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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

## removal ofCBIclaimsregarding identified studies) 

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

## while there is awaiverofconfidentialbusinessinformation (CBI)claimsin the identified studiesthere is aanassertion ofcopyrightprotections and some additional textthat providesin part
... "Reproduction,reference,summarization,retransmissionorotheruse ofthis copyrighteddocument.

..includingelectronictransmission,usingthe internetoremail

....is strictlyprohibited

...." Whileweunderstandthebasisforthecopyrightprotections, we have a concernthatthelanguage in the fax mightbeinterpretedto meanthatePMA isassertingthatthestudies submittedperthe TSCAactivityarenottobemadeavailableinan Agencye-docket (electronic docket) orbe includedin EPA websitesamongthecollectionofTSCAhealthand safety studies. Wearethinkingthat the identifiedlimitations noted inthefax arenotintendedto applyto EPA's normal proceduresforthehandlingofnon-CBI materials submittedby entities subjectto TSCA regulations(includingthepostingofsuchmaterials ongovernmentwebsites) and submittedby suchentitiesinorderto complywithaTSCArequirement, but are rather intendedto limit entitiesoutsidethe government from the unauthorizeduseorappropriation ofthe studies. Canyou pleaseconfirm CPMA'sintent? Manythanks.

ScottM. Sherlock,AttorneyAdvisor Environmental AssistanceDivision OfficeofPollutionPrevention andToxics

## Confidential Business Report 

# REPORT

Study Title 

# 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 M.E.W. Beekhuijzen, M.Sc. Study comletion date

08 December 2008
Test Facility NOTOX B.V. Hambakenwetering 7 5231 DD 's-Hertogenbosch The Netherlands Laboratory Study Identification NOTOX Project 488231 (488232) NOTOX Substance 183591 -Page 1of249- 

# TSCA CONFIDENTIAL SuSINESS INFORMATION DOES NOT CONTAIN NATIOASECUiRITY INFORMATION (E-0- '2%65)

## C.I. PIGMENT BLUE 61 NOTOX Project488231
1. CONTENTS 

## 1 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. Study schedule.............................................................................. 8 5.3. Project numbers............................................................................. 8 5.4. Aimsofstudy................................................................................ 8 5.5. Guidelines ................................................................................... 9 5.6. Storage and retention ofrecords and materials......................................... 9 6. MATERIALS AND METHODS.................................................................. 10 6.1. Testsubstance............................................................................. 10 6.

1.1. Test substance information......................................................... 10 6.1.2. Study specifictestsubstance information......................................... 10 6.1.3. Testsubstanceformulation......................................................... 10 6.1.4. Chemical analysisof dose preparations.......................................... J1 6.2. Testsystem................................................................................ 1 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. Organweights....................................................................... 19 6.8.4. Histotechnology..................................................................... 19 6.8.5. 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. Listof protocol deviations....................................................... 21 6.13.2. Listofstandard operating procedure deviations............................... 21 7. RESULTS...................................................................................... 22 7.1. Analysis ofDose Preparations (see also Appendix 3)................................... 22 7.2. Observations............................................................................... 22 7.2.1. Mortality ............................................................................. 22 7.2.2. Clinical Signs........................................................................ 22 7.2.3. BodyWeights....................................................................... 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. Organ Weights
.......................... :.......................................... 24

Page 2

## C.I. PIGMENT BLUE61 N OTOX Project 488231 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 I SUMMARY TABLES............................................................... 1-27 ClinicalSigns...................................................................... 2 BodyWeight....................................................................... 5 BodyWeightGain................................................................. 7 Food Consumption.............................................................. -9 Relative Food Consumption .................................................... 11 Functional Observations........................................................ 13 Motor Activitytest ............................................................... 14 Haemnatology..................................................................... 15 Clinical Biochemistry............................................................ 17 Macroscopic Findings........................................................... 19 Organ weights/Organ to Body Weight Ratios................................ 20 Reproduction Processes........................................................ 24 Mating Performance ............................................................ 25 Breeding Data per Group....................................................... 26 Mean BodyWeights ofPups ................................................... 27 APPENDIX 2 INDIVIDUAL TABLES.............................................................. 1-85 Mortality data...................................................................... 2 Clinical Signs...................................................................... 4 BodyWeight...................................................................... 12 BodyWeight Gain............................................................... 15 Food Consumption .............................................................. 18 Relative Food Consumption .................................................... 20 Functional Observations........................................................ 22 MotorActivitytest ............................................................... 24 Haemnatology..................................................................... 26 Clinical Biochemistry............................................................ 31 Macroscopic Findings........................................................... 35 Organweights/Organto Body Weight Ratios................................ 38 Breeding Data per Litter........................................................ 46 Mean BodyWeightsof 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 5 SUMMARY DOSE RANGE FINDING .............................................. 1-2
TOTAL NUMBER OF PAGES...................................................................... 249

Page 3

## C.l. 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 and was conducted incompliance with: The Organization forEconomic Cooperation and Development (OECD) Good Laboratory Practice Guidelines(1997). Which essentially conform to: The United States Food and Drug Administration Good Laboratory Practice Regulations. The United States Environmental Protection Agency Good Laboratory Practice Regulations. The sponsor isresponsible forGood Laboratory Practice (GLP) compliance foralltest substance information unlessdetermined by NOTOX The GLP 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: .... b0CC...

Page4

## ClI. PIGMENT BLUE 61 NOTOX Project488231 3. LEAD QUALITY ASSURANCE STATEMENT NOTOX B.V., 's-Hertogenbosch, The Netherlands This reportwas inspected by the NOTOX QualityAssurance Unit to confirmthat the methods and results accurately and completely reflect the raw data.The datesof QualityAssurance inspections are given below. Duringthe on-site process inspections procedures applicable to thistype ofstudywere inspected. The reporting date isthe date of reporting tothe Study Director. The QAU reportwas then forwarded to theTest Facility Management.

Type of haeroesStart End Reporting inspections PaerossInspection Inspection date date date Study Protocol 29-May-08 30-May-08 30-May-08 Amendment 1of Protocol 03-Jun-08 03-Jun-08 03-Jun-08 Amendment 2of Protocol 17-Jun-08 17-Jun-08 17-Jun-08 Amendment 3of Protocol 02-Jul-08 02-Jul-08 02-Jul-08 Amendment 4of Protocol 05-Aug-08 05-Aug-08 05-Aug-08 Report 19-Nov-08 24-Nov-08 24-Nov-08 Process Test substance unit 08-Apr-08 10-Apr-08 14-Apr-08 Testsubstance handling 21-Jul-08 23-Jul-08 28-Jul-08 SPF unit 11-Mar-08 17-Mar-08 17-Mar-08 16-Jun-08 19-Jun-08 19-Jun-08 Test substance handling Exposure Observations/Measurements Specimen handling Clinical pathology unit 13-May-08 16-May-08 19-May-08 Observations/Measurements Specimen handling Histology unit 03-Jun-08 10O-Jun-08 23-Jun-08 Specimen handling Pathology unit 30-Jun-08 04-Jul-08 07-Jul-08 Observations/Measurements Analytical and physical chemistry 19-May-08 27-May-08 03-Jun-08 Testsubstance handling Observations/Measurements The QualityAssurance programme for thedelegated phase histopathology was performed by the Quality Assurance appointed by thetest site management and aQualityAssurance statement isincluded inAppendix 4ofthis report. Head ofQuality Assurance ,C.J. Mitchell B.Sc.

Dat.
.......
.... Page5

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

## Combined repeated dose toxicitystudywith reproduction /developmental toxicityscreening test with ClI. Pigment Blue 61 administered byoral gavage inWistar rats. The purpose ofthisstudy was to evaluatethe potential toxic effectsof C.l. Pigment Blue 61 when administered to rats foraminimum of28 daysand toevaluate the potential of thetest substance toaffect male and female reproductive performance such asgonadal function, mating behaviour, conception, parturition and early postnatal development. The study was based on the following guideline: Organisation of Economic Co-operation and Development Guidelines (OECD) fortesting of Chemicals Guideline 422, Combined Repeated DoseToxicity Studywith the Reproduction/Developmental Toxicity Screening Test, March 1996. After acclimatisation, four groups often male andten femaleWistar Han ratswere exposed by oral gavageto the testsubstanceat 0,50, 250 and 1000mg/kg/day. Males were exposed for 30 days, i.e. 2weeks priorto mating, during mating, and upto termination. Females were exposed for41 to 45days, i.e.during 2weeks priorto mating, during mating, during post-coitum, and during at least4daysoflactation. The following parameters were evaluated: clinical signs,functional observations, bodyweights, food consumption, reproduction processes, observations offspring, clinical pathology, macroscopy, organ weights, and histopathology. Chemical analyses offormulations were conducted once duringthe study toassess accuracy, homogeneity and stability. RESULTS 

## Accuracy, homogeneity and stabilityofformulations were demonstrated by analyses. Notreatment-related toxicologically significantchanges were noted inanyof the parameters investigated inthisstudy (i.e. mortality, clinical appearance, functional observations,body weight, food consumption, clinical laboratory investigations, macroscopicexamination, organ weights, microscopicexamination, reproduction, breeding and pup development). CONCLUSION 

## From the results presented inthis report, the parental, reproduction, breeding and 

# 0 developmental NoObserved Adverse Effect Level (NOAEL) forC.I. PIGMENT BLUE61 ofat least 1000 mg/kg/day wasestablished.

Page 6

## C.l. PIGMENT BLUE 61 NOTOX Project488231 5. INTRODUCTION 5.1. Preface This isamulti-site study. Thestudy was performed according tothe OECD Consensus document Number 13 The Application ofthe OECD Principles ofGLPtothe Organization and Management ofMulti-Site studies. Information concerning the delegated phase isgiven inthe applicable appendix. Phase: Histopathology Principal Investigator: Dr. A.Pilling,ToxPath Consultancy Ltd, UK (seeAppendix4) Sponsor Color Pigments Manufacturers Association, Inc. 300 N.Washington St. Ste. 102, ALEXANDRIA, VA 22314 USA Study Monitor Mr. L.Robinson jir@cpma.com Study Director M.E.W. BeekhuiJzen, M.Sc. NOTOX B. Hambakenwetering 7 's-Hertogenbosch The Netherlands Tel: +31.73 640 6700 Fax: +31.73 6406799 Email: manon.beekhuilzen~cDnotox.ni Coordinating Biotechnician M.M.A. Rijkers (NOTOX B.V.) Clinical Pathology J.E. van Kesteren (NOTOX BYV) Analytical Chemistry Ir.M.J.C. Brekelmans (Principal Scientist, NOTOX B.V.) Histotechnology IngT.A. Mijnders; (NOTOX B.V.) Necropsy J.H.van den Brink, DVM (NOTOX B.V.) Lead QA C.J. Mitchell, B.Sc. (NOTOX B.V.): christine.mitchell(¬ox.nI Testfacility Management H.H. Emmen, M.Sc. (NOTOX B.V.): harry.emmen(~notox.nI Test Facility NOTOX B.V. Hambakenwetering 7 5231 DD 's-Hertogenbosch The Netherlands

Page7

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 5.2. Studyschedule Delivery ofanimals 11 June 2008 Experimental start date 12June 2008 (allocation) Starttreatment 17June 2008 Start mating 01 July2008 Necropsyof males 17July2008 Delivery oflitters 23 to27 July2008 Necropsy offemales and pups 28 Julyto 01 August 2008 Experimental completion date 01 August2008 (end in-life) 5.3. Project numbers Two project numberswere usedto collectonlinedata. Eventually, all datawere reported under NOTOX Project 488231 Project number Online data 488236 Parental animals: Clinical signs Functional observations Body weights and food consumption pre-mating Body weights mating period Clinical laboratory investigations Macroscopic findings and organ weights 488231 Allother data 5.4. Aims ofstudy The purpose ofthisstudy wasto evaluatethe potential toxic effectsofthe testsubstancewhen administered to ratsfor aminimum of28 daysand to evaluate the potential ofthetest 

# 0 substancetoaffect male and female reproductive performance such asgonadal function, mating behaviour,conception, parturition and early postnatal development. ANoObserved Adverse Effect Level (NOAEL) was evaluated. This study should provide partof arational basisfortoxicological risk assessment inman.

Page 8

## C.I. PIGMENT BLUE 61 NOTOX Project488231 5.5. Guidelines The protocol was reviewed and agreed bythe Laboratory AnimalWelfare Officer and the Ethical Committee of NOTOX (DEC NOTOX 07-13) as required by the Dutch Acton Animal Experimentation (February 1997). The study procedures described inthis report were based on thefollowing guideline: 1)Organisation of Economic Co-operation and Development Guidelines(OECD) fortesting of Chemicals Guideline 422, Combined Repeated DoseToxicity Studywith the Reproduction/Developmental Toxicity Screening Test, March 1996. 5.6. Storage and retention of records and materials Records and materials pertaining to the study, including protocol, raw data, specimens (except specimens requiring refrigeration orfreezing) and thefinal report, will be retained inthe NOTOX archives for aperiod ofat least2years afterfinalization ofthe report. After this period, the sponsorwill be contacted todetermine whether rawdata and specimens should be returned to them, retained or destroyed on their behalf. Those specimens requiring refrigeration orfreezing will be retained byNOTOX for as long as the qualityofthe specimens permits evaluation but no longerthan three months afterfinalization ofthe report. NOTOX will retain atestsubstance sample until theexpiry date, but no longerthan 10years afterfinalization ofthe report. Afterthis period thesample will be destroyed. Storage and retention of recordsand materials by thetest sites forthe delegated phasesare described inthe applicable appendices.

Page 9

## ClI. PIGMENT BLUE 61 NOTOX Project 488231 6. MATERIALS AND METHODS 6.1. Test substance 6.1.1. Test substance information Identification C.l. Pigment Blue61 Structure

HO 3 

# Molecularformula C3 7

### H 2 9

### N3

### 0 3

### S S3 

## Molecularweight 596 GAS Number 1324-76-1 Description Dark blue powderwith granules Batch 2084701 Composition 96.830% Testsubstance storage At room temperature inthe dark Stability under storage conditions Stable Expiry date 02January 2018 6.1.2. Studyspecifictestsubstance information pH 4.3 atconcentration of>1% Solubilityinvehicle: *Propylene glycol Yes, <1% 6.1.3. Testsubstance formulation Vehicle Propylene glycol, specificgravity 1.036 (Merck, Darmstadt, Germany). Rationale forvehicle Based ontrial formulations performed at NOTOX Stability oftestsubstance invehicle Formulations at 10 and 200 mg/mI were stable when stored at room temperature for at least6hours (determined during this project; see Appendix 3). Method offormulation Formulations (w/w) were prepared dailywithin 6hours priorto dosingand were homogenisedto avisuallyacceptable level. Adjustment was made for specificgravity ofthe vehicle (1.036). Storage conditions At ambient temperature.

Page 10

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 6.1.4. Chemical analysis ofdose preparations Analyses were performed on asingle occasion during thetreatment phase (30 June2008) according to avalidated method (NOTOX Project 488235) asspecified below: Group Analysis (type ofsample)
1 acc(M) 2 acc +

## hom +

## stabt,, 0 

## (TMB), stabl,6

## . RT 

### (S) 

## 3 acc(M) 4 acc +

## hom +

## stabt~,o(TMB), Stabt=, RT 

### (S) Duplicate samples were analysed acc--accuracy, hom=homogeneity, stab=stability (hours), T-lop, Mmiddle, B=bottom position ofcontainer S=stabilitysample taken at middle position ofcontainer RT=room temperature 

## The accuracy ofpreparation wasconsidered acceptable ifthe mean measured concentrations were90-1 10% ofthe targetconcentration for solutions,or 85-115% for suspensions. Homogeneitywas demonstrated ifthecoefficient ofvariation was :5 10%. Formulations were considered stable ifthe relativedifference before and after storagewas maximally 10%. 6.2. Test system Test System Rat: Crl:WI(Han) (outbred, SPF-Quality). Untreated animals and virgin femaleswere used at initiationofthe study. Rationale Thisspecies and strain of rathas been recognized as appropriate forgeneral and reproductive toxicitystudies. NOTOX BV has general and reproductive historical data inthis speciesfrom the same strain and source. This animal model has been proven to be susceptible tothe effects of reproductive toxicants. Source Charles River Deutschland, Sulzfeld, Germany. Age at start F 0

## -treatment Approximately 10weeks. Numberof F 0

## -animals 40females and 40 males. Acclimatisation F 0 

## At least5days priortostart oftreatment. Health check F 0 

## Ahealth inspection was performed priorto commencement of treatment to ensure thatthe animalswere inagood state of health. Randomisation F 0 

## Prior to commencement oftreatment, bycomputer-generated random algorithm according to bodyweight, with all animals within ±20% 

## ofthe sexmean. Identification F 0 

## Earmark and tattoo.

Page 11

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 Breeding procedures Following aminimum of 14 days of exposureforthe males and females, one female was cohabitated withone male ofthesame treatmentgroup, avoiding sibling mating (Charles Riversupplied non-littermates). Detection of mating wasconfirmed by evidence ofsperm inthevaginal lavageor bythe appearance of an intravaginal copulatory plug. This daywas designated Day0postcoitumn. Once mating had occurred, the males andfemales were separated. Amaximum of14 dayswas allowed for mating. Parturition F 0 

## Thefemales were allowed to litter normally. Day 1oflactationwas defined asthedaywhen alitterwas found completed (i.e. membranes, placentas cleaned up, nest build upand/or feeding of pupsstarted). Females thatwere litteringwere leftundisturbed. Lactation F 0 

## Deficiencies inmaternal care, such as inadequate construction or cleaning ofthe nest, pupsleftscattered and cold, physicalabuse of pups or apparently inadequate lactation orfeeding, were recorded. Number ofpups Approximately 480 pups (40 liters x12 pups). Identification offspring On Day 1oflactation, all pupswere individually identified by means of intracutaneous injection of Indian ink. 6.3. Allocation Group Dose level

- 

## Number of animals Animals numbers

_____-mg/kg 

### b.w./day -Fr) 

### males F 0

### females males -females 1___ 

### 0 10 10 01-10 41-50 

### 2 50 10 10

. 

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

_

*Dose 

## levelsfollowing were based on thedose rangefinding study (NOTOX Project488232; see Appendix 5). 

## 6.4. Animal husbandry * Animalswere housed inroom 16. Conditions Animals were housed inacontrolled environment, inwhich optimal conditionswere considered to be approximately 15 airchanges per hour, atemperature of21 ±

## 3*C (actual range: 19.8- 22.40C), 

## arelative humidity of 30

- 

## 70% (actual range: 41

-

## 87%) and 12 hoursartificial lightand 12 hours darkness perday.Temporary fluctuations from the light/dark cycle (with amaximum of 1hour) occurred due to performance offunctional observations inthe room. Cleaning procedures inthe room might have caused thetemporary fluctuationsabovetheoptimal levelof 70% forrelative humidity. Based on laboratory historical data theseconditions were considered notto haveaffected thestudy integrity.

Page 12

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

Accommodation Pre-mating Animalswere housed ingroups of 5animals/sex/cage inMacrolon cages (MIV type, height 18 cm). Mating Females were caged togetherwith males on aone-to-one-basis inMacrolon cages (MiII type, height 18 cm). Post-mating Maleswere housed intheir home cage (Macrolon cages, MIV type, height 18 cm) with amaximum of5animals/sex/cage. Females were individually housed inMacrolon cages (MIII type, height 18 cm). Lactation Offspring was keptwith thedam until termination. General Sterilised sawdust as 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 ofanalysiswere examined and then retained inthe NOTOX archives. During overnight activity monitoring, animals were housed individually inMacrolon cages (MIII type; height 15 cm) with sterilised sawdust asbedding material. No cage-enrichment was provided during overnight activity monitoring. Diet Free accessto pelleted rodent diet(SM RIM-Z from SSNIFF® Spezialdiaten GmbH, Soest, Germany). Results of analysesfor nutrients andcontaminants ofeach batch were examined and archived. Water Free accessto tap-water. Certificates ofanalysis (performed quarterly) were examined and archived. Analysis of bedding, paper, diet andwater did not reveal anyfindingsthat were considered to have affected study integrity. 6.5. Treatment 6.5.1. Parental animals Method Oral gavage, usingaplasticfeeding tube. Formulations were placed on amagnetic stirrer during dosing. Frequency Once dailyfor7days perweek, approximately thesame time each day with amaximum of6hours difference between theearliest and latest dose. Animals were dosed up to theday priorto scheduled necropsy. Exposure period Males were exposed for 30 days, i.e. 2weeks priorto mating, during mating, and up totermination. Femaleswere exposed for 41-45 days, i.e.during 2weeks priorto mating, during mating, during post-coitum, and during atleast4days oflactation. Female 78 was notdosed for one occasion as thisfemale was littering at thetime ofdosing. Dose volume 5m~lkg bodyweight. Actual dosevolumes were calculated according to the latest body weight. 6.5.2. Offspring Offspring was not treated.

Page 13

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 6.6. Observations Thefollowing animals were selected (first5mated males and first5femaleswith live offspring) for functional observations, clinical laboratory investigations, macroscopicexamination, determination oforganweights and histopathological examination.

Group Animals numbers
S_______Males_(n=5)_ 

## Females (n=5) 1 1 2 3 7 

## I7 10 42 43 47 48 1 50 2 11 12 

## 114 115 16 52 55 56 57 158 

## 3 21 22 

# 1 23 24 25 61 1 63 64 

# I65 167 14 3 34135 

## I36 i37 73 

## I74 75 76T 77 By mistake, notthefirst 5mated males butthe first males per groupwere selectedfor macroscopic examination, determination oforgan weights and histopathological examination.

Group Animals numbers

SMales 

### (n=5) 

## 1 1 213 4 5 2 11 I12 13 14 1 

## 3 21 22 123 

# 24 

# .. 25 4 31 132 33 

# 134135 6.6.1. Parental animals Mortality/Viability At leasttwice daily. Clinical signs At leastonce daily, detailedclinicalobservations were made inall animals. Once priorto start oftreatment and atweekly intervals thiswas also performed outsidethe home cage inastandard arena. Arena observations were notperformed when the animals were mating, or housed individually. Thetime ofonset, degree and duration was recorded. All symptoms were recorded and graded according tofixedscales: Maximum grade 1: grade0=

## absent, grade 1=

## present Maximum grade 3or4: grade 1=

## slight, grade 2=

## moderate, grade 3=

## severe, grade4 =

## verysevere Cage debris of pregnantfemales were examined to detectabortion orpremature birth, ifapplicable. Signs of difficultor prolonged parturition were recorded, ifapplicable. Functional Observations Thefollowing testswere performed inthe first5mated males and thefirst 5females with live offspring,from eachgroup:

- 

## 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 for individual animals, usingacomputerised monitoring system, Pearson Technical Services, Debenham, Stowmarket, England).

Page 14

## C.1, PIGMENT BLUE 61 NOTOX Project488231 During the motor activitytest, mates werecaged individuallyand females were caged with theiroffspring. The assigned males were tested during week 4oftreatment and the assignedfemales were testedduring lactation (all before blood sampling). Inorderto avoid hypothermia ofpups, dams were removed from the pupsfor not more than 30-40 minutes. Body weights Males andfemales were weighed on thefirstday ofexposure and weeklythereafter. Mated femaleswere weighed on Days 0,4,7, 11, 14,17 and 20 ofgestation, and during lactation on Days 1and 4. Food consumption Weekly,for males and females. Food consumption was not recorded during the breeding period. Food consumption of mated femaleswas measured on Days 0,4, 7,11, 14,17 and 20 postcoitum and afterdelivery on Days 1and4post-partumn. Water consumption Subjectiveappraisal was maintained during thestudy, but no quantitative investigation introduced as noeffectwassuspected. Reproduction processes Male number paired with, mating date, confirmation of pregnancy, and delivery daywas recorded. 6.6.2. Offspring Each litterwas examined todetermine the following, ifpractically possible: Mortality /Viability The numbers oflive and dead pups atthe First Litter Check ( 

## checkat Day 1of lactation) and dailythereafter weredetermined. If possible, defects or cause ofdeath were evaluated. Clinical signs At leastoncedaily, detailed clinicalobservations were made inall animals. Body weights Live pups were weighed during lactation on Days 1and 4. Sex Was determined for all pups on Days 1and 4oflactation (by assessmentofthe ano-genital distance).

Page 15

## ClI. PIGMENT BLUE 61 NOTOX Project488231 6.7. Clinical laboratory investigations Blood samples were collected from the first5mated males and thefirst 5females with live offspring from each group under iso-flurane (Abbott Laboratories Ltd., Zwolle,The Netherlands) anaesthesia immediately priorto scheduled postmortem examination, between 7.00 and 10.30 a.m. The animals were fasted overnight (with amaximum of20 hours) before blood sampling, butwaterwas provided. Blood samples weredrawn from the retro-orbitalsinus and collected intotubes (Greiner Bio-One, Bad Hailer, Austria) prepared with EDTA for haemnatological parameters (0.5 ml), with citrate forclotting tests (0.9 ml) and Li-heparin treated tubesforclinical biochemistry parameters (0.5 ml). The following parameters were determined: Parameter Abbreviation Unit Haematologya White bloodcells WBC 1109/LDifferential leucocyte count %WBC neutrophils, lymphocytes, monocytes, eosinophils, basophils 11 

## Red blood cells 1 2 

## /L Reticulocytes %RBC Red blood celldistribution width ROW% Haemoglobin mmol/L Haemnatocrit LL Mean corpuscularvolume MVCV flMean corpuscular haemnoglobin MCH fmol Mean corpuscularhaemnoglobin MCHC mmol/L concentration Platelets 10 9

## /LClottingPotentialb 

## Prothrombin time PT s Activated Partialthromboplastin time APTT s ClinicalBiochemistry c 

## Alanine aminotransferase ALAT U/L Aspartate aminotransferase ASAT U/L Alkaline phosphatase ALP U/L Total Protein g/L Albumin g/L Total Bilirubin pmol/L Urea mmol/L Creatinine pmol/L Glucose mmol/L Cholesterol mmol/L Sodium mmol/L Potassium mmol/L Chloride mmol/L Calcium mmol/L Inorganic Phosphate lnorg. Phos mmol/L

'Instrumentation: ADVIA 120 (Bayer Diagnostics). bInstrumentation: STA Compact (Roche Diagnostics). cInstrumentation: OlympusAU400 (Goffin Meyvis).



## Page 16

## ClI. PIGMENT BLUE 61 NOTOX Project 488231 6.8. Pathology Parental Animals 6.8.1. Termination All animalswere fasted overnight (with amaximum of 20 hours) priorto planned necropsy, but water was provided. Females Which Deliver On lactation Day 5,the F 0

## -females wereanaesthetised using iso-flurane (Abbott Laboratories Ltd., Zwolle, The Netherlands) and subsequently exsanguinated. Organ weights were collected andtissueswere preserved for possiblefuture histopathological examination as described in the following paragraphs. The number of former implantation siteswere counted and recorded. Corpora lutea were also counted and recorded. Gross lesionswere saved forpossible future histopathological examination. Females Which Failed to Deliver On post-mating Day 25-27 (females with evidence ofmating), the F 0

## -femaleswhich had not delivered were anaesthetised using iso-flurane (Abbott Laboratories Ltd., Zwolle, The Netherlands) and subsequently exsanguinated. No organ weights were collected. Tissueswere preserved for possible future histopathological examination as described inthe following paragraphs, withthe following exceptions: Nongravid uteri were stained using the Salewski technique (Salewski, 1964) inorderto determine any former implantation sites(Salewski staining prepared atNOTOX using AmmoniumsulIfide-solution 20% (Merck, Darmstadt, Germany) and Milli-Ro water (Millipore Corporation, Bedford, USA)). Ifevidence of macroscopic implantationswas present,the number ofimplantation sites and corpora lutea was recorded. Gross lesionswere savedfor possiblefuture histopathological examination. Termination Procedures for F 0

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

## -maleswere anaesthetised using iso-flurane (Abbott Laboratories Ltd., Zwolle, The Netherlands) and subsequently exsanguinated. Organweights werecollected and tissues were preserved for possible future histopathological examination asdescribed inthe following paragraphs.

Page 17

## C.!. PIGMENT BLUE 61 NOTOX Project488231 6.8.2. Macroscopic examination After sacrifice all parental animalswere subjected to macroscopic examination ofthe cranial, thoracic and abdominal tissues andorgans, with specialattention being paid tothe reproductive organs. Descriptions ofall macroscopic abnormalities was recorded. Samples of thefollowing tissuesand organs werecollected andfixed inneutral phosphate buffered 4%formaldehyde solution (Klinipath, Duiven, The Netherlands): From the first5males pergroup and the first5femaleswith live offspring pergroup:

Identification marks: notprocessed 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 (ifdetectable) and (Skin) Harderian gland) Spinal cord -cervical, midthoracic, lumbar (Female mammary gland area) Spleen (Femur including joint) Sternum with bone marrow Heart Stomach Ileum Testes* Jejunum Thymus Kidneys Thyroid including parathyroid (ifdetectable) (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 NOTO)( using Formaldehyde 37-40%, Ethanol,Aceticacid (glacial)(all Merck, Darmstadt, Germany) and Milli-Ro water (Millipore Corporation, Bedford, USA)) andtransferredto formalin after fixationforatleast 24 hours. Tissues/organs mentioned inparentheses were notexamined bythe pathologist,since no signs of toxicitywere noted atmacroscopicexamination.



## Page 18

## C.I. PIGMENT BLUE 61 NOTOX Project488231 6.8.3. Organ weights The following organ weights (and terminal bodyweight) was recorded: From the first 5males pergroup and thefirst 5females with liveoffspring pergroup: Adrenal glands Liver Brain Spleen Epididymides Testes Heart Thymus Kidneys From all remaining males: Epididymides Testes 6.8.4. Histotechnology All organ and tissue samples, as defined under Histopathology (following), were processed, embedded andcut atathicknessof 2-4 micrometers and stainedwith haematoxylin and eosin (Klinipath, Duiven, The Netherlands). Ofthe selected 5males/group ofthe controland high dose group, additional slides ofthe testes were prepared to examine stagingof spermatogenesis. The testeswas processed, sectioned at 3-4 microns, and stained with PAS/haematoxylin (Klinipath, Duiven, The Netherlands). 6.8.5. Histopathology The delegated phase was performed bythe Principal Investigator for Histopathology. See Appendix 4. Thefollowing slideswere examined by apathologist:

- 

## The preserved organs and tissues ofthe selectedanimals of Groups 1and 4.

- 

## The additionalslides ofthe testesofthe selected 5males/group of Groups 1and 4to examine staging ofspermatogenesis

- 

## Allgross lesions ofall animals (all dose groups)

- 

## The reproductive organs*ofall animalsthatfailed to mate, conceive, sire ordeliver healthy offspring. Theseanimals included:

- 

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

- 

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

- 

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

- 

## Group 4 Male 32and Female 72 (implantation sites only). Histological examination was extended tothe mesenteric lymph nodes oftheselected males andfemales ofGroup 2based on possible treatment-related changes noted formid and high dose animals. Allabnormalities were described and included inthe report. An attemptwas made tocorrelate gross observations with microscopicfindings.
* 

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

Page 19

## C.I. PIGMENT BLUE 61 NOTOX Project488231 6.9. Pathology Offspring 6.9.1. Termination Pups were killed bydecapitation on Day 5oflactation or shortly thereafter. 6.9.2. Macroscopic examination Alloffspring was sexed and externallyexamined ifpracticallypossible. The stomachwas examined forthe presence ofmilk. Descriptions ofall macroscopic abnormalities were recorded. Ifpossible, defects or causeofdeath was evaluated. Anyabnormal pup, organ ortissue was preserved in10% buffered formalin, for possiblefurther examination. 6.10. Calculations For each dose group reproduction parameters were expressed intwo ways:

- 

## As amean (with standard deviation)ofthe numberobserved for each litter

- -Relative toasecondparameterandcalculatedonatotalgroupbasis For each groupthe following calculationswere performed: Percentage mating Number offemales mated x 100 Number offemales paired Fertility index Number ofpregnant females x 100 Number offemales paired Conception rate Number ofpregnant females x 100 Number offemales mated Gestation index Number offemales bearing live Pups x 100 Number ofpregnant females Duration ofgestation Number ofdays between confirmation of mating andthe beginning of parturition Percentage live males Number of livemale Pups atFirst LitterCheck x 100 *at First Litter Check Number of livepups at First Litter Check Percentage livefemales Number oflivefemale pups at First LitterCheck x 100 at First Litter Check Number oflive pups at First Litter Check Percentage ofpostnatal Number ofdead pupson Day4postpnartumx 100 loss Days 0-4postpartum Number oflive pups at First Litter Check Viability index Number oflive pups on Day 4postpaitumx 100 Numberof pups born alive

Page20

## 0.1. 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.

- 

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

-Advia 120Version 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

- 

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

- 

## MAMS version 6.5 (Pearson Technical Services, Suffolk, Great Britain): Motor activity measurement.

- 

## Empower version 5.00 (Waters, Milford, MA, USA): System control, data acquisition and data processing for analyticalchemistry. 6.12. Interpretation Thefollowing statistical methodswill beused to analysethe data:

- 

## Ifthe variables could be assumed tofollow anormal distribution, the Dunnett-test (Dunnett, 1955) (many-to-one t-test) based on apooled variance estimate was appliedforthe comparison ofthe treited groups and the control groups foreach sex.

- 

## The Steel-test (Miller, 1 981) (many-to-one rank test)was applied ifthe data could not be assumed tofollow anormal distribution.

- 

## The Fisher Exact-test (Fisher, 1950) wasapplied tofrequencydata. Alltestswere two-sided and inall cases p<

## 0.05 wasaccepted asthe lowest level of significance. Nostatistical analysiswas performed on histopathology findings. 6.13. Listofdeviations 6.13.1. Listof protocol deviations 1 Noorgan weightwasdetermined ofthe testes and epididymides ofanimal 30. * Evaluation: Sufficientdata were availableforevaluation. 2 Afewtissueswere not availablefor histopathology. Reasonsfor this includedthat thesetissues were notdiscernable at necropsy ortrimming, orwere erroneously not collected at necropsy. Tissues are listed inraw dataand pathology report. Evaluation: Sufficient data was availablefor evaluation. 3 Tissues forhistopathology were preserved and organ weightswere determined for another subsetof animals (out of20 selected males, 5were different: Group I males 4 and 5instead of7and 10, Group 2male 13 instead of 16, and Group 4males 31 and 33 instead of36 and 37) than tested forfunctional observations and clinical laboratory investigations. Evaluation: No toxicitywas noted upto the high dose levelfor all parameters; therefore itwas not essential to have results ofthese parameters on the same subset of animals. The study integritywas notadversely affected by thedeviations. 6.13.2. List ofstandard operating procedure deviations There were no deviationsfrom standard operating procedures.

Page21

## ClI. PIGMENT BLUE 61 NOTOX Project488231 7. RESULTS 7.1. Analysis of Dose Preparations (see alsoAppendix3) The concentrations analysed inthe formulations ofGroup 2,Group 3and Group 4were in agreement with target concentrations (i.e. mean accuracies between 85% and 115%). Notest substance wasdetected intheGroup 1formulations. Theformulations ofGroup 2and Group 4 were homogeneous (i.e. coefficientofvariation :5 10%). Formulations ofGroup 2and Group 4 were stable when stored at room temperature for at least6hours. 7.2. Observations 7.2.1. Mortality No mortality occurred during thestudy period. 7.2.2. Clinical Signs Therewere noclinical signs oftoxicity noted during theobservation period. Blue discolouration ofthe faeces was noted inall animals of both sexes at50, 250, and 1000 mg/kg. Blue staining of severalbody parts (tail, back, mouth, snout,abdomen, forelegs, flank) was also observed atthese dose levels but atalower incidence.These findingswere considered due tothe staining properties ofthe test substance and notto be toxicologically relevant. Slightsalivation was noted for one lowdose male (1day), five mid dose males (1 -3days), six high dose males (2-18 days) and two high dose females (1day). Salivation wasconsidered to be aphysiological responsetothe tasteoftheformulation rather than asign ofsystemictoxicity, considering the nature, minorseverity ofthe effectand its time ofoccurrence (i.e. afterdosing). Therefore, this wasconsidered nottoxicologicallyrelevant. Incidental findingsthatwere noted included alopecia and scabbing on several body parts, rales, piloerectionand abrokentail apex.These findingsare commonly noted inrats ofthisage and strain which are housed and treated underthe conditions inthisstudy.At the incidence observed, these wereconsidered signs of notoxicological significance. 7.2.3. Body Weights Bodyweights and bodyweight gain oftreated animals remained inthe same range as controls overthe study period. 7.2.4. Food Consumption Food consumption (absoluteand relative) oftreated animals inall groups remained inthe same range ascontrols over thestudy period. Twofemale rats (numbers 55 and 60) ofthe low dosegroup showed very lowfood consumption on Days 11-14 post-coitum. As bodyweight gainsforthese damswere normal,these low valueswere probably caused byaweighing error rather than being true values and were therefore considered nottoxicologically relevant.

Page22

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 7.2.5. Functional observations No changes were observed inhearing ability, pupillary reflex, static righting reflex and grip strength inthe animalstreated with C.l. Pigment Blue 61, whencompared to control animals. The variation inmotor activitydid not indicate arelation withtreatment. 7.3. Clinical Laboratory Investigations 7.3.1. Haematology Haematological parameters oftreated ratswere notaffected bytreatment up to 1000 mg/kg. Females treated with 1000 mg/kg showed statistically significantly lower prothrombin time than controls. Thisfinding isconsidered tobe ofno toxicological significance sincethe valuewas within normal limits and no corroborative findings were noted. Inaddition,the statistically significantly increased haematocrit value forlow dosefemales was notconsidered treatment related as no dose response relationship was noted. 7.3.2. Clinical Biochemistry Notoxicologicallyrelevant changes occurred inclinical biochemistry parameters oftreated rats. Males treated with 1000 mg/kg had statistically significantly highercholesterol measures than controls. Thiswas considered to be ofnotoxicological significance asthe valuewaswithin normal limits and no corroborative findingswere noted. 7.4. Pathology 7.4.1. Macroscopic Examination Notoxicologically relevant macroscopic findingswere noted up to 1000 mg/kg bodyweight/day. Greenish or bluish discolouration ofthe mesenteric lymph nodes was noted for three lowdose animals, seventeen mid dose animalsand all high dose animals. Seven high dose females had bluish contents inthe gastro intestinal tract. Bluish discolouration of thetail wasobserved for two males. These findingswere considered duetothe staining properties ofthetest substance and notto be toxicologicallyrelevant. Incidentalfindings included bent tail, hard yellowish noduleon the tail ofthe epididymidis, alopecia at several body parts, reduced seminalvesicle size, dark red or reddish discolouration ofthe mandibular lymph nodes, reddish discolouration ofthe pancreas, and red-brown discolouration ofthe adrenal glands.These findings are occasionallyseen among rats used in these types of studies and intheabsence ofcorrelated microscopic findingstheywere not considered changes oftoxicological significance.

Page 23

## C.l. PIGMENT BLUE 61 NOTOX Project 488231 7.4.2. Organ Weights Notoxicologically relevant organ weight changeswere notedfor animalstreated up to 1000 mg/kg bodyweight/day. The statisticallysignificantly increased relative spleenweight noted forhigh dose males was considered not to be asignoftoxicity inthe absenceofcorroborative (microscopic) findings. 7.4.3. Microscopic Examination (see also Appendix 4) Notreatment related histopathological changeswere observed inthetissuesof any animal receiving C.l. Pigment Blue61. No abnormalities were noted on evaluation ofPAS stained sectionsoftestes ofF 0

## males. All semniniferous tubules were normal with respectto theirstage inthe spermatogenic cycle and the integrity ofthe variouscell types present within the different stages. No pathologicalfindingswere present inthe reproductiveorgans of maleand female animals thatfailed to produce offspring. Small amounts of blue material were observed inthe lumenoftheduodenum, ileum,jejunum, colon or rectum inafewgroup 4animals. Blue material wasalso noted inthesuperficial skin (keratin layer) ofthetail inone group 4male. These findings are considered to represent the presence ofthetestsubstanceand insome cases correlated withthe macroscopic observations. No microscopic findings were observed inthe mesenteric lymph nodes to accountfor the discolouration ofthese tissues recorded at necropsy. Itisconsidered thatthediscolouration is dueto the presence ofthetestsubstance and thatthe pigment was lostfrom thetissues inthe processing or staining procedures. Allother microscopic findingswere considered to be incidental and unrelated to treatmentwith the testsubstance. 7.5. Reproduction Reproduction parameters are described intables Iand II: Table 1.Reproduction Data. Number offemales Group 1 Group 2 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 sites only 0 0 0 1(F72) Number offemales with 9 9 9 9 living pups atfirst littercheck

Page24

## C.I. PIGMENT BLUE 61 NOTOX Project 488231 Table 11. 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) * 

### 100 

## 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 pupsatfirst littercheck/ Number offemales pregnant) *

### 100 

## Reproduction parameterswere unaffected by treatment up to 1000 mg/kg body weight/day. Notreatment related findingswere observed for mating performance, fertility parameters, duration ofgestation, number ofdead and livingpups atfirst litter check, number ofimplantation sitesand number ofcorpora lutea. 7.6. Breeding Data Breeding parameters were unaffected bytreatment up to 1000mg/kg body weight/day. Postnatal loss andviability indexwere similarfor thecontrol and treated groups. 7.7. Pup Development Development ofpups was unaffected by treatment up to 1000 mg/kg bodyweight/day. (Mean) bodyweights were similarfor thecontrol and treated groups. Incidental clinicalsymptoms consisted ofsmall size ofthe pup, red spoton the noseor head, and pale appearance. At macroscopic examination, autolysis and absence of milk inthe stomach was noted for afew dead pups. No relationship withtreatment was established for theseobservations and theywere considered to be within the normal biological variation for rats ofthis ageand strain.

Page 25

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 8. DISCUSSION AND CONCLUSION C.l. PIGMENT BLUE 61 was administered bydaily oral gavageto male and femaleWistar Han rats at dose levels of 50, 250 and 1000 mg/kg/day. The males were exposed for 2weeks prior to mating, during mating, and upto termination (for30 days). Thefemales were exposedfor 2 weeks priorto mating, during mating, during post-coitum, and at least4daysof lactation (for41- 45 days). Formulation analysisshowed thatthe formulations were prepared accurately, were homogeneous and were stableforat least6hours atroom temperature. Notreatment-related toxicologically significantchanges were noted inany ofthe parameters investigated inthis study (i.e. mortality, clinicalappearance, functional observations, body weight, food consumption, clinical laboratory investigations, macroscopicexamination, organ weights, microscopicexamination, reproduction, breeding and pup development). From the results presented inthis report, the parental, reproduction, breeding and developmental NoObserved Adverse Effect Level (NOAEL) for C.1. PIGMENT BLUE 61 of at * least 1000 mg/kg/day was established.

Page26

## ClI. PIGMENT BLUE 61 NOTOX Project 488231 9. REFERENCES Dunnett C.W., AMultiple Comparison Procedure for Comparing Several Treatments with a Control, J.Amer. Stat. Assoc. 50, 1096-1121 (1955). Fisher R.A., Statistical Methods for Research Workers, Oliver and Boyd, Edinburgh (1950). Miller R.G., Simultaneous Statistical Inference, Springer Verlag, New York (1981). Salewski, E.Farbemethode zumn makroskopischen Nachweisvon Implantationsstellen am Uterus der Ratte. [Staining method foramacroscopic test for implantation sites inthe uterus of the rat]. Naunyn

- 

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

Page 27

### 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1

APPENDIX 1 SUMMARY TABLES

-Page 1

### C.I. 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 I(CONTROL) Various

Broken(1) G:

... 1

(Tailapex) %:

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

Scabs(3) G:

. 1

(Neck) %:

.. 1 Blue staining (1) G: ill11 Blue discolouration (1) G: 11 ill

(Faeces) %: AAAA Secretion/excretion

Salivation(3) G:

.. 1 %:

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

Alopecia (3) G:

.. I.

(Forelegs) %

. . I Scabs (3) G:

.. 1

(Neck) %:

. 1 Bluestaining (1) G:

. .

(Back) %

. 1.. Blue staining(1) G:

... 1

(Tail) %

. . Blue discolouration (1) G: 11 ill

(Faeces) %: AA AA Secretion/excretion

Salivation (3) G:

. .11 %: .24 GROUP 4(1000 MG/KG) Skin/fur

Alopecia (3) G:

. 11

(Foreleg right) %:

.. 11 Alopecia (3) G:

.. 121

(Forelegs) %: Ill11 Scabs (3) G:

....

(Foreleg right) %:

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

(Back) %. 1. Bluestaining(1) G:

. 1 Bluediscolouration (1) G: 11 ill

(Faeces) %: AA Secretion/ excretion

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

G: Median value ofthe highest individualweekly grades %:Percentofaffected animals (O=lessthan 5%, 1 =between 5%and 15%,.. Amore than 95%) Observation performed, sign not present

- Page 2

- 24O0t08 151145

### C.I. 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 I(CONTROL) SkinI/fur

Piloerection (1) G:

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

(Cheekleft) %: 11 11. Alopecia (3) G: .11..

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

(Back) %: 23 32... Alopecia(3) G: 11 1111.

(Neck) %: 43 4441. Alopecia (3) G:

.. ... 22

(Abdomen) %:

.. .. 22 Alopecia (3) G:

... 1

(Flank left) %:

. .. 1 Alopecia (3) G:

. .. 1

(Thigh hind left) %:

.. ... 12 Alopecia (3) G:

... 1.

(Foreleg left) %:

... 1. Alopecia (3) G:

..... 33

(Forelegs) %:

..... 12 Scabs(3) G:

.... 1

(Mouth) %:

... 1. Scabs(3) G:

. 1.

(Snout) %:...1. GROUP 2(50 MG/KG) Skin/fur

Piloerection(1) G:

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

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

... 1

(Abdomen) %:

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

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

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

Rales (3) G:

... 2 %11 Skin/ fur

Piloerection (1) G:

.... 1 %.1 Alopecia(3) G: I1 I11I... (Cheekright) %: 12 11
... Bluestaining(1) G:

. 1

(Mouth) %:

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

(Snout) %: 1. Blue staining (1) G: 11 1
....

(Back) %: 1

.. Blue staining (1) G:

. 1

(Abdomen) %:

.. .1 Blue staining (1) G:

. .. 1

(Forelegs) %:

... 1 Bluediscolouration(1) G: 11 11111

(Faeces) %: AA AAAAA

G: Median value ofthe highestindividual weekly grades %:Percent ofaffected animals (O=less than5%, 1 =between 5%and 15%,.,A=more than 95%) Observation performed, sign notpresent

- Page 3 -' 24Oct08 15h45

### C.I. 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 MGIKG) Breathing

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

Alopecia (3) G:

... .

(Back) %:

. 1 Alopecia (3) G: ,1

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

... .

(Abdomen) %:....1 Alopecia (3) G:'

. . .. 333

(Forelegs) %

. . .112 Alopecia (3) G:

. . . 22

(Hindlegs) %

. ... 12 Scabs (3) G:

.... 1

(Mouth) %

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

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

... 1

(Flank right) %1 Bluediscolouration (1) G: 11111

(Faeces) %: AA AAAAA Secretion/ excretion

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

G: Medianvalue ofthehighest individualweekly grades %:Percent ofaffected animals (O=less;than 5%, 1 =between 5%and 15%, .,A=more than 95%) Observation performed, sign not present

- Page 4
- 

## ~"24Oct08 15h45

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


### BODY WEIGHTS (GRAM) SUMMARY MALES
GROUP 1 GROUP2 GROUP 3 GROUP 4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
PRE MATING DAY 1 MEAN 302 301 303 303 WEEK 1 ST.DEV 15.1 139 8.4 9.2 IN 10 10 10 10 DAY8 MEAN 325 322 328 321 WEEK2 ST.DEV 18.5 19.8 120 10.9 N 10 10 10 10 MATING PERIOD DAY 1 MEAN 346 345 352 340 WEEKi1 ST.DEV 21.0 23.1 146 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 GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50 MG/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 WEEK2 ST.DEV 8.5 9.1 6.4 5.6 N 10 10 10 10 MATING PERIOD DAY1 MEAN 208 212 211 208 WEEK 1 ST.DEV 7.6 6.5 7.1 10.1 N 10 10 10 10

I~Dunnett-test based on pooled variance significant at 5%()or 1%

### ()level

- Page5

- 00- 29Sep08 09h35

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 BODY WEIGHTS (GRAM) SUMMARY FEMALES FO-GENERATION
GROUP 1 GROUP2 GROUP 3 GROUP4 CONTROL 50MG/KG 250 MG/KG 1000 MG/KG
POST COITUM DAYO MEAN 209 214 212 210 ST.DEV. 9.7 7 3 44 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 78 6.7 N 9 9 9 10 DAY 11 MEAN 246 251 253 249 ST.DEV. 13.1 14.1 8&5 70 N 9 9 9 10 DAY 14 MEAN 258 263 265 259 ST.DEV. 18.5 15.8 101 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 1 MEAN 241 245 248 238 ST.DEV. 145 147 8.0 7.1 N 9 9 9 9 DAY 4 MEAN 252 250 256 251 ST.DEV. 153 17.6 6.8 10.6 N 9 9 9 9

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

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

- Page 6

- 0- 23Oct08 161107

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1
BODY WEIGHT GAIN I%) SUMMARY MALES
GROUP 1 GROUP2 GROUP3 GROUIP4 CONTROL 50MG/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 8 7 8 6 WEEK 2 ST.DEV 25 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 2.4 3.2 N 10 10 10 10 DAY8 MEAN 19 16 21 16 WEEK 2 ST.DEV 5.9 4.6 2.1 4 1 N 10 10 10 10 DAY15 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 GROUP 2 GROUP 3 GROUP4 CONTROL 50 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 00 N 10 10 10 10 DAY8 MEAN 3 4 4 3 WEEK2 ST.DEV 1.8 1.8 3.0 1.9 N 10 10 10 10 MATING PERIOD DAYl MEAN 7 9 10 8 WEEK 1 ST.DEV 2.2 3.0 5.0 4.4 N 10 10 10 10

P~Dunnett-test based on pooled variancesignificant at 5%()or 1%(**) level

- Page 7

- NO- 29Sep08 09h35

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

### (%)SUMMARY FEMALES FO-GENE RATION

GROUP 1 GROUP 2 GROUP 3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
POST COITUM DAY0 MEAN 0 0 0 0 ST.DEV. 0.0 0.0 00 00 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 3.4 2.9 2.8 N 9 9 9 10 DAY14 MEAN 23 23 25 23 ST.DEV. 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 6.4 8.2 N 9 9 9 10 DAY20 MEAN 50 48 50 49 ST.DEV. 10.4 65 9.1 14.6 N 9 9 9 10
LACTATION DAYl MEAN 0 0 0 0 ST.DEV. 0.0 0.0 0.0 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

/Dunnett-test basedon pooled variance significantat 5%

### ()or 1 %

### (**) level Explanations forexcluded dataare listed inthe tablesofthe individualvalues

- Page 8

- 23Oct08 16h07

### C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1 FOOD CONSUMPTION (G/ANIMALIDAY) SUMMARY MALES
GROUP 1 GROUP 2 GROUP 3 GROUP4 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 02 02 N(CAGE) 2 2 2 2 MEAN OF MEANS OVER PRE MATING MEAN 22 21 22 22 

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

PRE MATING DAYS 1-8 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 MATINGMEAN 14 14 14 15

/*Dunnet-test basedon pooled variance significant at 5%()or 1%()level

- Page 9

- 29Sep08 09h36

### C]I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1 FOOD CONSUMPTION (GIANIMAL/DAY) SUMMARY FEMALES FO-GENERATION
GROUP 1 GROUP 2 GROUIP3 GROUP 4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG

POST COITUM 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 0EV 1.9 3.0 2.1 0.8 N 9 9 9 10 DAYS 7-11 MEAN 19 18 20 19 ST.DEV 1.4 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 36 1.8 2.6 N 9 9 9 10 DAYS 17-20 MEAN 22 22 22 21 ST.DEV. 3.3 3.3 1.7 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 98



## rDunnet-test based on pooled variance significant at 5% ()or 1 % ()level Explanationsfor excluded data are listed in the tables ofthe individual values -Page 10 - 

## ~'23Oct08 16h07

### CI. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1 RELATIVE FOOD CONSUMPTION (G/KG BODY WEIGHT/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 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 GROUP 4 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 11 N(GAGE) 2 2 2 2 MEAN OF MEANS OVER PRE MATING MEAN 73 71 72 73

*/* Dunnett-test basedon pooled variance significant at 5%()or 1 %

### ()level

- Page 11
- 

### IS'29Sep08 09h35

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

### RELATIVE FOOD CONSUMPTION (G/KG BODYWEIGHTIDAY) SUMMARY FEMALES FO-GENERATION

GROUPi 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 5.9 5.7 N 9 9 9 10 DAYS 11-14 MEAN 79 64 85 78 ST.DEV. 9.3 27.5 17.9 67 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 0EV 24.8 26.3 21.7 39.1 N 9 9 9 8

*r Dunnett-test based on pooled variancesignificant at 5% ()or 1 % ()level Explanations for excluded data are listed in the tablesofthe individual values

- Page 12

- 23Oct08 16h07

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 FUNCTIONAL OBSERVATIONS SUMMARY MALES
GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50 MG/KG 250MG/KG 1000 MG/KG
AT WEEK 4 HEARING MEDIAN 0 0 0 0 SCOREO0[l N 5 5 5 5 PUPILL MEDIAN 0 0 0 0 SCORE 0/1 N 5 5 5 5 PUPIL R MEDIAN 0 0 0 0 SCOREO0/1 N 5 5 5 5 STATIC R MEDIAN 0 0 0 0 SCOREO0/1 N 5 5 5 5 GRIP MEDIAN 0 0 0 0 SCOREO0/1 N 5 5 5 5 

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

AT WEEK 4 HEARING MEDIAN 0 0 0 0 SCOREO0/1 N 5 5 5 5 PUPIL L MEDIAN 0 0 0 0 SCOREO0/1 N 5 5 5 5 PUPIL R MEDIAN 0 0 0 0 SCOREO0/1 N 5 5 5 5 STATIC R MEDIAN 0 0 0 0 SCOREO0/1 N 5 5 5 5 GRIP MEDIAN 0 0 0 0 SCOREO0/1 N 5 5 5 5

+/++ Steel-test significant at5% ()or 1%(++) level

-Page 13

- 

## ~08Oct08 12h21

### C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 1 MOTOR ACTIVITY MEASUREMENTS SUMMARY MALES WEEK 4
GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG 

### total high MEAN 3328 2361 2061 1777 sensor count ST.DEV 1736 578 573 581 N 5 5 5 5 total low MEAN 4533 4690 4204 4759 sensor count ST.DEV 1009 929 266 1030 N 5 5 5 5



### FEMALES LACTATION
GROUP 1 GROUP 2 GROUP 3 GROUP 4 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 5 5 total low MEAN 16563 8827 16471 18503 sensor count ST.DEV 3736 4455 6655 6789 N 5 5 5 5

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

- Page 14-

### 0.1. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 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 76 10OE9/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 %WBC 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 0EV 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 06 03 0.2 0.4 N 5 5 5 5 Basophils MEAN 0.4 0.4 0.4 0.4 %WBC ST.DEV 01 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.IJEV 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 mmol/L ST.DEV 0.4 0.6 0.4 0.2 N 5 5 5 5 Haematocrit MEAN 0.424 0.444 0.455 0.448 UlL ST.DEV 0.025 0.029 0.020 0.010 N 5 5 5 5 MCV MEAN 52.4 51.2 51.9 51.6 ft. 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 0.07 0.03 0.04 N 5 5 5 5 MCHC MEAN 22.91 22.27 22.12 22.09 mmol/L ST.DEV 0.68 0.66 0.30 0.48 N 5 5 5 5 Platelets MEAN 805 862 873 981 10OE9/L ST DEV 179 93 90 71 N 5 5 5 5 PT MEAN 179 18.1 174 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 N 4 5 5 5

+/++ Steel-testsignificant at 5%(+)or 1 %(++) level Dunnett-test based on pooled variance significant at 5%()or 1%(-) level

- Page 15

- OW~29Sep08 09h39

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 HAEMATOLOGY SUMMARY FEMALES
GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
END OF TREATMENT WBC MEAN 4.0 4.0 45 5.7 10OE9/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 %WCST.DEV 6.3 15.6 3.0 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 26 2.0 2.1 1.4 %WBC ST.DEV 0.9 0.6 0.6 1.0 N 5 5 5 5 Eosinophils MEAN 0.7 0.3 0.6 0.5 %WBC ST 0EV 0.6 0.4 0.5 0.5 N 5 5 5 5 Basophils MEAN 0.2 0.1 04 0.1 %WBC ST.DEV 0.2 0.2 0.4 0.1 N 5 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 15.8 14.0 %ST.DEV 1.8 37 1.5 1.2 N 5 5 5 5 Haemoglobin MEAN 8.9 9.3 88 9.2 mmol/L ST.DEV 0.3 0.1 0.4 0.2 N 5 5 5 5 Haemnatocrit MEAN 0.386 0.405 0.386 0.405 UJL ST.DEV 0.008 0.009 0.014 0.014 N 5 5 5 5 MCV MEAN 52.9 53.5 52.7 53.1 fL ST.DEV 1.2 2.2 0.4 1.1 N 5 5 5 5 MCH MEAN 1 .22 1 .23 1 .20 1.20 fmol ST.DEV 0.04 0.08 0.03 0.05 N 5 5 5 5 MCHC MEAN 23.09 22.94 22.76 22.64 mmol/L ST.DEV 0.58 0.55 0.64 0.58 N 5 5 5 5 Platelets MEAN 1117 1320 1385 1195 10OE9/L ST 0EV 315 95 274 322 N 5 5 5 5 PT MEAN 17.1 16.5 163 15.6 s ST.DEV 0.5 0.6 0.7 05 N 4 5 5 5 APTT MEAN 17.7 18.4 16.4 15.9 s ST.DEV 2.3 3.4 1.5 3.4 N 4 5 5 5

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

/~Dunnett-test based on pooled variance significant at 5%()or 1 %(-) level -Page 

### 16- 

## I 29Sep8091139

### C.I. PIGMENT BLUE61 NOTOX Project 488231 APPENDIX 1
CLINICAL BIOCHEMISTRY SUMMARY MALES
GROUP 1 GROUP2 GROUP 3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000MG/KG
END OF TREATMENT ALAT MEAN 44.7 52.0 46.8 47.8 U/L ST.DEV 7.5 14.1 6.7 6.2 N 5 5 5 5 ASAT MEAN 81.9 82.9 79.7 82.6 U/L 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 g/L 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 gtL 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 03 0.3 0.2 N 5 5 5 5 Urea MEAN 8.8 9.5 8.6 8.4 mmolIL ST.DEV 2.3 13 0.6 1.1 N 5 5 5 5 Creatinine MEAN 42.8 46.0 46.3 43.0 umolIL ST.DEV 1.7 6.8 4.7 1.5 N 5 5 5 5 Glucose MEAN 8.07 7.49 7.32 720 mmol/L ST.DEV 1.49 1.70 0.98 1.56 N 5 5 5 5 Cholesterol MEAN 1.65 197 1.88 2.13 mmol/L ST.DEV 0.31 0.20 027 0.33 N 5 5 5 5 Sodium MEAN 142.3 142.0 142.6 143.4 mmollL 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 mmol/L ST.DEV 0.23 0.16 0.07 0.26 0N 5 5 5 5 Chloride MEAN 104 104 105 105 mmolIL ST.DEV 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 lnorg.Phos MEAN 2.07 2.05 2.18 2.04 mmolIL ST.DEV 0.21 0.25 0.13 0.13 N 5 5 5 5

ID unnett-testbased on pooled variancesignificant 

### at 5% 

### ()or 1 %

### (**) level

-Page 17

- 

## IWP 29Sep08 09h39

### C.I. PIGMENT BLUE 61 NOTOX 

### Project488231 APPENDIX I CLINICAL BIOCHEMISTRY SUMMARY FEMALES
GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
END OF TREATMENT ALAT MEAN 58.1 67.8 66.6 46.2 U/L ST.OEV 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.DEV 11.7 12.0 10.2 5.9 N 5 5 5 5 ALP MEAN 65 72 70 69 U/L ST.DEV 22 19 20 44 N 5 5 5 5 Total protein MEAN 65.8 65.3 67.7 65.6 g/L ST.DEV 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 09 2.2 1.2 10 N 5 5 5 5 Total bilirubin MEAN 2.4 2.0 2.5 1.8 umol/L ST.DEV 0.4 0.4 0.6 0.3 N 5 5 5 5 Urea MEAN 9.4 10 1 10.0 9.7 mmol/L 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 35 1.6 5.0 3.0 N 5 5 5 5 Glucose MEAN 6.48 592 5.47 581 mmol/L ST.DEV 1.28 1.23 0.82 076 N 5 5 5 5 Cholesterol MEAN 1.45 169 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 55 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 5 5 5 lnorg.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 Dunnett-test based onpooled variancesignificant at 5% ()or 1 %

### (-) level

- Page 18

- 

## , 29Sep08 09h39

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

Reduced insize 0 0 0 1 Mesenteric Inode

Discolouration 0 2 9## 10## Skin

Alopecia 0 0 0 1 Discolouration 0 1 0 1 Bone

Talbent 1 0 0 0 

### FEMALES
GROUP 1 GROUP2 GROUP 3 GROUP 4 CONTROL 50 MG/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 0 7## Pancreas

Discolouration 0 1 0 0 Adrenal glands

Discolouration 0 0 0 1 MesentericLnode

Discolouration 0 1 8## 9 Mandibular [.node

Discolouration 1 2 0 1 

# 0SiAlopecia 1 0 0 1

#/ Fishers Exact testsignificant at 5%

### ()or 1%

### (##) level

- Page 19

- NW 29Sep08 09h39

### ClI. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 ORGAN WEIGHTS (GRAM) SUMMARY MALES
GROUP 1 GROUP 2 GROUP 3 GROUP4 CONTROL 50 MG/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 204 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 0991 1.005 (GRAM) ST.DEV 0.035 0053 0.083 0.083 N 5 5 5 5 LIVER MEAN 8.44 8.22 903 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.DEV 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 0.068 0065 0.075 (GRAM) ST.DEV 0.012 0.010 0005 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 0032 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 1139 1.138 1.207 1.182 (GRAM) ST.DEV 0.086 0.114 0.136 0.124 N 10 10 9 10

IDunnett-test basedon pooled variance significant at 5%()or 1 %

### (**) level -Page 20
- 010 29Sep0809h39

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

## (%)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 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 N 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.DEV 0.03 0.03 0.02 0.04 N 5 5 5 5 ADRENALS MEAN 0.020 0.020 0.019 0.021 ()ST.DEV 0.003 0.003 0.002 0.003 N 5 5 5 5 SPLEEN MEAN 0164 0.174 0.178 0.193* 

### ()ST.DEV 0.026 0.016 0.015 0007 N 5 5 5 5 TESTES MEAN 0.95 0.94 0.92 0.98 

### ()ST.DEV 0.09 0.14 0.08 0.10 N 10 10 9 10 EPIDIDYMIDES MEAN 0.323 0.327 0.339 0340 ()ST.DEV 0.040 0.036 0.035 0033 N 10 10 9 10

PDunnett-test based on pooled variance significant at 5%()or 1%()level -Page 21
- 011 29Sep08 091139

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1
ORGAN WEIGHTS (GRAM) SUMMARY FEMALES
GROUP 1 GROUP 2 GROUP 3 GROUP4 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 5 5 5 5 BRAIN MEAN 1. 84 1.79 1.83 186 (GRAM) ST.DEV 0.05 0.04 0.06 0.01 N 5 5 5 5 HEART MEAN 0.733 0.725 0.785 0.716 (GRAM) ST.DEV 0.073 0.055 0.041 0.043 N 5 5 5 5 LIVER MEAN 7.23 7.07 7.81 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.DEV 0.21 0.06 0.08 0.16 N 5 5 5 5 ADRENALS MEAN 0.079 0084 0.082 0.088 (GRAM) ST.DEV 0.005 0.009 0.008 0.014 N 5 5 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

I~Dunnett-test based on pooled variance significant at 5%()or I% 

### ()level

- Page 22

- 

## ~r29Sep08 091139

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 ORGANIBODY WEIGHT RATIOS I(%) SUMMARY FEMALES
GROUP 1 GROUP 2 GROUP 3 GROUP 4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG
NECROPSY BODYW. MEAN 225 220 228 221 (GRAM) ST.DEV 17 18 2 15 N 5 5 5 5 BRAIN MEAN 082 0.82 0.80 0.84 ()ST.DEV 0.08 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 0.016 0.021 N 5 5 5 5 LIVER MEAN 3.22 3.21 3.42 3.23 

### (%) ST.DEV 0.27 0.13 0.15 0.28 N 5 5 5 5 THYMUS MEAN 0.071 0.077 0.075 0.081 ()ST.DEV 0.023 0.010 0.010 0.014 N 5 5 5 5 KIDNEYS MEAN 073 0.74 0.76 0.74 

### ()ST.DEV 0.11 0.05 0.03 0.10 N 5 5 5 5 ADRENALS MEAN 0.035 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 

### ()ST.DEV 0.035 0.031 0.042 0.029 N 5 5 5 5

/~Dunnett-test based onpooled variance significant at 5%()or 1 % ()level

- Page 23

- 

## ~-29Sep08 09h39

### C.I. PIGMENT BLUE 61 NOTOX 

### Project 488231 APPENDIX 1 

### REPRODUCTION PROCESSES FO-GENERATION

- 

### POST COITUM

FEMALE MALE MATING PREGNANT SCHEDULE DELIVERY NECROPSY NUMBER NUMBER DATE RECORDED DATE

GROUP I(CONTROIL) 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 05JUL2008 YES BREEDING 27JUL2008 01AUG2008 46 6 05JUL2008 YES BREEDING 27JUL2008 01AUG2008 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 28JUL2008 57 17 04JUL2008 YES BREEDING 25JUL2008 30JUL2008 58 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 GROUP4(1000 MG/KG) 71 31 05JUL2008 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 02JUL2008 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> implantationsites only <NP> Non-pregnant

- Page 24

- -23Oct08 16h08

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1 

### MATING PERFORMANCE FO-GENERATION

- 

### POST COITUM

DAYOFTHE 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 significant at 5% ()or 1 % (++) level

-Page 25

- 011 23Oct08 16h08

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 1
BREEDING DATA PER GROUP FO-GENERATION
- 

### LACTATION

GROUP 1 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 21.2 211 21 2 ST.DEV. 0.5 04 03 04 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 0A1 0.2 ST.DEV. 00 00 0.3 04 N 9 9 9 9 LIVING PUPS AT FIRST LITTER CHECK

%OFMALES/FEMALES (#) 44/56 47/53 50/50 50/50 TOTAL 104 103 101 114 MEAN(+) 11.6 11.4 11.2 127 ST.DEV. 33 1.6 3.9 17 N 9 9 9 9 POSTNATALLOSS

%OFLIVING PUPS 1.0 1.0 0.0 1.8 LITTERS AFFECTED 

### (#) 1 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 04 N 9 9 9 9 VIABILITY INDEX 

### (#) 99.0 99.0 100.0 98.2

Viability index =(Number ofalive pups before planned necropsy/ Number of pups born alive) *100 +/++ Steel-test significantat 5%(+)or 1% (++) level #/##Fisher's Exact test significantat 5%

### (#)or 1 %

### (##) level

- Page26

- 

## ~'23Oct08 1008

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


### MEAN BODYWEIGHTS OF PUPS PER GROUP (GRAM) P0-GENERATION

- 

### LACTATION

DAY SEX GROUP 1 GROUP 2 GROUP 3 GROUP4 CONTROL 50 MG/KG 250 MG/KG 1000 MG/KG I M MEAN 6.0 6.1 6.1 5.9 ST.DEV. 06 0.7 0.4 0.4 N 9 9 9 9 F MEAN 5.8 57 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 0.4 0.4 N 9 9 9 9 4 M MEAN 8.6 9A1 9.0 8.5 ST.DEV. 0.9 11 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 8.7 8.3 ST.DEV. 0.9 1.1 1.0 0.7 N 9 9 9 9

/Dunnet-test based on pooled variance significant at 5% 

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

- Page 27
- 

## ~'23Oct08 16h08

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2

APPENDIX 2 INDIVIDUAL TABLES

-Page 1

-

### C.I. PIGMENT BLUE61 NOTOX Project 488231 APPENDIX 2 MORTALITY DATA MALES
ANIMALSCHEDULED TREATMENT SACRIFICE FROM TO
GROUP I(CONTROL) 1 17JUL08 17JUN08 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 16JUL08 GROUP 2(50 MGIKG) 11 17JUL08 17JUN08 16JUL08 12 17JUL08 17JUN08 16JUL08 13 17JUL08 17JUN08 16JUL08 14 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(250 MG/KG) 21 17JUL08 17JUN08 16JUL08 22 17JUL08 17JUN08 16JUL08 23 17JUL08 17JUN08 16JUL08 24 17JUL08 17JUN08 16JUL08 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 MGIKG) 31 17JUL08 17JUN08 16JUL08 32 17JUL08 17JUN08 16JUL08 33 17JUL08 17JUN08 16JUL08 34 17JUL08 17JUN08 16JUL08 35 17JUL08 17JUN08 16JUL08 36 17JUL08 17JUN08 16JUL08 37 17JUL08 17JUN08 16JUL08 38 17JUL08 17JUN08 16JUL08 39 17JUL08 17JUN08 16JUL08 40 17JUL08 17JUN08 16JUL08

-Page 2

- 29Sep08 101121

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2
MORTALITY DATA FEMALES
ANIMALSCHEDULED TREATMENT SACRIFICE FROM 

### TO
GROUP I (CONTROL) 41 29JUL08 17JUN08 28JUL08 42 28JUL08 1 7JUN08 27JUL08 43 28JUL08 17JUN08 27JUL08 44 01AUG08 17JUN08 31JUL08 45 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 17JUN0B 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(1000 MG/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

- Page3-If 29Sep08 I0h21

### 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 I(CONTROL) ANIMAL I No clinicalsigns noted ANIMAL 2 No clinicalsigns noted ANIMAL 3 Various

Broken(1) G: ........................111111

(Tailapex) ANIMAL 4 No clinicalsigns noted ANIMAL 5 No clinicalsigns noted ANIMAL 6 No clinical signs noted ANIMAL 7 No clinical signs noted ANIMAL 8 No clinical signsnoted ANIMAL 9 No clinical signsnoted ANIMAL 10 No clinicalsignsnoted GROUP 2(50 MG/KG) ANIMAL II Skin/fur

Bluediscolouration (1) G: ...... 1111111 lll1lllllllll1

(Faeces) Secretion /excretion

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

.... ANIMAL 12 Skin /fur

Bluediscolouration(1) G: .... 111111111111111111111111

(Faeces) ANIMAL 13 Skin/fur

Bluediscolouration(1) G: .... 111111111111111111111111

(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 Skin/fur

Bluediscolouration(1) G: .... 111111111111111111111111

(Faeces) ANIMAL 17 Skin/fur

Bluediscolouration(1) G: .... 111111111111111111111111

(Faeces)

G: Highestdaily grades

Observation performed, sign not present

- Page 4

- '24Oct08 15h46

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

GROUP 2(50 MG/KG) ANIMAL 18 Skin/fur

Scabs(3) G......................1111111i

(Neck) Bluediscolouration(1) G: 111111111111111111111111i

(Faeces) ANIMAL 19 Skin/fur

Bluediscolouration (1) G: 111111111111111111111111iiii

(Faeces) ANIMAL 20 Skin/fur

Bluediscolouration(1) G: 111111111111 * (Faeces) GROUP 3(250 MG/KG) ANIMAL 21 Skin/fur

Blue discolouration (1) G:

... 1111111111111

(Faeces) Secretion /excretion

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

Bluediscolouration (1) G: ... 111111111111111111111111111 (Faec-es) ANIMAL 23 Skin/ fur

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

(Forelegs) Bluediscolouration (1) G:

.. 11111111111111

(Faeces) Secretion/ excretion

Salivation (3) G:
........................... 111l ANIMAL 24 Skin/fur

Bluediscolouration (1) G: .111111111111111111111111111

(Faeces) ANIMAL 25 Skin/Ifur

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

Salivation(3) G:.................1...........1. ANIMAL 26 Skin /fur

Bluediscolouration (1) G:...111111111111111111111111111

(Faeces) ANIMAL 27 Skin/fur

Bluediscolouration (1) G:
. . .1111111111111

(Faeces) Secretion /excretion

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

G:Highest dailygrades

Observation performed, sign not present

-Page 5

- 24Oct08 15h46

### C.I. PIGMENT BLUE 61 NOTOX 

### Project488231 APPENDIX 2 

### CLINICAL SIGNS MALES PRE MATING REPRO PHASE SIGN (MAX GRADE) WEEK: 1
............. 1......................... (LOCATION) DAY: 123456712345671234567123456712345671234567123

GROUP 3(250 MG/KG) ANIMAL 28 Skin/fur

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

(Neck) Bluediscolouration (1) G:

.. 1111111111111

(Faeces) Secretion/excretion

Salivation (3) G...............................1. ANIMAL 29 Skin Ifur

Bluediscolouration (1) G: ... .11111111111111111111111111. (Faeces) ANIMAL 30 Skin/fur

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

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

(Tail) Bluediscolouration (1) G: ... .111111111111111111111111111 (Faeces) GROUP 4(1000 MG/KG) ANIMAL 31 Skin/fur

Alopecia(3) G:................... ...... 1111il

(Foreleg right) Scabs(3) G:.......................111ill

(Foreleg right) Bluediscolouration (1) G

.. 11111111111111

(Faeces) ANIMAL32 Skin/ fur

Bluediscolouration (1) G: . . 111111111111111111111111111

(Faeces) ANIMAL 33 Skin/fur

Blue discolouration (1) G: .11111111111111

(Faeces) Secretion /excretion ANIMAL 34 Skin /fur

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

(Forelegs) Bluediscolouration (1) G: .1111111111111111111111111111i

(Faeces) Secretion /excretion

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

Bluediscolouration (1) G: . .1111111111111111111111111111

(Faeces) ANIMAL36 Skin/fur

Bluediscolouration (1) G: . . 111111111111111111111111111

(Faeces) Secretion/ excretion

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

G: Highest daily grades

Observation performed, sign not present

- Page 6 -I'24Oct08 15h46

### C.I. PIGMENT BLUE 61 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:

. . 11111111111111111111111111

(Faeces) ANIMAL 38 Skin/fur

Bluediscolouration (1) G: -
. llllllllllliiiiiiiii (Faeces) Secretion / excretion

Salivation (3) G:................1... 1........ ANIMAL 39 Skin/fur

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

(Tail) Bluediscolouration (1) G: .11111111111111

(Faeces) Secretion Iexcretion

Salivation(3) G:...........1.111...11.1
.... 1 ANIMAL40 Skin/fur

Bluestaining(1) G: .. ill1........................

(Back) Blue discolouration (1) G .11111111111111

(Faeces) Secretion/ excretion

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

### FEMALES PRE MATING REPRO PHASE SIGN (MAX GRADE) WEEK: 1
......... I................... 4......... (LOCATION) DAY: 123456712345671234567123456712345671234567123

GROUP 1(CONTROL) ANIMAL 41 Skin/fur

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

(Abdomen) ANIMAL 42 Skin/fur

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

(Mouth) Scabs(3) G:......................111l...... ........

(Snout) ANIMAL 43 No clinicalsigns noted ANIMAL 4 No clinicalsigns noted ANIMAL45 No clinicalsigns noted ANIMAL46 Skin/fur

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

(Back) (Neck)

G: Highestdaily grades

Observation performed, sign not present

- Page 7

- 01" 24Oct08 15h46

### C.I. 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 I(CONTROL) ANIMAL 47 Skin/fur

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

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

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

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

.......... 111

(Thigh hind left) Alopecia(3) G:......................... 11......

(Foreleg left) Alopecia (3) G..................... ..... ....... .. 2222233

(Forelegs) ANIMAL48 Skin/fur

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

(Neck) ANIMAL 49 Skin/Ifur

Alopecia (3) G: .111111111111111111111111111... ........... (Back) (Neck) ANIMAL 50 Skin/fur

Alopecia(3) G: 11111111111..... ............

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

(Cheek right) Alopecia (3) G:........11111111111111111..............

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

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

Alopecia(3) G:
... .111111.............. ................... (Neck) (Faeces) ANIMAL52 Skin/fur

Blue discolouration (1) G:........1111111111111111111111111111111111111i

(Faeces) ANIMAL53 Skin/fur

Piloerection (1) G:........I.................................1. Blue discolouration (1) G:liilillllllllillilll

(Faeces) ANIMAL 54 Skin/fur

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

(Abdomen) Blue discolouration (1) G: .111111111111111111111111111111111111

(Faeces)

G:Highest dailygrades

Observation performed, sign not present

- Page8

- 24Oct08 15h46

### C.I. 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 2(50 MG/KG) ANIMAL 55 Skin /fur (Faeces) ANIMAL 56 Skin/fur

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

... 1111111.... .................. ..

(Neck) Bluediscolouration (1) G: 111111111111111111111

(Faeces) ANIMAL 57 Skin /fur

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

(Tail) Blue discolouration (1) G:1111111111111111111

(Faeces) ANIMAL 58 Skin/fur

Blue discolouration (1) G: 1111111111111111111111111111111111111iiii

(Faeces) ANIMAL 59 Skin /fur

Bluediscolouration (1) G: 1111111111111111111111

(Faeces) ANIMAL 60 Skin/fur

Bluediscolouration (1) G: 111111111111111111111

(Faeces) GROUP 3(250 MGIKG) ANIMAL 61 Skin /fur (Faeces) ANIMAL 62 Breathing

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

Bluestaining(1) G:.........11.........................

(Back) (Faeces) ANIMAL63 Skin/fur

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

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

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

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

(Forelegs) (Faeces) ANIMAL64 Skin/fur

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

(Cheek right) (Faeces)

G: Highest daily 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.......... I...1................. 4 .. . (LOCATION) DAY: 123456712345671234567123456712345671234567123

GROUP 3(250 MG/KG) ANIMAL 65 Skin/ fur (Faeces) ANIMAL66 Skin/fur

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

............. 1

(Faeces) ANIMAL67 Skin/fur

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

# 0 (Faeces) ANIMAL 68 Skin /fur

Bluestaining(1) G:
...il

_ .. . .. . . .. . .. .... .

(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:.........
... I........................ .11.

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

(Forelegs) Alopecia (3) G:...
...... ......................... 112222222

(Hindlegs) 

# Ble0soorain()G

(Faeces) ANIMAL 72 Skin/Ifur (Faeces) ANIMAL73 Skin/fur

Bluestaining (1) G:...........1111111111111111111111111111 ....

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

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

G: Highestdailygrades

Observationperformed, sign notpresent -Page 10- 24Oct:08 15h46

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

### CLINICAL SIGNS FEMALES PRE MATING REPRO PHASE SIGN (MAX. GRADE) WEEK: 1
...... ..... 1I..................4

. (LOCATION) DAY: 123456712345671234567123456712345671234567123

GROUP 4(1000 MG/KG) ANIMAL 75 Skin/fur

Blue discolouration (1) G:

.. 1111111111111111111

(Faeces) ANIMAL 76 Skin/fur (Faeces) ANIMAL 77 Skin/fur

Alopecia (3) G:.......................................11111 (Back) Bluediscolouration (1) G. 1111111111111111111

(Faeces) ANIMAL78 Skin/fur

Bluediscolouration (1) G. 111111111111111111111

(Faeces) ANIMAL 79 Breathing

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

Scabs(3) G:.......................................1111..

(Mouth) (Faeces) Secretion / excretion

Salivation (3) G...................... ANIMAL 80 Skin/fur

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

(Flank right) (Faeces)

G: Highest dailygrades

Observation performed, sign not present -Page 11- 24Oct08 15h46

### ClI. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2
BODYWEIGHTS (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
GROUP II(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 MGIKG) 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 28 312 340 368 373 395 29 300 321 345 362 375 30 301 329 359 372 397 GROUP 4(1000 MGlKG) 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 -fr29Sep08 101121

### C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2
BODY WEIGHTS (GRAM) FEMALES PRE 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 MGIKG) 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(250 MGIKG) 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 MGIKG) 71 198 205 203 72 201 203 221 73 186 196 201 74 196 204 220 75 190 193 214 76 183 195 207 77 196 197 218 78 195 200 203 79 184 187 190 80 195 201 201

-Page 13

- 

## i~ 29Sep08 101121

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2
BODY WEIGHTS (GRAM) FEMALES FO-GENERATION 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 MG/KG) 51 212 229 234 247 262 279 300 243 247 52 206 218 224 238 247 270 298 225 229 

# 053 223 249 262 277 290 316 352 269 276 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 MG/KG) 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 

# 076 205 223 228 244 250 283 316 232 238 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-P 230ct0816h10

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 BODY WEIGHT GAIN(% MALES PRE MATING MATING PERIOD DAYS 1 8 1 8 15 2229 36 WEEKS 1 2 1 2 3 4 5 6 ANIMAL

GROUP I(CONTROL) 1 0 10 20 2532 2 

### a 5 12 1824 3 0 10 17 2228 4 0 3 7 7 13 5 0 6 11 14 20 6 0 8 19 2735 7 0 9 18 2125 8 0 6 12 1621 9 0 6 14 20 26 10 0 11 18 2531 GROUP 2 (50 MGIKG) 11 0 5 13 15 20 12 0 5 14 19 23 13 0 7 13 16 20 14 0 6 12 15 18 15 0 12 18 21 27 16 0 8 15 1724 17 0 9 20 5 30 18 0 6 11 15 21 19 0 5 12 15 20 20 0 9 172233 GROUP 3 (250 MGIKG) 21 0 8 16 22 28 22 0 11 17 22 25 23 0 6 12 2025 24 0 8 15 1922 25 0 6 13 1722 26 0 11 19 24 30 27 0 9 16 21 27 28 0 9 18 2027 29 0 7 15 21 25 30 0 9 19 24 32 GROUP 4(1000 MGIKG) 31 0 8 14 1724 32 0 5 13 20 26 33 0 3 9 13 18 34 0 5 9 14 18 35 0 7 15 1924 36 0 8 17 22 26 37 0 9 172024 38 0 5 11 14 22 39 0 6 9 10 18 40 0 3 9 14 20

- Page 15

- 29Sep05 10h21

### ClI. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 BODY WEIGHT GAIN(% FEMALESPRE MATING MATING PERIOD DAYS 1 8 1 8 15 222936 WEEKS 1 2 1 2 3 4 5 6 ANIMAL
GROUP I(CONTROL) 41 0 2 5 42 C 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 MGIKG) 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 101121

### C.I. 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 GROUP1I(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 14 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 58 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 MGIKG) 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 28 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 

# 976 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-- 23Oct08 16h14

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 FOOD CONSUMPTION (GIANIMALJDAY) 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 MGIKG) 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) 11 15 15 12 14 14 GROUP 3(250 MG/KG) 13 14 15 14 14 14 GROUP 4(1000 MG/KG) 15 15 15 16 14 14

-Page 18

- 

## ~29Sep08 101121

### C.I. PIGMENT BLUE 61 NOTOX 

### Project 488231 APPENDIX 2
FOOD CONSUMPTION (G/ANIMALIDAY) FEMALES FO-GENERATION POST COITUM LACTATION DAYS 0-4 4-7 7-11 11-14 14-17 17-20 1-4 ANIMAL GROUP1I(CONTROL) 41 <NP> 14 16 16 18 15 17 -- 42 18 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 MGIKG) 51 16 16 17 20 19 20 21 52 17 16 20 17 19 19 21 53 22 24 15 25 28 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 MGIKG) 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 65 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 MGIKG) 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 18 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-' 230ct081014

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2
RELATIVE FOOD CONSUMPTION (GIKG BODY WEIGHTIDAY) 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 MGIKG) 3 66 68 4 64 68 GROUP 3(250 MG/KG) 5 67 69 6 65 67 GROUP 4(1000 MGIKG) 7 66 68 8 67 68 

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

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

-Page 20 -N'29Sep08 10h21

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2
RELATIVE FOOD CONSUMPTION 

### (GIKG BODY WEIGHT/DAY) FEMALES FO-GENERATION POST COITUM LACTATION DAYS 0-4 4-7 7-11 11-14 14-17 17-20 1-4 ANIMAL

GROUPl1(CONTROL) 41 <NP> 63 71 69 75 61 71 -- 42 83 88 85 92 77 76 ill 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 85 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 MGIKG) 61 70 80 73 81 74 69 109 62 68 74 73 70 69 69 49 63 80 87 78 83 72 72 i11 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 230ct0816h14

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 FUNCTIONAL OBSERVATIONS MALES AT WEEK 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 a 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 Oil SCORE 0/1 SCORE 0/1

GROUP 1(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 GROUP2 (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
- 0- 08Oct08 12h22

### C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 FUNCTIONAL OBSERVATIONS FEMALES ATWEEK 4
ANIMAL HEARING PUPIL L PUPIL R STATIC R GRIP SCORE 0/1 SCORE 0/1 SCORE 0i1 SCORE 011 SCORE 0/1
GROUP 4(1000 MG/KG) 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 -I'08Oct08 12h22

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 MOTOR ACTIVITY TEST MALES WEEK 4
Counts persample 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 556 766 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 High 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(50MG/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 98 246 216 111 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 16t 742 557 596 561 812 131 261 340 554 589 880 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 418 195 158 466 314 488 273 129 295 943 562 306 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 161 138 267 313 258 182 221 104 93 139 110 81 2067 25 LOW 341 439 402 513 397 223 512 251 211 326 354 218 4187 GROUP 4(500 MG/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 43 280 375 345 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 BLUE 61 NOTOX Project 488231 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 98 266 88 131 112 1733 43 LOW 1748 3127 3380 1556 1001 1599 2702 689 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 17896 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 72 666 228 235 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 91 594 315 427 981 7 244 118 354 760 552 5485 GROUP 3(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 GROUP4(500 MG/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

-Page 25-

### C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 HAEMATOLOGY MALES END OF TREATMENT
ANIMAL WBC Neutrophils Lymphocytes Monocytes EosinophIs 10E9/L %WBC %WBC %WBC %WBC
GROUP I(CONTROL) 1 5.2 1 9.0 761 2.2 2.3 2 6.9 14.1 828 1.9 0.7 3 9.2 133 82.6 2.8 1.0 7 6.9 13.2 835 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 85.5 1.7 0.6 14 8.1 16.0 81.0 2.2 0.5 15 7.9 15.7 80.2 2.7 1.2 16 8.7 10.7 856 2.6 0.7 GROUP 3(250 MG/KG) 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 1.7 0.9 25 10.4 12.4 84.8 1.7 0.7 GROUP 4(1000 MG/KG) 32 10.0 8.0 89.5 1.6 05 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 0.5 37 6.7 9.9 87.6 1.7 0.6 

### MALES END OF TREATMENT
ANIMAL Basophils Red blood cells Reticulocytes RDW Haemoglobin %WBC 10E12/1. %RBC %mmol/L

GROUP I (CONTROL) 1 0.3 7.43 2.8 132 9.0 2 0.5 842 3.3 13.6 10.1 3 0.2 8.57 2.6 11.8 9.7 7 0.3 8.05 2.7 13.0 9.7 10 0.5 8.06 2.8 11.7 10.1 GROUP 2(50MG/KG) 11 0.6 8.09 27 13.2 9.6 12 0.4 9.04 2.6 12.7 102 14 0.3 9.36 19 12.1 10.6 15 0.3 8.24 2.6 12.7 10.0 16 0.3 8.61 3.8 172 90 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 12.6 104 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 llh1O

### C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 HAEMATOLOGY MALES END OF TREATMENT
ANIMAL Basophils Red blood cells Reticulocytes RDW Haemoglobin %WBC 10E12/L %RBC %mmolIL
GROUP4(1000 MGIKG) 32 0.4 8.48 2.8 12.4 9.5 34 0.4 8.83 2.5 12.9 10.1 35 0.6 8.89 2.3 12.8 9.9 36 03 8.25 2.2 23.9 9.9 37 0.1 8.99 20 12.9 9.9 

### MALES END OF TREATMENT
ANIMAL Haematocrit MCV MCH- MOHO Platelets L/L ft. final mmol/L 1 0E9/L

GROUP I(CONTROL) 1 0.382 51.5 121 23.44 699 2 0.438 52.0 1.20 23.03 985 3 0.447 52.2 114 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 GROUP3(250 MGIKG) 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 MGIKG) 32 0.441 52.0 1.12 21.61 988 34 0.460 52.1 1.14 2192 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 5 5
GROUP I (CONTROL) 2 17.5 19.9 3 19.5 19.4 7 18.3 18.8 10 16.2 17.4

-Page 27 OW~29Sep08 llhlO

### C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 HAEMATOLOGY MALES END OF TREATMENT
ANIMAL PT APTT sS GROUP 2(50 MG/KG) 11 16.7 183 12 18.4 21.4 14 19.1 22.6 15 18.4 21.5 16 17.9 19.6 GROUP 3(250 MG/KG) 21 16.7 16.2 22 17.9 20.7 23 17.6 22.3 24 170 16.2 25 176 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 OFTREATMENT
ANIMAL WNBC Neutrophils Lymphocytes Monocytes Eosinophils 10E9/L /%WBC %WBC %WBC %WBC

GROUP I(CONTROL) 42 3.3 29.2 68.2 1.8 0.3 43 47 12.0 84.0 3.0 1.0 47 4.3 22.8 73.5 2.0 1.6 48 37 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 45.0 2.0 0.0 57 3.5 21.0 77.0 2.0 0.0 58 2.9 25.4 709 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 53 23.5 73.4 1.6 1.4 67 2.2 30.0 66.0 3.0 1.0 GROUP 4(1000 MG/KG) 73 52 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

-Page 28

- 

## ~29Sep08 lihiG

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 HAEMATOLOGY FEMALES END OF TREATMENT
ANIMAL Basophils Red blood cells Reticulocytes RDW Haemoglobin %WBC 10E12/L %RBC %mmolIL 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(50MG/KG) 52 0.3 7.42 4.1 12.6 9.3 55 0.1 7.81 3.6 12.3 9.2 56 0.0 706 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 7.6 172 8.4 63 0.4 7.58 4.3 13.7 9.2 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 MG/KG) 73 0.2 7.91 4.3 12.1 9.3 74 0.0 7.66 5.7 14.9 88 75 0.0 7.50 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 OFTREATMENT
ANIMAL Haematocrit MCV MCH MCHC Platelets L/L ft. final mmol/L 1 0E9/L

GROUP I(CONTROL) 42 0.377 52.9 1.27 24.01 1102 43 0.396 53.9 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 50 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 53.0 1.18 22.28 1328 56 0.395 55.9 1.30 2330 1443 57 0.405 55.1 1.28 23.17 1197 58 0.415 50.1 1.12 22.43 1262 GROUP 3 (250 MG/KG) 61 0.361 527 1.22 23.18 1650 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 0391 52.2 1.21 23.12 1022

-Page 29 NW- 29Sep08 1lhi0

### C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 HAEMATOLOGY FEMALES END OF TREATMENT
ANIMAL Haematocrit MCV MCH MCHC Platelets LJL fl- final mmolIL 10E9/L
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 0.414 535 1.20 22.4283 

### FEMALES END OF TREATMENT
ANIMAL PT APiT S S
GROUP I(CONTROL) 42 17.7 1 9.4 43 16.8 16.0 47 -- 48 16.7 19.9 50 17.2 15.4 GROUP 2(50 MGIKG) 52 16.8 22.0 55 17.4 21.9 56 16.1 14.8 57 15.9 16.9 58 16.4 16.2 GROUP 3(250 MG/KG) 61 16.6 18.8 63 16.2 16.0 64 17.3 170 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 -I 29Sep08 I1hlO

### C.l. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 CLINICAL BIOCHEMISTRY MALES END OFTREATMENT
ANIMAL ALAT ASAT ALP Total protein Albumin UIL UI/L UIL gIL gIL

GROUP I(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) 11 409 851 197 61.7 32.6 12 56.3 79.3 187 68.0 34.5 14 655 77.7 195 65.3 336 15 33.6 73.2 142 65.6 33.5 16 636 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 62.4 33.3 25 45.6 81.4 135 65.4 34.7 GROUP 4(1000 MVG/KG) 32 47.1 82.2 120 649 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 bilirubin Urea Creatinine Glucose Cholesterol umoilL mmolIL umol/L mmollL mmolIL
GROUP I(CONTROL) 1 2.6 6.7 42.1 705 1.48 2 1.9 6.4 40.6 1052 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 

## S 29Sep08ll1111-MEW



### C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 CLINICAL BIOCHEMISTRY MALES END OF TREATMENT
ANIMAL Total bilirubin Urea Creatinine Glucose Cholesterol umnol/L mmol/L umol/L mmol/L mmolIL
GROUP 4(1000 MG/KG) 32 2.1 78 43.6 9.76 2.10 34 2.1 8.3 44.3 6.36 1.72 35 2.0 8.2 41.3 7.61 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 mmolIL

GROUP11CONTROL) 1 142.1 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 MGIKG) 11 142.0 3.88 104 2.64 1.94 12 142 1 4.22 104 2.76 2.08 14 142.5 4.18 104 2.66 171 15 142.6 3.88 104 2.73 2.10 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 2.16 23 142.1 3.71 105 2.77 2.08 24 1421 3.80 105 2.61 2.10 25 143.5 3.79 106 2.76 2.15 GROUP 4(1000 MGIKG) 32 142.6 4.25 104 2.69 1.94 34 143.5 3.88 104 2.77 2.08 35 146.2 3.90 104 2.72 2.21 36 141.9 4.37 106 2.77 2.09 37 142.6 3.77 105 2.72 1.88 

### FEMALES END OF TREATMENT
ANIMAL ALAT ASAT ALP Total protein Albumin U/L U/L UI/L g/L g/L
GROUP 1(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 64 657 317

-Page 32 -I 29SepO8llhlO1

### C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 CLINICAL BIOCHEMISTRY FEMALES END OF TREATMENT
ANIMAL ALAT ASAT ALP Total protein Albumin UI/L U/1- U/1- g/L g/L
GROUP 2 (50 MGIKG) 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 29.5 GROUP 3(250 MG/KG) 61 724 74.6 86 69.2 35.4 63 58.7 83.2 61 68.2 34 1 64 77.5 76.2 87 67.3 32.5 65 65.1 70.1 76 64.1 328 67 59.4 96.3 39 69.5 34.1 GROUP 4(1000 MVG/KG) 73 38.8 62.5 36 64.3 31.7 74 42.3 769 113 62.6 31.8 75 554 72.1 120 66.5 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 mmol/L umol/L mmolIL mmol/L
GROUP I(CONTROL) 42 3.0 7.8 46.5 6.56 1.35 43 2.1 9.3 50.0 4.95 1.14 47 2.0 11.3 45.4 8.26 1.58 48 2.3 7.1 41.7 5.63 1.48 50 2.4 11.4 50.0 7.02 1.69 GROUP 2(50 MG/KG) 52 2.4 9.7 47.6 6.46 2.09 55 1.5 9.5 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 1.80 65 3.1 9.5 47.2 4.49 1.87 67 1.7 9.7 54.2 6.69 1. 59 GROUP 4(1000 MG/KG) 73 1.6 11.4 49.8 5.10 1.65 74 1.5 9.9 50.0 5.94 1.33 75 17 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 lihiC

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 CLINICAL BIOCHEMISTRY FEMALES END OF TREATMENT
ANIMAL Sodium 

### Potassium Chloride Calcium lnorg.Phos mmolIL mmol/L mmol/L mmol/L mmol/L
GROUP I(CONTROIL) 42 142.2 3.64 96 2.76 2.64 43 144.3 3.56 100 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 3.63 98 2.70 1.71 55 141.9 400 101 2.82 2.61 56 141.2 3.68 100 2.72 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 MGIKG) 61 140.3 3.49 96 2.88 2.63 63 140.1 3.65 97 2.91 2.47 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 MGIKG) 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

-Page34
- W* 29Sep08 I1hlo

### C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 MACROSCOPIC FINDINGS MALES ALL NECROPSIES
ANIMALORGAN FINDING DAY OF DEATH
GROUP I(CONTROL) INo findings noted Scheduled sacrifice, 17JuI2008 2 No findings noted Scheduled sacrifice, 17,12008 3 Bone Tail, tail bent. Scheduled sacrifice, 17JuI2008 4 Nofindings noted Scheduled sacrifice, 1 7Jul2008 5 No findings noted Scheduled sacrifice, 1 7JuI2008 6 No findings noted Scheduledsacrifice, i17JuI2008 7 No findings noted Scheduledsacrifice, 17JuI2008 8 No findings noted Scheduled sacrifice, 17JuI2008 9 No findings noted Scheduled sacrifice, 17.102008 10 No findings noted Scheduled sacrifice, 1 7Jul2008 GROUP 2(50 MGIKG) 11 No findings noted Scheduledsacrifice, 17JuI2008 12 Mesenteric Lnode Discolouration, greenish. Scheduled sacrifice, 17JuI2008 13 No findings noted Scheduledsacrifice, 1 7Jul2008 14 No findings noted Scheduled sacrifice, 1 7Ju12008 15 Skin Tail: discolouration, bluish. Scheduled sacrifice, 17Jul2008 18 Mesenteric Inode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 17 No findings noted Scheduled sacrifice, 17Jul20C18 18 No findings noted Scheduledsacrifice, 17Jul2008 19 No findings noted Scheduledsacrifice, 17Jul2008 20 No findings noted Scheduledsacrifice. 17Jul2008 GROUP 3 (250 MG/KG) 21 Mesenteric Lnode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 22 Epididymides Rightside, tail: nodule(s), d=7x5 mm. Scheduled sacrifice. 17JuI2008 Yellowish, hard. Mesenteric Lnode Discolouration, greenish 23 Mesenteric Lnode Discolouration, greenish. Scheduled sacrifice, 17Ju12008 24 Mesenteric Lnode Discolouration, greenish. Scheduledsacrifice, 17Jul2008 25 Mesenteric Lnode Discolouration, greenish. Scheduled sacrifice, 17,102008 26 No findings noted Scheduled sacrifice, 1 7Jul2008 27 Epididymides Rightside, tail: nodule(s), d=2x2 mm, Scheduled sacrifice, 17Jul2008 Yellowish, hard. Mesenteric Inode Discolouration, greenish 28 Mesenteric Lnode Discolouration, greenish Scheduled sacrifice, 17Jul2008 29 Mesenteric Lnode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 30 Mesenteric Inode Discolouration, greenish Scheduled sacrifice, 17.102008 GROUP 4(1000 MG/KG) 31 MesentericLnode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 32 Mesenteric Inode Discolouration, greenish. Scheduled sacrifice, 17.102008 33 Mesenteric Inode Discolouration, greenish. Scheduled sacrifice, 1 7Jul2008 34 Mesenteric Inode Discolouration, greenish. Scheduled sacrifice. 1 7Jul2008 Skin Foreleg, left side: alopecia. 35 Mesenteric Inode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 36 Mesenteric Inode Discolouration, greenish Scheduled sacrifice, 17Jul2008 37 Mesenteric Lnode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 38 Mesenteric Inode Discolouration, greenish. Scheduled sacrifice, 17Jul2008 39 Mesenteric Inode Discolouration, greenish Scheduled sacrifice, 17Jul2008 Skin Tail, discolouration, bluish. 40 Seminal vesicles Left side: reduced insize. Scheduled sacrifice, 1 7Jul2008 Mesenteric Inode Discolouration, greenish.

-Page 35

- 

### I-29SepO8 1111111

### C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 MACROSCOPIC FINDINGS FEMALES ALL NECROPSIES
ANIMAL ORGAN FINDING DAY OF DEATH
GROUP I(CONTROL) 41 No findings noted Scheduled sacrifice, 29JuI2008 42 Nofindings noted Scheduled sacrifice, 28Ju12008 43 Nofindings noted Scheduled sacrifice,28Jul2008 44 Nofindings noted Scheduled sacrifice, OlAug2008 45 Nofindings noted Scheduled sacrifice, OlAug2008 46 Nofindings noted Scheduled sacrifice, OlAug2008 47 Mandibular Inode Both sides: discolouration, dark red. Scheduled sacrifice, 30Jul2008 Skin Foreleg, leftside: alopecia. Abdominal region: alopecia. Foreleg, right side: alopecia. Flank, rightside: alopecia. Flank, leftside: alopecia. 48 Nofindings noted Scheduled sacrifice, 30JuI2008 49 Nofindings noted Scheduled sacrifice, 30Jul2008 50 Nofindings noted Scheduled sacrifice, 28Jul2008 GROUP 2(50 MGIKG) 51 No findings noted Scheduledsacrifice, 31Jul2008 52 Mandibular Inode Both sides: discolouration,reddish. Scheduled sacrifice, 30Jul2008 53 No findings noted Scheduled sacrifice, 01Aug2008 54 No findings noted Scheduled sacrifice, 29Jul2008 55 No findings noted Scheduled sacrifice, 30Ju12008 56 Pancreas Discolouration, reddish. Scheduledsacrifice, 28Jul2008 Mesenteric Inode Discolouration, bluish. Mandibular Lnode Both sides: discolouration,dark red. 57 No findings noted Scheduled sacrifice, 30Jul2008 58 No findings noted Scheduledsacrifice, 30Jul2008 59 No findings noted Scheduled sacrifice, 30.Jul2008 60 No findings noted Scheduled sacrifice, 30Jul2008 GROUP 3(250 MGIKG) 61 Mesenteric Inode Discolouration, bluish. Scheduled sacrifice, 28Jul2008 62 Nofindings noted Scheduled sacrifice, 31Jul2008 63 Mesenteric Lnode Discolouration, bluish. Scheduledsacrifice, 28Jul2008 64 Mesenteric Lnode Discolouration, bluish. Scheduled sacrifice, 28Jul2008 65 Mesenteric Inode Discolouration, bluish. Scheduled sacrifice, 28Jul2008 66 Nofindings noted Scheduled sacrifice, 29JuI2008 67 Mesenteric Inode Discolouration, bluish. Scheduled sacrifice, 3OJul2008 68 Mesenteric Inode Discolouration, bluish. Scheduled sacrifice, 30Jul2008 69 Mesenteric Inode Discolouration, bluish. Scheduled sacrifice, 30Jul2008 70 Mesenteric Lnode Discolouration, bluish Scheduled sacrifice, 31Jul2008 GROUP 4(1000 MGIKG) 71 General observationsG.i-tractus, contents: bluish. Scheduled sacrifice, 01Aug2008 Mesenteric Inode Discolouration, bluish. Skin Foreleg, rightside: alopecia. Hindleg, leftside: alopecia. Hindleg. right side: alopecia. Foreleg, left side: alopecia. 72 General observationsG.i-tractus, contents: bluish. Scheduled sacrifice, 29Jul2008 73 General observationsGi-tractus, contents: bluish. Scheduled sacrifice, 30Jul2008 Mesenteric Inode Discolouration, bluish. 74 General observationsGi-tractus, contents: bluish. Scheduled sacriffice, 28JuI2008 Adrenal glands Both sides: discolouration, red-brown. Mesenteric Lnode Discolouration, bluish. Mandibular Lnode Both sides: discolouration, reddish. 75 Mesenteric Lnode Discolouration, bluish. Scheduled sacrifice, 28Jul2008 76 General observationsG.i-tractus, contents: bluish. Scheduled sacrifice, 29Jul2008 Mesenteric Inode Discolouration, bluish. 77 Mesenteric Lnode Discolouration, bluish. Scheduled sacrifice, 28Jul2008 78 General observationsG.i-tractus, contents: bluish. Scheduled sacrifice, OlAug2008 Mesenteric Inode Discolouration, bluish. 79 Mesenteric Inode Discolouration, bluish. Scheduled sacrifice, 31Jul2008

- Page 36

- MW 29Sep08 IWhO

### 0.1. PIGMENT BLUE61 NOTOX Project488231 APPENDIX 2 MACROSCOPIC FINDINGS FEMALES ALL NECROPSIES
ANIMALORGAN FINDING DAY OF DEATH
GROUP 4(1000 MVG/KG) 80 General observationsGi-tractus, contents: bluish. Scheduled sacrifice, 31Jul2008 Mesenteric Lnode Discolouration, bluish.

-Page 37

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## S-29Sep08 llhlO

### C.I. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2
ORGAN WEIGHTS (GRAM) MALES NECROPSY
ANIMAL BODY W. BRAIN HEART LIVER TH-YMUS (GRAM) (GRAM) (GRAM) (GRAM) (GRAM)
GROUP I(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 7.25 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) 11 341 199 1.019 7.54 0.279 12 332 2.04 0.966 7.81 0.444 

# 913 337 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(250 MG/KG) 21 379 2.11 1.091 10.70 0.425 22 361 1.95 1.055 9.34 0.434 23 .340 2.08 0.889 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 0.337 34 340 2.10 0936 8.15 0.287 35 342 2.03 0941 9.10 0.347 36 352 --- --- ---- 37 343----- - 38 350------- - 39 348------- - 40 341 --- ----- -

-Page 38 -3 29Sep08 IlhIO

### 0.1. PIGMENT BLUE 61 NOTOX Project 488231 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 0.051 0.504 3.32 1.254 3 2.46 0.077 0.709 343 1.238 4 2.34 0.082 0434 3.45 1.163 5 2.43 0.066 0503 367 1.153 6 ------- 3.30 1.001 7 ----- 308 1.027 8 --- --- -- 336 1.187 9 - -- -- -329 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 0.586 347 1.067 13 2.46 0.071 0.540 2.60 0.969 14 2.21 0.052 0.495 3.40 1.206 15 2.42 0.073 0.703 3.24 1.095 16 --- ---- 266 0.951 17 - -- -- -3.67 1.177 18 - -- -- -3.21 1.183 19 --- ---- 3.64 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 2.36 1.079 26 ---- 3.33 1.078 27 ----- -329 1.323 28 ---- 3.40 1.195 29 - -- -- -3.47 1.283 30---------- GROUP 4(1000 MG/KG) 31 2.65 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 

# S35 2.17 0.077 0.625 3.38 1.165 36 --- --- -- 3.47 1.195 37 ------ 378 1.266 38 --- --- 2.61 0.959 39 ------- 346 1.273 40 ---- --- 344 1.143

-Page 39

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## ~29Sep08 lihiC

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 ORGANIBODY WEIGHT RATIOS(% 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 5 344 061 0.256 2.39 0.101 6 401 ------ -- - 7 345------- - 8 373------- - 9 346------- - 10 383------- - GROUP 2(50 MGIKG) 11 341 0.58 0.299 2.21 0.082 12 332 0.61 0,291 2.35 0.134 13 337 0.58 0.272 2.51 0.107 14 320 063 0.275 2.31 0.088 15 365 0.56 0.255 271 0.131 16 335------- - 17 398----- - 18 336----- - 19 335------- - 20 395------- - GROUP 3(250 MGIKG) 21 379 0.56 0.288 2.82 0112 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 MGIKG) 31 363 0.59 0.294 282 0.090 32 348 0.60 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

- 29Sep08 llhlO

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 ORGANIBODY WEIGHT RATIOS(% MALES NECROPSY
ANIMAL KIDNEYS ADRENALS SPLEEN TESTES EPIDIDYMIDES

GROUP I(CONTROL) 1 0.69 0019 0.179 0.95 0.323 2 0.77 0.016 0.159 1.05 0397 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 0335 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 MGIKG) 11 074 0.023 0.184 1.14 0.350 12 0.72 0.019 0177 1.05 0.321 13 0.73 0.021 0.160 0.77 0,288 14 0.69 0.016 0.155 1.06 0377 15 0.66 0.020 0.193 0.89 0.300 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 MGIKG) 21 0.67 0.017 0.183 0.98 0.322 22 0.65 0016 0.184 0.98 0.400 23 0.69 0.021 0.190 0.99 0362 24 0.69 0.020 0.182 0.93 0.291 25 0.65 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(1000 MG/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 1.05 0.327 34 0.68 0.019 0.199 095 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 ---- 0.99 0.366 40 ---- 1.01 0.335

-Page41

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## ISW 29SepO8111hlO

### C.I. PIGMENT BLUE61 NOTOX Project488231 APPENDIX 2 ORGAN WEIGHTS (GRAM) FEMALES NECROPSY
ANIMAL BODY W BRAIN HEART LIVER THYMUS (GRAM) (GRAM) (GRAM) (GRAM) (GRAM)
GROUP II(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 6.73 0.161 56 223 1.85 0.727 7.54 0.207 57 207 1.83 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 64 227 178 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 MGIKG) 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 0165 78---------- 79---------- 80----------

- Page42
- M*- 29Sep08 llhIO

### CI. 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 MGIKG) 51---- 52 1.65 0.075 0.391 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 MGIKG) 61 1.69 0,077 0693 62------ 63 1.88 0.096 0479 64 1.70 0.080 0.543 65 1.71 0.076 0.497 66-- 67 1.71 0.079 0.478 68-- 69------ 70------ GROUP 4(1000 MGIKG) 71------ 72------ 73 1.44 0.065 0.400 74 1.48 0.100 0.501 75 1.72 0.088 0.535 76 1.68 0.089 0510 77 1.81 0.098 0.559 78------ 79------ 80------

-Page 43 W#-~29Sep08 lihiC

### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 ORGANIBODY WEIGHT RATIOS(% 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 0.365 3.48 0.090 49---------- 50 222 0.87 0.343 3.50 0.052 GROUP 2(50 MG/KG) 51-------- - 52 202 0.87 0.374 3.11 0.075 53---------- 54---------- 55 220 0.81 0332 3.06 0.073 56 223 0.83 0326 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 0333 3.63 0.090 62---------- 63 230 0.81 0.327 3.53 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 0.83 0.349 3.30 0.069 68---------- 69---------- 70---------- GROUP 4(1000 MG/KG) 71 --------- 72---------- 73 211 0.89 0.316 2.91 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 0.104 77 220 0.84 0.339 3.59 0.075 78---------- 79---------- 80----------

-Page~ 44 29Sep08 lihiG

### C.I. PIGMENT BLUE61 NOTOX Project488231 APPENDIX 2 ORGANIBODY WEIGHT RATIOS 

## (%) FEMALES NECROPSY
ANIMAL KIDNEYS ADRENALS SPLEEN 

### N% N% N%
GROUP I(CONTROL) 41------ 42 0.65 0.032 0.186 43 0.63 0.032 0.207 44------ 45------ 46------ 47 0.74 0.036 0.155 48 0.91 0.038 0.249 49---- 50 0.74 0.038 0.215 GROUP 2(50 MGIKG) 51---- 52 0.82 0.037 0.194 53------ 54------ 55 0.74 0.038 0.216 56 0.73 0.043 0.236 57 0.74 0.037 0.243 58 0.68 0,035 0.167 59------ 60------ GROUP 3(250 MGIKG) 61 0.75 0.034 0.308 62------ 63 0.82 0.042 0.208 64 075 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 MGIKG) 71------ 72------ 73 0.68 0.031 0190 74 060 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 OW~29Sep08 llhlOLA



### C.I. 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 GROUP1I(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 14 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 25 3.3 0.0 0.3

-Page 46

- 010 23Oct08 16h12

### C.I. PIGMENT BLUE 61 NOTOX Project488231 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 MGIKG) 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 5i5 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 5.3 6.1 11.4 0.0 0.1 ST.DEV. 0.4 0.0 0.0 1.6 0.9 1.6 0.0 0.3

-Page 47

- 230ct0816h12

### C.I. PIGMENT BLUE 61 NOTOX Project488231 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 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 1 5 0 0 64 2 1 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 0.1 5.6 5.7 11.2 0.0 0.0 ST.DEV. 0.3 0.0 0.3 2.1 2.4 3.9 0.0 0.0

-Page 48

- 

## ~23Oct08 16h12

### C.I. 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 4(1000 MGIKG) 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 14 0 0 79 21 0 0 5 8 13 0 1 80 21 0 0 8 4 12 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 16h12

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

- 

### LACTATION
LITTER SEX D" 1 D" 4
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 8.9 44 M 5.6 7.9 F 5.7 7.8 M+F 5.6 79 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 5.1 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 6.2 8.5 F 5.9 8.0 M+F 6.1 8.4 GROUP 2(50 MGIKG) 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 10.3 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 55 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 8.2 M+F 5.5 8.4

-Page 50

- 

## ~ 29SepO8 I11h31

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

- 

### LACTATION
LITTER SEX DAY 1 DAY 4
GROUP 2(50 MGIKG) 60 M 5.6 8.6 F 5.0 7.8 M+F 5.2 8.1 GROUP 3 (250 MIGIKG) 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 6.6 10.3 63 M 59 8.3 F 5.4 7.5 M+F 5.7 8.0 64 M 5.8 7.9 F 5.0 7.2 M+F 5.2 74 65 M 5.6 8.0 F 5.6 8.2 M+F 5.6 8.1 67 M 68 10.3 F 6.2 9.8 M+F 6.5 10.1 68 m 6.0 9.4 F 5.8 9.3 MWF 59 93 69 M 6.1 9.5 F 5.7 90 MWF 5.9 9.,2 70 M 6.0 8.8 F 5.4 7.9 M+F 5.7 8.3 GROUP 4(1000 MGIKG) 71 M 6.4 9.1 F 6.2 8.7 M+F 6.3 8.9 73 M 6.0 9.1 F 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 86 F 5.8 8.3 M+F 60 8.5 78 M 6.4 9.2 F 6.4 9.2 M+F 64 9.2

-Page 51 010- 29Sep0811h3l

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

- 

### LACTATION

LITTER SEX DAY 1 DAY 4

GROUP 4(1000 MGIKG) 79 M 5.6 8.0 F 52 7.5 M+F 5.4 7.7 80 M 58 8.9 F 56 8.5 M+F 5.7 8.8

-Page 52 00,~29Sep081I1Ih31

### 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) 42 1 M 6.7 8.6 2 M 6.7 87 3 F 5.7 79 4 F 6.0 8.3 5 F 6.1 84 6 F 5.5 7.4 7 F 6.2 86 8 F 6.9 92 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 95 4 M 6.0 8. 8 5 F 6.0 90 6 F 5.5 7.9 7 F 6.1 8.6 8 F 5.7 8.4 9 F 6.3 9.3 10 F 5.8 8.6 44 1 M 6.0 8.0 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 78 7 M 5.3 7.6 8 M 5.6 83 9 M 4.8 6.6 10 M 5.7 8.3 11 F 5.4 75 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 6.5 9.3 10 F 6.1 8.8 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 6.8 10.9 3 F 6.9 10.8 4 F 6.9 10.3

- Page 53 -I 29Sep08 11h31

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

- 

### LACTATION
LITTER PUP SEX DAYl1 DAY4
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 6.7 6 F 5.1 7.3 7 F 5.0 7.5 8 IF 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 62 8.7 3 M 6.1 8.8 4 M 57 7.9 5 M 5.4 7.9 6 M 5.5 8.2 7 F 5.5 79 8 F 5.3 7.2 9 F 5.8 79 10 F 5.6 8.2 11 F 5.2 7.7 12 F 5.6 79 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 4.5
-- 7 F 5.3 84 8 F 5.2 8.5 9 IF 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 6.2 8.2 7 M 6.2 8.4 8 M 5.9 8.0 9 M 5.7 7.4 10 M 6.6 9.6 11 M 6.0 7.8 12 F 6.5 88 13 F 6.4 8.6 14 F 5.0 6.7 GROUP 2(50 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 92 7 F 6.8 10.8 8 F 4.8
-- 9 F 6.7 10.5

-Page 54

- 

## ~ ~29Sep08 11h31

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

- 

### LACTATION

LITTER PUP SEX DAYlI DAY4

GROUP 2(50 MG/KG) 52 1 M 5.9 8.7 2 M 5.6 8.3 3 M 6.2 8.7 4 M 5.6 8.0 5 F 5.7 8.4 6 F 5.9 8.3 7 F 5.8 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 11.6 3 M 7.0 9.9 4 M 7.8 109 5 M 77 11.1 6 F 7.8 11.0 7 F 7.7 103 8 F 7.1 103 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 5 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 5.6 8.6 56 1 M 6.2 8.3 2 M 6.3 7.9 3 M 6.2 7.9 4 M 6.3 8.0 5 M 6.1 8.3 6 M 64 81 7 M 6.0 8.1 8 M 6.5 8.1 9 F 5.9 8.0 10 F 62 7.9 11 F 6.2 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 8.0 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 53 8.1 10 M 61 8.8 11 M 57 8.4

-Page 55 00~S- 29Sep08 IIh31

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

- 

### LACTATION
LITTER PUP SEX DAYl 1 AY4
GROUP 2(50 MGIKG) 58 1 M 6.0 9.1 2 M 5.7 88 3 M 5.9 8.8 4 M 5.1 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 79 12 F 5.4 82 13 F 5.2 76 59 1 M 5.7 8.8 2 M 6.0 88 3 M 5.6 8.4 4 M 6.0 9.0 5 F 5.9 9.0 6 F 5.2 8.0 7 F 4.9 7.7 8 F 5.4 8.2 9 F 5.8 88 10 F 5.2 7.8 60 1 M 6.2 9.2 2 M 5.8 8.9 3 M 5.1 7.9 4 M 55 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 75 11 F 5.3 8.4 GROUP 3(250 MGIKG) 61 1 M 5.8 8.4 2 M 5.9 78 3 M 5.8 8.1 4 M 5.8 8.2 5 M 6.4 8.7 6 F 6.1 8.0 7 F 5.5 7.6 8 F 5.3 7.4 9 F 5.3 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 2 M 6.1 7.7 3 M 6.4 86 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 86 9 M 5.8 8.2 10 F 5.6 8.7 11 F 5.3 7.1 12 F 5.5 7.7 13 F 4.9 6.4

-Page 56

- , 29Sep08 111131

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

- 

### LACTATION
LITTER PUP SEX DAYl1 DAY4
GROUP 3(250 MGIKG) 14 F 55 7.4 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 6.3 6 F 4.9 70 7 F 51 6.8 8 F 56 8.0 9 F 5.2 7.6 10 F 4.9 7.2 11 F 5.8 8.5 12 F 4.9 7.0 13 F 4.5 64 65 1 M 5.5 8.1 2 M 5.0 7.1 3 M 4.6 6.3 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 6.4 99 7 F 65 10.3 8 F 5.9 9.4 9 F 6.0 9.5 10 F 6.5 10.0 11 F 6.1 9.7 68 1 M 6.2 97 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 8.9 7 F 5.8 9.2 8 F 5.8 93 9 F 5.8 92 10 F 57 9.3 11 F 6.2 9.8 12 F 5.8 8.8 13 F -- --

-Page 57 W*-~ 29Sep08 11h3l

### ClI. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 2 BODYWEIGHTS OF PUPS (GRAM) FO-GENERATION

- 

### LACTATION
LITTER PUP SEX DAY 1 DAY4
GROUP 3(250 MG/KG) 69 1 M 6.0 9.5 2 M 5.8 9.4 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 8.8 5 M 5.8 8.9 6 M 5.7 8.5 

# 07 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 7.8 14 F 4.8 6.9 GROUP4(1000 MG/KG) 71 1 M 6.5 9.4 2 M 6.3 9.1 3 M 6.2 8.9 4 M 65 9.1 5 M 6.6 9.1 6 M 6.7 9.4 7 M 6.1 8.9 8 F 6.0 8.3 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 

# 03 M 5.7 8.5 4 M -- - 5 F 5.6 8.7 6 F 5.8 9.5 7 F 5.6 8.5 8 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 8 F 5.2 7.2 9 F 5.5 8.0 10 F 5.3 7.1 11 F 4.0 5.8 12 F 5.1 7.0

- Page 58

- 

## P~29Sep08 11h3l

### 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 4(1000 MG/KG) 75 1 M 5.8 7.9 2 M 4.9 7.0 3 M 50 7.2 4 M 5.9 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 5.5 78 11 F 5.1 7. 3 12 F 5.3 7. 5 13 F 5.3 7.8 14 F 4.9 6.5 15 F 5.4 7.3 76 1 M 6.2 8.4 2 M 6.1 8.7 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 1 0 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 8.4 6 M 6.4 8.8 7 M 6.3 8.8 8 M 6.4 9.0 9 F 5.6 8.2 10 F 5.8 8.4 11 F 5.9 8.4 12 F 5.7 8.0 78 1 M 7.0 10.1 2 M 6.4 9.3 3 M 6.7 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 10 M

- - I1I F 65 9.2 12 F 67 9.7 13 F 6.1 8.6 14 F 6.4 9.6 15 F 63 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 51 7.0 7 F 5.1 7.8 8 F 56 8.1

-Page 59

- 

## ~29Sep08 11h3l

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

- 

### LACTATION
LITTER PUP SEX D" 1 DAY4
GROUP 4(1000 MG/KG) 9 F 5.1 7.4 10 F 5.0 7.1 11 F 5.2 7.3 1 2 F 55 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 82 5 M 57 9.3 6 M 60 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 11 F 5.8 8.4 12 F 5.6 8.4

-Page 60

- 29Sep08 llh31

### C.I. PIGMENT BLUE61 NOTOX Project488231 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 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 FLC No findings LLC No findings 

# 0 MACRO Nofindings 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 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 LITTER 43 1 M DAY 5 Planned Necropsy FLC No findings 23JUL2008 LLC Nofindings MACRO Nofindings 2 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings 

# 0 MACRO Nofindings 3 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 4 M DAY 5 Planned Necropsy FIC Nofindings LLC Nofindings MACRO Nofindings 5 F DAY 5 Planned Necropsy FLC Nofindings 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 FLC No findings LLC Nofindings MACRO No findings 8 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings

FLC

- FIRST LITTER CHECK, DAY P.P.
- CLINICAL SIGNS. LLC

- LASTLITTER CHECK, MACRO

- MACROSCOPIC FINDINGS -Page 61
- 00 240ct:081I5h47

### 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 DAYS5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 10 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings LITTER 44 1 M DAY 5 Planned Necropsy FLC Nofindings 27JUL2008 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 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 PLC No findings LLC No findings MACRO No findings 6 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 7 M DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 8 M DAYS5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 9 M 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 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 12 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 13 P DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 14 F DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 15 P DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings LITTER 45 1 M DAY 5 Planned Necropsy PLC No findings 27JUL2008 LLC No findings MACRO No findings 2 M DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings 3 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings

0 PLC

- FIRST LIT-TER CHECK, DAY P.P.

- CLINICAL SIGNS, LLC

- LAST LITTER CHECK, MACRO

- MACROSCOPIC FINDINGS -Page 62

- *, 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 DELtIVERY GROUP1I(CONTROL) 4 M DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 5 F DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 6 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO Nofindings 7 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 8 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 9 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 10 F DAYS5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings I1IF DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings 12 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 13 F DAY 5 Planned Necropsy PLC No findings LLC Nofindings 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 Nofindings LLC Nofindings MACRO Nofindings 4 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings LITTER 47 1 M DAY 5 Planned Necropsy PLC Nofindings 25JUL2008 LLC Nofindings MACRO Nofindings 2 M DAYS5 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 F DAY 5 Planned Necropsy PLC 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 No findings MACRO No findings

FLC
- FIRST LITTER CHECK, DAY P.P.

- CLINICAL SIGNS, LLC

- LAST LITTER CHECK, MACRO

- MACROSCOPIC FINDINGS

- Page63

- )ON 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 1(CONTROL) 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 finoings 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 LITTER 48 1 M DAY 5 Planned Necropsy FLC Nofindings 25JUL2008 LLC Nofindings MACRO Nofindings 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 No findings LLC No findings MACRO No findings 6 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 7 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 8 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 9 F DAYS5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 10 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 11 F DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 12 F DAY5 Planned Necropsy FLC No findings 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 FLC Nofindings LLC Nofindings MACRO Nofindings LITTER 49 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

FLC

- FIRST LITTER CHECK, DAY P.P.

- CLINICAL SIGNS, LLC

- LAST LITTER CHECK, MACRO

- MACROSCOPIC FINDINGS

- Page 64
- 0- 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 GROUP1I(CONTROL) 3 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 4 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 5 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 6 F DAY 2 Missing FLC No findings 7 F DAY 5 Planned Necropsy PLC Nofindings 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 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 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 LITTER 50 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 No findings LLC No findings MACRO Nofindings 4 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 5 M DAYS5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 6 M DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 7 M DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 8 M DAYS5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 9 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 10 M DAY5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 11 M DAY5 Planned Necropsy PLC No findings LLC No findings MACRO No findings

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- LAST LITTER CHECK, MACRO

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

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### LACTATION
LITTER PUP END OF PP.PHASE FINDINGS DELIVERY
GROUP II(CONTROL) 12 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 13 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 14 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings GROUP 2(50 MGIKG) LITTER 51 1 M DAY 5 Planned Necropsy FLC No findings 26JUL2008 LLC Nofindings 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 No findings LLC No findings MACRO No findings 6 F DAYS5 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 2 Missing FLC Small 9 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings LITTER 52 1 M DAY5 Planned Necropsy FLC Nofindings 25JUL2008 LLC Nofindings MACRO Nofindings 2 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 3 M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 4 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 5 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 6 F DAY5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 7 F DAYS5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 8 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings

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### LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 2(50 MGIKG) 9 F DAY 5 Planned Necropsy FLC Na findings LLC Nofindings MACRO Nofindings 10 F DAY 5 Planned Necropsy FLO No findings LLC No findings MACRO No findings 11 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings LITTER 53 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 Nofindings MACRO Nofindings 5 M DAY 5 Planned Necr-opsy FLC Nofindings LLC Nofindings MACRO Nofindings 6 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 7 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings 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 No findings LLC No findings MACRO Nofindings 11 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings LITTER 55 1 M DAY 5 Planned Necropsy FLC Nofindings 25JUL2008 LLC Nofindings MACRO Nofindings 2 M DAY 5 Planned Necropsy FLC; Nofindings LLC No findings MACRO No findings 3 M DAYS5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 4 M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 5 M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 6 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings

FLC

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- LAST LITTER CHECK, MACRO

- MACROSCOPIC FINDINGS

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

- 

### LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 2(50 MG/KG) 7 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 8 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 9 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 10 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 11 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 12 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 13 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings LITTER 56 1 M DAY 5 Planned Necropsy FLC No findings 23JUL2008 LLC Nofindings MACRO No findings 2 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy FLO No findings LLC Nofindings 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 No findings MACRO No findings 7 M DAY 5 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 Nofindings LLC No findings MACRO No findings 11 F DAYS5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 12 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 13 F DAY5 Planned Necropsy FLO No findings LLC No findings MACRO No findings

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### 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 MG/KG) 14 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings LITTER 57 1 M DAY 5 Planned Necropsy FLC Red spot nose 25JUL2008 DAY 2 Red spot nose LLC Nofindings MACRO Nofindings 2 M DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 3 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 4 F DAY 5 Planned Necropsy FLO Nofindings LLC No findings 

# 0 MACRO No findings 5 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 6 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 7 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings 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 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 11 M DAY 5 Planned Necropsy FLC Red spot nose DAY 2 Red spot nose LLC No findings MACRO No findings LITTER 58 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 Nofindings MACRO Nofindings 3 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 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 Nofindings LLC No findings MACRO No findings 7 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings

FLC

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- LAST LITTER CHECK, MACRO

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### LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 2(50MG/KG) 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 No findings LLC No findings MACRO No findings 11 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 12 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 13 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings LITTER 59 1 M DAY 5 Planned Necropsy FLC Nofindings 25JUL2008 LLC Nofindings MACRO Nofindings 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 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 FLC No findings LLC No findings MACRO Nofindings 8 F DAYS5 Planned Necropsy FLC Nofindings 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 LITTER 60 1 M DAY5 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 No findings 4 M DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings

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### 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) 5 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 6 F DAYS5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 7 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRD 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 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 GROUP 3(250 MGIKG) LITTER 61 1 M DAYS5 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 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 No findings MACRO No findings 5 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 6 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 7 F DAYS6 Planned Necropsy FLC No findings LLC No findings MACRONo 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 LLC No findings MACRO No findings 11 F DAY 5 PlannedNecropsy FLC No findings LLC Nofindings MACRO No findings LITTER 62 1 M DAY 5 Planned Necropsy FLC Nofindings 26JUL2008 LLC No findings MACRO Nofindings

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- CLINICAL SIGNS, LLC

- LAST LITTER CHECK, MACRO

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### 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) 2 M DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings LITTER 63 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 No findings MACRO No findings 4 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 

# S5 M DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 6 M DAY 5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 7 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 8 M DAY 5 Planned Necropsy PLC Nofindings LLC No findings MACRO Nofindings 9 M DAY 5 Planned Necropsy PLC Nofindings LLC No findings MACRO No findings 10 F DAY 5 Planned Necropsy PLC: No findings LLC No findings MACRO No findings 11 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 12 F 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 PLC Nofindings LLC Nofindings MACRO Nofindings 15 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings LITTER 64 1 M DAY 5 Planned Necropsy PLC No findings 23JUL2008 LLC No findings MACRO No findings 2 M DAY 5 Planned Necropsy PLC Nofindings LLC No findings MACRO No findings 3 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings 4 M DAY5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings

PLC

- FIRST LITTER CHECK, DAY P.P.

- CLINICAL SIGNS, LLC

- LAST LITTER CHECK, MACRO

- MACROSCOPIC FINDINGS

- Page 72

- -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 3(250 MGIKG) 5 F DAY 5 Planned Necropsy FLC Small DAY 2 Small DAY 3 Small DAY 4 Smal LLC Small MACRO Nofindings 6 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 7 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 8 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 9 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO Nofindings 10 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 11 F DAYS5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 12 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 13 F DAYS5 Planned Necropsy FLC Nofindings LLC No findings MACRO Nofindings LITTER 65 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 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 DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 6 M DAYS5 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 F DAYS5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 9 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 10 F DAY5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings

FLC

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- Page 73

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### 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 3(250 MGIKG) 11 F DAYS5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 12 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 13 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 14 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings LITTER 67 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 Nec-ropsy PLC No findings LLC No findings MACRO No findings 4 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 5 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 6 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings 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 No findings 9 F DAYS5 Planned Necropsy PLC Nofindings LLC No findings MACRO Nofindings 10 F DAYS5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 11 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings LITTER 68 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 Nofindings LLC Nofindings MACRO No findings 4 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 5 M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings

PLC

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- LASTLITTER CHECK, MACRO

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### 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 3(250 MG/KG) 6 M 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 Nofindings 8 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings 9 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 10 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 11 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 12 F DAYS5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 13 F DAY 1 Deadat FLC PLC Dead MACROAutolysis, no milk LITTER 69 1 M DAY 5 Planned Necropsy PLC Nofindings 25JUL2008 LLC Nofindings MACRO Nofindings 2 M DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 3 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 4 M DAY 5 Planned Necropsy PLC 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 Nofindings LLC Nofindings MACRO Nofindings 7 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO Nofindings 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 LITTER 70 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 PLC No findings LLC No findings MACRO No findings

PLC

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### C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 INDIVIDUAL PUP DATA P0-GENERATION

- 

### LACTATION
LITTER PUP END OF P.P. PHASE FINDINGS DELIVERY
GROUP 3(250 MGIKG) 4 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 5 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 6 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 7 M 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 

# 09 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 10 F DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 11 F DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 12 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO Nofindings 13 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 14 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings GROUP 4(1000 MGIKG) 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 No findings 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 No findings 5 M DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 6 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 7 M 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

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### 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 4(1000 MG/KG) 9 F DAY 5 Planned Necropsy FLO Nofindings LLC Nofindings MACRO Nofindings 10 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 11 F DAY 5 Planned Necropsy FLC: Nofindings LLC Nofindings MACRO Nofindings 12 F DAY 5 Planned Necropsy FLC No findings 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 FLC: Pale LLC No findings MACRO No findings LITTER 73 1 M DAY 5 Planned Necropsy FLC No findings 25JUL2008 LLC No findings MACRO No findings 2 M DAYS5 Planned Necropsy FLC No findings LLC Nofindings MACRO No findings 3 M DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 4 M DAY 1 Dead at FLC FLC Dead MACRO Autolysis,no milk 5 F DAY5 Planned Necropsy FLC 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 DAYS5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 8 F DAY 5 Planned Necropsy FLC Nofindings 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 LITTER 74 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 No findings 3 M DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO No findings 4 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO No findings

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### 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(11000 MG1KG) 5 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 6 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 7 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 8 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 9 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO Nofindings 10 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 11 F DAY 5 Planned Necropsy FLC Small DAY 2 Small DAY 3 Small DAY 4 Small LLC Small MACRO Nofindings 12 F DAY 5 Planned Necropsy FLC Na findings LLC No findings MACRO No findings LITTER 75 1 M DAY 5 Planned Necropsy FLC Nofindings 23JUL2008 LLC Nofindings MACRO Nofindings 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 Nofindings MACRO Nofindings 5 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 6 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings 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 Nofindings LLC No findings MACRO No findings 9 F DAY 5 Planned Necropsy FLC No findings 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

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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 4(1000 MG/KG) 12 F DAY 5 Planned Necropsy FLO Nofindings LLC Nofindings MACRO Nofindings 13 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 14 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 15 F DAY 5 Planned Necropsy FLO Nofindings LLC Nofindings MACRO Nofindings LITTER 76 1 M DAY 5 Planned Necropsy FLC Nofindings 24JUL2008 LLC Nofindings MACRO Nofindings 2 M DAY 5 Planned Necropsy FLC No findings LLC No findings MACRONo findings 3 M DAY 2 Missing FLC 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 No findings MACRO No findings 7 F DAY 5 Planned Necropsy FLC No findings LLC No findings MACRO Nofindings 8 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 9 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 10 F DAYS5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 11 F DAYS5 Planned Necropsy PLC No findings LLC Nofindings MACRO Nofindings 12 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings 13 F DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings LITTER 77 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 Nofindings 3 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 4 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings

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### 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 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 Nofindings LLC No findings MACRO Nofindings 9 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings 

# S MACRO Nofindings 10 F DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 11 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO Nofindings 12 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings 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 Nofindings LLC Nofindings MACRO Nofindings 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 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 7 M DAYS5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings 8 M DAY5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 9 M DAY5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings 10 M DAY 1 Dead atFLC FLC Dead MACRO Autolysis, no milk 11 F DAY 5 Planned Necropsy PLC No findings LLC No findings MACRO Nofindings 12 F DAY 5 Planned Necropsy PLC Nofindings LLC Nofindings MACRO No findings

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### 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 4(1000 MGIKG) 13 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 14 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 15 F DAY 5 Planned Necropsy FLC Nofindings LLC No findings MACRO No findings LITTER 79 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 DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings 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 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 Nofindings LLC No findings MACRO Nofindings 9 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 10 F DAY 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 

# S12 F DAYS5 Planned Necropsy FLC No findings LLC No findings MACRO No findings 13 F DAY 2 Spontaneous death FLC No findings LLC Dead MACRO Autolyse LITTER80 1 M DAY 5 Planned Necropsy FLC No findings 26JUL2008 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 Nofindings LLC No findings MACRO Nofindings 4 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO No findings

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### 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 4(1000 MG/KG) 5 M DAY 5 Planned Necropsy FLC No findings LLC Nofindings MACRO Nofindings 6 M DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 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 F DAY 5 Planned Necropsy FLC Nofindings LLC Nofindings MACRO Nofindings 

# 010 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

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### C.I. PIGMENT BLUE 61 NOTOX 

## Project488231 APPENDIX 2

CORPORA LUTEA FO-GENERATION GROUP 1 GROUP 2 GROUP 3 GROUP 4 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 Lutea

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 

# SMean 18.0 17.1 15.7 14.3 Stdev. 4.15 2.98 3.28 5.21 N 9 9 9 10

<NP> non-pregnant
*rI Dunnett-test based on pooled variance significantat 5% ()or 1 %(-) level

- Page 83

-

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

IMPLANTATION SITES FO-GENERATION GROUP 1 GROUP2 GROUP 3 GROUP4 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 44 15 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

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

### ()or 1%

### (**) level

- Page 84-

## C.I. PIGMENT BLUE 61 NOTOX Project488231 APPENDIX 2 KEY TO MISSING VALUES AND REMARKS CLINICAL LABORATORY INVESTIGATIONS End ofTe tn_

56

- =Technical 

### errorforWBC 1,47

___ ___=Citrate sample clotted 

### 43, 50, 56,57, 6-7, 74, 

### 1Differential leucocyte countwasalso performed manually because 175,77 1ofatechnicalerror/an abnormal plot inthe automated count andthese resultsare

______________- - - 

## ~reported_____

_______________ 

### 156

___ 

### Manual differentialleucoc te count revealed that1%ofneutrophilswere band cells.
C~mLncl 

### Biochemistry: No Remarks

______ __ _____

-Page 

### 85

-

# APPENDIX 3



# ANALYTICAL REPORT

Author Ir.M.J.C. Brekelmans Completion date analytical study November 25, 2008

Laboratory Protect Identification NOTOX Project 488231 NOTOX Substance 1835911A

-Page 

### Iofi 11

## 0.1. PIGMENT BLUE 61 NOTOX Project488231 1. CONTENTS 1. CONTENTS
...................................................................................... 2 2. REPORT APPROVAL ............................................................................ 3 3. SUMMARY..........................4 4. INTRODUCTION.........................5 4.

1. Preface
...................................................................................... 5 4.2. Aim ofthestudy ............................................................................. 5 5. MATERIALS AND METHODS ................................................................... 5 5.1. Reagents
................................................................................... S 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 Accuracyand homogeneity testforthe formulations preparedfor use during treatment................................................................................ Table 3 Stability testforthe formulations prepared for use during treatment............... 10
FIGURES Figure 1 Chromatograms of a15.1 mg/I test substance 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: n

-Page 

### 3

-

### C.l. PIGMENT BLUE 61 NOTOX 

## Project488231
3. SUMMARY 

## The purpose of this partof thestudy wasto determinethe accuracy ofpreparation, homogeneity and stabilityofthe test substance informulations. Accuracyofpreparation The concentrations analysed intheformulations ofGroup 2,Group 3and Group 4were in agreementwith target concentrations (i.e. mean accuracies between 85% and 115%). Notest substancewas detected inthe Group 1formulations. Homogeneity The formulations of Group 2and Group 4were homogeneous (i.e. coefficientofvariation :5 10%). Stability Formulations of Group2and Group 4were stable when stored at room temperature for at least * 

## ~6hours.

-Page4-

### C.l. PIGMENT BLUE 61 NOTOX 

## Project 488231
4. INTRODUCTION 4.1. 

## Preface Study plan Start 27 June 2008 (analytical study) Completion 01 July 2008 4.2. Aim of the study The purpose oftheanalytical study wasto determine theaccuracy of preparation, homogeneity and stability ofthe testsubstance informulations. 6. MATERIALS AND METHODS 5.1. Reagents Water Tap water purified by aMilli-Q water purification system (Millipore, Bedford, MA, USA) Methanol VWR International, Leuven, Belgium Sodium 1-heptanesulfonate Fluka Chemie, Buchs, Switzerland monohydrate (HSA) Acetic acid Merck, Darmstadt, Germany Dimethyl sulfoxide(DMSO) Merck All reagentswere of analyticalgrade, unless specified otherwise. 5.2. Samples Accuracy, homogeneity and stabilitywere determined forformulations preparedfor use during treatment. Duplicate samples (approximately 500 mg), which weretaken from theformulations using a pipette, wereaccurately weighed intovolumetric flasksof5,25 or50 ml. Fordetermination of accuracy, samples were taken at50% heightorat90%, 50% and 10% height.The latter setof samples wasalso usedforthe determination ofthe homogeneityofthe formulations. For determination ofstability, additional sampleswere taken at 50% heightand stored at room * temperaturefor6hours. Thevolumetric flaskswere filled up to the mark with DMS0. The solutionswere furtherdiluted with methanol byafactorof atleast 100, toobtain concentrationswithin the calibration range.

- 

### Page 5

-

## C.I. PIGMENT BLUE 61 NOTOX Project488231 5.3. Analytical method 5.3.1. Analytical conditions Quantitativeanalysiswas basedon the analytical method validated forthe test substance in NOTOX project488235. Analytical conditions: Instrument Alliance Separation Module 2695 (Waters, Milford, MA, USA) Detector Dual A Absorbance Detector2487 (Waters) Column Symmetry Shield C18, 150 mm x

## 4.6mm id., dp =5 

## pm (Waters) Column temperature 50 0

## C± 

## 10C Injection volume 10 PI Mobile phase A

- 

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

- 

## 5/0.5/94.5 (v/v/v) water/acetic acid/[50/50 (v/v) methanol/waiter] containing 1.2 g/l HSA Grdin C- methanol Time %A %B %C [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 2m1/min UV detection 313 nm 5.3.2. Preparation of thecalibration solutions 

# 0 Stock-andsikingi solutions Stocksolutionsofthetest substancewere prepared inDMS0 atconcentrations of 1108 and 1236 mg/I. Inordertodissolve thetestsubstance the solutionswere ultrasonicatedfor 10 minutes. Calibration solutions Calibration solutions inthe concentration range0.997

- 

## 15.1mg/I were preparedfrom two stock solutions. The endsolution ofthe calibration solutionswas methanol. Procedural recoverysamples Approximately 500mg blankvehiclewasspiked withthe testsubstanceat atargetconcentration of9.66 or 194 mg/g. The accuracysamples were treated similarlyas the testsamples (see paragraph 5.2 'Samples'). 5.3.3. Sample injections Calibration solutionswere injected induplicate.Test samples and procedural recoverysamples were analysed by single injection.

- 

### Page 6

-

## C.I. PIGMENT BLUE 61 NOTOX Project488231 5.4. Electronic data capture System control, data acquisition and dataprocessing were performed usingthe following programme:

- 

## Empower version 5.00 (Waters, Milford, MA, USA). 5.5. Formulas Response (R) Peakarea testsubstance [units]
Calibration curve R=a

## CN + 

## b where: CN = 

## nominal concentration [mg/] a = 

## slope [units x 

## I/mg] b = 

## intercept[units]
Analysed concentration (CA) 

## CA = (R-b)xV 

### xd 

## [mg/g] a w where: w =weight sample [mg] V = 

## volumevolumetric flask[ml] d = 

## dilution factor Recovery SAX100% CN where: CN = 

## nominal concentration [mgtg] Accuracy CAX100% CT where: CT = 

## target concentration [mg/g] Relativedifference (relative duff.) Ct -C0 

## x100% CO where:

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

0 

### = 

## mean concentration ofnon-stored samples [mglg] 5.6. Specifications Preparation offormulations was considered acceptable ifthe mean accuracywas inthe range 85%

- 

## 115% of thetargetconcentration and ifthe coefficientofvariation was _- 10%. Formulations were considered stable ifthe relative difference between the stored and freshlytaken samples was 10%.

-Page 

### 7-

## C.I. PIGMENT BLUE 61 NOTOX Project488231 6. RESULTS 6.1. Calibration curves Acalibrationcurve was constructed using four concentrations. Foreachconcentration, two responses were used.2Linear 

## regression analysis was performed usingthe least squares method with a1/concentration 2 

## weighting factor.The coefficientof correlation (r)was >

## 0.99. Representative chromatograms of atest substance solution and blanksolution are shown in Figure 1. 6.2. Samples 6.2.1. Procedural recovery samples The resultsforthe procedural recovery samples aregiven inTable 1. Mean recoveriesofthe procedural recovery sampleswere 100% and 101%. Becausethe criterion that mean recoveries should be between 85% and 115% was met,the resultsfor thetest samples were accepted. 6.2.2. Testsamples The resultsofthe testsamples are given inTables2and 3. Accuracy of preparation IntheGroup 1formulations, no testsubstancewas detected. The concentrations analysed intheformulations ofGroup 2,Group 3and Group 4were in agreement withtargetconcentrations (i.e. mean accuracies between 99% and 102%). Homogeneity Theformulations ofGroup 2and Group 4were homogeneous (i.e.coefficient ofvariation of2.4% and 0.92%, respectively). Stability * Analysis ofGroup 2and Group4formulations after storageyielded arelative difference of0.79% and 0.35%, respectively. Based on this,the formulations were found to be stable during storage at room temperature for at least 6hours.

-Page8-

### C.I PIGMENT BLUE 61 NOTOX Project 488231 TABLES
Table 1 Procedural recovery samples Date of Date of Target Nominal Analysed Recovery Mean recovery preparation analysis concentration concentration concentration (coefficient of [dd-mm-yy] [dd-mm-yyJ [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 Accuracy and homogeneity testforthe formulations prepared for use during treatment Group Date of Sample Concentration Accuracy Homogeneity analysis position (coefficient of

____[dd-mm-yy] 

### Target Analysed Individual Mean 

## [%I 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. na. Notapplicable.

-Page 

### 9

-

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



## Table 3 Stability testfortheformulations preparedfor useduring treatment

Group Date ofanalysis Analysed concentration Relative duff.

____ 

### [dd-mm-yy] t=0 I__t=6_hours__%_ 

## 2 30-06-08 9.889.6207 4 30-06-08 192 12303

Mean ofsixsamplesatt0 takenat 10%, 50% and 90% height (seeTable 2). 2 

### Mean oftwosamples att-6 hourstakenat 50% height. Individual resultswere 9.90 and 10.0 mg/g. 3 Mean of twosamplesat t-6hours takenat50% height. Individual resultswere 192 and 193 mglg.

-Page 

### 10

-

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

* -'-



## Figure 1 Chromatograms of a15.1 mg/ltestsubstance solution [top; res. id. 1261] and corresponding blank [bottom; res. id. 1297].

- 

### Page 11

-

# ToxPath Consultancy Ltd

APPENDIX 4



# * PATHOLOGY REPORT A Combined 28-Day Repeated Dose Toxicity Studywith the Reproduction/Developmental Toxicity Screening Test of C.l. Pigment Blue 61 inRats byOral Gavage NOTOX Project: 488231 NOTOX Substance 183591/A

DrAM Pilling PO Box 156 Sandy UK SG191WJ Tel: +44 (0)1767 681546 arnpilling~btinternet.corn

### Page 2of97 PATHOLOGY REPORT Testsubstance: C.lI. Pigment Blue61 Notox No.:488231 Testsystem: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor:Color Pigments Manufacturers Association Inc.



## Table ofContents Statement ofCompliance
............................................................ 3 QualityAssurance Statement
........................................................ 4 Summary
.............................................................................. 5 Materials and Methods.................................................................... 6 Results. ..................................................... .............................. 8 Conclusions
................................................................................ 9 *Table 1Incidence all Macroscopic Findings............................................ 10 Table2 Incidence all Microscopic Findings..................I......................... 11 Table 3 IndividualAnimal Data
......................................................... 18

## Page 3of97 PATHOLOGY REPORT Test substance: 

### C.I. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.



## Statement of Compliance The histopathology findings inthis report were compiledby the undersigned pathologist and accurately reflectthe raw data. The phase ofwork inthis report wascarried out incompliancewith the OECD Principles of Good Laboratory Practice, 1997 Revision (ENVIMC/CHEM (98) 17, Paris 1998 and the UK Good Laboratory Practice Regulations 1999 (Statutory Instrument 1999 No 3106) as amended by the Good Laboratory Practice (Codification Amendments etc) Regulations 2004 (Statutory Instrument 2004 No 994).



# 22 ~4J1? DrAM Pilling BVSc PhD RCVS FRCPath Principal Investigator ToxPath Consultancy Ltd PO Box 156 Sandy UK SG19 1WJ

## PATHOLOGY REPORT Pg f9 Testsubstance: C.I. Pigment Blue61 Notox No.: 488231 Testsystem: 28-Day Oral ToxicityStudy intheRat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc. 

## QualityAssurance Statement QualityAssurance surveillance ofthisstudy phase wascarriedout incompliancewiththe OECD Principles ofGood Laboratory Practice, 1997 Revision (ENV/MC/CHEM (98) 17, Paris 1998 and the UK Good Laboratory Practice Regulations 1999 (Statutory Instrument 1999 No 3106) as amended by the Good Laboratory Practice (Codification Amendments Etc) Regulations 2004 (Statutory Instrument2004 No994). The programme includes thefollowing inspections: StudySpecificInspections: Study planand report forthe abovestudy phase are reviewed and itisconfirmed that the report isatrueand accurate reflection ofthe rawdata. Findings are reported to P1, SD, Lead QAU and testfacilitymanagement. ProcessInspections: Processes are inspected atapproximately three monthly intervals, irrespective ofthe study being dealtwith at thetime. Findings are reported to P1. FacilityInspections: Thefacilities and operations ofToxPath Ltd are audited atapproximately six monthly intervals. Findings are reported to P1. Dates ofInspections The following inspections have been carried out overthe period ofthis study phase: Aspect inspected Date ofinspection Date of report StudySpecific Study Plan 4July 2008 4July 2008 Report 17 November2008 17 November 2008 Process Slide reading 18 July2008 24 July2008 Slide receipt/despatch 18 July 2008 24 July 2008 Facility 18 July2008 24 July2008

David JFord, PhD, FRQA Quality Assurance Consultant David Ford Consultancy Goodwyn House Bank Street Pulham Market Diss Norfolk IP21 4TG ENGLAND

### PATHOLOGY REPORT Pg f9 Test substance: 

### C.I. Pigment Blue 61 NotoxNo.: 488231 Test system: 28-Day Oral Toxicity Study intheRat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.


## Summary Histopathological examinations were performed ontissues from Han VWistar rats which had received 0.1. Pigment Blue61 orallyfor aminimum of28 days. Ratswere assigned tofour dosegroups each containing ten F 0 

### males and ten F 0

### females. The test substance was administered by oral gavage oncedaily for aminimum of28 days at doses of50, 250 or 1000 mg/kg. The control group receivedthe vehicle (propylene glycol) alone. Afull necropsywas performed on allanimals. Afull tissuelistwas retained for histopathological evaluation from five rats/sex/group with alimited listbeing retained from the remaining animals. Histopathological evaluation was performed on tissuesfrom the first five males, and thefirstfivefemaleswith liveoffspring, ingroups 1and 4and on macroscopic abnormalities from rats inall groups. Inaddition,the reproductive organsfrom all animals thatfailed to mate, conceive, sireor deliverhealthy offspringwere examined histologically. PAS stained sections oftestis from thefive selected F 0 

### males ingroups 1and 4were examined for abnormalities inspermatogenesis. Histological examination was extendedto the mesenteric lymph nodesofthe selected malesand females ofGroup2. Noanimals died, orwere killed, during thetreatment period. Grossfindings included blue/green discolouration ofthe mesenteric lymph nodes inrats inall dose groups and the presence ofbluegastrointestinal contents inhigh dose females. Noadverse histopathological changeswere observed inassociationwiththe gross findingsor inanyother tissuesofrats receiving thetest substance. On the basisofthe pathological findingsthe NOAEL may be regarded as 1000 mg/kg/day.

### PATHOLOGY REPORT Pg f9 Test substance: C.1. Pigment Blue 61 Notox No.: 488231 Testsystem: 28-Day OralToxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.



### Materials and Methods StudyDesign

Group Dose level Number ofAnimals 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 Test Substance The test substancewas administered orally once dailyfor at least 14 days priorto mating, during mating and uptothe day priorto necropsy at doses of0,50, 250 or 1000 mg/kg. For each animal thetotal exposure periodwas aminimum of28 days. The control group received the vehicle (propylene glycol) alone. NecropsyandHistopathology Atthe end ofthe studyperiod, the ratswere killed byexsanguination, following anaesthesia by isoflurane, and afullnecropsy was performed on allanimals. Afull tissue listwas retained for histopathological evaluation from five rats/sex/group with alimited listbeing retainedfrom the remaining animals. Organs were preserved in10% neutral buffered formalin (testes, epididymides and eyes were initiallyfixed inmodified Davidson's solution before being transferred toformalin). Histopathological evaluation was performed on representativesamples ofthe following tissues from the firstfive males, and thefirstfive femaleswith liveoffspring, ingroups 1and 4: adrenal glands; aorta; brain

- 

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

- 

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

- 

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

- 

### duodenum, jejunum, ileum; spinal cord

- 

### cervical, mid-thoracic, lumbar, spleen; sternumwith bone marrow; stomach; testes; thymus; thyroid with parathyroid; trachea; urinary bladder, uterus; vagina; allgross lesions (examined from all F 0

### rats on study). The reproductive organs (testis, epididymides, coagulating gland, seminal vesicles,prostate, preputial gland, ovaries, uterus, cervix, vagina,clitoral gland) from all animals that failedto mate, conceive, sire ordeliver healthy offspring were alsoexamined (male nos. 1,14, 26 &32; female nos. 41, 54, 66 &72). Histological examination was extended tothe mesenteric lymph nodes of the selected males andfemales of Group2. Following fixation, tissues weretrimmed, processed and embedded inparaffin wax. Sections 2-4 Ipm inthicknesswere prepared and stained with haemnatoxylin and eosin for routine evaluation. Additional sections oftestis from the fiveselected F 0 

### males ingroups 1and 4 were stained with PAS and examined forabnormalities inspermatogenesis.PATHOLOGY REPORT Pg f9 Test substance: ClI. Pigment Blue 61 Notox No.: 488231 Test system: 28-DayOral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc. 

## Data Compilation Details ofstudy design and macroscopic findingswere transferred electronically from the necropsy raw datafiles ofNOTOX B.V. into the computersystem ofToxPath Consultancy Ltd where the microscopic findingswere recorded using on-line input. Macroscopic findingsare summarised inTable 1

## (Incidence ofMacroscopic Findings) and listed inTable 3

## (IndividualAnimalData). Asfaras possible, macroscopic findingswere correlated with amicroscopicfinding. Microscopic findingsare listed for each animal, alongwith the severity grades, inTable 3and summarised inTable 2

## (Incidence ofMicroscopicFindings). Histological changeswere described according to their distribution and morphological character. Findings werealso graded for severity (or degree) on ascale of 1-5 and theseare described inTable3. 

## Archiving Thefinal pathology report (original hardcopy), all glass microscope slides, slideinventory sheets and acopyofthe electronic records (including final pathology report) ascaptured by the appropriate software on CD-ROM will be sentto NOTOX 8Vforarchiving. Anyother non-study related documents and electronic data, covered by GLP, will be archived by ToxPath Consultancy Ltd according to company standard operating procedures.

### Pages8of97 PATHOLOGY REPORT 

### Testsubstance: C.I. Pigment Blue61 Notox No.: 488231 restsystem:28-Day OralToxicityStudy in the Rat 27-Nov-08 Sponsor: ColorPigments Manufacturers Association Inc.
Results Mortality 

## All F 0

## rats survivedthe scheduled duration ofthestudy. Macroscopicfindings Green or blue discolouration ofthe mesenteric lymph nodes was observed inafew group2 rats and inthe majority of rats of both sexes ingroups 3and 4. Bluegastrointestinal tract contents were present inseveral group 4females. Blue discolouration ofthetail skinwas noted inone malegroup 4rat. Nootherfindingswere recorded at necropsy thatwere considered to be dueto the test substance. Microscopic indings Notreatment related histopathological changeswereobserved inthetissues ofany animal receivingC.l. Pigment Blue 61. No abnormalities were noted on evaluation of PASstained sections oftestes of 

## F 0

### males. All seminiferous tubuleswere normalwith respect totheir stage inthe spermatogenic cycleand the integrity ofthe variouscell types present withinthe different stages. Nopathological findingswere present inthe reproductive organs ofmale and female animals thatfailed to produce offspring. Small amounts ofblue material were observed inthe lumen of theduodenum, ileum, jejunum, colon or rectum 

## inafew group 4animals. Blue material was alsonoted inthe superficial skin (keratin layer)ofthe tailinone group 4male. These findingsare considered to represent the presence ofthetest substanceand insome cases correlated withthe macroscopic observations. Nomicroscopic findingswere observed inthe mesenteric lymph nodes to account forthe discolouration ofthesetissues recorded at necropsy. Itisconsideredthatthe discolouration isduetothe presence ofthe test substance and thatthe pigmentwas lostfrom the tissues in the processing orstaining procedures. All other microscopicfindings were consideredto be incidental and unrelated totreatment * withthe test substance.Page 9 of97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue61 Notox No.:488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.



## Conclusions Administration ofC.l. Pigment Blue 61 by oral gavage to Han Wstar rats at doses of50, 250 or 1000 mg/kg/day was associatedwith blue gastrointestinal contents inhigh dosefemales and blue/green discolouration ofthemesenteric lymph nodes in rats inall dose groups. No adverse histopathological changeswere observed inassociation withthe gross findings or inany othertissuesofrats receiving thetest substance. Onthe basis ofthe pathological findingsthe NOAEL may beregarded as 1000 mg/kg/day.

### Page 10 of97 PATHOLOGY REPORT Testsubstance: C.I. Pigment Blue 61 Notox No.:488231 Testsystem: 28-Day OralToxicity Study inthe Rat 27-Nov-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

G tract contents 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 IN SIZE 0 0 0 1 - - - -
SKIN
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 

# 4 indleg 0 0 0 0 0 0 0 1
TAIL

TAIL BENT 1 0 0 0 0 0 0 0Page 11 of97 PATHOLOGY REPORT Test substance: C.l. Pigment Blue61 Notox No.:488231 Test system: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 2
- INCIDENCE OF MICROSCOPIC FINDINGS

The grading systems used are as follows:

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

### Page 12 of 97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue61 Natox No.: 488231 Testsystem: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-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
- - 5 5 - - 5
AORTA

number examined 5 - - 5 5 - - 5
BRAIN numberexamined 5
- - 5 5 - - 5
CAECUM 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 blue material in

lumen present 0

- - 1 0

- - 1

DUODENUM

number examined 5

- - 5 5

- - 5 blue material in

lumen present 0
- - 1 0

- - 0 EPIDIDYMIS

number examined 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 of97 PATHOLOGY REPORT Testsubstance: 

### C.I. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 27-Nov-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
ILEUM numberexamined 5 - - 5 5
- - 5 blue material in

lumen present 0
- - 0 0
- - 1 JEJUNUM

number examined 5 - - 5 5
- - 5 * blue material in lumen present 0
- - 1 0

- - 1
KIDNEYS

number examined 5
- - 5 5
- - 5 basophilic tubules

(minimal) 1

- - 0 0

- - I interstitial

inflammatory cell infiltration (minimal) 1
- - 0 0
- - 0 tubular hyaline droplets (minimal) I - - 3 0
- - 0 tubular mineralization (minimal) 0
- - 0 2
- - I pelvic dilatation

(minimal) 2
- - 1 0
- - 0 

# 0 LIVER number examined 5
- - 5 5 - - 5 parenchymal

inflammatory cell infiltration (minimal) 3

- - 2 1

- - 0 periportal

inflammatory cell infiltration (minimal) 0

- - 1 0

- - 0Page 14 of 97 PATHOLOGY REPORT Testsubstance: 

### C.l. Pigment Blue 61 Notox No.: 488231 Testsystem: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 2

- INCIDENCE OF MICROSCOPIC FINDINGS

SFX: Male Female GROlUP: 1 2 3 4 1 2 3 4
number of animals 10 10 10 10 10 10 10 10
LUNG number examined 5
- - 5 5 - - 5 perivascular

inflammatory cell infiltration (minimal) 2
- - 1 0
- - 1 alveolitis

(minimal) 1
- - 0 0
- - 0
alveolar macrophage 

# 0 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 histiocytosis

(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
- - 1 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) 1I - 0 0 0
- 0 focal acinar atrophy

(minimal) 0

- - 0 1 0
- 0

### Page 15of 97 PATHOLOGY REPORT Testsubstance: 0.1. Pigment Blue 61 Notox No.:488231 Testsystem: 28-Day OralToxicityStudy inthe Rat 27-Nov-05 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
PARTHYROID numberexamined 5
- - 4 5 - - 5
PEYERS PATCHES number examined 5
- - 5 5 - - 5
PITUITARY number examined 5
- - 5 5

- . 5 cyst (s) present 0
- - 1 0

- - 0
PREPUTIAL 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 5 - - 5 blue material in

lumen present 0
- - 1 0 - - 1
SCIATIC NERVE 

# 0number examined 5 - - 4 5 - - 5
SEMINAL VESICLES
number examined 5 1 1 6 - - - reduced luminal

secretory contents present 0 0 0 1 - - -MER

### Page 16 of 97 PATHOLOGY REPORT Test substance: 0.1. Pigment Blue 61 Notox No.:485231 Test system: 28-Day Oral ToxicityStudy inthe Rat 27-Nov.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 

# S hyperplasia (minimal)

- 0
- 0 1. - - 1
flank left side: epidermal hyperplasia (minimal)

- 0
- 0 1

- - 0
blue material in

surface keratin present

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

- 0
- 0 0
- - I SPINAL CORD number examined 5

- - 5 5 - - 5 SPLEEN number examined 5

- - 5 5
- - 5 extramedullary haematopoiesis (slight) 0
- - 0 0
- - 1
STERNUM number examined S - - 5 5
- - 5
STOMACH number examined 5 - - 5 5 - - 5
TAIL number examined 1 - - - - - - - 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: ClI. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 27-Nov-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
THYMUS number examined 5
- - 5 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
VAGIA

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

present - - - - 0 0 1 1 dioestrum present - - - - 5 1 0 5 epithelial vacuolation (Minimal) - - - - 3 0 0 1 (slight) - - - - 1 0 0 3 (moderate)
- - - - 1 0 0 0 Total - - - - 5 0 0 4Page 18 of 97 PATHOLOGY REPORT Test substance:C.I. Pigment Blue 81 Notox No.:488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 1 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 

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

MICROSCOPIC FINDINGS LUNG perivascular inflammatory cell infiltration (minimal)
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, CASCUM, 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.

### Page 19 of97 PATHOLOGY REPORT Test substance: C.l. Pigment Blue 81 Notox No.:488231 Test system: 28-Day OralToxicity Studyin the Rat 27-Nov-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 data of end oftreatment: 16/07/08 data of necropsy : 17/07/08 No abnormality detected
MICROSCOPIC FINDINGS
KIDNEYS pelvic dilatation (minimal)
LUNG perivascular inflammatory cell infiltration (minimal)
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, CAECUN, COAGULATING

GLAND, COLON, DUODENUM, EPIDIDYMIS, HEART, ILEUM, JEJUNUM, 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.Page 20 of 97 PATHOLOGY REPORT Test substance: 

### 0.1. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study in the Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers AssociationInc.

TABLE 3
- INDIVIDUAL ANIMAL DATA

Animal No.: 3 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
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 no lesion to account for gross finding(s)
TESTIS PAS section: no abnormalities present
No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, COAGULATING

GLAND, COLON, DUODENUM, EPIDIDYMIS, HEART, ILEUM, JEJUNUM, LUNG, LYMPH NODE MESENTERIC, OESOPHAGUS, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, PREPUTIAL GLAND, RECTUN, SCIATIC NERVE, SEMINAL VESICLES, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER.Page 21 of97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue 61 Notox No.:488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 4 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 oftreatment: 16/07/08 date of necropsy 17/07/08 No abnormality detected
MICROSCOPIC FINDINGS
LIVER parenchymal inflammatory cell infiltration (minimal)
LUNG alveolijis (minimal)
PANCREAS acinar vacuolation (minimal)
TESTIS PAS section: no abnormalities present
Mo abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, COAGULATING GLAND, COLON, DUODENUM, EPIDIDYMIS, 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 Test substance: 0.1. Pigment Blue61 Notox No.: 488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 27-Nov-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 sacrifice group terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date Of end oftreatment; 16/07/08 date 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 tminimal)

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

GLAND,. COLON. DUODENUM, EPIDIDYMIS, HEART, ILEUM, JEJUNUM, K(IDNEYS, 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 of 97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue61 Notox No.: 488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 6 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 oftreatment: 16/07/08 data of necropsy :17/07/08 Mo abnormality detected
MICROSCOPIC FINDINGS
No tissues examinedPage 24 of 97 PATHOLOGY REPORT Test substance:

### C.l. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 7 SEX: Male DOSE LEVEL: 1
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 oftreatment: 16/07/08 date of necropsy : 17/07/08
No abnormality detected
MICROSCOPIC FINDINGS
No tissues examined

### Page 25 of97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue 61 Notox No.:488231 Test system: 28-Day OralToxicity Studyinthe Rat 27-Nov-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: 17/06/08 date of end of treatment: 16/07/08 date of necropsy : 17/07/08
No abnormality detected
MICROSCOPIC FINDINGS
No tissues examined

### Page26 of97 PATHOLOGY REPORT Testsubstance. 

### C.l. Pigment Blue61 Notox No.: 488231 Testsystem: 28-Day OralToxicityStudy inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 9 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 oftreatment: 16/07/08 date of necropsy : 17/07/08
No abnormality detected
MICROSCOPIC FINDINGS
No tissues examined

### Page27 of 97 PATHOLOGY REPORT Testsubstance: 

### ClI. Pigment Blue61 Notox No.:488231 Testsystem: 28-Day OralToxicity Studyinthe Rat 27-Nov-08 Sponsor: Color Pigments ManufacturersAssociation 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 oftreatment: 16/07/08 date of necropsy : 17/07/08
No abnormality detected
MICROSCOPIC FINDINGS
No tissues examined

### Page 28 of97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-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 oftreatment: 16/07/08 date of necropsy : 17/07108
No abnormality detected
MICROSCOPIC FINDINGS
No abnormality detected
No abnormalities found in: LYMPH NODE MESENTERIC.Page 29 of97 PATHOLOGY REPORT Testsubstance: 

### ClI. Pigment Blue61 Notox No.:488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- INDIVIDUAL ANIMAL DATA

Animal No. - 12 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 oftreatment: 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

### PATHOLOGY REPORTPae3of9 Testsubstance: 0.1. Pigment Blue61 Notox No.: 488231 Testsystem: 28-Day OralToxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments ManufacturersAssociationInc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 13 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 oftreatment; 16/07/08 date of necropsy : 17/07/08
No abnormality detected
MICROSCOPIC FINDINGS No tissues examinedPage 31 of 97 PATHOLOGY REPORT Testsubstance: 

### C.l. Pigment Blue61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 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: 16/07/08 date of necropsy : 17/07/08
No abnormality detected
MICROSCOPIC FINDINGS
LYMPH NODE macrophage aggregates tminimal) MESENTERIC No abnormalities found in: COAGULATING GLAND, EPIDIDYMIS, PREPUTIAL GLAND. PROSTATE, SEMINAL VESICLES, TESTIS.

### Page 32 of 97 PATHOLOGY REPORT 

### Testsubstance: 

### ClI. PigmentBlue 61 Notox No.:488231 Testsystem: 28-Day Oral ToxicityStudy inthe Rat 27-Nov-OS Sponsor:Color Pigments ManufacturersAssociation Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 15 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
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(s)
No abnormalities found in: LYMPH NODE MESENTERIC.

### Page 33of97 PATHOLOGY REPORT Test substance: 

### C.l. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-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 necropsy status :planned terminal date of start of treatment: 17/06/08 date of end oftreatment: 16/07/08 date of necropsy ; 17/07/08
MESENT. LYMPH DISCOLOURiATION 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) MESENTERICPage 34 of97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 1'? 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 oftreatment: 16/07/08 date of necropsy : 17/07/08
No abnormality detected
MICROSCOPIC FINDINGS
No tissues examinedPage 

### 35of 97 

### PATHOLOGY REPORT Testsubstance: 

### ClI. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-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: 17/06/08 date of end of treatment: 16/07/08 date of necropsy : 17/07/08
No abnormality detected
MICROSCOPIC FINDINGS No tissues examinedPage 36 of97 PATHOLOGY REPORT Test substance: 

### C.l. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association 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: 16107108 date of necropsy : 17/07/08
No abnormality detected
MICROSCOPIC FINDINGS
No tissues examinedPATHOLOGY REPORTPae3of9 Testsubstance: 0.1. Pigment Blue 61 Notox No.: 488231 Test system:28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- INDIVIDUAL ANIMAL DATA

Animal No.: 20 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 examinedPage 38 of

### 97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue61 Notox No.: 488231 Testsystem: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 21 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) MESENTERICPage 39 of 97 PATHOLOGY REPORT Test substance: 

### ClI. Pigment Blue61 Notox No.:488231 Test system:26-Day Oral ToxicityStudy inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association 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: 17/06/08 date of end of treatment: 16/07/08 date of necropsy : 17/07/08
EPIDIDYMIDES tail: right side NODULE(S) D=7X5 MM, 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) MESENTERICPage 40 of 97 PATHOLOGY REPORT Test substance: C.1.Pigment Blue 61 Notox No.: 488231 Test system: 25-Day Oral Toxicity Study inthe Rat 27-Nov-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

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 41 of 97 

### PATHOLOGY REPORT Test substance: 0.1I. Pigment Blue61 Notox No.:488231 Test system: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 24 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) MESENTERICPage 42 of 97 PATHOLOGY REPORT Testsubstance: 

### ClI. Pigment Blue61 Notox No.:488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-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 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

### Page43 of 97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue81 Notox No.: 488231 Test system: 28-Day Oral ToxicityStudy in the Rat 27-Nov-08 

### Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 26 SEX: Male DOSE LEVEL: 3
MACROSCOPIC FINDINGS days of traatment ;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.Page 44 of97 PATHOLOGY REPORT Testsubstance: ClI. Pigment Blue 61 Notox No.: 488231 Testsystem: 28-Day OralToxicity Study inthe Rat 27-Nov-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 0 date of necropsy : 17/07/08
EPIDIDYMIDES tail: right aide NODULE(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 granulomna (slight) tcorrelates to GROSS finding)
LYMPH NODE no lesion to account for gross finding(s) MESENTERIC

### Page 45of97 PATHOLOGY REPORT Test substance: ClI. 

### Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.


### TABLE 3
- 

### INDIVIDUAL ANIMAL DATA

Animal No.: 28 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 Ho abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE macrophage aggregates (minimal)

MESENTERIC no lesion to account for gross findingts)Page 46 of 97 PATHOLOGY REPORT Test substance: 

### C.l. Pigment Blue 61 Notox No.: 488231 Testsystem: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-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 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
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 47 of97 PATHOLOGY REPORT Test substance: 0.1. Pigment Blue61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association 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) MESENTERICPage 48 of 97 PATHOLOGY REPORT Test substance: 

### 0.1. Pigment Blue81 Notox No.: 488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 27-Nov-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 sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end oftreatment: 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
JEJUNUM 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, OESOPHAGUS, PANCREAS, PEYES PATCHES, PITUITARY,* PREPUTIAL GLAND, PROSTATE, RECTUM, SCIATIC NERVE, SEMINAL VESICLES, SPINAL CORD, SPLEEN, STERNUM, STOMACH, 

# 40 THYMUS, THYROID, TRACHEA, URINARY BLADDER.Page 49 of 97 PATHOLOGY REPORT Test substance: C.l. Pigment Blue61 Notox No.:488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-08 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 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
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, CAECUN, COAGULATING GLAND, DUODENUM, EPIDIDYMIS, HEART, ILEUM, JEJUNUM, 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 50 

### of 97 PATHOLOGY REPORT Test substance: 

### ClI. Pigment Blue61 Notox No.: 488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 33 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 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: ADREHAL, AORTA, BRAIN, CAECUM, COAGULATING

GLAND, COLON, DUODENUM, EPIDIDYNIS, 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 of97 PATHOLOGY REPORT Test substance: C.A. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 34 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/O8 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 finding(s) MESENTERIC MANDIBULAR plasma cell infiltration (minimal)

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

GLAND, COLON, DUODENUM, EPIDIDYMIS, HEART, ILEUM, JEJUNUM, LUNG, OESOPHAGUS, PANCREAS, PARATHYROID, PEYERS PATCHES, PREPUTIAL GLAND, SEMINAL VESICLES, SPINAL CORD, SPLEEN. STERNUM, STOMACH, THYMUS, THYROID: TRACHEA, URINARY BLADDER.Page 52 of97 PATHOLOGY REPORT Test substance:

### 0.1. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-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 : 17/07/08
NESENT. LYMPH DISCOLOURATION GREENISH

NODE No abnormalities ware found in any of the other tissues examined
MICROSCOPIC FINDINGS KIDNEYS tubular hyaline droplets (minimal)
LIVER parenchymal inflammatory cell infiltration (minimal)
LUNG perivascular inflammatory cell infiltration (minimal)
LYMPH NODE no lesion to account for gross finding(s) MESENTERIC
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, EPIDIDYMIS, HEART, ILEUM, JEJUNUM, OESOPHAGUS, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, PREPUTIAL GLAND, PROSTATE, RECTUM, SCIATIC NERVE, SEMINAL VESICLES, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER.

### Page53of97 PATHOLOGY REPORT Test substance: 

### C.I. Pigment Blue61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-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) MESENTrERIC no lesion to account for gross finding(s)

### Page 54 of97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.


### TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 37 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 MESEN'. LYMPH DISCOLOURATION GREENISH

NODE
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS No tissues examinedPage 55 of 97 PATHOLOGY REPORT Testsubstance:

### 0.1. Pigment Blue 61 Notox No.:488231 Testsystem: 28-Day OralToxicity Study inthe Rat 27-Nov-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 were found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE no lesion to account for gross finding(s) MESENTERICPage 56 of 97 PATHOLOGY REPORT Test substance: ClI. Pigment Blue61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-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 : 17/07/08 MESENT. LYMPH DISCOLOURATION GREENISH

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 of 97 PATHOLOGY REPORT Test substance: Cl
. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicityStudy inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association 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 GREENSH

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 finding(s) MESENTERIC
SEMINAL reduced luminal secretory contents present (correlates to GROSS finding) VESICLES

### Page58 of97 PATHOLOGY REPORT Test substance:0.1. Pigment Blue61 NatoxNo.:488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments ManufacturersAssociation Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 41 SEX: Female DOSE LEVEL: 1
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
No abnormality detected
MICROSCOPIC FINDINGS
CLITORAL GLAND interstitial inflammatory cell infiltration (minimal) VAGINA oestrus present No abnormalities found in: CERVIX, OVARIES, UTERUS.Page 59 of97 PATHOLOGY REPORT Test substance: 0.1. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal 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)
MANoIBULAR plasma cell infiltration (minimal)

LYMPH NODE
THYMUS involution/atrophy (minimal)
VAGINA dioestrus present epithelial 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, UTrERUS.Page 60 of 97 PATHOLOGY REPORT Testsubstance: 

### ClI. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 43 SEX: Female DOSE LEVEL: 1
MACROSCOPIC FINDINGS days of treatment 41 sacrifice group terminal aacrifice 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 dicestrus present epithelial vacuolation (moderate)
No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, 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, UTrERUS.Page 61 of97 

### PATHOLOGY REPORT Testsubstance: 0.1. Pigment Blue61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 44 SEX: Female DOSE LEVEL: 1
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

### Page62 of97 PATHOLOGY REPORT Test substance:

### C.l. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 45 SEX: Female DOSE LEVEL: 1
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 Blue61 Notox No.: 488231 

### Testsystem: 28-Day OralToxicity Studyinthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 46 SEX: Female DOSE LEVEL: 1
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 oftreatment: 31/07/08 date of necropsy : 01/08/08
No abnormality detected
MICROSCOPIC FINDINGS
No tissues examinedPage 64 of 97 PATHOLOGY REPORT Test substance: 0.1. Pigment Blue61 Notox No.: 488231 Test system: 28-Day oral ToxicityStudy inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3 - INDIVIDUAL ANIMAL DATA

Animal No.: 47 SEX: Female DOSE LEVEL: 1
M4ACROSCOPIC FINDINGS days of treatment :43 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end oftreatment: 29/07/08 date of necropsy :30/07/08 MANDIBULAR both aides DISCOLOURATION DARK RED LYMPH NODES
SKIN foreleg: left side ALOPECIA abdominal region: ALOPECIA foreleg: right aide ALOPECIA flank: right aide ALOPECIA flank: left aide 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 aide: epidermal hyperplasia (minimal) (correlates to GROSS finding) flank left side: epidermal hyperplasia (minimal) (correlates to GROSS

finding)
VAGINA diostrus 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: 

### C.l. Pigment Blue 61 Notox No.:488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 48 SEX: Female DOSE LEVEL: 1
MACROSCOPIC FINDINGS days of treatment :43 sacrifice group :terminal sacrifice necropsy statues 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
KIDNEYS tubular mineralization (minimal)
MANDIBULAR plasma cell infiltration (minimal)

LYMPH NODE VAGINA dioestrus 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. OBSOPHAGUS, OVARIES, PANCREAS, PARLATHYROID, PEYERS PATCHES, PITUITARY, RECTUM, SCIATIC NERVE, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, TRACHEA, URINARY BLADDER, UTERUS.

### Page 66 of 97 PATHOLOGY REPORT Testsubstance: ClI. PigmentBlue 61 Notox No.: 488231 Testsystem: 28-DayOral Toxicity Study intheRat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

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 of 97 PATHOLOGY REPORT Test substance: 0.1. Pigment Blue61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-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 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end of treatment: 27/07/08 

# 40 date of necropsy t 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, CAECU4, CERVIX, CLITORAL

GLAND, COLON, DUODENUM, HEART, ILEUM, JEJUNUM, KIDNEYS. LUNG, LYMPH NODE MESENTERIC, MANDIBULAR LYMPH NODE, OVARIES, PARATHYROID, 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: 

### ClI. Pigment Blue61 Notox No.:488231 Testsystem: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-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 oftreatment: 30/07/08 date of necropasy : 31/07/08
No abnormality detected
MICROSCOPIC FINDINGS
No tissues examinedPage 69 of 97 PATHOLOGY REPORT Testsubstance: 0.1. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments ManufacturersAssociation 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 10 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: 0.1. Pigment Blue 61 Notox No.:488231 

### Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments ManufacturersAssociation 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 data of start of treatment: 17/06/08 date of end oftreatment: 31/07/08 date of necropsy : 01/08/08
No abnormality detected
MICROSCOPIC FINDINGS No tissues examinedPage 71 of97 PATHOLOGY REPORT Test substance: 

### ClI. Pigment Blue 61 Notox No.:488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-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 :42 sacrifice group :terminal sacrifice necropsy statues planned terminal date of start of treatment: 17/06/08 date of end oftreatment: 28/07/08 date of necropsy : 29/07/08
No abnormality detected
MICROSCOPIC FINDINGS
VAGINA dioestrus present
No abnormalities found in: CERVIX, CLITORAL GLAND, OVARIES, UTERUS.

### Page 72 of97 PATHOLOGY REPORT Testsubstance: C.I. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day oral Toxicity Study inthe Rat 27-Nov-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 :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 MODE MESENTERIC.Page 73 of97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association 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 sides 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 MANDIBULAR congestion present (correlates to GROSS finding)

LYMPH NODE PANCREAS no lesion to account for gross finding(s)

### Page74 of97 PATHOLOGY REPORT Testsubstance:

### C.l. Pigment Blue61 Notox No.: 488231 Testsystem: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-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: 17/06/08 date of nd oftreatment: 29/07/08 date of enecropfsy : 30/07/08
No abnormality detected
MICROSCOPIC FINDINGS No abnormality detected
No abnormalities found in: LYMPH NODE MESENTERIC.Page 75 of 97 

### PATHOLOGY REPORT Test substance: C.l. Pigment Blue 61 Notox No.:

### 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-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 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/06/08 date of end oftreatment: 29/07/0a date of necropsy : 30/07/08 No abnormality detected
MICROSCOPIC FINDINGS
No abnormality detected
No abnormalities found in: LYMPH NODE MESENTERIC.PATHOLOGY REPORTPae7of9 Testsubstance: ClI. Pigment Blue 61 Notox No.: 488231 Testsystem: 28-Day OralToxicity Study inthe Rat 27-Nov-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 oftreatment: 29/07/08 date of necropsy : 30/07/08 No abnormality detected
MICROSCOPIC FINDINGS No tissues examinedPATHOLOGY REPORTPae7of9 Test substance: CJ. Pigment Blue61 Notox No.:488231 Test system: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association 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: 17/06/08 date of end of treatment: 29/07/08 date of necropsy ;30/07/08
No abnormality detected
MICROSCOPIC FINDINGS
No tissues examinedPage 78of 97 PATHOLOGY REPORT Testsubstance: C.I. Pigment Blue 61 Notox No.: 488231 Testsystem: 28-Day Oral ToxicityStudy inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.- 61 SEX: Female DOSE LEVEL: 3
MACROSCOPIC FINDINGS days of treatment :41 sacrifice group :terminal sacrifice nacropsy status :planned terminal date of start of treatment: 17/06/08 date of and oftreatment: 27/07/08 data of nacropsy :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

### PATHOLOGY REPORTPae7of9 Testsubstance: C.l. Pigment Blue61 Notox No.:488231 Testsystem: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-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 end of treatment: 30/07/08 date of necropsy : 31/07/08
No abnormality detected
MICROSCOPIC FINDINGS No tissues examined

### Page80of97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue 61 Notox No.: 488231 Test system: 28-DayOral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments 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 status :planned terminal date of start of treatment: 17/06/o8 date of end oftreatment: 27/07/08 40 date of necropsy : 28/07/08 NESENT. LYMPH DISCOLOURATION BLUISH

NODE
No abnormalities ware 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 Test substance: 

### C.l. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 

### Sponsor: Color Pigments ManufacturersAssociation Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 64 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 macrophage aggregates (minimal) MESENTERIC no lesion to account for gross finding(s)

### Page82 of97 PATHOLOGY REPORT Test substance: C. Pigment Blue 61 Notox No.:488231 Testsystem: 28-Day OralToxicity Study inthe Rat 27-Nov-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 oftreatment: 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) MESENTERICPage 83 of 97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue 61 Notox No.:488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-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 necropay 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 VAGINA metoestrus present
No abnormalities found in: CERVIX, CLITORAL GLAND, OVARIES, UTERUS.

### Page 84of97 PATHOLOGY REPORT Test substance: 

### ClI. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Studyinthe Rat 27-Nov-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 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 finding(s) MESENTERICPage 85 of 97 PATHOLOGY REPORT Test substance: 0.1. Pigment Blue61 Notox No.: 488231 Test system: 28-Day oral Toxicity Study in the Rat 27-Nov-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 necropsy status :planned terminal date of start of treatment: 17/06/08 date of end oftreatment: 29/07/08 date of necropsy : 30/07/08
MESENT. LYMPH DISCOLOURATION BLUISH NODE
No abnormalities ware found in any of the other tissues examined
MICROSCOPIC FINDINGS
LYMPH NODE sinus histiocytosis (minimal) MESENTERIC no lesion to account for gross finding Is)Page 86 of 97 PATHOLOGY REPORT Test substance: 

### C.l. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-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 oftreatment: 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(s) MESENTERIC

### Page 87 of97 PATHOLOGY REPORT Testsubstance: 0.1. Pigment Blue 61 Notox No.: 488231 Testsystem: 28-Day Oral Toxicity Study inthe Rat 27-Nov-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 oftreatment: 30/07/08 date of necropsy : 31/07/08
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)

MESENTERICPage 88 of 97 PATHOLOGY REPORT Test substance: 

### ClI. Pigment Blue61 Notox No.:488231 Test system: 28-Day OralToxicity Studyinthe Rat 27-Nov-08 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 necropsy status :planned terminal date of start of treatment: 17/06/08 date of end oftreatment: 31/07/08 date of necropfsy : 

### o/o8/08
GENERAL GI-TRACT, CONTENTS BLUISH

OBSERVATIONS
MESENT. LYMPH DISCOLOURATION BLUISH NODE SKIN FORELEG: right side ALOPECIA HINOLEG: 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(s) MESENTERIC SKIN foreleg right side: epidermal hyperplasia (minimal) (correlates to GROSS finding) foreleg left side: epidermal hyperplasia (minimal) (correlates to GROSS finding)Page 89 of 97 PATHOLOGY REPORT Test substance: 

### ClI. Pigment Blue61 Notox No.: 488231 Test system:28-Day Oral Toxicity Studyinthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- INDIVIDUAL ANIMAL DATA

Animal No.: 72 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 oftreatment: 28/07/08 date of necropsy : 29/07/08
GENERAL GI-TRACT, CONTENTS BLUISH

OBSERVATIONS
No abnormalities were found in any of the other tissues examined
MICROSCOPIC FINDINGS VAGINA metoestrus present
No abnormalities found in: CERVIX, CLITORAL GLAND, OVARIES, UTERUS.

### Page 90

### of 97 PATHOLOGY REPORT Test substance: C.I. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor: ColorPigments 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 GI 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(s)

MESENTERIC
MANDIBULAR plasma cell infiltration (minimal)

LYMPH NODE
VAGINA dioestrus present
No abnormalities found in: ADRENAL, AORTA, BRAIN, CARCUM, CERVIX, CLITORAL

GLAND, COLON, DUODENUM, HEART, ILEUM, KIDNEYS, LIVER, LUNG, OESOPHAGUS, OVARIES, PANCREAS, PARATHYROID, PEYERS PATCHES, PITUITARY, RECTUM, SCIATIC NERVE, SPINAL CORD, SPLEEN, STERNUM, STOMACH, THYMUS, THYROID, 0 TRACHEA, URINARY BLADDER, UTERUS.Page 91 of97 

### PATHOLOGY REPORT Test substance: 0.1. Pigment Blue 61 Notox No.: 488231 Testsystem: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.
TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 74 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 necropsy : 28/07/08
ADRENAL GLANDS both sides DISCOLOURATION RED-BROWN GENERAL GI TRACT, CONTENTS BLUISH

OBSERVATIONS MANDIBULAR both sides DISCOLOURATION REDDISH LYMPH NODES MESENT. LYMPH DISCOLOURATION BLUISH

NODE No 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 NODE
RECTUM blue material in lumen present (GENERAL OBSERVATIONS) (correlates to GROSS finding)
SPLEEN extramedullary haematopoiesis (slight)
THYMUS involution/atrophy (minimal)
VAGINA dioestrus present epithelial vacuolation (slight) 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: C.I. Pigment Blue 61 Notox No.: 488231 Test system: 28-Day Oral ToxicityStudy inthe Rat 27-Nov-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 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 ware found in any of the other tissues examined
MICROSCOPIC FINDINGS KIDNEYS basophilic tubules (minimal)
LYMPH NODE no lesion to account for gross finding(s)

MESENTERIC
MANDIBULAR plasma cell infiltration (slight)

LYMPH NODE VAGINA dioestrus present epithelial vacuolation (slight)
No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUN, 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 Test substance:

### ClI. Pigment Blue61 Notox No.: 488231 Testsystem: 28-Day Oral Toxicity Study in the Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE3

- INDIVIDUAL ANIMAL DATA

Animal No.: 76 SEX: Female DOSE LEVEL: 4
MACROSCOPIC FINDlINGS 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/Os date of necropsy :29/07/08
GENERAL GI-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 NODE no lesion to account for gross finding(s) MESENTERIC
MANDIBULAR plasma cell infiltration (minimal)

LYMPH NODE
VAGINA dioastrus present epithelial vacuolation (slight)
No abnormalities found in: ADRENAL, AORTA, BRAIN, CAECUM, CERVIX, CLITORAL

GLAND, DUODENUM, HEART, ILEUM, 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.

### PATHOLOGY REPORTPae9of7 Test substance:

### 0.1. Pigment Blue61 Notox No.:488231 Testsystem:28-Day Oral Toxicity Studyinthe Rat 27-Nov-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 necropsy : 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 perivascular 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, CAECUM, 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.

### Page95 of97 PATHOLOGY REPORT Test substance:ClI. PigmentBlue 61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments ManufacturersAssociation 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 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(s)

MESENTERICPage 96 of 97 PATHOLOGY REPORT Test substance: C.l. Pigment Blue 61 Notox No.:488231 Testsystem: 28-Day Oral Toxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3

- INDIVIDUAL ANIMAL DATA

Animal No.: 79 SEX: Female DOSE LEVEL: 4
MACROSCOPIC FINDINGS days of treatment :44 sacrifice group :terminal sacrifice necropsy status :planned terminal date of start of treatment: 17/OS/OB date of end oftreatment: 30/07/08 date of necropsy :31/07/08
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

### Page97 of97 PATHOLOGY REPORT Test substance: 

### C.I. Pigment Blue61 Notox No.: 488231 Test system: 28-Day OralToxicity Study inthe Rat 27-Nov-08 Sponsor: Color Pigments Manufacturers Association Inc.

TABLE 3
- INDIVIDUAL ANIMAL DATA

Animal No.: 80 SEX: FemalIe DOSE LEVEL: 4
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
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 Project 488231 APPENDIX 5

APPENDIX 5 SUMMARY DOSE RANGE FINDING STUDY

Page 1of2

## 0.1. PIGMENT BLUE 61 NOTOX Project 488231 APPENDIX 5 

## SUMMARY OF DOSE RANGE FINDING STUDY Inorder tosetthe dose levelsforthe main study (NOTOX Project 488231), apilotstudywas performed. Two groups of3females (10 weeks old) were dosed at500 or 1000 mg/kg bodyweight/day (Groups 1and 2)for 5days by oral gavagewith adose volume of 5mI/kg bodyweight. Dose levelswere based onthree non-GLP acute oral studies inwhich the LD50valueswere> 15,000 mg/kg (SPF Wistar rats), >15,000 mg/kg (SPF Wistar rats) and >5,000 mg/kg (Sprague Qawley rats). Inanon-GLP repeat dose study, mixed raceAlbino ratswere exposed for 14 days at 500 mg/kg: notoxicitywas noted. Allanimalswere housed 3percage, identified by ear-and tailmark andallocation was performed atrandom. Ifnot mentioned otherwise, testsystem and techniques were identical to those used during the main study. Data was collected under NOTOX Project488232. Planning Delivery: 28 May 2008 Treatment: 02to 06 June 2008 Terminal procedures: 06 June2008 Animal husbandry Animals were housed inacontrolled environment, inwhichoptimal conditionswere considered tobe approximately 15 air changes per hour, atemperature of21 ±

## 30C (actual range: 19.8

- 

## 21.10C), a relative humidity of30- 70% (actual range: 50

- 

## 81%)and 12 hours artificial lightand 12 hours darkness per day. Cleaning procedures inthe room might have caused the temporary fluctuations above theoptimal level of70% for relative humidity. Based on laboratory historical datathese conditionswere considered notto have affected the study integrity. Chemical analysis Not performed during the pilotstudy. Observations Clinicalsigns: At leastonce daily. Mortality: At leasttwice daily. Bodyweights: Daily. * Food consumption: Daily. Pathology Necropsy: On Day 5 Noorgans were fixed. Terminal body weight, organweights: kidney and liver. 

## RESULTS No mortality wasobserved inthe pilot study, andthere were no differences inbody weights (absolute or relative), food consumption (absoluteor relative), ororgan weights (absolute or relative). Foranimalstreated with 500 and 1000 mg/kg ofC. 1.Pigment Blue 61, blue coloration ofthe faecesand bluish contents inthe GI tractwere noted. 

## CONCLUSION There were no toxicologically-significant resultsfortreatment upto 1000 mglkg; based on the results ofthe pilot studythe doses selected for the main study are: 50, 250, and 1000 mg/kg.



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