BLUE 61 NOTOX 

## Project488231 APPENDIX 2 BODY WEIGHTS OF PUPS (GRAM) FO-GENERZATION
- 

## LACTATION
LITTER PUP SEX DAY 1 DAY 4
GROUP 1(CONTROL) 42 1 M 67 8.6 2 M 6.7 8.7 3 F 5.7 7.9 4 F 6.0 8.3 5 F 6.1 8.4 6 F 5.5 7.4 7 F 6.2 86 8 F 6.9 9.2 9 F 6.0 8.1 10 F 6.0 8.1 11 F 5.6 7.6 12 F 6.0 8.0 43 1 M 6.6 9.6 2 M 6.2 9,1 3 M 6.4 9.5 4 M 6.0 8.6 5 F 6.0 9.0 6 F 5.5 7.9 7 F 6.1 8.6 a F 6.7 8.4 9 F 6.3 9.3 10 F 5.8 8.6 44 1 M 6.0 60 2 M 6.1 8.6 3 M 6.0 8.2 4 M 5.3 7.7 5 M 5.6 8.1 6 M 5.5 7.8 7 M 5.3 7.6 8 M 5.6 8.3 9 M 4.8 6.6 10o M 5.7 8.3 11 F 5.4 7.5 12 F 6.0 8.3 13 F 5.7 7.4 14 F 5.7 7.7 15 F 5.6 7.9 45 1 M 5.6 7.8 2 M 6.6 8.9 3 M 5.7 8.3 4 M 6.5 9.0 5 F 6.5 9.3 6 F 6.0 8.7 7 F 5.8 8.5 8 F 6.0 9.1 9 F 65 9.3 10 F 6.1 8.6 11 F 6.3 8.6 12 F 5.5 8.2 13 F 5.6 7.9 46 1 M 6,9 10.7 2 F 6.6 10.9 3 F 69 10.8 4 F 6.9 10.3

- Page 53-- 29Sep08 11h31

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 BODY WEIGHTS OF PUPS (GRAM) FO-GENERATION
- 

## LACTATION
LITTER PUP SEX DAY 1 DAY 4
GROUP I (CONTROL) 47 1 M 5.4 7.8 2 M 5.1 7.3 3 M 5.3 7.3 4 F 5.3 7.8 5 F 4.7 67 6 F 6.1 7.3 7 F 5.0 7.5 8 F 5.2 7.5 9 F 5.4 7.7 10 F 4.8 6.9 48 1 M 5.8 8.3 2 M 6.2 8.7 3 M 6.1 8.8 4 M 5.7 7.9 5 M 5.4 7.9 6 M 5.5 8.2 7 F 5.5 7.9 8 F 5.3 7.2 9 F 5.8 7.9 10 F 5.6 8.2 11 F 5.2 7.7 12 F 5.6 7.9 13 F 5.6 7.8 14 F 5.5 8.2 49 1 M 5.4 8.8 2 M 4.4 7.0 3 M 5.5 8.3 4 M 5.4 8.5 5 M 5.3 7.8 6 F 45
-- 7 F 5.3 8.4 8 F 5.2 8.5 9 F 5.3 7.8 10 F 5.4 8.3 11 F 5.1 8.2 12 F 5.4 8.1 50 1 M 6.1 8.6 2 M 5.8 8.0 3 M 6.4 8.8 4 M 6.9 9.8 5 M 6.2 8.5 6 M 62 8.2 7 M 6.2 8.4 8 M 5.9 8.0 9 M 5.7 7.4 1 0 M 6.6 8.6 11 M 6.0 7.8 12 F 6.5 8.8 13 F 8.4 8.6 14 F 5.0 6.7 GROUP 2 150 MGIKG) 51 1 M 7.1 11.3 2 M 6.9 10.9 3 M 7.6 11.6 4 M 6.7 10.6 5 M 7.0 11.0 6 F 6.1 9.2 7 F 6.8 10.8 8 F 4.8
-- 9 F 6.7 10.5

-Page 54

- -29SepO8 I Ih3l

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 BODY WEIGHTS OF PUPS (GRAM) FO-GENERATION
- 

## LACTATION
LITTER PUP SEX DAY I DAY 4
GROUP 2(50 MGIKG) 52 1 M 5.9 8.7 2 M 5.6 8.3 3 M 6.2 8.7 4 M 5.6 6.0 5 F 5.7 8.4 6 F 5.9 83 7 F 58 8.4 8 F 5 .7 8.3 9 F 5.3 8 .1 10 F 5.8 8.3 11 F 5.8 8.4 53 1 M 7.5 10.5 2 M 8.0 116 3 M 7.0 99 4 M 7.8 10.9 5 M 7.7 11.1 6 F 7.8 11.0 7 F 7.7 10.3 8 F 7.1 10.3 9 F 7.2 10.4 10 F 7.3 10.0 11 F 7.0 9.9 55 1 M 5.5 8.6 2 M 6.2 9.4 3 M 5.9 8.8 4 M 5.3 7.9 6 M 5.4 8.4 6 M 5.4 8.2 7 M 5.4 8.1 8 F 5.3 8.5 9 F 5.8 8.5 10 F 5.3 8.5 11 F 5.6 8.4 12 F 5.6 8.5 13 F 56 8.6 56 1 M 62 8.3 2 M 6.3 7.9 3 M 6.2 7.9 4 M 6.3 80 5 M 6.1 83 6 M 6.4 8.1 7 M 6.0 8.1 8 M 6.5 8.1 9 F 5.9 8.0 10 F 6.2 7.9 11 F 62 7.7 12 F 5.9 7.3 13 F 5.7 7.4 14 F 5.4 6.6 57 1 M 5.2 80 2 M 6.2 9.1 3 F 5.6 8.2 4 F 4.7 7.0 5 F 5.7 8.6 6 F 5.9 9.0 7 F 5.0 7.8 8 F 5.6 8.4 9 F 5.3 8.1 10 M 61 8.8 11 M 57 8.4

- Page 55- 29Sep08 11ih31

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 BODY WEIGHTS OF PUPS (GRAM) FO-GENERATION
- 

## LACTATION
LITTER PUP SEX DAYl1 DAY4
GROUP 2(50 MGIKG) 58 1 M 6.0 9.1 2 M 5.7 8.8 3 M 5.9 8.8 

### 4 M 51 8.0 5 M 6.2 9.2 6 M 5.8 8.3 7 M 6.0 9.0 8 F 5.7 8.8 9 F 5.4 8.4 10 F 5.2 7.6 11 F 5.2 7.9 1 2 F 5.4 8.2 13 F 5,2 7.6 59 1 M 5.7 8.8 2 M 6.0 8.8 3 M 5.6 8.4 4 M 6.0 9.0 5 F 5.9 90 6 F 5.2 8.0 7 F 4.9 7.7 8 F 5.4 8.2 9 F 5.8 8.8 10 F 5.2 7.8 60 1 M 6.2 9.2 2 M 5.6 8.9 3 M 5.1 7.9 4 M 5.5 8.5 5 F 5.6 8.8 6 F 5.2 8.1 7 F 5.3 8.2 8 F 4.8 7.3 9 F 4.1 6.4 10 F 4.5 7.5 11 F 5.3 8.4 GROUP 3 (250 MOIKG) 61 1 M 5.8 8.4 2 M 5.9 7.8 3 M 5.8 8.1 4 M 58 8.2 5 M 64 8.7 6 F 6.1 8.0 7 F 5.5 7.6 8 F 5.3 7.4 9 F 53 7.4 10 F 5.6 7.9 11 F 5.2 7.3 62 1 M 6.5 10.4 2 M 6.6 10.3 63 1 M 5.9 8.8 2 M 6.1 7.7 3 M 6.4 8.6 4 M 6.0 8.7 5 M 5.6 7.7 6 M 5.9 8.2 7 M 5.9 8.3 8 M 5.7 8.6 9 M 5.8 6.2 10 F 5.6 8.7 1 1 F 5.3 71 12 F 5.5 7.7 13 F 4.9 6.4

-Page56- 29Sep05 11h31

## C.I. PIGMENT BLUE 61 NOTOX 

## Project 488231 APPENDIX 2 BODY WEIGHTS OF PUPS (GRAM) FO-GENERATION
- 

## LACTATION

LITTER PUP SEX DAY I DAY4
GROUP 3(250 MG/KG) 14 F 5.5 74 15 F 5.4 7.5 64 1 M 5.7 8.4 2 M 5.4 7.2 3 M 5.9 8.0 4 M 6.1 8.1 5 F 4.1 5.3 6 F 4.9 7.0 7 F 5.1 6.8 8 F 5.6 8.0 9 F 5.2 7.6 10 F 4.9 7.2 I1I F 5.8 8.5 12 F 4.9 7.0 13 F 4.5 6.4 65 1 M 5.5 81 2 M 5.0 7.1 3 M 4.6 63 4 M 5.9 8.7 5 M 6.0 8.6 6 M 6.0 8.3 7 M 6.2 8.9 8 F 5.5 8.4 9 F 5.8 8.6 10 F 5.4 8.0 11 F 5.9 8.6 12 F 5.5 8.1 13 F 5.2 7.7 14 F 5.7 8.2 67 1 M 7.0 10.6 2 M 6.9 10.4 3 M 6.7 10.3 4 M 7.0 10.5 5 M 6.5 10.1 6 M 64 99 7 F 6.5 10.3 8 F 5.9 9.4 9 F 6.0 9.5 10 F 65 10.0 11 F 6.1 9.7 68 1 M 6.2 9.7 2 M 5.9 9.5 3 M 6.2 9.7 4 M 6.0 9.0 5 M 6.0 9.4 6 M 5.8 89 7 F 6.8 9.2 8 F 5.8 9.3 9 F 5.8 9.2 10 F 5.7 9.3 11 F 6.2 9.8 12 F 5.8 8.8 13 F -- -

-Page 57

- 29Sep08 111131

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 BODY WEIGHTS OF PUPS (GRAM) FO-GENERATION
- 

## LACTATION
LITTER PUP SEX DAYl1 DAY4
GROUP 3 (250 MG/KG) 69 1 M 6.0 9.5 2 M 5.8 94 3 M 6.5 9.5 4 M 6.1 9.5 5 F 5.8 9.0 6 F 5.7 9.1 7 F 5.8 9.1 8 F 6.1 9.7 9 F 5.2 8.2 70 1 M 6.1 9.4 2 M 6.0 8.2 3 M 6.1 8.7 4 M 6.1 6.8 5 M 5.6 6.9 6 M 5.7 8.5 7 M 6.2 8.9 8 F 5.8 8.5 9 F 5.7 8.1 10 F 5.5 8.3 11 F 5.0 7.4 12 F 5.7 8.3 13 F 5.1 7.6 14 F 4.8 6.9 GROUP 4(1000MG/KG) 71 1 M 6.5 9.4 2 M 6.3 9.1 3 M 6.2 8.9 4 M 6.5 91 5 M 6.6 91 6 M 6.7 9.4 7 M 6.1 8.9 8 F 6.0 6.3 9 F 6.5 9.2 10 F 6.5 9.3 11 F 5.4 7.7 12 F 6.8 9.3 13 F 6.1 8.5 14 F 6.4 8.9 73 1 M 6.1 9.9 2 M 6.1 8.9 3 M 5.7 6.5 5 F 5.6 8.7 6 F 5.8 9.5 7 F 5.6 8.5 a F 5.9 9.4 9 F 5.9 9,4 10 F 6.0 9.4 74 1 M 5.4 8.0 2 M 4.8 7.1 3 M 5.8 7.7 4 M 5.6 7.7 5 M 5.1 7.3 6 M 5.5 7.4 7 F 5.7 7.8 6 F 5.2 72 9 F 5.5 8.0 1 0 F 5.3 7.1 11 F 4.0 5.6 12 F 5.1 7.0

-Page 58 -l* M 9ep06 11h31

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2
BODY WEIGHTS OF PUPS (GRAM) FO-GENERATION
- 

## LACTATION
LITTER PUP SEX DAYl1 DAY4
GROUP 4(1000 MGIKG) 75 1 M 5.8 7.9 2 M 4.9 7.0 3 M 50 7.2 4 M 59 8.0 5 M 6.1 8.4 6 F 4.9 6.2 7 F 5.3 7.8 8 F 5.6 7.7 9 F 5.6 7.9 10 F 55 7.8 11 F 5.1 73 12 F 5.3 7.5 13 F 5.3 78 14 F 4.9 6.5 15 F 54 7.3 76 1 M 62 8.4 2 M 61 87 3 M 5.7

-- 4 M 5.4 7.9 5 M 5.3 6.8 6 M 6.3 8.9 7 F 5.2 7.5 8 F 5.3 7.8 9 F 5.3 7.6 10 F 5.5 8.1 11 F 5.7 8.6 12 F 5.5 7.5 13 F 5.2 7.6 77 1 M 5.9 8.9 2 M 6.0 ,8.2 3 M 5.7 8.2 4 M 6.1 8.9 5 M 5.9 6.4 6 M 6.4 8.8 7 M 6.3 8.8 8 M 64 9.0 9 F 5.6 82 10 F 5.8 8.4 1 1 F 5.9 8.4 12 F 5.7 8.0 78 1 M 7.0 10.1 2 M 6.4 9.3 4 M 6.2 9.0 5 M 6.4 9.4 6 M 5.9 8.5 7 M 6.3 8.7 8 M 5.8 8.9 9 M 66 9.7 1 0 M -- - 11 F 6.5 9.2 12 F 6.7 9.7 13 F 6.1 8.6 14 F 6.4 9.6 15 F 6.3 8.8 79 1 M 5.5 8.2 2 M 5.7 7.8 3 M 5.4 7.8 4 M 5.7 7.8 5 M 5.6 8.2 6 F 5.1 7.0 7 F 5.1 7.8 8 F 5.6 8.1

- Page 59 29Sep08 llh~l

## 0.1. PIGMENT BLUE61 NOTOX Project 488231 APPENDIX 2 BODY WEIGHTS OF PUPS (GRAM) FO-GENERATION - 

## LACTATION
LITTER PUP SEX DAY I DAY 4
GROUP 41(1000 MGIKG) 9 F 5.1 7.4 10 F 5.0 7.1 11 F 5.2 7.3 12 F 5.5 7.8 13 F 5.1
-- 80 1 M 6.3 9.6 2 M 5.5 8.2 3 M 6.2 9.5 4 M 5.4 8.2 5 M 5.7 9.3 6 M 6.0 9.1 7 M 6.0 8.9 8 M 5.4 8.3 9 F 5.5 8.7 10 F 5.6 8.6 1 1 F 5.8 8.4 12 F 5.6 8.4

-Page 60

- - ep0811h3l

## C.!. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP I (CONTROL) LITTER 42 1 M DAY 5 Planned Necropsy FLC No findings 23JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 3 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 4 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 5 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 6 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 F DAY 5 Planned Necropsy FILC No findings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings g F DAY 5 Planned Necropsy FLC No findings LLC No findings 10 F MACRO No findings 10FDAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 11 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 12 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings LITTER 43 1 M DAY 5 Planned Necropsy FLC No findings 23JUL2008 LLC No findings MACRO Nofindings 2 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 3 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings 4 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 5 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 6 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 7 F DAYS5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings

FLC - FIRST LITTER CHECK, DAY PP.- CLINICAL SIGNS, LLC
- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS -Page 61

- "24Oct08 15h47

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2
INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP I (CONTROL) 9 F DAY 5 Planned Necropsy FLO No findings LLC No findings MACRO No findings 10 F DAY5 Planned Necropsy FLO Nofindings LLC No findings MACRO Nofindings LITTER 44 1 M DAY 5 Planned Necropsy FLC No findings 27JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 3 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 4 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings S M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 6 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 8 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 9 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 1D M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 11 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 12 

### F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 13 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 14 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 15 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings LITTER 45 1 M DAY 5 Planned Necropsy PLC No findings 27JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 3 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings

FLC
- FIRST LITTER CHECK, DAY P.P. - CLINICAL SIGNS, LLC
- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

- Page 62

- ON- 24Oct08 15h47

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP I (CONTROL) 4 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 5 F DAY 5 Planned Necropsy PLC Nofindings LLC No findings MACRO No findings 6 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 P DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 10 P DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 11 F DAY 5 Planned Necropsy PLC Nofindings LLC No findings MACRO No findings 12 P DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 13 P DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings LITTER 46 1 M DAY 5 Planned Necropsy PLC No findings 27JUL2008 LLC No findings MACRO No findings 2 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 3 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 4 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings LITTER 47 1 M DAY 5 Planned Necropsy PLC No findings 25JUL2008 LLC No findings MACRO Nofindings 2 M DAYS5 Planned Necropsy PLC Nofindings LLC No findings MACRO No findings 3 M DAYS5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 4 F DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO No findings 5 P DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 6 P DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings

PLC
- FIRST LITTER CHECK, DAY P.P. - CLINICAL SIGNS, LLC
- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

- Page 63

- 24O0t08 15h47

## ClI. PIGMENT BLUE61 NOTOX Project 488231 APPENDIX 2
INDIVIDUAL PUP DATA FO-GENERATION
- 

### LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP I (CONTROL) 7 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 10 F DAY S Planned Necropsy FLC No findings LLC No findings MACRO No findings LITTER 48 1 M DAY 5 Planned Necropsy FLC No findings 25JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 3 M DAY5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 4 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRD No findings 5 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 6 M DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 7 P DAY 5 Planned Necropsy PLC No findinga LLC No findings MACRO Nofindings 8 F DAYS5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO No findings 10 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings I1 P DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 12 P DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 13 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 14 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings LITTER 49 1 M DAY 5 Planned Necropsy PLC No findings 25JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings

PLC
- PIRST LITTER CHECK, DAY P.P. - CLINICAL SIGNS, LLC
- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

- Page 64

- 24Oct08 15h47

## ClI. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2
INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF PP. PHASE FINDINGS DELIVERY
GROUP I (CONTROL) 3 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 4 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 5 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 6 F DAY 2 Missing FLC No findings 7 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 10 F DAYS5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 11 F DAYS5 Planned Necropsy FLO No findings LLC No findings MACRO No findings 12 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings LITTER 50 1 M DAY 5 Planned Necropsy FLC No findings 23JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 3 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 4 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 5 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 6 M DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 7 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 8 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 9 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRD No findings 10 M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 11 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings

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- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

- Page 65 -. 240cl08 15h47

## C. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2
INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY GROUP1I(CONTROL) 12 F DAY 5 Planned Necropsy FLO No findings LLC No findings MACRO No findings 13 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 14 F DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings GROUP 2(50 MG/KG) LITTER 51 1 M DAY 5 Planned Necropsy PLC No findings 26JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 4 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 5 M DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO No findings 6 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 7 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 8 F DAY 2 Missing PLC Small 9 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Ho findings LITTER 52 1 M DAY 5 Planned Necropsy PLC No findings 25JUL2008 LLC Nofindings MACRO Nofindings 2 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 3 M DAY 5 Planned Necropsy PLC Nofindings LLC No findings MACRO No findings 4 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings 5 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 6 F DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO No findings 7 F DAY 5 Planned Necropsy PLC Nofindings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings

PLC
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- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

- Page 66 - 0111 24O0t08 15h47

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 2(50 MG/KG) 9 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 10 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 11 F DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings LITTER 53 1 M DAY S Planned Necropsy PLC No findings 27JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 4 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings S M DAY S Planned Necropsy PLC No findings LLC No findings MACRO No findings 6 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 7 P DAY S Planned Necropsy PLC No findings LLC No findings MACRO No findings 6 F DAY S Planned Necropsy PLC No findings LLC No findings MACRO No findings 9 P DAYS5 Planned Necropsy PLC No findings LLC Nofindings MACRO No findings 10 P DAY S Planned Necropsy PLC No findings LLC No findings MACRO No findings 11 F DAY S Planned Necropsy PLC No findings LLC No findings MACRO No findings LITTER 55 1 M DAYS5 Planned Necropsy PLC No findings 25JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 3 M DAYS5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 4 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings S M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 6 M DAYS Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings

PLC
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- MACROSCOPIC FINDINGS -Page 67 - 

# ~I 24Oct08 15h47

## C.I. PIGMENT BLUE61 NOTOX 

## Project488231 APPENDIX 2 INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY GROUP 2(50 MG/KG) 7 M DAY 5 Planned Necropsy FLO No findings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 9 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 10 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 11 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 12 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 13 F DAY 5 Planned Necropsy FL-C No findings LLC No findings MACRO No findings LITTER 56 1 M DAY 5 Planned Necropsy FLC No findings 23JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 4 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 5 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 6 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 7 M DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 8 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy PLC No findings LLC: No findings MACRO No findings 10 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 11 P DAY 5 Planned Necropsy PLC; Nofindings LLC Nofindings MACRO No findings 12 P DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 13 P DAY 5 Planned Necropsy PLC No findings LLC: No findings MACRO No findings

PLC
- FIRST LITTER CHECK. DAY P.P. - CLINICAL SIGNS, LLC
- LAST LITTER CHECK. MACRO
- MACROSCOPIC FINDINGS

- Page 68 - *- 24Oct08 15h47

## ClI. PIGMENT BLUE 61 NOTOX 

## Project488231 APPENDIX 2 INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 2(50 MG)KG) 14 F DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO No findings LITTER 57 1 M DAY 5 Planned Necropsy FLC Red spot nose 25JUL2008 DAY 2 Red spot nose LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 3 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 4 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 5 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 6 F DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 7 P DAY 5 Planned Necropsy FLO Nofindings LLC Nofindings MACRO Nofindings 8 F DAY S Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 9 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 10 M DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 11 M DAY 5 Planned Necropsy PLC Red spot nose DAY 2 Red spot nose LLC Nofindings MACRO Nofindings LITTER 58 1 M DAYS5 Planned Necropsy PLC Nofindings 25JUL2008 LLC Nofindings MACRO No findings 2 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 4 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 5 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 6 M DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO No findings 7 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings

PLC
- FIRST LITTER CHECK, DAY P.P. - CLINICAL SIGNS, LLC
- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

- Page 69

- b-24Oct08 15h47

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2
INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 2(50 MGIKG) 8 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 9 F DAY 5 Planned Nerropsy FLC No findings LLC Nofindings MACRO Nofindings 10 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 11 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 12 F DAY 5 Planned Necropsy FLO Nofindings LLC Nofindings MACRO Nofindings 13 F DAYS5 Planned Necropsy FLC .No findings LLC Nofindings MACRO No findings LITTER 59 1 M DAY 5 Planned Necropsy FLC No findings 25JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO Nofindings 3 M DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 4 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 5 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 6 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 10 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings LITTER 60 1 M DAYS5 Planned Necropsy FLC No findings 25JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 4 M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings

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- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

- Page 70

- 

# ~"24Oct08 15h47

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 INDIVIDUAL PUP DATA P0-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 2(50 MG/KG) 5 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 6 P DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 8 F DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 9 F DAYS5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 10 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 11 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings GROUP 3(250 MG/KG) LITTER 61 1 M DAY 5 Planned Necropsy PLC Nofindings 23JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy FLO No findings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 4 M DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 5 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO Nofindings 6 P DAY 5 Planned Necropsy FLO Nofindings LLC Nofindings MACRO Nofindings 7 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRD No findings 8 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 9 F DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 10 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 11 F DAYS5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings LITTER 62 1 M DAY 5 Planned Necropsy PLC No findings 26JUL2008 LLC No findings MACRO No findings

PLC
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- LASTLITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

- Page 71
- 

## Ir24Oct08 15h47

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF PP. PHASE FINDINGS DELIVERY
GROUP 3(250 MG/KG) 2 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings LITTER 63 1 M DAY 5 Planned Necropsy PLC; No findings 23JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy FLC: Nofindings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 4 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 5 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 6 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 8 M DAY 5 Planned Necropsy FLO No findings LLC No findings MACRO No findings g M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 10 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 11 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 12 F DAY 5 Planned Necropsy FLC Na findings LLC No findings MACRO No findings 13 F DAYS5 Planned Necropsy FLC No findings LLC Na findings MACRO No findings 14 F DAYS5 Planned Necropsy FLC Na findings LLC No findings MACRO No findings 15 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings LITTER 64 1 M DAY 5 Planned Necropsy FLC No findings 23JUL2008 LLC Na findings MACRO No findings 2 M DAY 5 Planned Necropsy FLC Na findings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 4 M DAY 5 Planned Necropsy PLC Na findings LLC No findings MACRO No findings

FLC
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- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

- Page 72 -fr24Oct08 15h,47

## ClI. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 3(250 MG/KG) 5 F DAY 5 Planned Necropsy FLC Small DAY 2 Sma DAY 3 Smal DAY 4 Small 

## LLC Small MACRO No findings 6 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 F DAY 5 Planned Nec-ropsy FLC No findings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 10 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 11 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 12 F DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 13 F DAY 5 Planned Necropsy FLC No findings LLC Na findings MACRO No findings LITTER 65 1 M DAY 5 Planned Necropsy FLC No findings 23JUL2008 LLC No findings MACRO No findings 2 M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 3 M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 4 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 5 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 6 M DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 7 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings 8 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 10 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings

PLC
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- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

- Page 73

- 0- 24Oct08 15h47

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2
INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
UITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 3 (250 MG/KG) 11 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 12 F DAYS5 Planned Necropsy PLC Nofindings LLC No findings MACRO No findings 13 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 14 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings LITTER 67 1 M DAY 5 Planned Necropsy PLC No findings 25JUL2008 LLC No findings MACRO No findings 2 M DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 4 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 5 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 6 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 7 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy PLC Nofindings LLC No findings MACRO Nofindings 9 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 10 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings 11 F DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings LITTER 68 1 M DAY 5 Planned Necropsy PLC Nofindings 25JUL2008 LLC Nofindings MACRO Nofindings 2 M DAY 5 Planned Necropsy PLC Nofindings LLC No findings MACRO Nofindings 3 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 4 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 5 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings

PLC
- FIRST LITTER CHECK, DAY P.P. - CLINICAL SIGNS, LLC
- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

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- '24Oct08 15h47

## C.I. PIGMENT BLUE 61 NOTOX 

## Project 488231 APPENDIX 2
INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P PHASE FINDINGS DELIVERY
GROUP 3(250 MGIKG) 6 MDAY Plnne Neropy LLC Nofindings 6 MDAYS Pannd Ncrosy LC No findings MACRO No findings 7 P DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 10 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 11IF DAY 5 Planned Nacropsy FLC Nofindings LLC Nofindings MACRO Nofindings 12 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 13 F DAY 1 Dead at FLC PLC Dead MACRO Autolysis, no milk LITTER 69 1 M DAY 5 Planned Necropsy FLC No findings 25JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings 4 M DAY 5 Planned Necropsy PLC: Nofindings LLC Nofindings MACRO Nofindings 5 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 6 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRD No findings 7 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 8 P DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 9 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings LITTER 70 1 M DAY 5 Planned Necropsy FLC No findings 26JUL2008 LLC: No findings MACRO No findings 2 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 3 M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings

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- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS
-Page 75

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## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2
INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY GROUP 3(250MG/KG) 4 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 5 M DAY 5 Planned Necropsy FLC; No findings LLC Nofindings MACRO No findings 6 M DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 7 M DAY 5 Planned Necropsy FLC: No findings LLC No findings MACRO No findings 8 F DAY S Planned Necropsy FLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy FLO No findings LLC No findings MACRO No findings 10 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 11 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 12 F DAYS Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 13 F DAYS Planned Necropsy FLC No findings LLC No findings MACRO No findings 14 F DAYS5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings GROUP 4(1000 MG/KG) LITTER 71 1 M DAY 5 Planned Necropsy FLC No findings 27JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 3 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 4 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO Nofindings 5 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 6 M DAY S Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 M DAY 5 Planned Necropsy FLO: No findings LLC No findings MACRO No findings 8 F DAY S Planned Necropsy PLC No findings LLC No findings MACRO No findings

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- MACROSCOPIC FINDINGS

- Page 76

- -24Oct08 15h47

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 4(1000 MGIKG) 9 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 10 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 11 F DAYS5 Planned Necropsy FLC: No findings LLC No findings MACRO No findings 12 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 13 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 14 F DAYS5 Planned Necropsy FLC Pale LLC No findings MACRO No findings LITTER 73 1 M DAYS5 Planned Necropsy PLC No findings 25JUL2008 LLC No findings MACRO No findings 2 M DAYS Planned Necropsy PLC Nofindings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 4 M DAY 1 Dead at FLC PLC Dead MACROAutolysis, no milk 5 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 6 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 F DAY 5 Planned Necropsy FLC: No findings LLC No findings MACRO No findings 8 F DAYS5 Planned Necropsy FLC; No findings LLC No findings MACRO No findings g F DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 10 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings LITTER 74 1 M DAY S Planned Necropsy FLC No findings 23JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 3 M DAYS5 Planned Necropsy PLC No findings LLC Nofindings MACRO No findings 4 M DAYS5 Planned Necropsy FLC Nofindings LLC No findings MACRO Nofindings

FLC
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- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

- Page 77

- 

# ~-24Oct08 15h47

## 0,1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 4(1000 MG)KG) 5 M DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 6 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 9 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 10 F DAYS5 Planned Necropey PLC No findings LLC No findings MACRO No findings 11 F DAY 5 Planned Necropsy FLC Small DAY 2 Small DAY 3 Small DAY 4 Smell LLC Small MACRO No findings 12 F DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO No findings LITTER 75 1 M DAY 5 Planned Necropsy PLC No findings 23JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings 3 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 4 M DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 5 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 6 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 7 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings 10 F DAY 5 Planned Necropsy PLC Nofindings LLC No findings MACRO No findings 11 F DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings

PLC
- FIRST LITTER CHECK, DAY P.P. - CLINICAL SIGNS, LLC
- LAST LITTER CHECK. MACRO
- MACROSCOPIC FINDINGS

- Page 78

- 24Oct08 15h47

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2
INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 4(1000 MGIKG) 12 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 13 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 14 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 15 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings LITTER 76 1 M DAY 5 Planned Necropsy PLC No findings 24JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings 3 M DAY 2 Missing PLC No findings 4 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 5 M4 DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 6 M DAY 5 Planned Necropsy PLC Nofindings LLC No findings MACRO No findings 7 F DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 10 F DAYS5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 11 P DAY 5 Planned Necropsy PLC Nofindings LLC No findings MACRO No findings 12 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 13 P DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings LITTER 77 1 M DAY 5 Planned Necropsy PLC No findings 23JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 4 M DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO No findings

PLC
- FIRST LITTER CHECK, DAY PP.- CLINICALSIGNS, LLC
- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS -Page 79

- No 24Oct08 15h47

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 INDIVIDUAL PUP DATA FO0-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY GROUP 4(1000 MGIKG) 5 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 6 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 M DAY 5 Planned Necropsy FLC Red spot head DAY 2 Red spot head LLC Red spot head MACRO No Findings 8 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 10 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 11 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 12 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings LITTER 78 1 M DAY 5 Planned Necropsy FLC No findings 27JUL2008 LLC No findings MACRO Nofindings 2 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 3 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 4 M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 5 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 6 M DAY 5 Planned Necropsy FLC No Findings LLC No findings MACRO No findings 7 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 8 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 9 M DAY 5 Planned Necropsy FLO No findings LLC No findings MACRO No findings 10 M DAYl1 Dead atFLC FLC Dead MACRO Autolysis, no milk 11 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 12 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings

FLC - FIRST LITTER CHECK, DAY P.P. - CLINICAL SIGNS, LLC
- LASTLITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

- Page 50

- '24Oct08 15h47

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 INDIVIDUAL PUP DATA FO-GENERATION
- 

## LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 4(1000 MG/KG) 13 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 14 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 15 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings LITTER 79 1 M DAY 5 Planned Necropsy FLO No findings 26JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 4 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 5 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 6 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 10 F DAYS5 Planned Necropsy FLC No findings LLG No findings MACRO No findings 11 F DAY 5 Planned Necropsy FLO No findings LLC No findings MACRO No findings 12 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 13 F DAY 2 Spontaneous death FLC No findings LLC Dead MACRO Autolyse LITTER 80 1 M DAY 5 Planned Necropsy FLO No findings 26JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 4 M DAY 5 Planned Nacropsy FLC Nofindings LLC No findings MACRO No findings

FLC
- FIRST LITTER CHECK. DAY P.P. - CLINICAL SIGNS, LLC
- LAST LITTER CHECK, MACRO
- MACROSCOPIC FINDINGS -Page 81 - 24Oct08 15h47

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2
INDIVIDUAL PUP DATA FO-GENERATION
- 

### LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 4 (1000 MG/KG) 5 M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 6 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 8 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 10 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 11 F DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 12 F DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings

FLC
- FIRST LITTER CHECK, DAY P.P. - CLINICAL SIGNS, LLC
- LASTLITTER CHECK, MACRO
- MACROSCOPIC FINDINGS

- Page 82 

# ~-24Oct08 15h47

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2
CORPORA LUTEA FO-GENERATION GROUP 1 GROUP2 GROUP 3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG Female Corpora Female Corpora Female Corpora Female Corpora Number Lutea Number Lutea Number Lutea Number Luta

41<NP> -- 51 15 61 16 71 22 42 15 52 21 62 11 72 2 43 11 53 15 63 17 73 14 44 15 54<NP>

-- 

### 64 15 74 15 45 22 55 19 65 15 75 19 46 22 56 15 66<NP> -- 76 15 47 20 57 12 67 21 77 12 48 23 58 18 68 20 78 16 49 19 59 19 69 13 79 13 50 15 60 20 70 13 80 15
Mean 18.0 17.1 15.7 14.3 St.dev. 4.15 2.96 3.28 5.21 N 9 9 9 10

<NP> non-pregnant DI

unnett-test based on pooled variance significant at 5%()or 1%

## ()level

- 

### Page 83
-

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2

IMPLANTATION SITES FO-GENERATION GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG Female Implantation Female Implantation Female Implantation Female Implantation Number Sites Number Sites Number Sites Number Sites

41 <NP> -- 51 8 61 12 71 16 42 12 52 10 62 2 72 1 43 11 53 11 63 16 73 13 4415 

### 54<NP> --- 

### 64 14 74 10 45 14 55 12 65 13 75 15 46 7 56 14 66<NP> --- 

### 76 14 47 14 57 12 67 12 77 11 48 14 58 13 68 12 78 16 49 12 59 10 69 11 79 13 50 14 60 11 70 13 80 12

Mean 12.6 11.2 11.7 12.1 St.dev. 2.46 1.79 3.91 4.38 N 9 9 9 10

<NP> non-pregnant

1Dunnett-test 

### based on pooled variancesignificant at 5%()or 1 %()level

- 

### Page 84-

## C.I. PIGMENT BLUE61 NOTOX Project488231 APPENDIX 2 KEY TO MISSING VALUES AND REMARKS CLINICAL LABORATORY INVESTIGATIONS

56 = 

### Technicalerror forWBC

_______ 

### Citatesample clotted 43, 50,56, 57, 67,74. Differential leucocytecount was also performed manually because 75, 77 of atechnicalerror/Ian abnormal plot inthe automated count and these resultsare 56 

# ~~~~Manual differential leucocytecount revealed that 1%of netohl eandcels

*Page 85
-

# APPENDIX 3



# ANALYTICAL REPORT

Author Ir.M.J.C. Brekelmans Completion date analyticalstudy

Laboratory Project Identification NOTOX Project 488231 NOTOX Substance 183591/A

-Page 

## 1

## ofl 11

## 0.1. PIGMENT BLUE61 NOTOX Project488231 1. CONTENTS 1. CONTENTS........................................................................................... 2 2. REPORT APPROVAL.............................................................................. 3 3. SUMMARY
......................................................................................... 4 

## 4. INTRODUCTION .................................................................................. 5 4.1. Preface...................................................................................... 5 4.2. Aim ofthe study .............................................................................. 5 5. MATERIALS AND METHODS .................................................................... 5 5.1. Reagents................................................................................... 5 5.2. Samples..................................................................................... 5 5.3. Analytical method............................................................................. 6 5.3.1. Analytical conditions.................................................................. 6 5.3.2. Preparation ofthe calibration solutions.............................................. 6 5.3.3. Sample injections..................................................................... 6 5.4. Electronic data capture ...................................................................... 7 5.5. Formulas.................................................................................... 7 5.6. Specifications................................................................................. 7 6. RESULTS .......................................................................................... 8 6.1. Calibration curves ............................................................................ 8 6.2. Samples..................................................................................... 8 6.2.1. Procedural recoverysamples........................................................ 8 6.2.2. Test samples........................................................ ............. ... 8
TABLES 

## Table 1 Procedural recovery samples............................................................ 9 Table 2 Accuracy and homogeneity testfortheformulations prepared foruse during treatment................................................................................. 9 Table 3 Stability testfortheformulations prepared for useduring treatment............... 10
FIGURES Figure 1 Chromatograms ofa15.1 mg/I testsubstance solution [top; res. id.1261]and corresponding blank [bottom; res. id.1282]........................................... 11

-Page 

## 2

-

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 2. REPORT APPROVAL
PRINCIPAL SCIENTIST: Ir.M.J.C. Brekelmans (Analytical Chemistry)

Date:

-Page 

## 3

-

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 3. SUMMARY The purpose ofthis part ofthe studywas to determine theaccuracy ofpreparation, homogeneity and stability ofthe testsubstance informulations. Accuracy of preparation Theconcentrations analysed inthe formulations of Group 2,Group 3and Group 4were in agreement with target concentrations (i.e. mean accuracies between 85% and 115%). Notest substance wasdetected inthe Group 1formulations. Homogeneity Theformulations ofGroup 2and Group 4were homogeneous (i.e. coefficientofvariation 5 10%). Stability Formulations ofGroup 2and Group 4were stablewhen stored at room temperature for atleast 6hours.

-Page4-

# C.I. PIGMENT BLUE 61 NOTOX Project 488231 4. INTRODUCTION 4.1. Preface Study plan Start 27 June200B (analytical study) Completion 01 July2008 4.2. Aim ofthestudy The purpose oftheanalytical studywasto determine theaccuracyof preparation, homogeneity and stabilityofthe testsubstance informulations. 5. MATERIALS AND METHODS 5.1. Reagents Water Tapwater purified by aMilli-Q water purification system (Millipore, Bedford, MA, USA) Methanol VWR Intemnational, Leuven, Belgium Sodium 1-heptanesulfonate Fluka Chemie, Buchs, Switzerland monohydrate (HSA) Aceticacid Merck, Djarmstadt, Germany Dimethyl sulfoxide (DMSO) Merck All reagents were ofanalytical grade, unless specifiedotherwise. 5.2. Samples Accuracy, homogeneityand stabilitywere determined forformulations prepared for use during treatment. Duplicatesamples (approximately 500 mg),which weretaken from the formulations using a pipette,were accurately weighed intovolumetric flasksof5,25or50 ml. Fordetermination of accuracy, sampleswere taken at 50% heightorat 90%, 50% and 10% height. The latter setof samples was alsoused forthe determination ofthe homogeneity oftheformulations. For determination ofstability, additional sampleswere taken at50% heightand stored at room temperature for6hours. The volumetricflaskswerefilled up to themark with DMVSO. The solutionswere further diluted with methanol byafactorofat least 100, toobtain concentrationswithin the calibration range.

-Page 

## 5

-

## CJI. PIGMENT BLUE 61 NOTOX Project488231 5.3. Analytical method 5.3.1. Analyticalconditions Quantitative analysiswas based on theanalytical method validated forthetest substance in NOTOX project488235. Analyticalconditions: Instrument Alliance Separation Module 2695 (Waters, Milford, MA, USA) Detector Dual A Absorbance Detector 2487 (Waters) Column Symmetry Shield Ci1B, 150mm x4.8 mm i.d., dp =5 

## pm (Waters) Column temperature 50*C ±I*C Injection volume 10 P1 Mobile phase A

- 

## 5/0.5/94.5 (v/vlv)water/acetic acid/methanol containing 1.2 g/l HSA B

- 

## 5/0.5/94.5 (v/v/v)water/acetic acid/[50/50 (v/v) methanol/water] containing 1.2g/l HSA GradientC - 

## methanol Time %A %B % [minutes) 0 0 100 0 20 100 0 0 20.1 0 100 0 23 0 100 0 23.1 0 0 100

28 0 0 100 28.1 0 100 0

35 0 100 0
Flow 2mI/mmn UV detection 313 nm 5.3.2. Preparation ofthe calibration solutions Stock- and spiking solutions Stocksolutionsofthe testsubstancewere prepared inDMSO at concentrations of 1108 and 1236 mg/I. Inorderto dissolvethetest substance thesolutions were ultrasonicatedfor 10 minutes. Calibration solutions Calibration solutions intheconcentration range 0.997

- 

## 15.1mg/I were prepared fromtwo stock solutions,The end solution ofthe calibration solutionswas methanol. Procedural recovery samples Approximately500 mg blankvehicle wasspiked withthe test substance atatarget concentration of9.66 or 194 mg/g. The accuracysamples were treated similarlyasthe testsamples (see paragraph 5.2 'Samples'). 5.3.3. Sample Injections Calibration solutionswere injected induplicate.Testsamples and procedural recoverysamples were analysed bysingle injection.

- 

## Page 6

-

## ClI. PIGMENT BLUE61 NOTOX Project488231 5.4. Electronic data capture System control, data acquisitionand data processing were performed using the following programme:

- 

## Empower version 5.00 (Waters, Milford, MA, USA). 5.5. Formulas Response (R) Peak areatest substance[units]
Calibration curve R=aCN +b where: CN = 

## nominal concentration [mg/I] a = 

## slope [units x

## I/mg] b = 

## intercept[units]
Analysed concentration (CA) CA = 

## ( b)x [mg/g] a w where: w = 

## weight sample [mg] V = 

## volume volumetricflask [ml] d = 

## dilution factor Recovery A X100

## % CN where: CN = 

## nominal concentration [mg/g] Accuracy EA X10O% CT where: CT = 

## targetconcentration [mg/g]
Relativedifference (relativeduff.) Ct -C0 

## x1100% 

## CO where:

Ct= mean concentration ofstored samples [mg/g] C

0 

### = 

## mean concentration of non-stored samplesfmg/g] 5.6. Specifications Preparation offormulationswas considered acceptable ifthe mean accuracy was inthe range 85%

- 

## 115% ofthe targetconcentration and ifthe coefficientofvariationwas 10%. Formulations were considered stable ifthe relative difference between the stored and freshlytaken samples was 510%.

- 

## Page 7

-

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 

## 6. RESULTS 6.1. Calibration curves Acalibration curve was constructed using four concentrations. Foreach concentration, two responseswere used.2Linear 

## regression analysis was performed using the least squares method with a1/concentration weighting factor. Thecoefficient ofcorrelation (r)was>0.99. Representative chromatograms ofatest substance solutionand blanksolutionare shown in Figure 1. 5.2. Samples 6.2.1. Procedural recovery samples The resultsfor the procedural recovery samples aregiven inTable 1. Mean recoveries ofthe procedural recovery sampleswere 100% and 101%.Because the criterion that mean recoveries should be between 85% and 115% was met, the results forthe test samples were accepted. 6.2.2. Test samples The results ofthetest samples are given inTables 2and 3. Accuracy of preparation Inthe Group 1formulations, no test substance was detected. The concentrations analysed inthe formulations ofGroup 2,Group 3and Group4were in agreement withtargetconcentrations (i.e. mean accuracies between 99% and 102%). Homogeneity The formulations ofGroup 2and Group 4were homogeneous (i.e. coefficient ofvariation of2.4% and 0.92%, respectively). Stability Analysis ofGroup 2and Group 4formulations afterstorage yielded arelative difference of0.79% and 0.35%, respectively. Based onthis, theformulations were foundto be stable during storage at room temperature for atleast 6hours.

-Page 

## 8

-

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 TABLES Table 1 Procedural recovery samples Date of Date of Target Nominal Analysed Recovery Mean recovery preparation analysis concentration concentration concentration (coefficientof [dd-mm-yy] [dd-mm-yy] [mg/g] [mg/g] [mg/g] [%] variation)
30-06-08 30-06-08 9.66 9.81 9.86 100 101 9.65 9.63 100 (1.9) 10.4 10.3 99 9.66 9.63 100 9.68 10.1 104 30-06-08 30-06-08 194 194 193 100 100 197 198 100

Table 2 Accuracyand homogeneity testforthe formulations prepared foruse duringtreatment Group Date of Sample Concentration Accuracy Homogeneity analysis position (coefficientof 

# I 

# Jg/....... variation) [dd-mm-yy] Target Analysed Individual Mean % 1 30-06-08 50% height 0.00 n.d. n.a. 0.00 n.d. n.a. 2 30-06-08 90% height 9.66 10.0 104 102 2.4 9.66 10.3 106 50% height 9.66 9.79 101 9.66 9.87 102 10% height 9.66 9.54 99 9.66 9.82 102 3 30-06-08 50% height 48.3 48.1 99 100 48.3 48.3 100 4 30-06-08 90% height 194 190 98 99 0.92 194 191 98 50% height 194 194 100 194 190 97 10% height 194 192 99 194 193 99
n.d. Notdetected. n.

a. Notapplicable.

- 

## Page 9

-

## 0.1. PIGMENT BLUE61 NOTOX Project488231

Table 3 Stabilitytestforthe formulations prepared for use duringtreatment

Group Date ofanalysis Analysed concentration Relativeduff. [M /g] [dd-mm-yy] t=0 1t=6 hours 

# (%J 2 30-06-08 9.88 9.962 

## 0.79 4 30-06-08 192 192 3 

## 0.35

Mean of sixsamples at

## t=0 taken at 10%, 50% and 90% height (seeTable2). 2 

### Menotwsapeatt6hustkna50hehtInviulrslswr9.0ad1.mgg Mean of two samples att=6 hours taken at 50%height. Individual results were 929 and 1930mg/g.

-Page 

## 10

-

## C.I. PIGMENT BLUE 61 NOTOX Project488231
FIGURES

Figure 1 Chromatograms of a 15.1 mgI testsubstance solution [top: res. Id.1261] and corresponding blank [bottom; res. id.1297].

-Page 

## 11

-

# ToxPath Consultancy Ltd

APPENDIX 4



# PATHOLOGY REPORT ACombined 28-Day Repeated DoseToxicity Studywiththe Reproduction/Developmental ToxicityScreening Test ofC.I. Pigment Blue 61 inRats byOral Gavage NOTOX Project: 488231 NOTOX Substance 183591/A

Dr AM Pilling Po

## Box 16 Sandy UK SG19l1WJ Tel: +44 (0)1767 681546 arnpillinp0abtinternet.corn

### Page 2of 97 PATHOLOGY REPORT Testsubstance: 

## 0.1. Pigment Blue 61 Notox No.: 488231 Testsystem: 28-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.



# Table ofContents Statement of Compliance
............................................................ 3 QualityAssurance Statement
........................................................ 4 Summary
.............................................................................. 5 Materials and Methods .................................................................... 6 Results ................................................................................ 8 Conclusions
................................................................................ 9 Table I Incidence all Macroscopic Findings ........................................... 10 Table 2Incidence all Microscopic Findings............................................ 11 Table 3Individual Animal Data
......................................................... 18

### Page 3 of97 PATHOLOGY REPORT Test substance:

## C.l. Pigment Blue61 Notox No.:488231 Test system: 28-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments ManufacturersAssociation Inc.



# Statement of Compliance The histopathologyfindings inthis report were compiled bythe undersigned pathologistand accurately reflect the rawdata. The phaseof work inthis report was carried out incompliance with the OECD Principles of Good Laboratory Practice, 1997 Revision (ENV/MC/CHEM (98) 17, Paris 1998 andthe UK Good Laboratory Practice Regulations 1999 (Statutory Instrument 1999 No3106) as amended by theGood Laboratory Practice (Codification Amendments etc) Regulations2004 (Statutory Instrument 2004 No

# 994).

DrAM Pilling BVSc PhD MRCVS FRCPath Principal Investigator ToxPath Consultancy Ltd 

# PO Box156 Sandy UK SG19I1WJ

## Page4of97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association 

## Inc. 

## Quality Assurance Statement Quality Assurance surveillanceofthis study phasewas carried out incompliance with the OECD Principlesof Good Laboratory Practice, 1997 Revision (ENVIMCICHEM (98) 17, Paris 1998 and the UK 

# Good Laboratory Practice Regulations 1999 (Statutory Instrument 1999 No 3106) as amended bythe Good Laboratory Practice (Codification Amendments Etc) Regulations 2004 (Statutory Instrument 2004 No994). The programme includes thefollowing inspections: StudySpecific Inspections: Study plan and reportforthe above study phase are reviewed and itisconfirmed thatthe report isatrue and accurate reflection ofthe rawdata. Findings are reported to PI, SD, Lead QAU and testfacility management. Process Inspections: Processes are inspected atapproximately three monthly intervals, irrespective ofthe study being dealtwith atthetime. Findings are reported to Pl. FacilityInspections: The facilities and operations ofToxPath Ltd are audited at approximately sixmonthly intervals. Findings are reported to Pl. Dates ofInspections The following inspections have been carried outoverthe period ofthisstudy phase: Aspect inspected Date of inspection Date of report Study Specific Study Plan Report Process Slide reading 18 July2008 124 July2008 Slide receiptidespatch 18 July2008 24July 2008 Facility 18 July 2008 24 July 2008

David JFord, PhD, FRQA QualityAssurance Consultant David Ford Consultancy Goodwyn House Bank Street Puihamn Market Diss NorfolkIP21 4TG ENGLAND

## Page 5of97 PATHOLOGY REPORT Testsubstance: C.
. Pigment Blue 61 Notox No.:488231 Testsystem: 28-Day Oral ToxicityStudy Ini the Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.


## Summary Histopathologicalexaminations were performed on tissuesfrom HanWistar rats which had received 

# C.I. Pigment Blue 61 orallyfor aminimum of28 days. Ratswere assigned to fourdose groups each containing ten F 0 

## males andten F 0

## females. The testsubstance was administered by oralgavage once daily for aminimum of 28 days at doses of50, 250 or 1000 mg/kg. The control group receivedthe vehicle (propylene glycol) alone. Afull necropsywas performed on five rats/sex/group. Alimited necropsy was performed on the remaining animals. Histopathological evaluation was performed ontissues from the firstfive males, and thefirstfivefemales with live offspring, ingroups 1and 4and on macroscopic abnormalities from rats inall groups. Inaddition, the reproductive organs from all animals thatfailed to mate, conceive, sireordeliver healthyoffspring were examined histologically. PAS stained sections oftestis from thefive selected F 0

## males ingroups 1and 4were examined for abnormalities inspermatogenesis. Histological examinationwas extended to the mesenteric lymph nodes oftheselected males andfemales of Group 2. No animals died, orwere killed, duringthe treatment period. Gross findings included blue/green discolouration ofthe mesenteric lymph nodes inrats inall dose groupsand the presenceof blue gastrointestinal contents inhigh dose females. No histopathological changeswere observed inassociation with the gross findingsor inany othertissuesof rats receiving thetest substance. On the basis ofthe pathologicalfindings the NOAEL may be regarded as 1000 mg/kg/day.

## Page6of97 PATHOLOGY REPORT Testsubstance: 0.1. Pigment Blue61 Notox No.- 488231 Testsystem: 28-Day OralToxicityStudy In the Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.



# Materials and Methods StudyDesign

Group Dose level NumberofAnimals Animal Numbers (mglkg) F 0 

## Males F 0 

## Females Males Females
1 0 10 10 1-10 41-50 2 50 10 10 11-20 51-60 

# 3 250 10 10 21-30 61-70 4 1000 10 10 31-40 71-80

Administration ofthe TestSubstance 

# Thetest substancewasadministered orallyoncedailyforaminimum of28days atdoses of 0,50, 250or 1000 mg/kg. The controlgroup received thevehicle (propylene glycol) alone. NecropsyandHistopatho/ogy 

# Attheendofthe study period,the ratswere killedby exsanguinationfollowing anaesthesia by isoflurane. Afull necropsywas performed onfive rats/sex/group. Alimited necropsywas performed onthe remaininganimals. Organswere preserved in10% neutral buffered formalin (testes, epididymides and eyeswere initiallyfixed inmodified Davidson's solution before being transferred to formalin). Histopathological evaluation was performed on representative samples of thefollowing 

## tissuesfrom thefirst fivemales, and the firstfivefemales with liveoffspring, ingroups 1and 4: adrenal glands; aorta; brain

- 

# cerebellum, midbrain, cortex; cervix; epididymides; heart, kidneys; large intestine

- 

# caecum,clitoral gland: coagulating gland; colon, rectum; liver; lung; lymph nodes

- 

# mandibular, mesenteric; oesophagus; ovaries; pancreas; peyer's patches; pituitary; preputial gland; prostate gland; sciatic nerve; seminalvesicles;small intestine

- 

# duodenum,jejunum, ileum; spinalcord

- 

# cervical, mid-thoracic, lumbar; spleen; sternumwith bone marrow; stomach; testes; thymus; thyroidwith parathyroid; trachea; urinary bladder, uterus; vagina:all gross lesions (examinedfrom all F 0

# rats onstudy). The reproductive organs (testis, epididymides, coagulating gland, seminalvesicles, prostate, preputial gland, ovaries, uterus,cervix,vagina, clitoral gland)from allanimals thatfailedto mate, conceive, sireordeliverhealthy offspring were alsoexamined. Histologicalexamination was extended tothe mesenteric lymph nodes ofthe selected malesand femalesofGroup2. Followingfixation, tissuesweretrimmed, processed and embedded inparaffinwax. Sections 2-4 tm in

# thicknesswere prepared and stainedwith haemnatoxylin and eosin for routine evaluation. Additional sectionsoftestisfrom the fiveselected F 0

# males ingroups 1and4 were stainedwith PAS and examined forabnormalities inspermatogenesis.

## Page7of97 PATHOLOGY REPORT Test substance: C.i. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 24-Oct.08 Sponsor: Color Pigments Manufacturers Association Inc.


## Data Compilation Details of study design and macroscopic findings weretransferred electronically from the necropsy rawdata files of NOTOX By,.intothe computer system ofToxPath Consultancy Ltd wherethe microscopicfindings were recorded using on-line input. Macroscopic findingsare summarised inTable 1(Incidence ofMacroscopic Findings) and listed inTable 3 (IndividualAnimallData). As faras possible, macroscopic findingswere correlatedwith amicroscopicfinding. Microscopicfindings are listed foreach animal, alongwith the severitygrades, inTable 3and summarised inTable 2(incidence ofMicroscopicFindings). Histological changeswere described according to theirdistribution and morphological character. Findings were also gradedfor severity (or degree) on ascale of 1-5 andthese are described inTable 3. Archiving The final pathologyreport (original hardcopy), all glass microscopeslides, slide inventory sheetsand acopy ofthe electronic records (includingfinal pathology report) ascaptured by theappropriate software on CDROM will besent to NOTOX BVfor archiving. Anyother non-study related documents an~d electronic data, covered byGLP,will bearchived by ToxPath Consultancy Ltd according to company standard operating procedures.

## Page 8of97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc. 

# Results Mortality AllF 0 

## rats survived the scheduled duration ofthe study. Macroscopicfindings Green orblue discolouration ofthe mesenteric lymph nodeswas observed inafew group 2 rats and inthe majority ofrats of both sexes ingroups 3and 4. Blue gastrointestinaltract contentswere present inseveral group 4females. Blue discolouration ofthe tail skinwas noted inone male group 4rat. No other findings were recorded at necropsy that were considered to bedue to the test substance. Microscopic findings Notreatment related histopathological changeswere observed inthetissues ofany animal receiving 0.1. Pigment Blue 61. Noabnormalities were notedon evaluation of PAS stained sections oftestesof F 0

## males. All seminiferous tubuleswere normal with respect totheir stage inthe spermatogenic cycle and the integrity ofthevarious cell types presentwithin the differentstages. No pathologicalfindings were present inthe reproductive organs of male and female animals thatfailedto produce offspring. Small amounts of blue material wereobserved inthe lumen ofthe duodenum, ileum,jejunum, colon or rectum inafewgroup4animals. Blue material was also noted inthe superficial skin (keratin layer)ofthetail inone group 4male. Thesefindings are considered to represent the presence ofthe testsubstance and insome cases correlated withthe macroscopic observations. No microscopicfindings wereobserved inthe mesenteric lymph nodesto account forthe discolourationofthese tissues recorded atnecropsy. Itisconsidered thatthediscolouration isdue to the presence ofthetest substance and thatthe pigment was lostfrom the tissues in the processing orstaining procedures. Allother microscopic findingswere considered to be incidental and unrelated to treatment with the testsubstance.

## Page 9of97 PATHOLOGY REPORT Testsubstance: 0.1. Pigment Blue 61 NotoxNo.:488231 Testsystem; 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments ManufacturersAssociation Inc.



# Conclusions Administration ofClI. Pigment Blue 61 by oralgavage to HanWistar rats at doses of50, 250 or 1000 mg/kg/daywasassociated with bluegastrointestinal contents inhigh dosefemales and blue/green discolouration ofthe mesenteric lymph nodes inrats inall dose groups. No histopathological changeswere observed inassociation with the gross findings or inany 

# othertissues of rats receiving thetest substance. On the basis ofthe pathological findings the NOAEL may be regarded as 1000 mg/kg/day.

### PATHOLOGY REPORTPae1of9 Test substance: C.

## 1.Pigment Blue 61 Notox No.:488231 Test system: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 1
- 

### INCIDENCE OF MACROSCOPIC FINDINGS
SEX: Male Female GROUP: 1 2 3 4 1 2 3 4 number of animals 10 10 10 10 10 10 10 10
ADRENAL GLANDS
DISCOLOURATION 0 0 0 0 0 0 0 1
EPIDIDYMIDES NODULE(S)

tail 0 0 2 0

- - - - GENERAL OBSERVATIONS BLUISH

Gltract contenlts 0 0 0 0 0 0 0 7
MANDIBULAR LYMPH NODES
DISCOLOURATION 0 0 0 0 1 2 0 1
MESENT. LYMPH NODE
DISCOLOURATION 0 2 9 10 0 1 8 9
PANCREAS
DISCOLOURATION 0 0 0 0 0 1 0 0
SEMINAL VESICLES
REDUCED INSIZE 0 0 0 1
- - - - SKIM DISCOLOURATION tail 0 1 0 1 0 0 0 0
ALOPECIA

abdominal 

### region 0 0 0 0 1 0 0 0 flank 0 0 0 0 1 0 0 0 foreleg 0 0 0 1 1 0 0 1 hindleg 0 0 0 0 0 0 0 1
TAIL
TAIL BENT 1 0 0 0 0 0 0 0

## Page 11 of97 PATHOLOGY REPORT Testsubstance- C.1. Pigment Blue61 Notox No.:488231 Testsystem: 28-Day Oral ToxicityStudy inthe Rat 24-Oct-08 

## Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 2
- 

### INCIDENCE OF MICROSCOPIC FINDINGS

The grading systems used are as follows:

1 (minimal) 2 (alight) 3 (moderate) 4 (marked) 5 (severe)

## Page 12 

## of97 PATHOLOGY REPORT Testsubstance:

## C.l. PigmentBlue 81 Notox No.:488231 Testsystem: 28-Day Oral Toxicity Study 

## inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 2
- 

### INCIDENCE OF MICROSCOPIC FINDINGS

SEX: Male Female GROUP: 1 2 3 4 1 2 3 4 number of animals 10 10 10 10 10 10 10 10
ADRENAL

number examined 5

- - S 5 - - 5
AORTA

numberexamined 5 - 55 - - BRAIN

number examined 5
- 5 5 - - CASCUM number examined 5 - 5 5

- - 5
CERVIX number examined
- - - 6 1 1 6 CLITORAL GLAND

number examined

- - - -6 1 1 6 interstitial

inflammatory cell infiltration (minimal)

- - -1 0 0 0
COAGULATING GLAND

number examined 5 1 1 5 -- COLON number examined 5
- - 5 5 5 blue material in

lumen present 0

- - 1 0

- - 1 DUJODENUM4 number examined 5 5 blue material in

lumen present 0

- - 1 0

- - 0
EPIDIDYMIS numberexamined 5 1 3 5
- - -
sperm granuloma

(slight) 0 0 1 0

- - - (moderate) 0 0 1 0-

- - Total 0 0 2 0
- - - HEART number examined 5

- - 5 5

- - 5 epicardial fibrosis

(minimal) 0
- - 0 0

- - 1

## Page 13

## of 97 PATHOLOGY REPORT Test substance: 

## C.I. Pigment Blue61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study Inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 2
- 

### ZNCIDENCE 

## OF MICROSCOPIC FINDINGS

SEX: Male Female GROUP: 1 2 3 4 1 2 3 4 number of animals 10 10 10 10 10 10 10 10 ILEUM

number examined 5 5 -5 blue material in

lumen present 0

- - 0 0

- - 1
JEJUNUM

number examined 5

- - 5 5

- - 5 blue eaterial in

lumen present 0

- - 1 

## o0 1
KIDNEYS

number examined 5 - - 5 5

- - 5 basophilic tubules

(minimal) 1

- - 0 0

- - 1 interstitial

inflammatory call infiltration (minimal) 1

- 0 0

- - 0 tubular hyaline droplets (minimal) 1

- - 3 0

- - 0 tubular mineralization (minimal) 0

- - 0 2

- - 1 pelvic dilatation

(minimal) 2

- - 1 0

- - 0
LIVER number examined 5 5 5

- parenchymal

inflammatory cell infiltration (minimal) 3

- - 2 1

- - 0 periportal

inflammatory call infiltration (minimal) 0

- - 1 0

- - 0

### Page 14 of97 PATHOLOGY REPORT Test substance: 0.1. Pigment Blue 61 Notox No.:488231 Test system: 28-Day Oral Toxicity Study Inthe Rat 24-Oct-08 Sponsor Color Pigments Manufacturers Association Inc.

TABLE 2 - INCIDENCE 

### OF MICROSCOPIC FINDINGS
SEX: Male Female GROUP: 1 2 3 4 1 2 3 4 number of animals 10 10 10 10 10 10 10 10 LUNG

number examined 5

- 5 perivascular

inflammatory cell infiltration (minimal) 2

- - 1 0

- - 1 alveolitis

(minimal) 1

- - 0 0

- - 0 alveolar macrophage

infiltration (minimal) 0

- - 1 0

- - 1 LYMPH NODE MESENTERIC

number examined 5 5 9 9 5 5 8 9 macrophage aggregates

(minimal) 1 1 3 2 0 0 1 0 sinus histiocytosia

(minimal) 0 0 0 0 0 0 1 0 (slight) 0 0 0 0 0 1 0 0 Total 0 0 0 0 0 1 1 0
MANDIBULAR LYMPH NODE

number examined 5

- - 5 5 2

- 5 plasma cell

infiltration (minimal) 3

- - 4 0

- 2 (slight) 0

- - 3 0 0

- 1 Total 3

- - 4 4 0

- 3 congestion present 0

- - 0 1 2

- 1
OESOPHAGUS

number examined 5
- - 5 5
- - 5 muscularis

inflammation (minimal) 0

- - 0 2

- - 0 OVARIES number examined

- - - -6 1 1 6
PANCREAS number examined 5 5 5 1

- 5 acinar vacuolation

(minimal) 1

- - 0 0 0

- 0 focal acinar atrophy

(minimal) 0

- - 0 1 0

- 0

### PATHOLOGY REPORT Pglog Testsubstance: 0.1. Pigment Blue 61 Notox No.: 488231 Testsystem: 28-Day OralToxicityStudy Inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 2
- 

### INCIDENCE OF MICROSCOPIC FINDINGS

SEX: Male Female GROUP: 1 2 3 4 1 2 3 4
number of animals 10 10 10 10 10 10 10 10
PARATHYROID number examined 5

- 4 5

- PEYERS PATCHES number examined 5

- - 5 5

- 5
PITUITARY number examined 5 5 5 5 cyst(s) present 0

- - 1 0

- - 0 PREPIIAL GLAND number examined 5 1 1 5-

- - PROSTATE number examined 5 1 1 5-

- - interstitial inflammatory cell infiltration (minimal) 3 0 1 2-

- - - (slight) 1 0 0 0-

- - - Total 4 0 1 2-

- - RECTUM

number examined 5

- 5 - - blue material in

lumen present 0

- - 1 0
- - 1
SCIATIC NERVE

number examined 5

- - 4 5

- - 5 SEMINAL VESICLES number examined 5 1 1 6-

- - reduced luminal

secretory contents present 0 0 0 1--

-

### Page 16 of97 PATHOLOGY REPORT Test substance: C. 1.Pigment Blue61 Notox No.:488231 Test system: 28-Day OralToxicity Study intheRat 24-Oct-08 Sponsor. Color Pigments Manufacturers Association Inc.

TABLE 2
- 

### INCIDENCE OF MICROSCOPIC FINDINGS SEX: Male Female GROUP: 1 2 3 4 1 2 3 4 number of animals 10 10 10 10 10 10 10 10
SKIN number examined
- I - 2 1
- - I epidermal hyperplasia

(slight) 0

- 1 0

- - 0 abdomen: epidermal hyperplasia (minimal)

- 0
- 0 1
- - 0 foreleg right side: epidermal hyperplasia (minimal) 0

- 0 1

- - flank left aide: epidermal hyperplasia (minimal)

- 0

- 0 1

- - 0 blue material in

surface keratin present

- 0

- 1 0
- - 0 foreleg left side: epidermal hyperplesia (minimal)

- 0

- 0 0

- - 1
SPINAL CORD number examined 

### 5

- - 5 5 - - 5 SPLEEN number examined 5 - - 5 5 - - 5 extramedullary haematopoiesis (slight) 0

- - 0 0

- - STER1-UM number examined

- - - 5
STOMACH number examined 5 5

- TAIL number examined 1I -- - - - TESTIS number examined 5 1 1 5

- - - PAS section: no

abnormalities present 5 0 0 4

- - - hypospermatogenesis

(minimal) 0 0 0 1-

- -

## Page 17

## of97 PATHOLOGY REPORT Test substance: 0.1. Pigment Blue 61 Notox No.: 488231 Testsystem: 28-Day OralToxicity Study inthe Rat 24-Oct-OS Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 2
- 

### INCIDENCE OF MICROSCOPIC FINDINGS
SEX: Male Female GROUP: 1 2 3 4 1 2 3 4
number of animals 10 10 10 10 10 10 10 10
THYMUS

number examined 5
- - 

## s 5
- - 5 involution/atrophy

(minimal) 0 - - 0 1

- - 1
THYROID

number examined 5

- - 5 5 - - 5
TRACHEA

number examined 5

- - 5 5 - 5
URINARY BLADDER

number examined 5

- - 5 5 - - 5 UTERUS

number examined
- - - - 6 1 1 6
VAGINA

number examined
- - - 6 1 1 6
oestrus present - - - - 1 0 0 0 metoestrus

present - - 0 0 1 1 dioestrua present - - -5 1 0 5 epithelial1 vacuoletion (minimal) - 3 0 0 1 (slight) - 1 0 0 3 (moderate) - - -1 0 0 0 Total

- - - -5 0 0 4

### Page 18 of97 PATHOLOGY REPORT Testsubstance: C.1. Pigment Blue61 Notox No.:488231 Testsystem: 28-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: SEX: male DOSE LEVEL: 1
M AC

ROSCOPIC FI N DlI N

GS da s o tr t m n 30 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 

### 17/06/08 date of end of treatment: 16/07/08 date of necropsy 17/07/08
No abnormality detected
MICROSCOPIC FINDINGS LUNG perivascular inflammatory call infiltration (ainimal)
MANDIBULAR plasma cell infiltration (minimal)

LYMPH NODE
PROSTATE interstitial inflammatory cell infiltration (minimal)
TESTIS PAS section: no abnormalities present 

### No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, COAGULATING

GLAND, COLON, DUODENUM, EPIDIDYMIS, HEART, ILEUM, JEJUNUM, KIDNEYS, LIVER, LYMPH NODE MESENTERIC, OESOPHAGUS, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, PREPUTIAL GLAND, RECTUM, SCIATIC NERVE, SEMINAL VESICLES, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER.

### PATHOLOGY REPORTPae1of9 Test substance: C.I. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicityStudy Inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 2 SEX; Male DOSE LEVEL: 1
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatent: 16/07/08 date of necropsy :17/07/Os
Mo abnormality detected
MICROSCOPIC FINDINGS KIDNEYS pelvic dilatation (minimal)
LUNG perivascular inflammatory cell infiltration (minimal)
MANDIBULAR plasma cell infiltration (minimal)

LYMPM NODE PROSTATE interstitial inflammatory cell infiltration (minimal)
TESTIS PAS section: no abnormalities present
No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, COAGULATING

GLAND, COLON, DUODENUM, EPIDIDYMIS, HEART, ILEUM, JEJUNUM, LIVER, LYMPH MODE MESENTERIC, OESDPHAGUS, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, PREPUTIAL GLAND, RECTUM, SCIATIC NERVE, SEMINAL VESICLES, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER.

## Page20 of97 PATHOLOGY REPORT Test substance:C.l. PigmentBlue 61 NotoxNo.:488231 Testsystem:28-Day OralToxicityStudy In the Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.:i 3 SEX: Male DOSS LEVEL: 1
MACROSCOPIC FINDINGS days of treatment :30 sacrifice group :terminal sacrifice necropsy status :planned terminal data of start of treatment: 17/06/08 date of end of treetment: 

### 16/07/08 date of necropsy :17/07/08
TAIL BENT
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS KIDNEYS basophilic tubules (minimal) interstitial inflammatory cell infiltration (minimal) tubular hyaline droplets (minimal) pelvic dilatation (minimal)
LIVER parenchymal inflammatory cell infiltration (minimal)
MANDIBULAR plasma cell infiltration (minimal)

LYMPH NODE
PROSTATE interstitial inflammatory cell infiltration (slight)
TAIL to0lesion to account for gross finding(s)
TESTIS PAS section: no abnormalities present
Ho abnormalities found in: ADRENAL, AORTA, DRAIN, CAECUM, COAGULATING GLAND, COLON, DUODENUM, EPIDIDYMIS, HEART, ILEUM, JEJUNUM, LUNG, LYMPH MODE MESENTERIC, OESOPHAGUS, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, PREPUTIAL GLAND, RECTUM, SCIATIC NERVE, SEMINAL VESICLES, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER.

### Page21 of97 PATHOLOGY REPORT Test substance: 

## C.I. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments ManufacturersAssociation Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. 4 SEX: Male DOSE LEVEL: 1
MACROSCOPIC FINDlINGS daya of treatment :30 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 data of end of treatment: 16/07/08 date of necropsy :17/07/08 No abnormality detected
MICROSCOPIC FINDINGS
LIVER parenchymal inflammatory cell infiltration (minimal) LUNG alveolitis (minimal)
PANCREAS acinar vacuolation (minimal)
TESTIS PAS section: no abnormalities present No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, COAGULATING

GLAND, COLON, DUODENUM, EPIDIDYNIS, HEART, ILEUM, JEJUNUM, KIDNEYS, LYMPH NODE MESENTERIC, MANDIBULAR LYMPH NODE, OESOPHAGUS, PARATHYROID, PEYERS PATCHES, PITUITARY, PREPUTIAL GLAND, PROSTATE, RECTUM, SCIATIC NERVE, SEMINAL VESICLES, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER.

## Page 22 of97 PATHOLOGY REPORT Testsubstance: C.I. Pigment Blue 61 Notox No.: 488231 Testsystem: 28-Day Oral ToxicityStudy Inthe Rat 24-Oct-08 Sponsor. Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 5 SEX: Male DOSE LEVEL: 1 MACROSCOPIC FINDINGS days of treatment :30 sacrif ice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 data of end of treatment: 16/07/08 data of necropsy :17/07/08
No abnormality detected
MICROSCOPIC FINDINGS
LIVER parenchymal inflammatory cell infiltration (minimal)
LYMPH NODE macrophage aggregates (minimal) MESENTERIC PROSTATE interstitial inflammatory cell infiltration (minimal)
TESTIS PAS section: no abnormalities present No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, COAGULATING GLAND, COLON, DUODENUM, EPIDIDYMIS, HEART, ILEUM, JEJUNUM, KIDNEYS, LUNG, MANDIBULAR LYMPH NODE, OESOPHAGUS, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, PREPUTIAL GLAND, RECTUM, SCIATIC NERVE, SEMINAL VESICLES, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER.

## Page 23 of97 PATHOLOGY REPORT Test substance: C.!. Pigment Blue 61 Notox No.:488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: S SEX: Male DOSE LEVEL: I
MACROSCOPIC FINDINGS days of treatment 30 sacrifice 

### group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy :17/07/08
No abnormality detected MICROSCOPIC FINDINGS No tissues examined

## Page 24of97 PATHOLOGY REPORT Testsubstance: C. 1.Pigment Blue 61 Notox No.:488231 Testsystem: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments ManufacturersAssociation Inc.

TABLE 3 - INDIVIDUAL ANIMAL DATA

Animal No.: 7 SEX: Male DOSE LEVEL: 1
MACROSCOPIC FINDINGS days of treatment :30 sacrifice group :terminal sacrifice necropay status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 data of necropsy : 17/07/08
No abnormality detacted
MICROSCOPIC FINDINGS No tissues examined

### Page 25 of97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue61 Notox No.: 488231 Test system: 26-Day Oral ToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. : 8 

### SEX; Male 

### DOSE LEVEL: I 

### MACROSCOPIC FINDINGS days of treatment :30 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/O6/OS date of end of treatment: 16/07/08 date of necropsy :17/07/08 No abnormality detected 

### MICROSCOPIC FINDINGS No tissues examined

### PATHOLOGY REPORTPae2of9 Test substance: C.1. Pigment Blue 61 Notox No-.488231 Test system: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.


## TABLE 3

- 

### INDIVIDUAL ANIMAL 

## DATA

Animal No.: 9 SEX: Male DOSE LEVEL: I
MACROSCOPIC FINDINGS days of treatment :30 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 

### 17/06/08 date of end of treatment! 16/07/08 date of necropsy : 17/07/08
No abnormality detected

MICROSCOPIC FINDINGS No tissues examined

### PATHOLOGY REPORTPae2of9 Test substance: 

## C.I. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study Inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. 10 SEX: 

### Male DOSE LEVEL: 1
MACROSCOPIC FINDINGS days of treatment :30 sacrifice group terminal sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of end of treatment; 16/07/08 date of necropsy 17/07/08
No abnormality detected
MICROSCOPIC FINDINGS No tissues examined

### Page 28 of97 PATHOLOGY REPORT Test substance: 

## C.I. Pigment Blue 61 Notox No.:488231 Test system: 28-Day OralToxicity Study inthe Rat 24.Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3 - INDIVIDUAL ANIMAL DATA

Animal No. 11 SEX: Male DOSE LEVEL: 2
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group terminal sacrifice necropsy status :planned terminal date of start of treatment: 

### 17/06/08 date of end of treatment: 16/07/08 date of necropsy :17/07/08
No abnormality detected
MICROSCOPIC FINDINGS No abnormality detected
No abnormalities found in: LYMPH NODE MESENTERIC,

### PATHOLOGY REPORTPae2of9 Test substance: 

## C.l. Pigment Blue 81 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. 12 SEX: Male DOSE LEVEL: 2 M4ACROSCOPIC FINDINGS days of treatment 30 sacrifice group terminal sacrifice necropsy 

### atatua planned terminal date of atart of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropay : 17/07/08
MESENT. LYMPH DISCOLOURATION GREENISH NODE
No abnormalities were found in any of the other tiasues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding Is) MESENTERIC

### Page 30 of97 PATHOLOGY REPORT Testsubstance: C.1.Pigment Blue61 Notox No.:488231 Test system: 28-Day Oral ToxicityStudy in the Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 

### 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. 13 SEX; Male DOSE LEVEL; 2
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group terminal sacrifice necropsy statue planned terminal date of start of treatment; 17/06/08 date of end of treatment: 16/07/08 date of necropsy 17/07/08
No abnormality detected
MICROSCOPIC FINDINGS No tissues examined

## PATHOLOGY REPORTPae3of9 Testsubstance: C.I. PigmentBlue 61 Notox No.: 488231 Testsystem: 28-Day OralToxicityStudy in the Rat 24-Oct-OS Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. 14 SEX: Male DOSE LEVEL: 2
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/Os date of necropsy :17/07/08
No abnormality detected
MICROSCOPIC FINDINGS LYMPH MODE macrophage aggregates (minimal) MESENTERIC No abnormalities found in: COAGULATING GLAND, EPIDIDYMIS, PREPUTIAL GLAND, PROSTATE, SEMINAL VESICLES, TESTIS.

### Page32 of 97 PATHOLOGY REPORT Testsubstance:

## C.1.Pigment Blue61 Notox No.:488231 Testsystem: 28-Day Oral ToxicityStudy in the Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. is1 SEX Hale DOSE LEVEL: 2
MACROSCOPIC FINDINGS days of treatment :30 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/ce date of end of treatment: 16/07/08 date of necropsy ;17/07/08
SKIN tail: DISCOLOURATION BLUISH
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS SKIN no lesion to account for gross finding Is) No abnormalities found in: LYMPN NODE MESENTERIC.

### Page 33 of97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue61 Notox No.: 488231 Test system: 26-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 16 SEX: Male DOSE LEVEL: 2 MACROSCOPIC FINDINGS days of treatment :30 sacrifice group :terminal sacrifice nacropsy status :planned terminal date of start of treatment: 

### 17/06/08 date of end of treatment: 16/07/08 date of necropsy : 17/07/08 MESENT. LYMPH DIScOLOURATION GREENISH

NODE
Ho abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding(s) MESENTERIC

## Page 34of 97 PATHOLOGY REPORT Testsubstance: C.l. Pigment Blue 61 Notox No.:488231 Test system: 28-Day OralToxicity Studyinthe Rat 24-Oct-08 Sponsor: Color PigmentsManufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. : 

### 17 SEX: Male DOSE LEVEL: 2
MACROSCOPIC FINDINGS 

### days of treatment :30 

### sacrifice group :terminal 

### sacrifice necropsy status :planned 

### terminal data Of start of treatment: 17/06/08 data of end of treatment: 16/07/08 date of necropsy : 17/07/08
No abnormality detected

MICROSCOPIC FINDINGS No tissues examined

### PATHOLOGY REPORTPae3of9 Test substance: 

## C.
. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 18 SEX: Male DOSE LEVEL: 2
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/05 date of end of treatment: 16/07/08 date of necropsy : 17/07/08
No abnormality detected
MICROSCOPIC FINDINGS No tisaues examined

## Page 36of 97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue61 Notox No.: 488231 Test system: 28-Day OralToxicityStudyinthe Rat 24-Oct-08 Sponsor: Color Pigments ManufacturersAssociation Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. 19 SEX; Male DOSE LEVEL: 2
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group :terminal sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy 17/07/08 No abnormality detected
MICROSCOPIC FINDINGS No tissues examined

## Page37 

## of97 PATHOLOGY REPORT Test substance: C.!. Pigment Blue61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments ManufacturersAssociation Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 20 SEX: Male DOSE LEVEL; 2
MACROSCOPIC FINDINGS days of treatment :30 eacrifice group :terminal sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy :17/07/08
No abnormality detected
MICROSCOPIC FINDINGS No tissues examined

## Page 38 of97 PATHOLOGY REPORT Test substance. CtI. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.


## TABLE 3

- 

### INDIVIDUAL ANIMAL 

## DATA

Animal No.: 21 SEX: Male DOSE LEVEL: 

### 3
MACROSCOPIC FINDlINGS days of treatment 30 sacrifice group terminal sacrif ice necropsy status planned terminal date of 

### start of treatment: 17/06/08 date of end of treatment: 

### 16/07/08 date of necropay :17/07/08
MESENT. LYMPH DISCOLOURATION GREENISH

NODE No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding Is) MESENTERIC

### Page 39 of 97 PATHOLOGY REPORT Test substance: 0.1I. Pigment Blue 61 Notox No.:468231 Test system: 28-Day Oral ToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments ManufacturersAssociation Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 22 SEX: Male DOSE LEVEL: 3 MACROSCOPIC FINDINGS days of treatment 30 sacrifice group terminal sacrifice necropsy status planned terminal date of start of treatment; 

### 17/06/0e date of end of treatment: 16/07/08 date of necropsy 17/07/08 EPIDIDYNIDES tail: right side NODULE(S) D=7X5 NM, YELLOWISH, HARD MESENT. LYMPH DISCOLOURATION GREENISH

NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS EPIDIDYMIS sperm granuloma (moderate) (correlates to GROSS finding)
LYMPH NODE no lesion to account for gross finding(s) MESENTERIC

## Page 40 of97 PATHOLOGY REPORT Testsubstance: 

## ClI. Pigment Blue61 NotoxNo.: 488231 Test system: 28-DayOral ToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3 - INDIVIDUAL ANIMAL DATA

Animal No,: 23 SEX: Male DOSE LEVEL; 3
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy 17/07/08
MESENT. LYMPH DISCOLOURATION GREENISH

MODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE macrophage aggregates (minimal) MESENTERIC no lesion to account for gross finding 1s1

### Page 41 of97 PATHOLOGY REPORT Test substance: 

## C.I. Pigment Blue 61 Notox No.:488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 24-OctO8 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.; 24 SEX: Male DOSE LEVELi 3
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group terminal sacrifice necropsy status planned terminal date of start of treatment; 17/06/08 date of end of treatment: 16/07/08 date of necropsy 17/07/08 MESENT. LYMPH DISCOLOURATION GREENISH

NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS LYMPH NODE no lesion to account for gross findingis) MESENTERIC

## Page 42of97 PATHOLOGY REPORT Testsubstance: ClI. Pigment Blue 61 Notox No.:488231 Testsystem: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 25 SEX: Male DOSE LEVEL: 

### 3
MACROSCOPIC FINDINGS days of treatment :30 sacrif ice group terminal sacrifice necropsy status planned terminal date of start of treatment; 17/06/08 date of end of treatment: 

### 16/07/08 date of necropsy : 17/07/08
MESENT. LYMPH DISCOLOURATION GREENISH

NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding Is) MESENTERIC

## Page43of 97 PATHOLOGY REPORT Testsubstance: C.I. Pigment Blue 61 NotoxNo.: 488231 Testsystem: 28-Day Ora(ToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- INDIVIDUAL ANIMAL DATA

Animal No.: 26 SEX: Male DOSE LEVEL: 3
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group :terminal sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy :17/07/08
No abnormality detected
MICROSCOPIC FINDINGS PROSTATE interstitial inflammatory cell infiltration (minimal)
No abnormalities found in: COAGULATING GLAND, EPIDIDYMIS, PREPUTIAL GLAND,

SEMINAL VESICLES, TESTIS.

### Page44 of 97 PATHOLOGY REPORT Test substance: 

## C.I. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 27 SEX: Male DOSE LEVEL: 3
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group :terminal sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy : 17/07/08
EPIDIDYMIDES tail: right side NODTULE(S) D=2X2 MM, YELLOWISH, HARD
MESENT. LYMPH DISCOLOURATION GREENISH NODE No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS EPIDIDYMIS sperm granuloma (alight) (correlates to GROSS finding)
LYMPH NODE no lesion to account for gross findinglal MESENTERIC

## Page 45

## of 97 PATHOLOGY REPORT Test substance: C.l. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 28 SEX: Male DOSE LEVEL: 3
MACROSCOPIC FINDINGS daya of treatment :30 sacrifice group :terminal sacrifice necropsy status :planned terminal date of atart of treatment; 17/06/08 date of end of treatment: 16/07/08 date of necropsy :17/07/08 mEsENT. LYMPH DISCOLOURATION GREENISH NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS LYMPH NODE macrophage aggregates (minimal) MESENTERIC no lesion to account for gross finding(s)

### Page 46 of97 PATHOLOGY REPORT Test substance: ClI. Figment Blue61 Notox No.: 488231 Test system: 26-Day Oral Toxicity Studyinthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.; 29 SEX: Male DOSE LEVEL: 3
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group terminal sacrifice necropsy atatus :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy ;17/07/08 MESENT. LYMPH DISCOLOURATION GREENISH NODE No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE macrophage aggregates iminimal) MESENTERIC no lesion to account for gross findingis)

### Page 47 of97 PATHOLOGY REPORT Test substance: C. Pigment Blue 61 Notox No.:488231 Test system: 28-Day Oral Toxicity Study in the Rat 24-Oct-08 Sponsor: Color Pigments ManufacturersAssociation Inc.

TABLE 3 - 

### INDIVIDUAL ANIMAL DATA

Animal No.: 30 SEX: Male DOSE LEVEL: 3
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 

### 16/07/08 date of necropsy :17/07/08 MESENT. LYMPH DISCOLOURATION GREENISH

NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding(s) MESENTERIC

### Page48 of 97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue 81 Notox No.: 488231 Test system: 28-Day Owal Toxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 31 SEX: Male DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment :30 sacrifice group :terminal saecrifice necropsy status planned terminal date of start of treatment: 17/06/08 data of end of treatment: 16/07/08 date of necropsy 

### 17/07/08 MESENT. LYMPH DISCOLOURATION GREENISH NODE No 

### abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS DUODENUM blue material in lumen present
JEJUNUN blue material in lumen present
KIDNEYS pelvic dilatation (minimal)
LYMPH NODE macrophage aggregates (minimal) MESENTERIC no lesion to account for gross finding(s)
MANDIBULAR plasma cell infiltration (slight)

LYMPH NODE TESTIS PAS section: no abnormalities present
No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, COAGULATING

GLAND, COLON, EPIDIDYMIS, HEART, ILEUM, LIVER, LUNG, CESOPHAGUS, PANCREAS, PEYERS PATCHES, PITUITARY, PREPUTIAL GLAND, PROSTATE, RECTUM, SCIATIC NERVE, SEMINAL VESICLES, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER.

### Page 49 of 97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue 61 Notox No.:488231 Test system: 28-Day OralToxicity Study inthe Rat 24-Oct-OS Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3 -INDIVIDUAL ANIMAL DATA

Animal No.; 32 SEX; Male DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group terminal sacrifice nacropay status planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy : 17/07/08
MESENT. LYMPH DISCOLOURATION GREENISH

NODE No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS COLON blue material in lumen present
KIDNEYS tubular hyaline droplets (minimal)
LIVER parenchymal inflammatory cell infiltration (minimal)
LUNG alveolar macrophage infiltration (minimal)
LYMPH NODE no lesion to account for gross finding(s) MESENTERIC No abnormalities found in: ADRENAL, AORTA, BRAIN, CASCUN, COAGULATING GLAND, DUODENUM, EPIDIDYMIS, HEART, ILEUM, JEJUJNUM, MANDIBULAR LYMPH NODE, OESOPHAGUS, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, PREPUTIAL GLAND, PROSTATE, RECTUM, SCIATIC NERVE, SEMINAL VESICLES, SPINAL CORD, SPLEEN, STERNUM, STOMACH, TESTIS, THYMUS, THYROID, TRACHEA, URINARY BLADDER.

## Page 50of97 PATHOLOGY REPORT Test substance: 

## C.I. Pigment Blue 61 NotoxNo.:488231 Test system: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments ManufacturersAssociation Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. 33 SEX: Male DOSE LEVEL: 4
MACROSCOPIC FIND)INGS days of treatment :30 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy :17/07/08
MESENT. LYMPH DISCOLOURATION GREENISH

NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS LYMPH NODE no lesion to account for gross finding(s) MESENTERIC
MANDIBULAR plasma cell infiltration (slight)

LYMPH NODE
PROSTATE interstitial inflammatory cell infiltration (minimal)
TESTIS PAS section: no abnormalities present hypospermatogenesis (minimal)
No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, COAGULATING

GLAND, COLON, DUODENUM, EPIDIDYMIS, HEART, ILEUM, JEJUNUM, KIDNEYS, LIVER, LUNG, OESOPHAGUS, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, PREPUTIAL GLAND, RECTUM, SCIATIC NERVE, SEMINAL VESICLES, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER.

### Page 51 of197 PATHOLOGY REPORT Testsubstance: C.I. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study Inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 34 SEX: Hale DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment :30 aacrifice group :terminal sacrifice necropsy 

### status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy 17/07/08 MESENT. LYMPH DISCOLOURATION GREENISH

NODE
SKIN FORELEG: left side ALOPECIA
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS KIDNEYS tubular hyaline droplets (minimal)
LIVER periportal inflammatory cell infiltration (minimal)
LYMPH NODE no lesion to account for gross findingis) MESENTERIC MANDIBULAR plasma cell infiltration (minimal)

LYMPH NODE
PITUITARY cyst Is) present
PROSTATE interstitial inflammatory call infiltration (minimal)
RECTUM blue meaterial in lumen present
SKIN epidermal hyperplasia (slight) (correlates to GROSS finding)
TESTIS PAS section: no abnormalities present
No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUN, COAGULATING

GLAND, COLON, DUODENUM, EPrDIDYNIS, HEART, ILEUN, JIEJUNUM, LUNG, OESOPHAGUS, PANCREAS, PARATHYROID, PEYERS PATCHES, PREPUTIAL GLAND, SEMINAL VESICLES, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER.

## Page 52of 97 PATHOLOGY REPORT Testsubstance: 

## C.l. Pigment Blue 61 NotoxNo.: 488231 Testsystem: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 

### 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 35 SEX: Male DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment :30 sacrifice group :terminal sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy i17/07/08
MESENT. LYMPH DISCOLOURATION GREENISH

NODE No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS KIDNEYS tubular hyaline droplets (minimal)
LIVER parenchymal inflammatory cell infiltration (minimal)
LUNG perivasculer inflammatory cell infiltration (minimal)
LYMPH HODE no lesion to account for gross finding Is) NEsENTERIC
MANDIBULAR plasma cell infiltration (slight)

LYMPH NODE TESTIS PAS section: no abnormalities present No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, COAGULATING

GLAND, COLON, DUODENUM, EPIDIDYNIS, HEART, ILEUM, JEJUNUM, ORSOPHAGUS, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, PREPUTIAL GLAND, PROSTATE, RECTUN, SCIATIC NERVE, SEMINAL VESICLES, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER.

### PATHOLOGY REPORTPae5of9 Test substance: 

## C.I. Pigment Blue 61 Notox No.: 488231 Test 

## system: 28-Day Oral ToxicityStudy Inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 36 SEX; Male DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment 30 sacrifice group :terminal sacrifice necropsy status planned terminal date of start of treatment; 17/06/08 date of end of treatment: 16/07/08 date of necropsy : 17/07/08 MESENT. LYMPH DISCOLOURATION GREENISH NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE macrophage aggregates (minimal) MESENTERIC no lesion to account for gross finding Is)

### Page 54 of 97 PATHOLOGY REPORT Test substance: 0.1. Pigment Biue 61 Notox No.: 488231 Testsystem: 28-Day Oral Toxicity Study in the Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc,

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 37 SEX: 

### Male DOSE LEVEL: 4
MACROSCOPIC FINDlINGS days of treatment :30 sacrifice group terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 1s/07/08 date of necropsy :17/07/08
MESENT. LYMPH DISCOLOURATION GREENISH NODE No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
No tissues examined

### Page 55 of 97 PATHOLOGY REPORT Testsubstance: 0. 1.Pigment Blue61 Notox No.:488231 Testsystem: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 38 SEX: Male DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment .30 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy ; 17/07/08
MESENT. LYMPH DISCOLOURATION GREENISH NODE
No abnormalities ware found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding(s) MESENTERIC

## Page 56of97 PATHOLOGY REPORT Testsubstance: 

## ClI. Pigment Blue61 Notox No.: 488231 Testsystem: 28-Day Oral Toxicity Study Inthe Rat 24-Oct-08 

## Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 39 SEX: male DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment :30 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy :17/07/0s MESENT. LYNPH DISCOLOURATION GREEMISM

NODE
SKIN tail: DISCOLOURATION BLUISH
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS LYMPH NODE no lesion to account for gross finding(s) MESENTERIC SKIN blue material in Surface keratin present (correlates to GROSS finding)

### Page 57 of97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue61 Notox No.:488231 Test system: 28-Day Oral Toxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments ManufacturersAssociation Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.! 40 SEX: Male DOSE LEVEL: 4 MACROSCOPIC FINDINGS days of treatment : 30 sacrifice group : terminal sacrifice necropsy status : planned terminal date of start of treatment: 17/06/08 date of end of treatment: 16/07/08 date of necropsy :17/07/08 MESENT. LYMPH DISCOLOURATION GREENISH

NODE
SEMINAL left side REDUCED IN SIZE VESICLES No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross findingis) MESENTERIC
SEMINAL reduced luminal secretory contents present (correlates to GROSS finding) VESICLES

### Page 58 of 97 PATHOLOGY REPORT Test substance: Ctf. Pigment Blue61 Notox No.:488231 Test system: 28-Day Oral ToxicityStudy In the Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 41 SEX; Female DOSE LEVEL; 1

MACROSCOPIC FINDINGS days of treatment 42 aacrifice group terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment; 

### 28/07/08 date of necropsy :29/07/08
No abnormality detected
MICROSCOPIC FINDINGS CLITORAL GLAND interstitial inflammatory cell infiltration (minimal)
VAGINA oestrus present No abnormalities found in: CERVIX, OVARIES, UTERUS.

### Page 59

## of 97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue 61 Notox No.:488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 24-Oct-08 Sponsor: ColorPigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Anlimal No.: 42 SEX: Female DOSE LEVEL: 1

MACROSCOPIC FINDINGS days of treatment 41 sacrifice group :terminal sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of end of treatment: 27/07/08 date of necropsy ;28/07/08
No abnormality detected
MICROSCOPIC FINDINGS KIDNEYS tubular mineralization (minimal)
MANDIBUJLAR plasma cell Infiltration (minimal)

LYMPH NODE THYMUS involution/atrophy (minimal)
VAGINA dioestrus present epithel~ial vacuolation (slight) No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, CERVIX, CLITORAL

GLAND, COLON, DUODENUM, HEART, ILEUM, JEJUNUM, LIVER, LUNG, LYMPH NODE MESENTERIC, OESOPHAGUS, OVARIES, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, RECTUM, SCIATIC NERVE, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYROID, TRACHEA, URINARY BLADDER, UTERUS.

### Page60 of 97 

## PATHOLOGY REPORT Test substance: 0.1. Pigment Blue 61 Notox No.:488231 Test system: 28-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 43 SEX: Female DOSE LEVEL: 1
MACROSCOPIC FINDlINGS days of treatment 41 sacrifice group terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 27/07/08 date of necropsy :28/07/08
No abnormality detected
MICROSCOPIC FINDINGS
MANDIBULAR plasma cell infiltration (minimal)

LYMPH NODE OESOPHAGUS muscularis inflammation (minimal)
VAGINA dioestrus present epithelial vacuolation (moderate)
Ho abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUH, CERVIX, CLITORAL GLAND, COLON, DUODENUM, HEART, ILEUM, JEJUNUM, KIDNEYS, LIVER, LUNG, LYMPH NODE MESENTERIC, OVARIES, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, RECTUM, SCIATIC NERVE, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID), TRACHEA, URINARY BLADDER, UTERUS.

### Page61 of 97 PATHOLOGY REPORT Testsubstance: C.l. Pigment Blue 61 Notox No.:488231 Testsystem: 28-Day OralToxicity Study inthe Rat 24-Oct-O8 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 44 SEX: Female DOSE LEVEL; I MACROSCOPIC FINDINGS days 

### of treatment :45 sacrifice group :terminal 

### sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of end of treatment: 31/07/08 date of necropsy : 01/08/08
No abnormality detected
MICROSCOPIC FINDINGS
No tissues examined

### Page 62 of 97 PATHOLOGY REPORT Testsubstance: ClI. Pigment Blue 61 Notox No.:488231 Test system: 28-Day oral Toxicity Studyinthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.; 45 SEX: Female DOSE LEVEL: I MACROSCOPIC FINDINGS days of treatment :45 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 31/07/08 date of necropsy :01/08/08 No abnormality detected
MICROSCOPIC FINDINGS No tissues examined

## Page 63 of97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 46 SEX: Female DOSE LEVEL: I MACROSCOPIC FINDINGS days of treatment :45 sacrifice group terminal sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of end of treatment; 31/07/08 date of necropsy : 

### 01/08/08
No abnormality detected
MICROSCOPIC FINDINGS No tissues examined

## Page 64 of97 PATHOLOGY REPORT Testsubstance: 

## C.I. Pigment Blue 61 Notox No.:488231 Testsystem: 28-DayOral ToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3 - INDIVIDUAL ANIMAL DATA

Animal No.: 47 SEX: Female DOSE LEVEL: 1
MACROSCOPIC FINDINGS days of treatment 4 43 sacrifice group terminal sacrifice necropsy status planned terminal data of start of treatment: 17/06/08 date of end of treatment: 29/07/ca date of nacropsy : 

### 30/07/08
MANDIBULAR both sides DISCOLOURATION DARK RED

LYMPH NODES SKIN foreleg: left side ALOPECIA abdominal region: ALOPECIA foreleg: right side ALOPECIA flank; right side ALOPECIA flank; left side ALOPECIA
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS MANDIBULAR plasma cell infiltration (minimal)

LYMPH NODE congestion present (correlates to GROSS finding)
SKIN abdomen: epidermal hyperplasia (minimal) (correlates to GROSS finding) foreleg right side: epidermal hyperplasia (minimal) (correlates to GROSS finding) flank left side: epidermal hyperplasia (minimal) (correlates to GROSS finding)
VAGINA dioestrus present epithelial vacuolation (minimal)
No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, CERVIX, CLITORAL

GLAND, COLON, DUODENUM, HEART, ILEUM, JEJUNUM, KIDNEYS, LIVER, LUNG, LYMPH NODE MESENTERIC, OESOPHAGUS, OVARIES, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, RECTUM, SCIATIC NERVE, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER, UTERUS.

## Page65 of97 PATHOLOGY REPORT Testsubstance:

## ClI. Pigment Blue 61 NotoxNo.: 488231 Testsystem: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Aniimal No.: 4 SEX: Female DOSE LEVEL: 1
MACROSCOPIC FINDINGS days of treatment :43 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 29/07/08 

### date of necropsy :

## 30/07/08
No ebnormality detected
MICROSCOPIC FINDINGS KIDNEYS tubular mineralization (minimal)
MANDIBULAR plasma cell infiltration (minimal)

LYMPH NODE
VAGINA dinestrus present epithelial.vacuolation (minimal)
No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, CERVIX, CLITORAL

GLAND, COLON, DUODENUM, HEART, ILEUM, JEJUNUM, LIVER, LUNG, LYMPH NODE MESENTERIC, OESOPHAGUS, OVARIES, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, RECTUM, SCIATIC NERVE, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER, UTERUS.

### Page 66 of97 PATHOLOGY REPORT Test substance: 

## C.I. Pigment Blue61 Notox No.:488231 Test system: 28-Day Oral Toxicity Study in the Rat 24-Oct-06 Sponsor: Color Pigments ManufacturersAssociation tnc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 49 SEX: Female DOSE LEVEL: I
MACROSCOPIC FINDINGS days of treatment 43 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 

### 17/06/08 date of end of treatment; 29/07/08 date of necropsy : 30/07/08
No abnormality detected
MICROSCOPIC FINDINGS
No tissues examined

### Page 67 of97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue 81 Notox No.:488231 Test system: 28-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 50 SEX: Female DOSE LEVEL: 1
MACROSCOPIC FINDINGS days of treatment 41 aacrifice group terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 27/07/08 data of necropsy : 

### 28/07/08
No abnormality detected
MICROSCOPIC FINDINGS
LIVER parenchymal inflammatory cell infiltration (minimal)
OESOPHAGUS muscularis inflammation (minimal)
PANCREAS focal acinar atrophy (minimal)
VAGINA dioestrus present epithelial vacuolation (minimal)
No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, CERVIX, CLITORAL

GLAND, COLON, DUODENUM, HEART, ILEUM, JEJUNUM, KIDNEYS, LUNG, LYMPH NODE MESENTERIC, MANDIBULAR LYMPH NODE, OVARIES, PARATNYROID, PEYERS PATCHES, PITUITARY, RECTUM, SCIATIC NERVE, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER, UTERUS.

### Page 68 of 97 PATHOLOGY REPORT Test substance: 0.5. Pigment Blue61 Notox No.:488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 24-Oct.08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 51 SEX: Female DOSE LEVEL: 2

MACROSCOPIC FINDINGS days of treatment 44 sacrifice group terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 30/07/08 date of necropsy :31/07/08
No abnormality detected

MICROSCOPIC FINDINGS No tissues examined

## Page69 of 97 PATHOLOGY REPORT Test substance: C.I. 

## PigmentBlue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Studyinthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3 - INDIVIDUAL ANIMAL DATA

Animal No.: 52 SEX: Female DOSE LEVEL: 2
MACROSCOPIC FINDINGS days of treatment 43 sacrifice group :terminal sacrifice necropsy status planned terminal date of start of treatment: 

### 17/06/08 date of end of treatment: 

### 29/07/08 date of necropsy :30/07/08
MANDIBULAR both sides DISCOLOURATION REDDISH

LYMPH NODES
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
MANDIBULAR congestion present (correlates to GROSS finding)

LYMPH NODE
No abnormalities found in: LYMPH NODE MESENTERIC,

### PATHOLOGY REPORTPae7of9 Test substance: C.I. Pigment Blue 81 Notox No.:488231 Test system: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 53 SEX: Female DOSE LEVEL: 2
MACROSCOPIC FINDINGS days of treatment :45 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 

### date of end of treatment: 31/07/08 

### date of necropsy ; 01/08/08 No abnormality detected MICROSCOPIC FINDINGS No tissues examined

### Page 71 of97 PATHOLOGY REPORT Test substance: C.l. Pigment Blue61 Notox No.:488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 54 SEX: Female DOSE LEVEL: 2
MACROSCOPIC FINDINGS days of treatment t 42 sacrifice group terminal sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of end of treatment: 28/07/08 date of necropsy 29/07/01s
No abnormality detected
MICROSCOPIC FINDINGS VAGINA dioeatrua present
No abnormalities found in: CERVIX, CLITORAL GLAND, OVARIES, UTERUS.

### Page 72 of 97 PATHOLOGY REPORT Testsubstance: C.I. Pigment Blue61 Notox No.: 488231 Testsystem: 28-Day Oral Toxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 55 SEX: Female DOSE LEVEL: 2
MACROSCOPIC FINDINGS days of treatment :43 sacrifice group terminal sacrifice necropsy status i planned terminal date of start of treatment: 17/06/08 date of end of treatment: 29/07/08 date of necropsy :30/07/08
No abnormality detected MICROSCOPIC FINDINGS
No abnormality detected
No abnormalities found in: LYMPH NODE MESENTERIC.

## Page73 of 97 PATHOLOGY REPORT Testsubstance: C.I. Pigment Blue61 Notox No.: 488231 Testsystem: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments ManufacturersAssociation Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 56 SEX: Female DOSE LEVEL: 2
MACROSCOPIC FINDINGS days of treatment :41 sacrifice group terminal sacrifice necropsy status :planned terminal date of start of treatment; 

### 17/06/08 date of end of treatment: 

### 27/07/08 date of necropsy 

### 28/07/08
MANDIBULAR both aides DISCOLOURATION DARK RED

LYMPH NODES
MESENT. LYMPH DISCOLOURATION BLUISH

NODE
PANCREAS DISCOLOURATION REDDISH
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE sinus histiocytosis (slight) MESENTERIC
MANDIBUL.AR congestion present (correlates to GROSS finding)

LYMPH NODE
PANCREAS no lesion to account for gross finding Is)

### Page 74 of97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue61 Notox No.:488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.! 57 SEX: Female DOSE LEVEL: 2 MACROSCOPIC FINDINGS days of treatment 43 sacrifice group terminal sacrifice necropsy status planned terminal date of start of treatment; 17/06/0a date of end of treatment: 

### 29/07/08 date of necropsy :30/07/08
No abnormality detected
MICROSCOPIC FINDINGS No abnormality detected
Mo abnormalities found in: LYMPH NODE MESENTERIC.

### Page 75 of 97 PATHOLOGY REPORT Test substance: C.?. Pigment Blue61 Notox No.:488231 Test system: 28-DayOral ToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal 

### No.: 58 SEX: Female DOSE LEVEL: 2
MACROSCOPIC FINDINGS days of treatment 43 eacrifice group :terminal sacrifice necropsy status planned terminal date of start of treatment: 

### 17/06/08 date of end of treatment: 29/07/08 date of necropay 30/07/08
No abnormality detected
MICROSCOPIC FINDINGS No abnormality detected
No abnormalities found in: LYMPH NODE MESENTERIC.

### Page 76 of97 PATHOLOGY REPORT Test substance: 0.1. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study in the Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 59 SEX: Female DOSE LEVEL: 2 MACROSCOPIC FINDINGS days of treatment :43 sacrifice group terminal sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of end of treatment: 29/07/08 date of necropsy :30/07/08 No abnormality detected
MICROSCOPIC FINDINGS No tissues exami.ned

### Page 77 of97 PATHOLOGY REPORT Test substance: C.l. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicityStudy Inthe Rat 24-Oct-08 Sponsor~Color Pigments ManufacturersAssociation Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. 60 SEX: Female DOSE LEVEL: 2
MACROSCOPIC FINDINGS days of treatment :43 sacrifice group terminal sacrifice necropsy status planned terminal date of start of treatment: 27/06/08 date of end of treatment; 29/07/08 date of necropsy :30/07/08
No abnormality detected
MICROSCOPIC FINDINGS
No tissues examined

## Page 78of97 PATHOLOGY REPORT Test substance: 

## ClI. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments ManufacturersAssociation Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.; 61 SEX: Female DOSE LEVEL: 3
MACROSCOPIC FINDINGS days of treatment 41 sacrifice group terminal aacrif ice necropay status planned terminal date of 

### start of treatment: 17/06/08 date of end of treatment: 27/07/08 date of necropsy :28/07/08
MESENT. LYMPH DISCOLOURATION BLUISH NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding 

### Is) MESENTERIC

## Page 79 of97 PATHOLOGY REPORT Testsubstance: C.I. Pigment Blue 61 Notox No.:488231 Testsystem: 28-Day OralToxicityStudy intheRat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. : 62 SEX: Female DOSE LEVEL: 3
MACROSCOPIC FINDINGS days of treatment 44 sacrifice group terminal sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of and of treatment; 30/07/08 date of necropsy :31/07/08
No abnormality detected
MICROSCOPIC FINDINGS No tissues examined

### Page 80 of97 PATHOLOGY REPORT Test substance: C.t. Pigment Blue 81 Notox No.: 488231 Test system: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: ColorPigments Manufacturers Association Inc.

TABLE 3 - INDIVIDUAL ANIMAL DATA

Animal No.: 63 SEX: Female DOSE LEVEL: 3
MACROSCOPIC FINDINGS days of treatment :41 sacrifice group :terminal sacrifice necropsy statues planned terminal date of start of treatment: 17/06/08 date of end of treatment: 27/07/08 date of necropsy :28/07/08
MESENT. LYMPH DISCOLOURATION BLUISH NODE
Ho abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding Is) MESENTERIC

## Page 81 of97 PATHOLOGY REPORT Testsubstance: ClI. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Studyinthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.


## TABLE 3

- 

### INDIVIDUAL ANIMAL 

## DATA

Animal No.: 64 

### SEX: Female DOSE LEVEL: 3 MACROSCOPIC FINDINGS days of treatment 41 sacrif ice group :terminal sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of end of treatment: 27/07/08 date of necropsy : 28/07/08 MESENT. LYMPH DISCOLOURATION BLUJISH NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE macrophage aggregates (minimal) MESENTERIC no lesion to account for gross finding Is)

### Page 82 of97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 65 SEX: Female DOSE LEVEL: 3
MACROSCOPIC FINDINGS days of treatment :41 sacrifice group :terminal sacrifice necropsy status planned terminal date of start of treatment: 17/06/08 date of end of treatment: 

### 27/07/08 date of necropsy 28/07/08
MESENT. LYMPH DISCOLOURATION BLUISH

NODE No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding(s) MESENTERIC

### Page 83 

## of 97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue 61 Notox No.:488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. 66 SEX: Female DOSE LEVEL: 3
MACROSCOPIC FINDINGS days 

### of treatment 42 sacrifice group terminal sacrifice necropsy status planned terminal date of 

### start of treatment: 17/06/08 date of end of treatment: 28/07/08 date of necropsy :2 9/07/08
No abnormality detected MICROSCOPIC FINDINGS VAGINA metoestrus present
No abnormalities found in: CERVIX, CLITORAL GLAND, OVARIES, UTERUS.

## Page 84 of97 PATHOLOGY REPORT Testsubstance:C.I. Pigment Blue61 Notox No.: 488231 Testsystem: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3 - INDIVIDUAL ANIMAL DATA

Animal No.: 67 SEX: Female DOSE LEVEL: 3
MACROSCOPIC FINDINGS days of treatment 43 sacrifice group terminal sacrifice necropsy status planned terminal date of start of treatmenti 17/06/08 data of end of treatment: 

### 29/07/08 date of necropsy :30/07/08 MESENT. LYMPH DISCOLOURATION BLUISH

NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding ls) MESENTERIC

### Page 85 of 97 PATHOLOGY REPORT Test substance;

## C.I. Pigment Blue 61 Notox No.:488231 Testsystem: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. 68 SEX! Female DOSE LEVEL: 3
MACROSCOPIC FINDINGS days of treatment :43 sacrifice group terminal sacrifice necropay atatus planned terminal date of start of treatment: 17/06/08 date of end of treatment; 29/07/08 date of necropsy :30/07/08
MESENT. LYMPH DISCOLOURATION BLUISH

NODE No abnormalities were found in any of the other tissues examined

MICROSCOPIC FINDINGS LYMPH NODE sinus histiocytosia (minimal) MESENTERIC no lesion to account for gross finding Is)

### Page 86 

## of 97 PATHOLOGY REPORT Test substance: C.LPigment Blue 61 Notox No.:488231 Test system: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.! 69 SEX: Female DOSE LEVEL: 3 MACROSCOPIC FINDINGS days of treatment 43 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 29/07/08 date of necropsy 30/07/08 MESENT. LYMPH DISCOLOURATION BLUISH

NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross findingls) MESENTERIC

## Page87 of 97 PATHOLOGY REPORT Testsubstance:

## C.I. Pigment Blue61 NotoxNo.: 488231 Testsystem: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 70 SEX: 

### Female DOSE LEVEL: 3
MACROSCOPIC FINDINGS days of treatment 44 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 

### 17/06/08 date of end of treatment: 30/07/08 date of necropsy :31/07/08 MESENT. LYMPH DISCOLOU1RATION BLUISH

NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding Is) MESENTERIC

### Page 88 of

## 97 PATHOLOGY REPORT Test substance: 

## ClI. Pigment Blue 81 Notox No.: 488231 Test system: 25-Day Oral ToxicityStudy inthe Rat 24-Oct-OS Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.; 71 SEX: Female DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment :45 sacrifice group :terminal sacrifice necropey status :planned terminal date of start of treatment: 

### 17/06/08 date of end of treatment: 31/07/08 date of necropsy :01/08/08 GENERAL GI-TRACT, CONTENTS BLUISH OBSERVATIONS MESENT. LYMPH DISCOLOURATION BLUISH

NODE SKIN FORELEG: right side ALOPECIA HINDLEG: left side ALOPECIA FORELEG: left side ALOPECIA HINDLEG: right side ALOPECIA
No abnormalities were found in any of the other tissues examined MICROSCOPIC FINDINGS LYMPH NODE no lesion to account for gross finding Is) MESENTERIC SKIN foreleg right side: epidermal hyperplasia (minimal) (correlates to GROSS finding) foreleg left side: epidermal hyperplasia (minimal) (correlates to GROSS

finding)

## Page 89 of97 PATHOLOGY REPORT Test substance:0.1. Pigment Blue61 Notox No.:488231 Test system: 28-Day Oral Toxicity Study in the Rat 24-Oct-08 Sponsor: Color Pigments ManufacturersAssociation Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 72 SEX: Female DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment :42 sacrifice group :terminal sacrif ice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 28/07/08 date of necropay :29/07/08 GENERAL GI-TRACT, CONTENTS BLUISH

OBSERVATIONS No abnormalities were found in any of the other tissues examined MICROSCOPIC FINDINGS VAGINA mtoestrue preaent No abnormalities found in: CERVIX, CLITORAL GLAND, OVARIES, UTERUS.

### Page 90 of97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue 61 Notox No.:488231 Test system: 28-Day OralToxicity Study inthe Rat 24-Oct-OS Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 73 SEX: Female DOSE LEVEL: 4 

### MACROSCOPIC FINDINGS days of treatment :43 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 29/07/08 date of necropsy :30/07/08 GENERAL 01 TRACT, CONTENTS BLUISH OBSERVATIONS
MESENT. LYMPH DISCOLOURATION BLUISH

NODE No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
JEJUNUM blue material in lumen present (GENERAL OBSERVATIONS) (correlates to GROSS finding)
LYMPH NODE no lesion to account for gross finding Is) MESENTERIC
MANDIBULAR plasma cell infiltration (minimal)

LYMPH NODE VAGINA dioestrus present
No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, CERVIX, CLITORAL

GLAND, COLON, DUODENUM, HEART, ILEUM, KIDNEYS, LIVER, LUNG, OESOPHAGUS, OVARIES, PANCREAS, PARATNYROID, PEYERS PATCHES, PITUITARY, RECTUM, SCIATIC HERVE, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER, UTERUS.

## Page91 of97 PATHOLOGY REPORT Test substance: 

## C.l. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 74 SEX: Female DOSE LEVEL: 4 MACROSCOPIC FINDINGS daya of treataent :41 sacrifice group i terminal sacrifice necropay status :planned terminal date of atart of treatment: 17/06/08 date of end of treatment; 27/07/08 date of necropay 28/07/08 ADRENAL GLANDS both aides DISCOLOURATION RED-BROWN GENERAL GI TRACT, CONTENTS BLUISH

OBSERVATIONS
MANDIBULAR both sides DISCOLOURATION REDDISH

LYMPH NODES MESENT. LYMPH DISCOLOURATION BLUISH NODE Ho abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS

ADRENAL no lesion to account for gross finding~s)

HEART 

### epicardial fibrosis (minimal)
ILEUM 

## blue material in lumen present (GENERAL OBSERVATIONS) (correlates to GROSS finding) LYMPH NODE no lesion to account for gross finding (s) MESENTERIC MANDIBULAR congestion present (Correlates to GROSS finding)

LYMPH NUDE RECTUM blue material in lumen present (GENERAL OBSERVATIONS) (correlates to GROSS finding)
SPLEEN extramedullary haematopoiesis (slight)
THYMUS involution/atrophy (minimal)
VAGINA dioeatrus present epithelial vacuolation (alight)
No abnormalities found in: AORTA, BRAIN, CAECUM, CERVIX, CLITORAL GLAND,

COLON, DUODENUM, JEJUNUM, KIDNEYS, LIVER, LUNG, OESOPHAGUS, OVARIES, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, SCIATIC NERVE, SPINAL CORD, STERNUM, STOMACH, THYROID, TRACHEA, URINARY BLADDER, UTERUS.

## Page 92 of97 PATHOLOGY REPORT Test substance: 0.1. Pigment Blue61 Notox No.:488231 Test system: 28-DayOral Toxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 75 SEX: Female DOSE LEVEL: 4 MACROSCOPIC FINDINGS days of treatment 41 sacrifice group :terminal secrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 

### 27/07/08 date of necropsy :28/07/08 MESENT. LYMPH DISCOLOURATION BLUISH

NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
KIDNEYS basophilic tubules (minimal)
LYMPH MODE no lesion to account for gross finding Is) MESENTERIC
MANDIBULAR plasma cell infiltration (slight)

LYMPH NODE
VAGINA dioestrus present epithelial vacuolation (slight)
No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, CERVIX, CLITORAL

GLAND, COLON, DUODENUM, HEART, ILEUM, JEJUNUM, LIVER, LUNG, OESOPHAGUS, OVARIES, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, RECTUM, SCIATIC NERVE, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER, UTERUS.

## Page 93 of97 PATHOLOGY REPORT Testsubstance: 0.1. 

## Pigment Blue61 NotoxNo.: 488231 Testsystem: 28-Day OralToxicityStudy In the Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 76 SEX: Female DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment :42 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 28/07/08 date of necropsy :29/07/08 GENERAL 01 -TRACT, CONTENTS BLUISH

OBSERVATIONS MESENT. LYMPH DISCOLOURATION BLUISH

NODE No abnormalities were found in any of the other tissues examined MICROSCOPIC FINDINGS COLON blue material in lumen present (GENERAL OBSERVATIONS) (correlates to GROSS finding) LYMPH NODS no lesion to account for gross finding Is) MESENTERIC
MANDIBULAR plasma cell infiltration (minimal)

LYMPH NODE
VAGINA dioestrus present epithelial vacuclation (slight)
Ho abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, CERVIX, CLITORAL GLAND, DUODENUM, HEART, ILEUN, JEJUNUM, KIDNEYS, LIVER, LUNG, OESOPHAGUS, OVARIES, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, RECTUM, SCIATIC NERVE, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER, UTERUS.

### Page 94 of97 PATHOLOGY REPORT Testsubstance: 

## 0.1. Pigment Blue61 Notox No.: 488231 Testsystem: 28-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No. : 77 SEX: Female DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment :41 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 27/07/08 date of necropay :28/07/08 MESENT. LYMPH DISCOLOURATION BLUISH

NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS KIDNEYS tubular mineralization (minimal) LUNG perivescular inflammatory cell infiltration (minimal) alveolar macrophage infiltration (minimal) LYMPH NODE no lesion to account for gross finding(s) MESENTERIC VAGINA dioestrus present epithelial vacuolation (minimal)
No abnormalities found in: ADRENAL, AORTA, BRAIN, CARCUN, CERVIX, CLITORAL

GLAND, COLON, DUODENUM, HEART, ILEUM, JEJUNUM, LIVER, MANDIBULAR LYMPH NODE, OESOPHAGUS, OVARIES, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, RECTUM, SCIATIC NERVE, SPINAL CORD, SPLEEN, STERNUM, STOMACH, 'THYMUS, THYROID, TRACHEA, URINARY BLADDER, UTERUS.

### Page 95 of 97 PATHOLOGY REPORT Test substance: C.LPigment Blue81 Notox No.:488231 Testsystem: 28-Day OralToxicity Study inthe Rat 24-Oct-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3
- INDIVIDUAL ANIMAL DATA

Animal No.: 78 SEX: Female DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment :45 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 31/07/08 date of necropsy :01/08/08
GENERAL GE-TRACT, CONTENTS BLUISH

OBSERVATIONS MESENT. LYMPH DISCOLOURATION BLUISH

NODE No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding ls) MESENTERIC

### Page 96 of97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue 61 Notox No.:488231 Test system: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor: ColorPigments ManufacturersAssociation Inc.

TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 79 SEX: Female DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment :44 aacrifice group :terminal sacrifice necropsy atatus planned terminal date of atart of treatment: 17/06/08 date of end of treatment: 30/07/08 date of necropay :31/07/08 MESENT. LYMPH DISCOLOURATION BLUISH

NODE
No abnormalities were found in any of the other tiasues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding 1s) MESENTERIC

## Page 97 of97 PATHOLOGY REPORT Testsubstance: C.I. Pigment Blue 61 Notox No.: 488231 Testsystem: 28-Day OralToxicityStudy inthe Rat 24-Oct-08 Sponsor:Color PigmentsManufacturers Association Inc.
TABLE 3
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### INDIVIDUAL ANIMAL DATA

Animal No.: 80 SEX: Female DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment :44 sacrifice group :terminal sacrifice necropay status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 30/07/Ga date of necropsy 31/07/0a
GENERAL GI TRACT, CONTENTS BLUISH OBSERVATIONS MESENT. LYMPH DISCOLOURATION BLUISH

NODE No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding is) MESENTERIC

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 5

APPENDIX 5 SUMMARY DOSE RANGE FINDING STUDY

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# C. PIGMENT BLUE61 NOTOX Project488231 APPENDIX 5 SUMMARY OF DOSE RANGE FINDING STUDY Inorder to set thedose levelsfor the main study (NOTOX Project488231), apilot studywas performed. Two groups of3females (10 

# weeks old) weredosed at500 or 1000 

# mg/kg bodyweight/day (Groups 1and 2)for 5days by oral gavagewith adosevolume of5mI/kg bodyweight. Dose levels were basedonthree non-GLP acuteoral studies inwhich the LD50valueswere>15,000 mg/kg (SPF Wistar rats), >15,000 mg/kg (SPF Wistar rats) and >5,000 mg/kg (Sprague Dawley rats). Ina non-GLP repeat dose study, mixed raceAlbino rats were exposed for 14 days at500 mg/kg: no toxicitywas noted. Allanimals were housed 3percage, identified by ear-and tailmark and allocation was performed at random. Ifnot mentioned otherwise, testsystem and techniques were identical to those used during the main study. Datawas collected under NOTOX Project 488232. Planning Delivery: 28 May 2008 Treatment: 02 to 06 June 2008 Terminal procedures: 06 June 2008 Animal husbandry Animals were housed inacontrolled environment, inwhich optimal conditionswere considered to be approximately 15 air changes per hour, atemperature of21 ±

## 3 0

## C(actual range: 19.8- 2 1.1*0), a relative humidity of30- 70% (actual range: 50

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## 81%)and 12 hours artificial lightand 12 hours darkness perday. Cleaning procedures inthe room might havecaused the temporary fluctuationsabovethe optimal levelof70% for relative humidity. Based on laboratory historical datathese conditionswere considered notto have affected the study integrity. Chemical analysis Notperformed during the pilot study. Observations Clinicalsigns: At least once daily. Mortality: At least twice daily. Bodyweights: Daily. Food consumption: Daily. Pathology Necropsy: On Day5 Noorganswere fixed. Terminal body weight, organ weights: kidney and liver. 

# RESULTS No mortality wasobserved inthe pilotstudy, andthere were nodifferences inbodyweights (absolute orrelative), food consumption (absoluteor relative), ororgan weights (absolute or relative). For animals treated with 500and 1000 mg/kg of 0.1. Pigment Blue 61, bluecoloration ofthefaeces and bluish contents inthe GI tract were noted. 

# CONCLUSION There were notoxicologically-significant results fortreatment upto 1000 mg/kg; based on the results ofthe pilotstudythe doses selected forthe main study are: 50, 250, and 1000 mg/kg.

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