# Dossier: Leuco Sulfur Black 1 UUID IUC5-292149d1-a677-4fab-9692-1fad973281 ea Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea 

### Author Date Remarks Wilhelm Dietz 

### I Clariant Iberica Producci6n SA. 

### I El Prat de Uobregat (Barcelona) 

### I Spain 2010-07-1310:24:57 CEST Created the dossier 

# Dossier header Dossiertemplate Name Version REACH Registration above 1000 tonnes 2010-02-10 Name (given 

### by user) Leuco Sulfur Black 1 

## Dossiersubject Name phenol 2 4-dinitro- sulfurized leuco denvatives LR ROB 00011687 TLP JR I phenol 2 4-dinitra= sulfurized Jeuco derivatives /66241-11:0 1Clariant lb9rica Producci6n SA. I El Prat 

### de Uobregat !Barcelona\ I Soain Submitting legal 

### entity Clariant Iberica Producci6n SA I El Prat de Uobregat !Barcelona\ I Spain 

### Doulercreation date/lime 2010-07-1310:24:57 CEST 

# Type ofsubmission Joint submission Tonnage

### band Over 1000tonneslyear 

# Registration dossierspecific Information Phase-in



# coNTA\NS NO CB\

# Legal entity: Clariant Iberica Producci6n S.A. UUID IUC5-e16fc18b-79b0-41f2-977c-96407714ccc3 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author REACH-IT Date 2010-03-29 12:54:55 CEST Remarks 

# General information Legal entity n- Clariant Iberica Producci6n S.A. 

### Legal entity type company 

# Identifiers Legal entity identifiers llclentlflartypa 

## VAT



# Contact Information Contactaddress Address ZAl

- Prat 

### Address c/ Cal Coracero, 46-56 Post.sl code 

### 08820 T-n El Prat de Llobregat (Barcelona) Country Spain 

### Phone +34 93 479 8200 

### Fu +34 93 479 8211 

### E-mail raach.es71@clarlant.com 

### Web aile http://www.clariant.es ESA60597473

# Contact persons Title Mr. 

### Flnst name Product Safety Last 

### name Pigments & Additives (Pfr) Phone +49 223 348 615 2 E-mail SIEF.PAPfr@dariant.com Address lndustriestrasse Postal code 50351 Town HOrth 

### I Knapsack Country Germany

Title Mr. 

### Flnst name Product Safety Last name Pigments & Additives (Raw) Phone +49 619 675 783 04 E-mail SIEF.PARaw@clariant.com Address 

### Am Unisyspark 1 Postal code 65643 

### Town Sulzbach am Taunus Country Germany



### Title Mr. 

### Flnst name Product Safety Last name Textile, Leather & Paper Chemicals (IT) Phone +39 023 631 728 4 E-mail SIEF.TLPIT@clariant.comAddress Via Cappelinl Postal 

### code 20030 Town Country



### Title First name Last name Phone E-mail Address Postal 

### code Town Country Palazzolo Milanese Italy

Mr. Product Safety Pigments & Additives (Dye) +49 619 675 782 75 SIEF.PADye@clariant.com 

### Am Unisyspark 1 65843 Sulzbach 

### am Taunus Germany

Mr. First name Product Safety Last name Pigments 

### &Additives 

### (Wad) Phone +49 821 479 295 1 E-mail SIEF.PAWad@clariant.com Address Ludwlg-Harmann-Stralle 100 Postal 

### code 

### 86366 Town Country



### Tille Gersthofen Germany

Mr.

### First name Product Safety Last name Textile, leather& 

### Paper Chemicals (FR) Phone +33 146 969 782 E-mail SIEF.TlPFR@clariant.com Address Avenue des Champs Pierreux 52 

### Postal code 92000 Town 

### Country

Title First name Last name Phone E-mail Add""'s Nanterre 

### France

Mr. Team 01 REACH Center +39 029 918 753 8 SIEF.MB01@Ciarlant.com 

### Via lainate 26 Postal code 20010 Town Pogliano Milanese 

### Country Italy

Title Mr. First name Product Safety Last nama Textile, leather

### & Paper Chemicals (DE2) Phona +41 614 695 634 E-mail SIEF.TlPDE2@clariant.com Add,.... Rothausstrasse 61 Postal code 4132 Town Muttenz

### Country Switzerland

Title Mr. 

### First name Product Sefety Last name Textile. leather& 

### Paper Chemicals (GB) 

### Phone 

### +44 113239 830 8 E-mail SIEF.TlPGB@clariant.oom Address**Celve~ey** Lane 

### Postal code 

### l$184RP Town 

### Country

Title Horsfooh France

Mr. 

### First name Product Sefety Last name Textile, leather & 

### Paper Chemicals (ES) 

### Phone +34 934 798 235 E-mail SIEF.TlPES@clariant.oom Address ZAl-Prat. c/Cel Coracero 

### 46-56 Postal code 

### 08820 Town El Prat 

### de Uobregat Country Spain

First name Product Sefety 

### Laet name Pigments & 

### Additives (lnt) 

### Phone +49 61 96 75 76 14 5 E-mail SIEF.PAint@clarlant.oomAddress Am Unisyspar1< 1 Posllllcode 65843 Town Sulzbach am Taunus Country Germany

Title Mr. First name Product Safety Last name Pigments & Additives (Azo) Phone +49 619 675 761 52 

### E-mail SIEF.PAAzo@clariant.com Address Am Unisyspar1< 1 Posllll 

### code 65843 Town Sulzbach am Taunus Country Germany

Title Mr. First name Product Safety Last name Pigments & Additives (Hpp) Phone +49 619 675 761 51 

### E-mail SIEF.PAHpp@clariant.com Address Am Unlsyspar1< 1 Posllll 

### code 65843 Town Sulzbach am Taunus Country Germany

Title Mr.

### Firat n•m• Product Safety 

### L.,.tn.me FUN Phone +49 693 051 664 3 E-mail SIEF.FUN@clariant.com Address lndustrleparl< HOchst C 671 Posl81 

### code 65926 

## Town Country


# Sites Frankfurt am Main 

### Germany

Clariant Castellbisbal

# Site: Clariant Castellbisbal UUID IUC5-d513f60b-9cc7-4068-b192-d4987bef7e2b Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281 ea 

### Author Date Remarl<s mkuehn I Clariant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-04-09 14:08:56 CEST 

# Site flags General information Site name Clariant Castetlblsbel Legal entity

### owner Clariant Iberica Producci6n SA I Ei Prat de Llobregat 1Barceloni!l.LSJ1lilll 

# Contact address Address Postal code 

### Town Region 

### I State Country Poilgono lnduatrial Can Estepe sin 08755 Castetlbisbal Barcek>na Spain

# Substance: phenol,2,4-dinitro-,sulfurized. leuco derivatives LR ROB 00011687 TLP JR UUID IUC5-69271110-0636-4fd0-&4ac-30509507d07e Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author pbasto 

### IClartant Quimicos (Portugal) Lda 

### I Le98 do Balio 

### I Portugal Date 2010-07-08 15:51:48 CEST Remarks 

# 0 Related Information 0.1 Templates 0.2 Categories 0.3 Mixtures 1 General Information 1.1 Identification Substance identification Chemtc.l name phenol,2,4-dinttro-,sulfurized,leuoo dertvatives_LR_RDB_00011687 _TLP_JR Public name Leuoo Sulphur Black 1or Leuoo Sulfur Black 1 Legal entity C!ariant fbBrjca producci6n SA I El 

### Pratde Uobregat (Barcelona) I Spain 

# Role in the supply chain Role: Manufadurer; Importer 

# Reference substance phenol2 4:djoitro- su!fudzed leuco derivatjves/66241·11::0 

### EC number 

### EC name 266-273-5 phenol, 2,4-dinltro-, sulfurized, leuco derivatives 

### CAS number 

### CAS name 66241-11-0IUPACname 

# Type ofsubstance Composition UVCB Origin organic 

# Contact person Organisation Clariantlbi>rica Producci6n S.A. Department Product Safety TLP First name Product Safety Last name TLP Phone +34 934 798 235 E-mail SiEF.TLPES@clariant.com Address ZAL-Prat. c/Cal Coracaro 46-56 Postal 

### code 08820 Town El Prat de Llobregat Region 

### I State Barcelona Country Spain 

# 1.2 Composition Substance composition**phen~.2.4-dinitro-.sulfurized.** leuc:o derivatives



## Degree of purity

ca. 52.75 

### % (w/w) 

## ConstituentsReference substance ohenol 2 4-dinitro· sulfurized leuco derivatives 

### I66241-1 1-0



### Typical concentration 

### EC number EC name 266-273-5 phenol, 2,4-dinitro-, sulfurized, leuco derivatives 

### CAS CASname 

### number 66241-11..() 

### IUPACname

ca. 52.75% (wlw) Concentration range >45- < 

### 55 % (wlw) 

### Remarl<a Cartfficate 

### of analysis C43441 

# Impurities

Reference substance water I water I Water /7732-18=5 

### EC number EC name 231-791-2 water 

### CAS number CAS name 7732·18-5 Water 

### IUPAC name 

### water Typlc.ol concenlnlllon ca. 45.75 % (wlw) Concenlnlllon range > 

### 43.5- < 

### 53.5 % (wlw) Reference sodium chloride I

### sodium chloride I 

### Sodium ch!oride/7647-14-substance EC number EC name 231-598-3 sodium chloride CAS number CAS name Sodium chloride 7647-14-5 IUPAC name sodium chloride Concenlnltlon range >

## o- < 1.5% (wlw)
Reference 

### aub&tllnce

Concenlnltlon range sodium sulphate I disodium sulfate I Sutfyric acid sodium salt (1:2) /7757-82-<l EC number EC name 231-820-9 sodium sulphate CAS number CAS name Sulfuric acid sodium salt (1 :2) 7757-82-<l IUPAC name disodium sulfate

>

## o- < 1.5% (w/w)

Reference substance 2A:Qirlitrophenol I 2 4-dinitrophenol/ 51-28-5 EC number EC name 200-087-7 2,4-dinitrophenol CAS number CAS name 51-28-5 IUPAC name 2,4-dinitrophenolConcentration range <20 ppm Remerko Certifocate 

### of analysis C43441
Reference oubotance

Concentration range Remarka 1-chloro-2 4=dinitrobenzene I 1-chlorcr2 4-dlnjtrobenzene I 97- 

### QQ:I EC number EC name 202-551-4 1-chloro-2.4-<linnrobenzene CAS number CAS name**97~0-7** IUPAC name 1-chloro-2,4-<linitrobenzene

< 20 ppm Certificate ofanalysis , C43441 

# 1.3 Identifiers Identifiers Regulatory programme identifiers**~**j Regul.tory programmejtD 

# JRemarka REACH preregistration 05-2114286843-37- IUC5-e16fc18b-79b0-4112-9ncnumber 0000 9640n14ccc3 

# 1.4Analytical information Analytical information Analytical rnethodo and spectral Determination of NMR, IR, MS, HPLC/MS, UV-VIS, Physical measurements and Stability in Organic Solvents: see "CoA and 

### dota spactra_200B102B09104B.pdr Purity 

### of test item 

### Iwater content see 'C43441_CoA.pdr (amendment to CoA)CoAand spectra _20081028091048.pdf I 187.28 KB 

## Results ofanalysis

Analysis type H(1)-NMR, HPLC/MS, UV-VIS and Stability in Organic Solvents Tested Leuko Sulfur Black 1 (press cake) substance Method used not measurable, test ttem insotubie Remarks Results not available, because 

### ofthe insolubility

### ofthe test substanca in solvents used forthe corresponding measurements Results see attached "CoA and spectra_20081028091048.pdf" Analysts type Watercontent/ Purity ofthe test substance (amendment to CoA) Tested Leuko Sulfur Black 1 (press cake) 

### substalnce Method used Kart-Fischer-nration I Calculation C43441_CoA.pdf/3.98 MB Remarks new determination ofthe water content and calculation 

### ofthe purity of the test item Analysis MS type Tested Leuko Sulfur Black 1 (press cake) substance Method MALDI-MS used Remarks The test subsstance is a mixture of relative high molecular compounds with no defintte structure. Because 

### offragmentation, only fragments in the range of 100 to 500 Da could 

### ba detected. Results see also attached "CoA and spectra_20081028091048.pdf" Analysis type IR Tested substance Leuko Sulfur Black 1 (press cake) Method used KBr-peUet Remarks The test subsstance was dried overnightResults seealso attached "CoA and spectra_20081028091048.pdl" Analysis Physical measurements 

### type Tnted Leuko Sulfur Black 1 (press cake) subatllnce Method visual method 

### used Remarks The test substance Is insoluble in DMSO, DMSO + NH3, chlorofonn, methanol, water, water+ NaOH, DMF, THF, sulfuric acid and trifluoroaclic acid. Othersolvents were nottested. Because of**insolubi~ty** 

### of the test substance in trifluoroaclic acid, sulfuric acid and sodium hydroxyde, no calibration curve is available. Therefore it Is not possible to determine 

### lha exact solubility in water and octanol (and 

### lha log pow). Results sea also attached "CoA and spectra_20081028091048.pdl" 

# 1.5 Jointsubmission Joint submission

General lnfonnation Jointsubmission name Reguletoty programme 

# Lead Leuco_Sulphur_Biack_1 EU: REACH

Ciariantlb<irica Producci6n S.A I El Prat de Llobregat !Barcelona) I Spain 

# 1.6 Sponsors 1.7 Suppliers 1.8 Recipients 1.9 Productand process oriented research and development

# 2 Classification and Labelling 2.1 GHS Classification and Labelling according to GHS

General Information Not classified lmplement.tion State 

### Iform of

### the substance 

# Classification Physical hazards EU solid

Haz.ord Ruson for no statement classlflcatlon Explosives conclusive but not sullident for classification Flammable gases conclusive but not suffiCient for classification Flammable aerosols data 

### lacking Oxidising gases data 

### lacking Gaaes underpressure data 

### lacking Flammable liquids data 

### lacking Flammable solids conclusive but not 

### suffiCient for classification Self-rsactlve substancaa and data 

### lacking mixtures Pyrophortc liquids data 

### lacking

### Pyrophorlc solids**Self~eatlng** 

### substances and mixtures Subshlnces and mixtureswhich In conhlc:t

### with w.ler

### emitflammable 

### goes Oxldlslngllqulds Oxidising solids Organic peroxides Corrosive to metals 

# Health hazards

Acute toxicity • oral Acute toxicity •dennal Acute toxicity • Inhalation Skin corrosion/Irritation Serious eye damaga/ eye Irritation Respiratory sansltlutlon Skin sansltlutlon Aspiration hazard Hazard -nt data lacking data lacking data lacking

data lacking conclusive but not sufficientfor classification data lacking data lacking



### Reason for no classification
condustve but not sufficient for dasslfication conduslve but not sufficientfor dasslfication 

### condusive but not sufficient for dassification conduslve but not suffident for dasslficalion condusive but not sufficientfor 

### dasslfication condusive but not sufficient for 

### dasslf1C8tion conduslve but not sufficientfor dasslfiC8tion conduslve but not sufficientforclassification 

## Reproductive toxicity Reproductive toxicity condusive 

### but not sufficient for classification Specific effect Route ofexposu"' Effects on orvia lactation conclusive 

### but not sufficient for classification 

## Germ cell mutagenicity

Hazard Reason 

### for no cluaiflcatlon statement 

### Germ 

### cell conclusive but not suflicient for mutagenicity classification Route ofexposure 

## Carcinogenicity

Hazard Reason for noclassification statement Carcinogenicity conclusive but not sufficient for classification Route of exposure 

## Specific target organ toxicity • single 

# I Hazard Reason for no classification 

# I

### statement Specific target organ conclusive but 

### not sufficient for toxicity
- olngle classification Alleclldorgans Roull olexpooure 

## Specific target organ toxicity • repeated

HIIZIIrd Reason for noclautflcatlon 

### llallment Specific llrgetorgantoxicity conclusive but not sufficientfor 

### -repeated classification Affeclld organs Roull olaxpooure 

## Environmental hazards

H11Z1rd Reason for no classification 

### -mant HIIZIIrdous tothe aquatic conclusive but not sufficientfor environment 

### classification HIIZIIrdous tothe atmospheric data lacking environment 

# Labelling Signal word No signal word 

# 2.2 DSD- DPD Classification and Labelling according 67/548/EEC (DSD)

# General Information
Leuco Sulfur Black 1 Not classified Status 67/548/EEC settclassification 

# Classification

Explosiveness Oxidising propertleo

Thermal stability 

### Acuta toxicity Acute toxicity • Irreversible damage after single exposurs Repeated dosetoxicity lrrltallon 

### I Corrosion
Sensltlsatlon 

### Carcinogenicity Mutagenicity• Genetic Toxlclty Toxicity

### to rspnoductlon ·fertility Classification Reason for no classification conclusive but not sufficient for classification conclusive but not suffiCient for classification conclusive but not sufficient for classification data lacking conclusive but not sufftcient for classification conclusive but not sufficient for classification conclusive but not suffiCient for classification conclusive but not sufficient for classification conclusive but not sufficient for classification conclusive but not sufficient for classification conclusive but not sufficient for classlfocation conclusive but not suffiCient

### for classification 

### Toxicity

### to reproduction
- development condusive but not sufficient for classification 

### Toxicity

### to reproduction
- breastfed 

### b8bin condusive but

### not sufficient 

### for classification 

### Environment condusive but not sufficient 

### for classification 

# 3 Manufacture, use and exposure 3.1 Technological process Technological process

Methods of 

### manufacture Reaction of2,4-dinitrochlorobenzene with Inorganic alkalis and further reaction ofobtained dinltrophenolate with reducing agents such as sodium sulfhydrate, sodium sulphide and sulfur. Reactions ofhydrolisis, reduction and thlonatlon are involved in the manufacturing process. The reactions take place per batch In a dosed reactor connected to a water reflux with water recirculation and keeping constantthe temperature. The gases exit is connected

### to a scrubber. 

### The reaction is controlled bytemperature, time and pressure
. At the end the coloured pigment is isolated by acid precipitation , filtrated and washed. 

# 3.2 Estimated quantities Estimated quantities

Year 2010 

# Total tonnage Manufloctured 3000 tonneslyear 

### im.,-d 

# Remarks 150tonneslyear

# ITonnage=3150.0 

# 3.3 Sites Sites

Site Clariant Castellbisbal/ Castellbisbal/ Spain Legal entity 

### owner Clariant Iberica Producci6n 

### SA I El Prat de Uobregat !Barcelona) I Spain Production sHe; Use site 

# 3.4 Form in the supply chain Form in the supply chain

Available as substance Substance in mixture Substance in mixture
Trede name

### of mixture Products of Diresul selling range Type 

### ofmixture Liquid formulation Typical concanlrlltlon >= 20- <= 40 % (wlw) 

# Substance in article 

# 3.5 Identified uses Identified uses Information on uses

## Uses by workers in Industrial settings IUnumber Identified uu name 

### Process category

Environmental release category Substance supplied 

### to that uselnfonn of
Market sector

### by 

### type of chemical product Sector ofend use Subsequentservice life relevant forthat use? Article category related 

### to subeaquent service life

IU number Identified use name Process catagory

Environmental release category Substance supplied tothat use In fonn of Market sector by

### type of Manufacture ofdye preparation PROC 5: Mixing 

### or blending In batch processes for lonnulation ofpreparations and articles (multistage and/or significant contact) ERC 2: Formulation 

### of preparations As such 

### In a mixture PC 

### 34: Textile dyes, finishing and impregnating products; Including bleaches and other processing 

### aids SU 

### 10: Formulation [mixing] of preparations and/or

### re- packaging (excluding alloys) yes
AC 

### 01: Other (non intended to 

### be released): package material


### 2 Manufacture oftextile and paper PROC 6: Calendering operations PROC 2: Use in dosed, continuous process whh occasional controUed exposure ERC 

### 5: Industrial use resulting In Inclusion into or onto 

### a matrix In a mixturechemical product



### Sectorofend use
Subsequent service life relevant

### lorthat use? Article cetegory related

### to subsequentservice life PC 34: Textile dyes, finishing and impregnating products; inCluding bleaches and other processing aids PC 26: Paper and board dye, finishing and impregnation products: including bleaches and other processing aids SU 

### 5: Manufacture of textiles, leather, fur SU 6b: Manufacture ofpulp, paper and paper products yes
AC 

### 5: Fabrics, textiles and apparel AC 

### 8: Paper articles 

## Uses by professional workers

IU number 

### Identified use name 

### Proceucategory

Environmental release category Subetance supplied 

### to that 

### use In form of Merklll sectorby

### type of chemical product



### Sectorofend use Manufacture oftextile and paper PROC 6: Calendering operations PROC 2: Use in dosed, continuous process with occasional controlled exposure ERC 5: Industrial use resulting in indusk>n intoor onto a matrix In a mixture
PC 34: Textile dyes, finishing and impregnating products; induding bleaches and other processing aids PC 26; Paperand board dye, finishing and impregnation products: including bleaches and other processing aids SU 5: Manufacture 

### oftextiles, leather, fur SU 6b: Manufacture ofpulp, paperand paper productsSubuquentservice life 

### yes relevant lorthat use? Article cslegoty related to AC 5: Fabrics, textiles and apparel subsequent service life AC 8: Papararticles 

# Mostcommon technical functions ofthe substance

Technical function ofsubstance (what 

### Itdoes) Colouring agents. dyes Remarks coloring agent 

# Significant routes ofexposure

Human exposure Dermal 

## Environmental exposure Water 

# Pattern ofexposure Accidental/ infrequent 

# 3.6 Uses advised against 3.7Waste from production and use Waste from production and use

Remarks Waste waterdoes not balong to 

### the waste. Other waste not Identified during production process. 

# 3.8 Exposure estimates 3.9 Biocidal information 3.10 Application for authorisation ofuses

# 4 Physical and chemical properties Endpointsummary: Physicalandchemicalproperties
UUID IUC5-c3ad99aa-Oc79-47 44-8b8d-Oc1688008384 Dossier UUID IUC5-292149d1-a677-41eb-9692-1fad973281ea Author pbasto I Clariant Quimicos (Portugal) Lda I**L~** do Balio I Portugal Date 2010-05-28 16:23:16 CEST Remarks 

# Administrative Data Discussion IUCLIDPhysical -chemical data



## 4.1 State ofthe substance at 20'C and 101kPa 4.2 Melting/freezing point 4.3 Boiling point 4.4 Relative density 4.5 Particle size distribution 4.6 Vapour pressure 4.7 Partition coefficient n-octanol/water 4.8 Water solubility Reference/ Report Results No 

## Harlan, C30920 Harlan, C30931 Harlan, C30942 Waiving Harlan, C30953 Harlan, C88384 Harlan,C75560 wet paste -1,7 °C 99,9 °C 0,5091 at 20"C only sold as liquid 1,8x1o-7

## Pa, 250C impossible to determine 62 

# mg/1

## 4.10 Surface tension Harlan, C30964 negligible 4.11 Flash point waiving not needed , Column 2, Annex VII 4.12 Autoflammability Harlan 0047/3108 348° C, 20 °C 4.13 Flammability , for solids Harlan, C30986 not highly inflammable 4.14 Explosive properties Harlan,C30997 not explosive 4.15 Oxidizing properties Harlan, C31010 no oxidizing properties 4.17 Stability in organic solvents and identity of relevant waiving not needed , column 2, degradation products Annex VII 4.21 Dissociation constant wiaving not needed , column 2, Annex VII 4.22 Viscosity Harlan, C31032 thixotrop

# 4.1 Appearance/physical state/colour Endpoint summary: Appearance/physicalstate/colour
UUID IUC5-a02788a5-13e9-4dd9-Be37-27eb020afflf Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author pbaslo I Clariant Quimicos (Portugal) Lda I Leo;a do Balio I Portugal Date 2010-04-30 15:17:22 CEST Remarks 

# Administrative Data Short description of key information Physical state at 20 "C solid (wet paste) Colour : black Odour :odour1ess

# Endpointstudyrecord: Appearance/physicalstate/colour
UUID IUC5-580e072b-40a3-49bd-8fd4-00b856a0a7ec Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks pbasto 

### I Clariant Quimicos (Portugal) Lda 

### I Le<;a do Balio 

### I Portugal 2010-04-0811:58:38 CEST


# Administrative Data Purpose flag weight ofevidence Study result

### type experimental result Reliability 1 (reliable 

### without restriction) 

# Materials and methods GLP compliance

no data 

# Test materials Identity oftest material same as forsubstance defined In section 1 (If not read-across) yes 

## Test material Identity

Identifier Identity CAS number 66241-11-0 EC number 266-273-5 

# Results and discussion Physical state at20"C and 1013 hPa solid (wet paste)

## Form paste 

## Colour black 

## Odour odourless 

## Substance 

## type organic 

# Applicant's summary and conclusion Conclusions black wet paste

# 4.2 Melting point/freezing point Endpointsummary: Meltingpoint/freezingpoint UUID IUC5-2c666c2d-b30c-4735-924f-!60a45fb2d87 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author pbasto I Clariant Quimicos (Portugal) Lda I**Le~** 

### do Balio I Portugal Dale 2010-04-1611:03:04 CEST 

### Remar1<s 

# Administrative Data Short description of key information

The freezing point 

### of Leuco Sulfur Black 1was determined 

### to be -1.7 

### 'C 

### ±0.2 

### 'C which is equal to 271 .7 K ± 0.2 K. 

# Discussion The freezing point of Leuco Sulfur Black 1 is influenced by its high water content. It can be assumed that the measured value corresponds to the waterfraction only.

# Endpointstudyrecord: KEY_OECD 102_2009_Har/an_C30920_Habeck
UUID IUC5-93aab045-c437-4355-af70-82908d9e2931 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks jregano I Clariant lbllfica Producci6n S.A. I El Pral de Llobregat (Barcelona) I Spain 2010-05-1815:17:33 CEST



# Administrative Data key study; robust study summary Purpose 

### flag Study result

### type Reliability experimental result Study period 2009-01-12 ti112009-01-13 1 (reliable without restriction) Rationale for reliability Incl. deficiencies Guideline conform study under GLP without deviations 

# Data source Reference

Reference Author

type study report Habeck 

# c Data access Year
2009

data submitter Is data owner 

## Data protection claimed yas 

### Title
Determination 

### ofthe Freezing Point 

### I Freezing Range



# Materials and methods Test guideline BlbNographlc Testing laboratory 1oun::e Hanan Labomtories Ltd. ltingen (Switzerland)

Guideline Report 

### Owner 

### no. company C30920 Clariant Company Report 

### study no. 

### deta 2009-07- 03

# IDev~~n•laccording EU Method A.1 (Melting 

### I Freezing Temperature) to aa::ording other guideline: EEC Directive 92/69, Part A, Methodsforthe determination ofphysico-chemical properties, A.1 "Melting/Freezing lempereture", EEC to Publication No. L383, December 1992. according OECD Guideline 102 (Melting point/**Me~ing** Range) to 

### Type ofmethod capillary method 

### GLP compliance yes (ind. oertificate) 

## Test materials Identity oftest material same as for substance defined In section 1 (If not read-across)
yes 

### Test material identity

Identifier Identity CAS number 66241-11-0 ECnumber 266-273-5 

### Details on test matenal

- Name 

### oftest material : Leuco Sulfur Black 1

- Physical 

### state: lhixotrophic, black, 

### wet paste -Composition: Leuco Sulfur Black 1:52.75% (w/w) (caicuiatad by amountof by-compounds) 2,4-Dinitrochlorobenzene: < 20 ppm 2,4-Dinitrophenole: < 20 ppm Sulfate: 970 

### ppm Chloride: 750 ppm Sodium: 1.3 

### o/o (w/w) Water. 45.75 

### % (wiw) by**Ka~-Fischer** titration

- Lot/batch No.: ESA0007595

-**Exp~ation** date 

### ofthe lot/batch: 2011-07-11

-**Stabll~** undertest 

### conditions: Stable under storage conditions. -Storage condition 

### of test material: AI roomtemperatura (range 

### of20

### ± 5 

### 'C), light protected. no

no

no

# Results and discussions MeHing 

## Ifreezing point

Melt./Freez. 

### Atm. Decomposition 

### pt. pnaaure Decamp. Sublimation Subl. Remart.a temp. temp. 1.7 

### '1:: ca. 100 kPa no 

### no The study was parfonned under atmospheric pressurewhich was a11hat time ca. 100kPa. 

### The exact value 

### ofthe pressure was not

### recorded. 

## Any other Information on results Incl. tables 

## Detennination ofthe Freezing Point of Leuco Sulfur Black 1 (Summary) Freezln 

# ['C) Preliminary test Above -22 

# 'C 1stmain test 

# ·1.8 2ndmain test 

# ·1.5 Mean value* -1.7 Standard deviation 

## (±) 0.2 *The mean freezmg po1nt was calculated from the bold values 

# Applicant's summary and conclusion Conclusions 

## Point

The freezing point of Leuco Sulfur Black 1was determined to be -1.7 

### '1:: 

### ±0.2 

### '1:: which is equal to 271 . 7 K

### ±0.2 K. 

# [K] Above 251.2 271.4 271.7 271.6

0.2

The freezing point ofLeuco SuWur Black 1 is inftuencad by Ks high water content. It can be assumed thatthe measured value corresponds to water 

### fraction only. 

## Executive summary 

## The determination ofthe freezing point 

## Ifreezing range ofthe test item was performed according to the EEC Directive 92169, December 1992 and Council Regulation (EC) No. 44012008, A.1 "Melting/Freezing temperature" and to the OECD Guideline No. 102: "Melting point 

# I melting range", adopted July27, 1995.

## A preliminarytestwas performed to check the approximate freezing temperature of Leuco Sulfur Black 1. The test item was stored about 19 hours at-22 

# 'C in a freezer. As the sample was frozen afterthis time the freezing temperature of Leuco Sulfur Black 1 is above -22 

# 'C. During the main tests, about 1 ml ofthe test item placed in a small testtube were cooled down from room temperature to about-2 

# 'C using a dry ice and acetone mixture. At -1.8 

# 'C (1st test) and -1.5 

# 'C (2nd test), respe ctively, the test item was frozen byvisual judgement. The freezing point of Leuco Sulfur Black 1 was determined to be -1.7

# •c ±0.2 

# 'C which is equal to 271.7 K 

# ±0.2 K. The freezing point of Leuco Sulfur Black 1 is influenced by its high water content. Itcan be assumed thatthe measured value corresponds to the waterfraction only.

# 4.3 Boiling point Endpointsummary: Boilingpoint UUID IUC5-5eef9db9-723c-42a4-81fe-247c125d49ce Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author pbesto I.Ciariant Ouimicos (Portugal) Lda I Le.;a do Balio I Portugal Date 2010-04-28 16:28:19 CEST Remarks 

# Administrative Data Short description of key information

The boiling point of Leuco Sulfur Black 1 is determined 

### to be 99.9 

### 'C ± 0.1 

### 'C which is equal to 373.1 K ±0.1 

### K. 

# Discussion The boiling point of Leuco Sulfur Black 1 is determined to be 99.9 

# 'C 

# ±0.1 

# ·cwhich is equal to 373. 1 K 

# ±0.1 

# K. The atmospheric pressure during the measurements was 100.4 kPa (preliminarytest) and 99.8 kPa (main tests). The boiling point of Leuco Sulfur Black 1 is influenced by its high water content. It can be assumed thatthe measured value corresponds to water only.

# Endpointstudyrecord: KEY_OECD 103_2009_Harlan_C30931_Habeck
UUID IUC5-55a7ec7d-25a3-41d9-8755-183757971a48 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks jregano 

### IClariant Iberica Producci6n SA 

### I El Prat de Llobregat (Barcelona) 

### I Spain 2011l-05-1815:17:57 CEST



# Administrative Data key study; robust study summary Purpose flag Study result

### type experimental result Study period 2009-01-12 till 2009-01-13 Reliability 1 (reliable without restriction) Ratlo""le

### for reliability Incl. deficiencies Guideline conform study under GLP without deviations 

# Data source Reference

Rttfwtrence Author



### type Year Title study report Habeck 2009 Determination ofthe BoHing Point 

### I 

# c Data access data submitter is data owner 

### Data protection claimed yes Bolling Range



# Materials and methods Test guideline Bibliographic Testing laboratory source Harlan Laboretortas Ltd. ltingen**(Switze~and)**

Guideline Report Owner no. company C30931 Clariant Company Report study no. date 2009-07- 03



# I Deviationalaccording EU Method A.2 (Boiling Temperature) 

### to according OECD Guideline 103 (Boiling point/boiling range) 

### to according other guideline: EEC Dlnective 92/69, PartA, Methods forthe determination 

### of physico-chemical 

### properties. A.2 "Boiling temperature", EEC Publication 

### No. 

### to L383, December 1992. 

### Type ofmethod differential scanning catorimetry 

### GLP compliance yes (incl. certificate) 

## Test materials Identity oftest material same as for substance defined in section 1 

### (If not read-across) yes 

### Test material identity

Identifier Identity CAS number 66241-11-0 EC number 266-273-5 

### Details on test material

- Name 

### oftest material : Leuco Sulfur Black 1

- Physical state: thlxotrophlc, black, wet paste -Composition: Leuco Sulfur Black 1:52.75% (wlw) (calculated by amount of by-compounds) 2,4-Dinitrochlorobenzene: 

### < 20 ppm 2,4-0initrophenole: < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3% (wlw) Water. 45.75 % (wlw) by Karl-Fischer titration

- Lotlbatch No.: ESA0007595 -Expiration date 

### ofthe lotlbatch: 2011-07-11

- Stability under test conditions: Stable understorage conditions. -Storage condition oftest material: At room temperature (range 

### of20 ± 5 'C), light protected. no

no

no

# Results and discussions Boiling point



## ThermalAnalysis: Determination ofthe Boiling Point of Leuco Sulfur Black 1 (Summary) Heating Range Heating Rate Boiling Point 

# rCJ Boiling Point 

# ['C] ['C/min] PreliminaryTest 25-400 20 99.8 1stMain Test 25-215 10 

# 100.0 2ndMain Test 25-215 10 

# 99.8 Mean ofboiling point 99.9 Standard deviation (±): 0.1



# .. The mean bolimg pomt was calculated from the bold values (onset pomt ofthe endothermic reaction). 

# [K] 373.0 

# 373.2 373.0 373.1

0.1

The tabulated values represent rounded results obtained by calculation using the exact raw data In this study the DSC showed an endothermic heat effect at 99.9'C. This temperature is notthe true boiling point ofthe test item as it only reflects the boiling point ofthe water contained bythe test item. The DSC shows no further endothermic heat effect up to 400'C. Thus, the boiling point of the Leuco Sulfur Black 1 can be regarded as being above 400'C( see: Weissenfeld (2009) Vapour Pressure, Harlan Laboratories Ltd. ltingen (Switzerland), Report C30953) 

# Applicant's summary and conclusion Conclusions The boiling point 

### of Leuco Sulfur Black 1 is determined 

### to be 99.9 

### 'C 

### ±0.1 

### 'C which is equel

### to 373.1 K 

### ±0.1 K. The boiling point 

### of Leuco Sulfur Black 1 is influenced by its high water content. It can be assumed that the measured value corresponds to waterfraction only. 

## Executive summary

## The determination ofthe boiling point 

# I boiling range ofthe test item was performed according to the EEC Directive 92/69 and Council Regulation (EC) No. 440/2008, A.2 "Boiling temperature" and to the OECD Guideline No. 103: "Boiling point", adopted July27, 1995. Preliminary Test The DSC-run ofthe preliminary test was recorded from 25 'C to 400 'C with a heating rate of20 'C/min

.An endothermic heat effect, the boiling, was observed at 99.8 'C. Afterthe experiment, the sample had lost 55% of its mass and the colour ofthe sample was black. Main Test In order to determine the boiling point more precisely, further DSC-runs were recorded between 25 'C and 215 'C with a heating rate of 10 'C/min. During these runs, the boiling point was determined to be 100.0 'C and 99.8 'C. The samples lost 51% and 50% oftheir mass, respectively. After the measurements the sample was a black powder.

The boiling point of Leuco Sulfur Black 1 is determined to be 99.9 'C 

## ± 0.1 'C which is equal to 373.1 K 

## ± 0.1 K. The atmospheric pressure during the measurements was 100.4 kPa (preliminarytest) and 99.8 kPa (main tests). The boiling point of Leuco Sulfur Black 1 is influenced by its high water content. It can be assumed that the measured value corresponds to water only. Cross-reference tootherstudy Weissanfefd (2009)Vapour Pressure, Harlan Laboratories Ltd. ltingen (Switzerland), Report C30953. Basis for calculation ofthe vapor pressure; further interpretation ofthe OSC

# 4.4 Density Endpointsummary: Density
UUID IUCS-69998325-7dbe-40fa-a9dc-551b451aa275 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea 

### Author Date Remarks pbaslo I Clarian1 Quimicos (Portugal) Lda I**Le~** 

### do Balio I Portugal 2010-04-28 16:44:18 CEST



# Administrative Data Short description of key information

The relative densily

### of Leuco SuHur Black 1 was detemined

### to be 0.5091 g/cm'(= 0.5091 · 10'kg/m') at20.4 

### 'C ± 0.1 

### 'C

# Endpointstudyrecord: KEY_OECD 109_2009_Harlan_C30942_Weissenfeld
UUID IUC5-8818132c-510d-496d-a8be-Odf73142a7a 7 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remari<s jregano I Clariant Iberica Producci6n SA I El Prat de Llobregat (Barcalona) I Spain 2010-05-20 16:35:17 CEST



# Administrative Data Purposeflag Study rasult

### type Reliability key study; robust study summary experimental resun Study pariod 

### 2009-03-19 till 

### 2009-03-30 

### 1 (reliable without restriction) Rationale

### for rallablllty Incl. cleflclancles Guideline conform study under GLP wnhout deviations 

# Data source Reference

Referance Author



### type study repori Weissenfeld M 

## Data access data submitter is data owner 

## Data protection claimed yes Year TIUe


### 2009 Determination 

### ofthe Relative Density



# Materials and methods Test guideline 

# IQualifier

# I Bibliographic Testing laboratory oource Harlan Laboratories Ltd. ltingen (Switzerland)

Guideline Report Owner 

### no. company 

### C30942 Ctariant Company Report otudy no. date 

### 2009-08- 

### 06



# I**Davia!~•**

# Iaccording EU Method A.3 (Relative Density) 

### to according OECD Guideline 109 (Density of Liquids and Solids) 

### to according otherguideline: EEC Directive 92169, 

### Part A, Methods for 

### the detennination 

### ofphysico-chemical properties, A.3 "Relative density", EEC Publication No. 

### to L383, December 1992 

### Type of method aircomparison pycnometer (forsolids) 

### GLP compliance yes (Ind. certificate) 

## Test materials Identity oftest material same as for substance defined in section 1 (if not read-across)
yes 

### Test material identity

Identifier Identity CAS number 86241-11-0 EC number 266-273-5 

### Details on test material

• Name oftest material : Leuco Sulfur Black 1

- Physical state: lhlxotrophic, black, wet paste -Composition: Leuco**Su~ur** Black 1:52.75% (wlw) (calculated by amountofby-compounds) 2,4-0initrochlorobenzene: < 20 ppm 2,4-0initrophenole: < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3 % (wlw) Water. 45.75 % (wlw) by**Ka~-Fischer** titration • Lot/balch 

### No.: ESA0007595 -Expiration dele of

### the lot/belch: 2011-07-11

- Stablllty undertest conditions: Stable under storage conditions. -Storage condition 

### oftest material: AI room temperatura (range 

### of20 

### ±5 'C), light protected. 

### no

no

no

## Any other infonnation on materials and methods incl. tables 

## Due to the properties ofthe test item the handling using the pycnometerwas difficult. Instead a measuring cylinder (volumetric flask) ofknown volume is filled with an accurately weighed amount ofthe test item. Test Item Treatment The test item was not dried before use, because the density ofthe whole test item mixture should be determined. General Test Procedure A volumetric flask was filled with a known amount oftest item and the volume ofthe test item was read. This procedure was performed in triplicate. During the performance ofthe test, the temperature was kept constant and measured by a thermohygrograph. 

# Results and discussion Density Type Denalty 

### Temp. Remerl<a relative density 0.51 20.4 

### 'C density 0.51 g/cm' 20.4 

### 'C 

## Any other infonnatlon on results Incl. tables 

## Detennination ofthe Density of Leuco Sulfur Black 1

Sample No. Temperature Sample Weight Sample Volume Density Relative ['C) 

## [g) [ml] [g/cm3] Density 

# 1 20.5 2.54327 5.0 0.5087 2 20.5 3.48919 6.8 0.5131

# 13 20.5 2.62940 

# Applicant's summary and conclusion Conclusions The relative density 

### of Leuco Sulfur Black 1was determined

### to be 0(20/4) 

### =0.5091 

## Executive summary 

## 5.2 0.5057 

# 0.5091 0.5091

The determination ofthe relative density ofthe test item was performed in accordance with the EEC Directive 92/69 and Council Regulation (EC) No. 440/2008, PartA, Methodsforthe determination ofphysico-chemical properties, A.3 "Relative density" and the OECD Guideline for Testing of Chemicals, No. 109, "Density of Liquids and Solids". The densitywas determined ata temperature of20.4 

# 'C 

# ± 0.1 

# 'C with three independentaliquots ofthe test item. The density ofthe test item Leuco Sulfur Black 1 was determined to be 0.5091 g/cm3 

## (= 0.5091 · 103 

## kg/m 3) 

## at20.4 

# 'C 

# ± 0.1 

# 'C using measuring cylinder. The relative density of Leuco Sulfur Black 1 was determined to be D(20/4) 

# =0.5091

# 4.5 Particle size distribution (Granulometry) Endpointstudyrecord: waiving_Particle size distribution (Granulometry)
UUID IUC5-b830d8fa-9e6c-40b1-beb4-0e2a65a18838 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author pbaslo I Clariant Quimioos (Portugal) Lda I**Le~** 

### do Ballo I Portugal Date 2010-03-23 13:25:28 GET 

### Remart<s 

# Administrative Data otherjustification JustltlcaUon fordataw•lvlng waived according lo column 2 of REACH Annex VII

. study nol required 

### W the substance is 

### sold under liquid form. Study result

### type other. 

# Applicant's summary and conclusion Conclusions Substance is sold only in liquid form, particle size distribution not required

. aooording to column 2 ofAnnex VII of REACH 

## Executive summary 

## Substance is sold only in liquid form, particle size distribution not required , according to column 2 ofAnnex VII of REACH

# 4.6 Vapour pressure Endpointstudyrecord: KEY_Calculation_2009 _Harlan_C30953_Weissenfeld
UUID IUC5-e9b1b80d-4a9d-4858-9bb8-56e8837f19fa Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks XML Transformation V1.0 Plug-In 2009-12-1117:14:09 GET Successfully migrated to IUCLID 5.2 format. 

# Administrative Data Purposeflag Dllblwalvlng Justificationfor

### data waiving Study result

### type Reliability used for dassification otherjustification Calculation using Modified Watson Correlation estimated by calculation 1 (reliable without restriction) 

### Rationale for reliability Incl. deflclancles Using the boiling point of Leuco Sulfur Black 1 was determined by expertmentel study C30931, the 

### vapor pressure can be calculated using Modified Watson Correlation 

# Data source Reference

Reference Author



### type study report Mlrjam Weissenfetd 

## Data access data submitter Is data owner 

## Data protection claimedYear Title Bibliographic source
2009 Leuco Sulfur Black 1
- W.J. Lyman, W.F. Reahland D.H. Rosenblatt Calculation ofthe Vapor 

### (ad); Handbookof Chemical Property Estimation 

### Pressure Methods, McGraw-Hill, NY (1990) Testing Report Owner Company Report laboratory 

### no. company study no. date Hanan C30953 Clartant 

### 2009- Laboratortas, 07-16 Ltd

### yes 

# Materials and methods Principles ofmethod 

## ifotherthan guideline Hthe vapor pressure ofLeuco Sulfur Black 1 is expected lo be so small that the value is below the measurement range 

### ofan experimental detanninalion (e.g. the gas saturation method), then 

### the vapor pressure can also be calculated, using the Modified Watson Correlation. This method is described by Lyman etal. (1990). This method calculates the vapor pressure 

### from the boiling point ofthe test item. 

# Test materials Details on test material Identification: Leuco Sulfur Black 1 CAS 

### No.: 66241-11-0 Description: Thixothrophic, black, wet paste Balch Number: ESA0007595 Composition: Leuco Sulfur Black 1: 52.75% (calculated by amount 

### ofby-compounds) 2,4-Dinitrochlorobenzene: < 20 ppm 2,4-Dinitrophenol: 

### < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3% (w/w) Water: 45.75% (wlw) by Karl-Fischer titration Stability of

### the Test Hem: Stable under storage condHions 

### Expiry date: July 11,2011 Storage Conditions: At 

### room temperature (range 20+I- 5 'C). light protected 

# Results and discussions Vapour pressure

< 0.0000002 

### Pa**25~** 

## Any other Information on results incl. tables 

## The estimation ofthe vapor pressure of leuco Sulfur Black 1 was based on the EEC directive 92/69, Part A, Methods for the determination of physico-chemical properties, A.4 "Vapour Pressure", EEC Publication No. l383, December 1992 and the OECD Guideline No. 104, "Vapour Pressure", adopted July 27, 1995.

# Applicant's summary and conclusion Conclusions The vapor pressure of Leuco Sulfur Blacl 1 is below 

### 1.8 

## x 10 (-7) Paat25

## •c Executive summary 

## The boiling point of leuco Sulfur Black 1was determined in study C30931. In this studythe DSC showed an endothermic heat at 99.9 °C. This temperature is notthe true boilng point ofthe test item as it only reflects the boiling point ofthe water contained

. The DSC shows no further endothermic heat effect up to 400°C. Thus, the boiling point ofthe Leuco Sulfur Black 1 can be regarded as being above 4000C Based on this boiling pointthe vapor pressure of Leuco Sulfur Black 1 was calculated being below : 1.8 x 10·7

## Pa at 25

# oc calcuted using Modified Watson Correlation

# 4.7 Partition coefficient Endpointsummary: Partition coefficient
UUID IUC5-2bccdaa6-24c1-4fad-94b6-5fbd39ac087b Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author pbaslo I Clariant Quimicos (Portugal) Lda I**Le~** do Balio I Portugal Date 2010-06-1111:59:21 CEST Remarks 

# Administrative Data Short description of key information
Impossible to determine the log Pow 

# Discussion No chromatographic method is available to estimate the solubility of Leuco Sulfur Black 1 in n-octanol

_ Therefore the log Pow cannot be determined

# Endpointstudyrecord: Harlan Laboratories (2010) /K2 KS!Partition coefficient_statement UUID**IUC5~0d7160c-5fe4-4930-89d5-7e65a039849a** Dossier UUID IUC5-292149d1-a677-41eb-9692-1fad973281ea Author Date Remal't(s jregano I Clariant Iberica Producci6n S.A. I El Prat 

### de Llobregat (Barcelona) I Spain 2010-00-01 10:49:04 CEST



# Administrative Data Purpoaeftag key study; robust study summary Justification fordatawelvlng Study ruult

### type No chromatographic method is applicable to determine the solubility ofLeuco Suffur Black 1 in n octanol. estimated by calculation Study

### period Reliability 2 (refiable with restrictions) April 14, 2010

R.UOnele

### for reliability Incl. deficiencies Study conducted in accordance with generally accepted scientific principles, possibly with incomplete reporting or methodological deficiencies, 

### which do not affectthe quality 

### of the relevant results. 

# Data source Reference

Ref11rence Author



### type study report Dr Martin KOhne 

## Data access Year
2010

data submitter is data owner 

## Data protection claimed yes Title
Calculation 

### ofthe Partition Coefficient (n-Octanoll Water



# Materials and methods Partition coefficient

## type Bibliographic 

### source Testlnglaboretory Report Owner Company Report no. company study no. date**Ha~an** Laboretorias Ltd. Zelgllweg 1 C88384 Clariant 2010-04- 4452 ltingen 

### I Switzerland 14octanol-water 

### Test guideline

noguideline followed 

### Type of method other: no method was applicable 

### Principles ofmethod Ifotherthan guideline The test was not feasible to complete 

### GLP compliance

no (not mandantory) 

## Test materials Test material Identity Identifier Identity CAS number 66241-11-0 

### Common name Leuco Sulfur Black 1 

### Details ontest material Identification: Leuco Sulfur Black 1 CAS Nr.: 66241-11-0 Description: Thlxolrophlc, black, wet presscake Batch Number: ESA0007595**Compos~lon:** Leuco**Su~ur** Black 1: 52.75% (calculated by amount 

### ofby-compounds) 2.4-Dinltrochlorobenzene: < 20 ppm 2.4-Dinltrophenole: < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3% (w/w) Water. 45.75% (w/w) by Karl-Fischer tilration Stability 

### ofthe test**~ern:** Stable under storage conditions Stability 

### ofthe last Hem dilution: > 4 h In waterExpiry date: July 11, 2011 Storage Conditions: At room temperature (range 20 +I- 5 

### 'C). light protected 

## Analyticalmethod

no data (Itwas notfeasible to complete) 

## Any other Information on materials and methods Incl. tables No data available. 

# Results and discussions Details on results No chromatographic method 

### is applicable to datermine the solubility of Leuco Sulfur Black 1 in n octanol. Therefore, no Partition coefficient could be darived. 

## Any other Information on results Incl. tables 

## The partition coefficient (P

# owl is defined as the ratio ofthe solubility ofthe test item in octanol and in water. No chromatographic method is applicable to determine the solubility of Leuco Sulfur Black 1 in n-octanol. Although it is possible to estimate the water solubility of Leuco Sulfur Black 1 (62.4 mg/L as determined in Harlan Laboratories Study C75560) by measurementofthe Total Carbon (TC) content this analytical method is inapplicable in case of n-octanol as a solvent.

Therefore it is impossible to determine the log PowofLeuco Sulfur Black I according to the guideline. 

# Overall remarks, attachments Overall remarks No determination of partition coefficient by Method A8 

### ofCommission Directive 92169/EEC (which constijutes Annex V ofCouncil Directiva 671548/EEC) was possible. 

# Applicant's summary and conclusion ConclusionsNo chromatographic method is available to estimate 

### the solubility of Leuco Sulfur Black 1 in n-octanol. Therefore the log POW cannot 

### be determined according 

### to the guideline. 

## Executive summary 

## No chromatographic method is available to estimate the solubility of Leuco Sulfur Black 1 in n-octanol. Therefore the log P0 

## wcannot be determined according to the guideline.

# 4.8 Watersolubility Endpointsummary: Watersolubility UUID IUCS-492eba04-b1 ef-4ab4-b62b-3d9affdl54b1 Dossier UUID IUCS-292149d1-a677-4feb-9692-1fad97328 1 ea Author pbasto 

### I Clariant Quimicos (Portugal) Lda 

### I Leca do Balio 

### I Portugal Date 2010-04-28 17:37:06 CEST Remarks 

# Administrative Data Short description of key information

The water solubility Is 62.4 mg/l at 20.0 'C and a pH of7.9 

# Discussion The watersolubility of Leuco Sulfur Black 1 was determined to be 62.4 mg/L at 20.0 

# 'C and a pH of7.9 based on a measured concentration of 11.5 mg/L ofcarbon

# Endpointstudyrecord: Harlan Laboratories (2010) I K1 KS/watersolubilityLeuco Sulfur Black 1.CiariantProducts C75560
UUID IUC5-8afebae3-bd7a-4e44-89db-43f0b4485180 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks jregano I Clariant Iberica Producci6n SA I El Prat de Llobregat (Barcelona) I Spain 2010-06-01 10:38:47 CEST



# Administrative Data Purpose 

### flag Study result

### type Reliability key study: robust study summary experimental result 1 (reHable without restricllon) Study period 15-02-2010111-03-2010

Rationale for reliability Incl. deficiencies Study conducted 

### in compliance 

### with egreed protocols, 

### with no or minordeviationsfrom standard test guidelines and/or minor methodological deficiencies, which do

### not affect

### the quality 

### ofthe relevant results. 

# Data source Reference

Reference Author



### type study report Dr Martin KOhne 

## Data access Year
2010

data submitter is data owner 

## Data protection claimed yes, butwHiing to share TIUe
Leuco Sulfur Black 1: Determination of

### the Water Solubility



# Materials and methods Bibliographic 

### source Testing laboratory Report Owner Company 

### Report no. 

### compeny study no. 

### date Hanan Laboratories Ltd. ZelgHweg C75560 Clariant 2010-03- 14452 ltingen I Switzerland 11

### Test guideline Qualifier Guideline Dev!Mions according to EU MethodA.6 (WaterSolubility) 

## no according to OECD Guideline 105 (Water Solubility) 

## no Type ofmethod ftask method 

### GLP compliance
yes (ind. certificate) (Date 

### of Signature: November 12, 2008) 

## Test materials Test material Identity
Identifier Identity 

### Common name Leuco Sulfur Black 1 CAS number 66241-11-0 

### Details on test matenal Identification: Leuco Sulfur Black 1 Description: Thixotrophic, black, wet presscake 

### Batch Number: ESA0007595 Composition: Leuco Sulfur Black 1: 52.75% (calculated by amount ofby-compounds) 2,4-Dinitrochlorobenzene: < 20 ppm 2,4-Dinilrophenole: 

### < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3% (wlw) Water: 45.75% (w/w) by Kart-fischertitration Stability of

### the lest**~em:** Stable under storage conditions StabHity ofthe test**~em** dilution: > 4 h in water Expirydale: July 11, 2011 Storage Conditions: AI room temperature (range 20 +/- 5 'C), light protected 

### Details on methodsThe determination was carried out using

### the flask melhod, Method AS Water Solubility ofCommission Regulation (EC) No 440/2008 of 30 May 2008. 

## Any other information on materials and methods Incl. tables Test Perfonnance Test Solutions About1.5- 1.7 g oftest item were weighed into each of6 Erlenmeyerflasks and 50 mL ofdouble distilled waterwere edded 

### to each flask. The tighUy dosedflasks were shaken at aboul30 

### 'C for 24 hours, 48 hours and 72 hours, respectively. Aftarwards, 

### the flasks were equHibrated for another 24 hours at20.0 

### 'C. The pH was measured to be aboul7.9. Afterwards, the supernatant solutions were centrifuged (-2900 g for 10 minutes), filtered (0.2**~m.** Nylon)and 5 mL ofthis solution was diluted upto 50 mL with water prior

### to analysis. The quantification was performed using TC measurement as described below. Details of sample treatment are given inTable 1 below. 

### Analytical Method The concentration of the test item was analyzed using TC measurement. The test samples were quantified 

### wtth reference to a calibration curve, 

### which was generated automatically. The last calibration was performed on January 06, 2010. 

### System Suitability check A system suitability check was performed before sample measurement in order

### to check 

### the reliabilrty of the measurement. 

### Preparation of 1,4-Benzochlnone Solutions 75.02 mg of 1,4-banzochlnone were filled upto a volume of 1 L with MiiiK:lwater

### to get a stock solution of nominal conoantration of50 ppm of carbon. 2 mL ofthis stock solution was diluted 

### to a volume of200 mL wrth Mllli-Q waterto get a solution ofa nominal ooncentration of0.5 ppm ofcarbon. A seoond solution ofa nominal conoantration of 10 ppm ofcarbon was prepared by diluting 20 mL oftha stock solution upto 100 mL with MilK:!water.For both solutions, 

### the TOC was measured. The system surtabillly checkwas sucoassfully as all TOC 

### values were wHhin the specifications. 

### Prepar.tlon of

### Saccharose SoluUona 118.99 mg ofsaccharose were filled upto a volume 

### of 1 L with Milll-0 waterto 

### get a stock solution ofnominal concantretion of 50 ppm ofcarbon. 2 

### mL ofthis stock solution was dButed 

### to a volume of200 mL with**Miii~Q** water

### to get a solution ofa nominal oonoantration of 0.5 ppm ofcarbon.Afterwards, the TOC was measured. The system suitability 

### check was sucoassfuUy as 

### the TOC value was within 

### the specifications. 

### Data Handling The samples were quantified by Total Carbon (TC) measurement with referenoa 

### to a calibration curve.The calibration curve was calculated using the software. The concentration of carbon ofthe injected samples was quantified by peak area wrth referenoa 

### to the calibration curve. From the results obtained bythe software the conoantration ofcarbon in the analyzed solutions was calculated bythe following equation: (1)Where C1 Measured concentration ofcarbon in the sample solution [ppm= mgll] 

## c2 Concentration of carbon in the test item solution before dilution (mgll] I OHution factor From lhe concenlration ofcarbon the watersolubility ofthe test Hem was calculated bythe following equation: (2) Where C3 = Water solubility ofthe test Item 

### (mgll] Cc Percentage of carbon in lhe test Hem as obtained by elementaranalysis (here: 

### 18.5%) T

### able1 Data ofthe Performance 

### of the Water SolubililvTest

Sample Solution No. 

### 1 2 3 4 5 6
Weighed sample 

### of lhetest Hem [mg] 

### 1482.52 1674.48 1473.65 1621.87 1734.69 1563.06

Added volume ofwater (ml] 

### 50 50 50 50 50 50

Agitation time at 30 

### 'C (hours] 

### 72 72 48 48 24 24

Equilibration time at 

### 20 'C (hours] 

### 24 24 24 24 24 24

pH value 

### oftest item solution 

### 7.60 7.52 7.68 7.76 8.32 8.29

DUution factor before quantification 

### 10 10 10 10 10 10

# Results and discussions Water solubility 62.4 mgll Temp. 20 

### 'C

pH 7.9 

### Details 

### on results Please see section Remarl<s on resulls including tables and figures. 

### Any other Information on results Incl. tables The delermination ofthewatar solubility of Leuco Sulfur Black 1was performed according to the Directive 92169. A.6 "Water Solubility" (1992) and the OECD guideline No. 105. "Water Solubility" (1995) using theflask shaking method. After a shaking time of24 hours. 48 hours and 72 hours. respectively. at 

### 30 'C. 

### the samples were equilibrated for another24 hours at 20 

### 'C. Afterwards. 

### the samples were centrifuged. filtered. diluted and analyzed by TC measurement. Tha water solubility of Leuco Sulfur Black 1 at 20

## •c was determined to be 62.4 mg/L. Tha measured results diller by less than 15%. Therefore. 

### the study is considered to be valid.The resulls of

### the Individual measurements of

### the main test are summarized in Table 2 below.The tabulated values represent rounded results. which were obtained by calculation using 

### the exact 

### raw data. 

### T ble2 a 

### R esults ofth 

## w e ater Sol 

### Ill ub 1tyo Leuco**Su~** Bla ur cl<1 Concentration oftest item. Sample 

### No. Shaklngtime at30 

### 'C Concentration ofcarbon. Concentration of carbon. calculated Water solubility calculated analyzed calculated (rounded) [mg/L] (mg/L] [mg/L] 

### (mg/L] 1 1.094 10.94 59.14 72 2 1.400 14.00 75.68 3 1.147 11.47 62.00 62.35 48 4 1.104 11.04 59.68 

### so= 6.90 5 1.143 11.43 61.78 24 6 1.033 10.33 55.84

### The saturation mass concentration is calculated from the boldvalues SO 

### =standard deviation 

# Applicant's summary and conclusion Interpretation of results**sligh~y** soluble (0.1-100 mg/L} 

### Conclusions During this studythe watar solubility of Leuco Sulfur Black 1 was detennined and found to be 62.4 mg/l at 20 

### 'C and a pH of7.9 

### based on a measured cone en!ration of 11.5 

### mgJL of carbon using the flask shaking method. 

### Executive summary During this study 

### the water solubility of Leuco Sulfur Black 1 

### was determined. Vessels containing water and test Item were shaken for 24 hours, 48 hours and 72 hours, respectively, at 30 

### 'C. Afterwards the samplas were equilibrated for another24 hours at20.0 

### 'C. After dilution with water

### the concentrations in the aqueous solutions were determined bya TC-malhod. 

### The watersolubility ofLeuco Sulfur Black 1 was detennlned to

### ba

based on a measured concentration 

### of

using 

### the flask shaking method. 

### Cross-reference to otherstudy System Suitabilitytestwas 

### performed in Study C79744 

### 62A 

### mg/L at 20.0 

### 'C and 

### a pH 

### of 7.9



### 11.5 mgiL 

### of carbon

# 4.10 Surface tension Endpoint studyrecord: Waiving_Surface tension
UUID IUC5-4c623182-907b-4871-8672-d85c85dc5463 Dossier UUID IUC5-292149d1-a6n -4feb-9692-1fad973281ea Author Date Remarks jregano I Clariant Iberica Producci6n S.A. I El Prat de Llobregat (Barcelona) I Spain 2010-05-18 14:38:20 CEST



# Administrative Data Data waiving 

### Justification for data waiving otherjusttficatk>n waived according to column 2 of REACH Annex VII: 

### the surface activity is nota desired property 

### ofthe material. Thare is no hydrophylic haad and no hydrophobic tail in tha stnucture oftha substanca.

# 4.11 Flash point Endpointstudyrecord: Waiving_F/ash point UUID IUCS-37401ac0-953b-413c-9c37-379871eee02c Dossier UUID IUC5-292149d1-a6n-4feb-9892-1fad973281ea Author Dale Remarks pbasto I Clariant Ouimicos (Portugal) Lda I**Le~** do Balio I Portugal 2010-03-23 13:17:23 

### GET



# Administrative Data Data 

### waiving study scientifically unjustified 

### Justificationfor

### datawaiving Waived 

### accordig to column 2 

### of REACH Annex VII ,

### test will 

### not be preformed as the substance is

### a solid end 

### does not contain any organic 

### solvent 

# Results and discussions Flash point

Remarks Substance it a solid, without organic solvents 

# Applicant's summary and conclusion Conclusions Solid substance Doesn't contain organic solvent 

## Executive summary 

## Data not required , according to column 2 of REACH Annex VII , as it is a solid substance ,thatdoesn't contain any organic solvent

# 4.12 Auto flammability Endpointsummary: Auto flammability UUID IUC5-a55c7ca4-20c1-4399-8355-f1cb24d69906 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks pbasto I Clariant Quimicos (Portugal) Lda I Lei'S do Balio I Portugal 2010-D4-28 17:09:53 CEST



# Administrative Data Short description of key information

tt has been detenmined 

### to have an auto-ignition temperature of 348 

### +1- S'C.

# Endpointstudyrecord: KEY_EUA.15_2009_Harlan_0047/310B_Tremain/Atwal UUID IUC5-afcfd8ff-193b-4453-932d-d1699fd8f71a Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date 

### Remarks jragano I Clariant Iberica Producci6n SA I El Prat de Llobragat (Barcelona) I Spain 2010-05-1815:26:23 CEST



# Administrative Data Purpose 

### !lag Study result

### type Reliability key study; robust study summary experimental result Study period 2009-02-11 till2009-02-17 1 (reliable 

### without restriction) Rationale for reliability Incl. deficiencies Guideline conform study under GLPwithout deviations 

# Data source Reference

Reference Author Year

type study report Tremain SP, 2009 Alwal SS 

## Data access data submitter is data owner 

## Data protection claimed yes 

### Title
Leuco Sulfur Black 1: Determination of Auto-Ignition Temperature (Liquids and Gases)



# Materials and methods Test guideline Bl>llographlc Tasting laboratory 

### source Harlan Laboratories Lid. Shardlow (United Kingdom) Report 

### Owner Compeny

### study no. no. company 004713108 Clariant Harlan Laboratories Ltd. (Switzerland) C31008 

### Report dele 2009- 06-15Qualifier 

# J Guideline 

# lo.vtmlona according lEU Method A.15 (Auto-Ignition Temperature (Liquids and Gases)) (Council R&gUatlon (EC) No 44012008 

### of 30 May 2008, PartA: Methodsforthe 

# lno to delerminatlon ofphysico-chemical properties.) 

## GLP compliance yes (ind. certificate) 

# Test materials Identity

## oftest material same as forsubstance defined In section 1 (If not read-across)
yes 

## Test material identity

Identifier Identity CAS number 66241-11-0 ECnumber 266-273-5 

## Details ontest material

- Name 

### oftest material : Leuco Sulfur Black 1

- Physical state: thixotrophic, black, wet paste

- Composrtion: Leuco Sulfur Black 1:52.75% (wlw) (calculated 

### by amount of by-compounds) 2,4-Dinitrochlorobenzene: < 20 ppm 2,4-Dinilrophenote: < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3% (wlw) Water: 45.75% (w/w) by**Ka~-Fischer** titration -Lot/batch No.: ESA0007595 -Expiration date

### ofthe lot/batch: 2011-07-11 -Stabilityundertest conditions: Stable understorage conditions. -Storagecondition oftest material: At room temperature (range of20 :t 5 'C), tight 

### protected. 

## Any other Information on materials and methods Incl. tables



## The auto-ignition temperature was determined by heating aliquots ofthe test material in a flask and observing for any

## ignition. Procedure The test flask was heated in a flask heater. Aliquots oftest material (tested as received) were dropped into the flask, which was observed for signs of ignition over a 300 second period. This procedure was repeated, varying the sample size, as necessary, until the lowesttemperature atwhich ignition, ifany, occurred within 300 seconds of insertion, was determined. 

# Results and discussions Autoflammablllty I 

## Self-Ignition temperature 348't: at >= 

### 101.46- <= 

### 102.46 kPa 

## Any other Information on results Incl. tables 

## The test material has been determined to have an auto-ignition temperature of348 

## +/- 5'C Results Test Temperature TestWeight Ignition Delay Observations 

## (oCj_ Ig]_ rsl 50 (clean flask) 0.2 >300 No Ignition 100 (clean flask) 0.2 >300 No lanition 150 0.2 >300 No Ignition 200 0.2 >300 No Ignition 250 0.2 >300 No!gnition 300 0.2 >300 No Ignition 350 0.2 >300 No lonition 400 (clean flask) 0.2 >300 No Ignition 400 (clean flask) 0.4 >300 No Ignition

## 400 clean flask) 0.6 182 An oranoe flame 396 clean flask) 0.6 74 An orange flame 392 clean flask) 0.6 >300 No lonition 394 clean flask 0.6 164 

# An orange flame 392 clean flask) 0.8 47 

# An orangeflame 388 (clean flask) 0.8 163 

# An orange flame 384{clean flask) 0.8 154 

# An orange flame 380 (clean flask 0.8 97 

# An orangeflame 376 (clean flask 0.8 151 

# An orange flame 372 (clean flask 0.8 86 

# An orang_e flame 368 (clean flask 0.8 134 An orange flame 360 (clean flask 0.8 97 

# An oranoe flame 352 (clean flask) 0.8 >300 No Ignition 356(clean flask 0.8 138 An orange flame 354 (clean flask) 0.8 >300 No Ignition 354 (clean flask) 1.0 67 

# An orange flame 350 (clean flask 1.0 >300 No Ignition 352 (clean flask) 1.0 264 An orange flame 350 (clean flask 1.2 201 

# An orange flame 348 (clean flask 1.2 >300 No Ignition 348 (clean flask) 1.4 >300 No Ignition 348 (clean flask) 1.6 >300 No Ignition 348_(clean flask 1.8 >300 No Ignition 348 (clean flask) 2.0 75 

# An orange flame 346 _(_clean flask 2.0 >300 No lonition 346 (clean flask 2.2 >300 No Ignition 346 (clean flask) 2.4 >300 No lonition 346 (clean flask 2.6 >300 No Ignition

# I346 (clean flask) 2.8 

# Applicant's summary and conclusion Conclusions
The test material has been determined

### to have an aut<Hgnition temperature 

### of 348 +I· 5"C. 

## Executive summary 

## >300 

# I No Ignition

The auto-ignition temperature was determined using Method A15 specified in Council Regulation (EC) No 44012008 of30 May 2008, Part A: Methods forthe determination of physico-chemical properties. The test material has been determined to have an auto-ignition temperature of348 +/- 5'C.

# 4.13 Flammability Endpointsummary: Flammability
UUID IUC5-c59109d0-bfe84caf-9122-5b4ccb2bf665 Dossier UUID IUC5-292149d1-a6n 4feb-9692-1fad973281ea Author pbaslo I Clariant Quimioos (Portugal) Lda I**Le~** do Balio I Portugal Date 2010-04-28 17:14:47 CEST Remarks 

# Administrative Data Short description of key information

Leuoo Sulfur 

### Black 1 Is not "highlyflammable" 

# Discussion Leuco Sulfur Black 1 could not be ignited with a flame during the preliminary test

# Endpointstudyrecord: KEY_2009_Harlan_C30986_Weissfeld
UUID IUC5-<:2b41d2a-f5a8-441d-8a12-<!65bfc89c86e Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973261ea 

### Author Date 

### Remart<s jregano I Clariant Iberica Producci6n S.A. I El Prat de Llobregat (Barcelona) I Spain 2010-05-16 12:32:03 CEST



# Administrative Data Purpose flag Study result

### type Reliability key study; robust study summary experimental result 1 (reMable without restriction) Studyparlod March 24,2009- August4, 2009

This study followed the procedures indicated by 

### the following intematlonally accepted guidelines and recommendations: 

### Rlllionale for reliability Incl. deflclanclea • EEC Directive 92169, Part A, Methods forthe detennination ofphysico-chemical pnopartias, A.10 "Flammability (solids)". EEC Publication No. L363, December 1992. 

# Data source Reference

Relwence Author



### type study report Welssenfeid M 

### Data access data submitter is data owner 

### Data protection claimed
yes, 

### bul willing Ia shere 

### Year
2009 • Council Regulation (EC) No. 44012006, 30 May 2008, PartA, Methods lorthe detannination of physico-chemical properties, A.10 "Flammability (solids)"

Title Bibliographic Testing laboratory 

### Report 

### Owner Company study 

### Report source no. company no. date Leuco Sulfur Black 1
- Detennination of Harlan C30986 Clariant 2009-08- 

### the FtammabiUiy Laboralorias, Ltd 

### 04



# Materials and methods

## Test guideline Guideline EU Method A.10 (Flammability (Solids))



### yas (ind. certificate) 

## Test materials Identity oftest material same as for substance defined In section 1 (Ifnot read-across)
yes 

## Test material Identity

Identifier 

### Identity CAS number 66241-11-0 EC number 266-273-5 

## Details on test material Identification: Leuco Sulfur Black 1 CAS No.:66241-11-0 Description: Thlxothrophic, black, wet paste Batch Number: ESA0007595 ComposHion: Leuco Sulfur Black 1: 52.75% (calculated by amount ofby-compounds) 2,4-0in"rochlorobenzene: < 20 ppm 2,4-Dinitrophenol: < 20ppm**Su~ate:** 970 ppm Chloride: 750 ppm Sodium: 1.3% (w/W) Water: 45.75% (w/W) by Kar1-Fischar titration Stability

### of

### the Test Item: Stable under

### storage conditions Expiry data: July 11, 2011 Storage Conditions: At room temperatura (range 20 +/- 5 'C), light protected 

## Any other Information on materials and methods Incl. tables 

## Preparations beforethe TestThe test item was homogenized by kneading and an aliquot was put onto an aluminium plate to form a pile with a length of250 mm and a height ofabout 10 mm.

## PreliminaryTestAn attemptwas made to ignite the sample with the aim to determinethe burning time over a distance of 200 mm, 

# ifapplicable. The flame ofa gas burnerwas used as ignition source. The hotflame was applied for a maximum of two minutes. The test was conducted in a fume cupboard with constant air speed. Additionally, a draught screen was erected around the apparatus. Main TestNofurther tests were performed according to the results obtained in the preliminary test. 

# Results and discussions Solidniquid: Ignition on contactwith air

no 

# Applicant's summary and conclusion Interpretation ofresults other: not highly flammable 

## Conclusions Leuoo Sulfur Black 1 is not "highlyflammable" according to the cr1teria ofthe guideline. 

## Executive summary 

## The determination ofthe flammability ofthe test item was performed in accordance with the EEC Directive 92/69, EEC Publication No. L383, December 1992 and Council Regulation (EC) No. 440/2008, 30 May 2008, PartA, Methods for the determination ofphysico-chemical properties, A. 10 "Flammability (solids)". In contact with the ignition source, the flame ofthe gas burner turned orange. Leuco Sulfur Black 1 emitted some glowing drops. Glowing sustained for about 1 minute butdid notexpand.Leuco Sulfur Black 1 could not 

# be Ignitedwith a flame during the preliminarytest (contacttime ofabout 2 minutes). Therefore, no main testwas performed.

# 4.14 Explosiveness Endpointsummary: Explosiveness UUID IUC5-aa8e5b5a-7b46-4efc-bccc-<la9a15273b5d Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author pbasto I Clarianl Quimicos (Portugal) Lda I Le99 do Balio I Portugal Date 2010-04-28 17:19:21 CEST Remarks 

# Administrative Data Short description of key information

Leuoo Sulfur Black 1 is notexplosive 

# Discussion Leuco Sulfur Black 1 is not a potential explosive and does not have a potential for rapid energy release.

# Endpointstudyrecord: KEY_ECA.14_2009_Harlan_C30997 _Weissenfeld
UUID IUC5-3776037a-1d18-4fb6-b86!>-583abca215d1 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks jregano I Clariant Iberica Producci6n S.A. I El Prat de Llobregat (Barcelona) I Spain 201Q-05-1815:28:43 CEST



# Administrative Data Purposeflag Study result

### type key study; robust study summary experimental resutt Study 

### period 2009-03-18 Reliability 1 (reliable 

### without restriction) Rationale lor reliability Incl. deficiencies Guideline conform study 

# Data source Reference

Reference Author Year Title Blbllogrephlc



### type source Testlngle-ry study report Weissenfeld 

### 2009 Determination 

### ofthe Hanan Laboratories Ltd. ltingen 

### M 

### Data access data submitter Is data owner 

## Data protection claimed
yes Explosive Properties



# Materials and methods Test guideline (Swilzerland)

Guideline 

### Report Ownar 

### no. company C30997 Ciariant Company Report study no. 

### data 2009-07- 16



# I**Devlet~n•l**otherguideline: UN Recommendations on the Transport of Dangerous Goods (Manual ofTests and Criteria, Fourlh revised edilioo, 2003, Appendix 

### 6, "Orange Bool<"



### no (not required) 

## Test materials Identity oftest material same as for substance defined In section 1 

### (If not read,.cross) yes 

### Test material identity

Identifier Identity CAS number 66241-11-0 ECnumber 266·273--5 

### Details ontest material

• Name oftest material : Leuco Sulfur Black 1 ·Physical state: thixotrophic, black, -tpaste • ComposHion: Leuco Sulfur Black 1:52.75 % (wlw) (calculated by amount of by-compounds) 2,4-Dinitrochlorobenzene: < 20 ppm 2,4·Dinitrophenote: < 

### 20 ppm Sulfate: 970 ppm Chloride: 

### 750 ppm Sodium: 1.3% (wlw) Water: 45.75% (wlw) by Kart-Fischer titralioo • Lot/batch 

### No.: ESA0007595 ·Expiration date ofthe lot/batch: 2011-07·11 • StabiiHy undertest conditions: Stable under storage condilioos. ·StoragecondHion 

### of test material: At room temperatura (range of

### 20 ± 5 

## q, light protected. 

# Results and discussions Explosive under influence offlame

no More sensitive to shock than m-dinltrobenzene not determined

## More sensitive tofriction than m-dlnltrobenzene not determined 

## Explosive (not specified)

no 

## Any other Information on results Incl. tables 

## Calorimetric Tests The exothermic decomposition energywas determined using Differential Scanning Calorimetry in a closed, gold plated high pressure vessel (DSC). The calorimetric test is ofspecial importancefor the evaluation of compounds containing chemical groups associated with explosive properties or 

## if no appraisal ofthe molecularstructure could be performed. There was no exothermic peakfound. Thus the decomposition energy does not exceed the UN limitof 500 

# Jig (AHDec determined between room temperature and 500 

# 'C ). 

# Applicant's summary and conclusion Conclusions Leuco Sulfur Black 1is not to be classified as explosive material and no experimental determination according 

### to the EC test guideline A.14 has to be performed. 

## Executive summary 

## Reactive Groups and Oxygen Balance No molecular structure was given for Leuco Sulfur Black 1. Thus, the appraisal ofthe molecular structure and the calculation ofthe oxygen balance could not be performed. Calorimetric Tests The exothermic decomposition energy was determined using Differential Scanning Calorimetry in a closed, gold plated high pressure vessel (DSC). The calorimetric test is ofspecial importance forthe evaluation ofcompounds containing chemical groups associated with explosive properties or if no appraisal ofthe molecular structure could be performed. There was no exothermic peak found. Thus the decomposition energy does not exceed the UN limit of 500 

# Jig (AHDec determined between room temperature and 500 

# 'C).

## Leuco Sulfur Black 1 is notto be classified as explosive material and no experimental determination according to the EC test guideline A.14 has to be performed. Thus it can be concluded beyond a reasonable doubt, that Leuco Sulfur Black 1 is not a potential explosive and does not have a potential for rapid energy release. Nevertheless, thisjudgment is in respect to classification; it does not necessarily mean that any processing hazards can be discounted.

# 4.15 Oxidising properties Endpointsummary: Oxidisingproperties UUID IUCS-0948cf5e-1ade-4e34-b673-4087945048a4 Dossier UUID IUCS-292149d1-a677-4feb-9692-1fad973281ea Author pbasto 

### IClariant Quimicos (Portugal) Lda 

### I**Le~** 

### do Balio 

### I Portugal Date 201CHJ4-28 17:20:40 CEST Remarks 

# Administrative Data Short description of key information

Leuco Sulfur Black 1has no oxidizing properties.

# Endpointstudyrecord: KEY_EU.A17_2009_Harlan_C31010_Weissenfeld
UUID IUC5-2c948e76-d6b1-4015-89d3-db14c11ba980 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarl<s jregano I Clariant Iberica Producci6n SA I El Prat de Llobregat (Bart:elona) I Spain 2010-05-1815:29:24 CEST



# Administrative Data key study; robust study summary Purpose 

### flag 

### Study resun type Rllllabllhy experimental**resu~** Study period 2009-{)4-03 till 2009-{)4-07 1 (reHabie without restriction) Rillionaie for rellabllhy Incl.deficiencies Guideline conform study underGLP without deviations 

# Data source Reference

Referencs Author



### type study report Weissenfeld M 

## Data access data submitter is data owner 

## Data protection claimed yes Year Title
2009 Determination 

### ofthe Oxidizing Properties (Solids)



# Materials and methods Test guideline Bibliographic Testlng laboratory source Harlan Laboratories Ltd. ltingen (Switzarland)

Guideline Report Owner no. 

### company C31010 Clariant 

### Company Report study

### no. data 2()()9..()8- 

### 06



# I**Devmt~n•l**according EU Method A.17 (Oxidising Properties 

### (Solids)) 

### to according 

### otherguideline: EEC Directive 92169, PartA, Methods forthe determination ofphysico-chemical properties, A.17 "Oxidizing Properties (solids)", EEC to Publication 

### No. L383, December 1992. 

### GLP compliance yes (incl. oertlficate) 

## Test materials Identity oftest material same as for substance defined in section 1 (If not read-across)
yes 

### Test material Identity

Identifier 

### Identity CAS number 66241-11-0 EC number 266-273-5 

### Details on test material

- Name

### of

### test meterial : Leuco Sulfur Black 1

- Physical state:thlxotrophic, black, wetpaste

- Composition: Leuco Sulfur Black 1:52.75 

### % (w/w) (calculated by amount ofby-compounds) 2,4-0initrochlorobenzena: < 20 ppm 2,4-Dinltrophenola: < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3 

### % (w/w) Water. 45.75% (w/w) by Karl-Fischertitration -Lot/batch No.: ESA0007595 -Expiration date ofthe lot/batch: 2011-07-11 -Stability undertest conditions: Stable understorage conditions. -Storage condition 

### oftest material: At room temperature (range of 20 

### t 5 'C), light protected. 

### Contact 

### with powdered cellulose 

### Any other information on materials and methods incl. tables no



### no

## The preliminarytestwas performed to determine ifoxidizing properties were to be expected (as conduded from the structural formula ofthe test item). The preliminary test was also conducted to ascertain adequate safety precautions during the main study. A barium nitrate/cellulose mixture (at a ratio 2:1 (w/w))was prepared with dried and sieved(< 0.125 mm) material. The test item is a paste containing water. Asthe test item should be analyzed as delivered, according to sponsor's request, the test item was not dried at 105'C prior to thetest. The test item was kneaded and an aliquot was then immediately mixed with the cellulose (at a ratio 2:1 (w/w)) in a dosedjar. Both mixtures were filled into a glass funnel. The mixtures were ignited with the flame ofa gas burner. The vigor and duration ofthe reactions were observed and recorded. Due to the unmistakable results obtained during the preliminarytest, no main-test was performed to confirm the results. 

# Results and discussions Test result maximum burning rate 

### oftest mixture 

### Os maximum burning rate 

### ofreference mixture 

### 77s 

## Any other information on results incl. tables 

## During the preliminary test the Leuco Sulfur Black 1/cellulose mixture ignited poorly and the flame became orange. Without contact to the ignition source, the flame extinguished. Only glowing was observed for 3 minutes. The surface was carbonized but underneath unchanged material remained. Since the test item mixture did not burn without contact to the ignition source and the handling ofthe test item was difficult (contact to air should be avoided according to sponsor's request), a main test was not performed.

## The barium nitrate/cellulose mixture during the same test burned for 77 secondswith a blazing, lightyellowish flame, emitting sparks. 

# Applicant's summary and conclusion Interpretation of

## results

no oxidising 

### properties 

## Conclusions Leuco Sulfur Black 1 has no oxidizing properties. 

## Executive summary 

## The determination ofthe oxidizing properties of Leuco Sulfur Black 1was performed in accordance with the EEC Directive 92/69, EEC Publication No. L383, December 1992 and Council Regulation (EC) No. 440/2008, Methods for the determination ofphysico-chemical properties, A.17 "Oxidizing Properties (solids)". During the preliminary testthe Leuco Sulfur Black 1/cellulose mixture ignited poorly and the flame became orange. Without contactto the ignition source, the flame extinguished. Only glowing was observed for 3 minutes. The surface was carbonized but underneath unchanged material remained. The barium nitrate/cellulose mixture during the same test burned for 77 secondswith a blazing, light yellowish flame, emitting sparks. Since the test item mixture did not burn without contact to the ignition source and the handling ofthe test item was difficult (contactto air should be avoided according to sponsor's request), a main testwas not performed. In conclusion, Leuco Sulfur Black 1 has no oxidizing properties.

# 4.17 Stability in organic solvents and identity of relevant degradation products Endpointstudyrecord: Waiving_Stabilityin organic solvents andidentityofrelevant degradation products UUID IUC5-128cf15b-174a-4de2-ad2c-f80c6e2cfe57 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author jregano I Clariant Iberica Producci6n S.A I El Prat de Llobregat (Barcelona) I Spain Date 2010-05-1815:30:09 CEST Remarks 

# Administrative Data Data waiving Justification lordata 

### waiving study technically not feasible According to Reach Regulation annex Xllhe test does not need to 

### be performed because lhe substance is 

### not soluble in organic solvents like DMSO, chloroform, methanol, ethanol and 1,2-propylene glycol

# 4.21 Dissociation constant Endpointstudyrecord: Waiving UUID IUC5-16b2f4ad-463&-4ab6-9ad2-929f7dd73998 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks jregano I Clariant Iberica Producci6n SA I El Prat de Llobregat (Barcelona) I Spain 201Q-05-1812:20:04 CEST



# Administrative Data Data waiving study technically not feasible Justlftcatlon 

### for datawaiving waived according to column 2 

### of REACH Annax IX:**~is** scientifically not possible to perform the test, the analytical method 

### is not sensitive enough.

# 4.22 Viscosity Endpointsummary: Viscosity UUID IUC5-{)62973fe-8268-4048-89bc-1990d633d39a Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author pbaslo I Clariant Quimicos (Portugal) Lda I Lei'" do Balio I Portugal Date 2010-04-28 17:22:28 CEST Remarks 

# Administrative Data Short description of key information lhlxotrophic 

# Discussion As the viscosity ofthe test item became lowerduring stirring, Leuco Sulfur Black 1 can be stated to being thixotrophic.

# Endpointstudyrecord: KEY_OECD 114_2009_Harlan_C31032_Weissenfeld
UUID IUC5-c371d7a7-a5a9-483e-a58b-e7e1f71920c8 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Aulhor Date Remarks jregano

### IClarian11benca Producci6n S.A. 

### 1El Prat 

### de Llobregal (Barcelona) I Spain 2010-05-18 15:30:24 CEST



# Administrative Data Purpose

### flag key study; robust study summary Study result

### type experimental**resu~** Study period 2009-03-19 Reliability 1 (reliable without restriction) 

# Data source Reference

Refersnce Author

type 

### Year Title study report Welssenfeld 2009 Determination 

### of

### the M 

## Data access data submitter is data owner 

## Data protection claimed
yes Viscosity



# Materials and methods Test guideline Guideline Bibliographic Testing laboratory 

### source Harlan 

### Laboratories Ltd. IUngen (Switzerland)

Deviations Report Owner no. compeny C31032 Clar1ant Compeny study Report 

### no. 

# - 2009-08- 

### 06

OECD Test Guideline 114 yes (Due 10

### thetexture 

### ofthe test llem, II was not possible 10 fill an aliquot into the rotaUonal viscometer and

### to determine 

### the viscosity.(Viscosity of Liquids) Thus, 

### the test Item wasjudged visually for any changes In viscosity whie stilling.)

other: Due to the texture oflhetest item, itwas not possible to fin an aliquot into

### the rotational viscometer and to detennlne 

### the viscosity. Thus, lhe test**~em** wasjudged visually for any changes in viscositywhile stilling. 

### Principles ofmethod Ifotherthan guideline Due to 

### the texture 

### ofthetest Item, itwas not possible tofill an aliquot into the rotational viscometer and to determine 

### the viscosity. Thus, the test item wasjudgedvisually for any changes In viscositywhile stilling. 

### GLP compliance
yes (ind. certificate) 

## Test materials Identity oftest material same as for substance defined in section 1 

### (if not read-across)
yes 

### Test material identity

Identifier Identity CAS number 66241-11-0 EC number 266-273-5 

### Details on test matenal

- Name

### oftest material : Leuco SuWur Black 1

- Physical stata: lhixotrophic, black, wet paste -Composition: Leuco Sulfur Black 1:52.75% (wlw) (calculated by amount

### of by-compounds) 2,4-Dinitrochlorobenzene: 

### < 20 ppm 2,4-0inltrophenole: < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3% (wlw) Water. 45.75 % (wlw) by Kari-Fischer titration -Lot/batch 

### No.: ESA0007595 -Expiration date

### ofthe lot/batch: 201 1-07-1 1

- Stability undertest conditions: Stable under storage conditions. -Storage condition oflest material: At room temperature (range of20 

### ± 5 'C), light protected.

## Any other Information on materialsand methods Incl. tables 

## Principle ofthe Method The viscosity ofthe test item was determined by visual observation. Test Conditions The determination was carried out at room temperature. General Test Procedure About 30 g oftest item were weighed into a 250 mL beaker. The test item was stirred for aboutone hour by a external stirrerwith a metal blade at 150 rpm. The beaker was sealed with parafilm and the test item in the beakerwas hold under nitrogen atmosphere. The texture ofthe test item wasjudged visually. 

# Results and discussions Viscosity

> 10000 mPa s (dynamic) Temp. 22 'C Remarks theviscosity oould not

### be exactly measured 

## Any other Information on results Incl. tables 

## Observation Results

StirringTlme_[minJ Phy15ical state oftheTest Item 

# 0 Coarse grained black powder 5 Powder crushed 10 Powder crushed 15 Powder, substance climbs up on the beakers glass

## 30 Substance black, paste-like, slobbery, oily,climbs upto 1/3 ofthe beaker's glass 55 Substance made one big black clump Substance defines to be thixotrophic. 

# Applicant's summary and conclusion Conclusions As the viscosity 

### ofthe test ttem became lowerdurtng stirring. Leuco Sulfur Black 1 can be stated

### to being thixotrop 

## Executive summary 

## The determination ofthe viscosity of Leuco Sulfur Black 1 deviated from the OECD guideline No. 114, "Viscosity of Liquids" using the rotational viscometer method. Due to the texture ofthe test item, itwas not possible to fill an aliquot into the rotational viscometer and to determine the viscosity. Thus, the test item wasjudged visually for any changes in viscosity while stirring. As the viscosity ofthe test item became lowerduring stirring, Leuco Sulfur Black 1 can be stated to being thixotrophic.

# 5 Environmental fate and pathways Endpointsummary: Environmental fate andpathways UUID IUC5-fflc36e3-clbc-47c3-9ada-77ed614d43e5 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author uensenba I Clarianl Produkte (Deutschland) GmbH 

### I Sulzbach am Taunus 

### I Germany Date 2010-05-{)4 12:06:06 CEST Remarks 

# Administrative Data Discussion The test item Leuco Sulfur Black 1 was investigated for its ready biodegradability in a manometric respirometry test over 28 days according tothe EU Commission Directive 92/69 EEC, C.4-D Commission Regulation (EC) No 440/2008, C.4-D and the OECD Guideline for Testing of Chemicals No. 301 F. The biochemical oxygen demand (BOD) ofthe test item Leuco Sulfur Black 1 in the test media was in the normal range found for the inoculum controls. Consequently, Leuco Sulfur Black 1was not biodegradable underthe test conditions within 28 days. In the toxicity control, containing both Leuco Sulfur Black 1 and the reference item sodium benzoate, Leuco Sulfur Black 1 had no inhibitory effect on the activity ofactivated sludge microorganisms atthe tested concentration of 142 mg/L. In the procedure controls, the reference item sodium benzoate was degraded by an average of 80% by Exposure Day 14, and reached an average biodegradation of82% by the end ofthe test (Day 28), thus confirming suitability ofthe activated sludge.

# 5.1 Stability 5.1.2 Hydrolysis Endpointstudyrecord: Waiving_Hydrolysis
UUID IUC5-84523700-e2ef-4bc1-81d4-cd0427894ef9 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks uensenba I Clariant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-05-21 14:56:46 CEST



# Administrative Data Data 

### waiving JustlflCIItlon 

### for data 

### waiving study technically notfeasible Due to the physico-<:hernical properties 

### ofthe UVCB substance no analytical method is available. According to Reach Regulation Annex XI, section 2 the study is not required.

# 5.2 Biodegradation 5.2.1 Biodegradation in water: screening tests Endpointstudyrecord: Memmert (2009) 

# I Biodegradation In water: screening tests
UUID IUC5-a3f5e34a-e689-4b5e-e3cd-3145768dfc6e Dosaler UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarl<s jregano 

### IClariant Iberica Producci6n SA 

### I El Prat de Llobregat (Barcelona) 

### I Spain 2010-05-20 16:34:30 CEST



# Administrative Data key study Purpose flag Study result

### type Reliability experimental resutt Study

### period 2009-03-23 tiH 2009-04-23 1 (reliable without restriction) Rationale for reliability Incl. deflclenclaa Guideline conform study under GLPwithout deviations 

# Data source Reference

Reference Author 

### Year



### type study report 

### Mernmert 

### 2009 

## u Data access data submitter is data owner 

### Data protection claimed
yes 

### Title Ready Biodegradability

### of Leuco Sullur Black 1 in a Manometric Respirometry Test



# Materials and methods Bibliographic source Tastinglaboratory
Harlan Laboratories Ltd. ltingen (Switzerland) Report Owner Company 

### no. company study no. c311n Clariant Report date 

### 2009-08- 19

### Testtype ready biodegradability 

### Test guideline

Qualifier Guideline aCCOfding EU Method C.4-0 (Detennlnation of

### tha "Ready" Biodagradability- Manometric Respirometry Test) (Com. Rag. (EC) No44012008 of30 May 20081aying 

### to down test methods pursuant to Ragulation (EC) No 1907/2006ofthe European Partiament and ofthe Council on

### tha Raglstration, Evaluation, Authorisation and Restriction ofChemicals (REACH), C.4·D, Manometri) aCCOfding EU Method C.4-D (Detennination of

### tha "Raady" Biodagradabllity- Manometric RespirometryTest) (EU Commission Oirectiva 92169 EEC, C.4-D, Manometric to Respirometry Test, 1992.) according OECD Guideline 301 F (Ready Blodagradabiity: Manometric Respirometry Test) (OECD Guidelinefor Testing 

### ofChemicals No. 301 F, Ready 

### to Biodagradability: Manometric Respirometry Test, 1992.) 

### GLP compliance yes (incl. certificate) 

## Test materials Identity oftest material same as for substance defined in section 1 (If not read-ecross)
yes 

### Test material identity

Identifier Identity CAS number 66241-11-0 ECnumber 266-273-5 

### Details on test material

- Name oftest material : Lauco Sulfur Black 1

- Physical state: thlxotrophic, black, wet paste -Composition: Leuco Sulfur Black 1:52.75 % (wlw) (calculated by amount of by-compounds) 2,4-Dinitrochlorobenzene: < 20 ppm 2,4-Dinitrophanole: < 20 ppm Suffate: 970 ppm Chloride: 750 ppm Sodium: 1.3 % (wlw) Water. 45.75% (wlw) by Kart-Fischertitration 

### Deviations
no



### no

no-Lot/batch 

### No.: ESA0007595 -Expiration date ofthe lot/batch: 2011-07-11

- Stability undertest conditions: Stable under storage conditions. -Storage condition oftest material: AI room temperature (range of20 ± 5 'C), light protected. 

## Study design Oxygen conditions aerobic 

### Inoculum ortest system activated sludge, non-adapted 

### Details on Inoculum -Source of inoculum/activated sludge: aerobic activated sludge from a wastewatertraatment plant 

### (ARA Ergolz 

### II, FOilinsdorf, Switzerland) treating predominantly domesticwastewater

- LaboraiOfY culture: 

### -Method ofcultivation: The sludge waswashed twice with tap water by centrltugation and the supernatant liquid phase was decanted. 

### Ahomogenized aliquot ofthe final sludge suspension

### was weighed, thereafterdried and the ratio ofwetto dry weightwas calculated. Based on this ratio, calculated amounts ofwet sludge wera suspended In testwaterto obtain a concentration equivalentto 

### 4 

### g (±10%) dry material per liter.- Storage conditions/length: During the holding period ofthree days priorto use, the sludge was aerated at 

### room temperature.

- Preparation of Inoculum lorexposure: Priorto use, the sludgewas first thoroughly mixed and then dHuted with testwater -Concentration ofsludge: Priorto use, the sludge was first thoroughly mixed and then diluted with testwaterto a concentration of1 

### g per liter (dryweightbasis). Basedon the determined dry weight ofthis diluted activated sludge, defined amounts were added to testwaterto obtain a final concentration of30 mg dry material per liter.



### Duration oftest (contacttime) 28d 

### Initial test substance concentration Initial 

## cone. Ba-on 140 mgiL test mat. 141 mgiL tes1 

### mat. 

### Parameter followed

## for biodegradation estimation

# 1 02 consumption

# 1 Details on studydesign Apparatus 

### The test flasks (500 

### mL reactionvassals, labeled with the necessary information to ensure unmistakable Identification) were incubeted under continuous stining In a SAPROMA T Dt2 {Voith GmbH,**~nhaim,** Garmany). 

### Oxygen consumption was recorded manually by taking a daily reading et

### least on each working day. 

### The final testvolume was 250ml partestflask Principle: 

### Electro-chemical analysis process: 

### The biodegradation process 

### consumes the dissolved oxygen in the test medium and generates C02. The C02 is adsofbed by 

### soda lime, which results in a d8CI8ase ofthe total pressure in 

### the airtight testflasks. The pressure drop Is detected and converted intoan electrical signal by means ofan electrode 

### type manometer. The consumed oxygen is replaced byetectrolytically generated oxygen 

### from a coppersulfate solution. Test duration: 

### 28days Light conditions: Dari<ness Testtemperature: 22 'C, maintained with 

### a built4n thermostat and checked once parweek. pH: Priorto test start, the pH was measured In each test flask before the eddRion ofthe activated sludge inoculum. Attha end 

### of incubation, tha pH was measured again in each test flask. Testwater prepared according to the testing guidelines. Analytical grade salts were dissolved in purified water to obtain the following stock solullons: 1) KH2P04 8.50 giL K2HP04 21.75giL Na2HP04 x 2H20 33.40 giL NH4CI 0.50 giL 

### The pl4 ofthis solution was 7.4. 2) MgS04 x 7H20 22.50 giL 3) CaCI2 x 2H20 36.40 giL 4) FeCI3 x 6H20 0.25 giL, stabilized wnh one drop ofconcentrated HCI par liter To obtain the final testwater, 10 mL

### ofstock solution No. 1 and 1 mL each ofstock solution Nos. 2, 3 and 4 were combined and made up to 1000 mL with purified water. The pH was adjusted

### from 7.7 to 7.4 wRh a diluted hydrochloric acid solution. Control and Blank System Inoculum blank: 

### yey Toxicity control: yes Reference substance: yes 

### Reference substance benzoic add, sodium salt

## Any other Information on materials and methods Incl. tables 

## Due to the complex structure (composition) ofthe test item, its chemical oxygen demand (COD) was determined according to the EU Commission Directive 92/69/EEC, C.6 following DIN 38414-59. The COD was determined at BMG Engineering AG, lfangstrasse 11, 8952 Schlieren 

# I Switzerland. The COD determination was not performed in compliance with the GLP regulations. Therefore, it is excluded from the Statement ofCompliance in this report. However, itwas performed in compliance with ISO/IEC 17025. The chemical oxygen demand (COD) ofthe test item Leuco Sulfur Black 1was determined according tothe EU Commission Directive 92/69/EEC, Part C.6 following DIN 38414-S9. The duplicate values were 0.78 and 0.81 mg 02/mg test item. The mean COD was 0.795 mg 02/mg test item. Test Concentrations

Identification Replicate Amount oftest item 1 

## Amount of reference item1 

## No. mg/L C002 mg/L Test item 1 140 111 

# - Test item 2 141 112 

# - Inoculum 1 

# --- n.a. 

# -- control Inoculum 2 

# --- n.a. 

# - control Procedure 1 

# -- n.a. 100 control Procedure 2 

# --- n.a. 100 control Toxicitycontrol• 1 142 113 100
1 

## The tabulated values represent rounded results obtained by calculation usmg the exact raw data. 2Chemical oxygen demand in mg 02/L test solution. Calculation ofthe ThOD was not possible due to the 

## Th003 

## n.a. n.a. n.a.
n.a. 167 167 167

### complex structure (composition) ofthe test item. The COD was not determined in compliance with the GLP regulations and is therefore excluded from the Statement ofCompliance in this report. However, the results will be archived underthis study number. 3Theoretical oxygen demand in mg 0211. test solution. •The test item can be assumed to be inhibitory ifin the toxicity control less than 25% degradation 

### (based on total ThOD) occurs within 14 days. n.a. not applicable. 

# Results and discussions Testperformance The percent biodegradation ofthe reference**~em** sodium benzoate was calculated besed on the theoretical oxygen demand 

### of 1.67 

### mg 02/mg. In the procedure controls, the reference Mm was degraded by an average of80% by Exposure Day 14, thus confinning suMbilily ofthe activated 

### sludge. At the end ofthe test (Day 28), the reference**~em** was degraded by an average of82%. 

## % Degradation oftest substance Remark• St.dev. not applicable

The percent biodegradation in the toxicity control, containing both the test**~em** and the reference**~em,** was calculated 

### based on the COD ofthe test**~em** and the ThOD ofthe reference item. In the toxicity control, the 

### course of biodegradation over the 28-<lay exposure period was similar to the 

### two procedure oontrols, oontainlng onlythe reference**~em.** WMin 14 days of exposure, biodegredetion amounted to 50%. Thus, according tothe test guidelines. the test 

### Mm had no Inhibitory effect on activated sludge microorganisms at thetested concentration of 142 

### mg/L because biodegradation in the toxicity control was >25% within 14 days. 

### The 

### pH measured in all flasks atthe start ofthe test was 7.4. At the end of exposure (Day 28), pH values of7.1-7.8

### were measured. 

## BODS/COD results 8005/COD BOD5 ca. 0 

### mg 02/gtest mat. 

### Results 

## with reference substance The percent biodegradation ofthe reference Kern sodium benzoate was calculated based on the theoretical oxygen demand of 1.67 

### mg 02/mg. In the procedure controls, the reference Hem was degraded by an average of80% by Exposure Day 14, thus confinning**suilabil~** of

### the activated 

### sludge. At the end ofthe test (Day 28), 

### the reference**~em** wes degraded by an average of82%.

## Any other Information on results Incl. tables 

# Biodegradation ofthe Test Item The percent biodegradation ofthe test item was calculated based on the chemical oxygen demand (COD) of

# 0.795 mg 02/mg test item. The biochemical oxygen demand (BOD) of the test item Leuco Sulfur Black 1 in the test media was in the normal range found for the inoculum controls throughoutthe study period of28 days. Consequently, Leuco Sulfur Black 1was not biodegradable underthe test conditions within 28 days. 

# Blod

# eara 

# dati h T 

# t Fl k 

# on nt e es as s Percentage Biodegradation1 Test item Procedure control Toxicity control Time Replicate No. Replicate No. Replicate No. (days) 

## 1 2 1 2 1



## 0 0 0 0 

# 0 0 1 0 0 19 21 14 2 

# -- - - - 

# - 3 

# - - 

# - - - 4 -6 

# -3 63 68 

# 41 5 -4 

# -3 68 72 44 6 -4 

# -4 70 74 45 7 -4 

# -4 72 77 

# 47 8 -5 

## -4 

# 73 78 48

# 9 

# - 

# - - 10 

# - - -- 11 -6 -6 77 

# 12 -9 -8 76 13 -9 -8 78 14 -10 -9 79 15


# - 

# -- - 16 

# - 

# -- -- 17


# - 

# -- -- 18 -12 -11 80 19 -13 -11 80 20 -14 -12 80 21 -16 -12 78 22 -16 -12 80 23 

# - - 

# - 24


# - - - 25 -18 -14 78 26 -18 -14 80 27 -19 -15 79 28 -20 -15 80 Mean (Day

# 28) (-17). 

# 82 ,correctedforthe mean oxygen uptake ofthe 1noculum controls (see Sect1on 3.5)

- Not determined n.a. not applicable 

# -- -

- - 80 49 81 49 81 50 81 50



# -- -



# - -

- - 84 

# 51 84 

# 51 84 

# 51 84 

# 50 84 

# 51



# - -

- 

# - 84 

# 51 84 

# 51 84 

# 51 84 

# 51 n.a.

• Negative value due to higher oxygen consumption in the inoculum controls than in the test flasks with test item. 

# Applicant's summary and conclusion Validity 

## criteria fulfilled yes 

## Interpretation ofresultsundertest conditions no biodegradation observed 

## Conclusions Leuco Sulfur Black 1 is not biodegradable under thetest**cond~ions** within 28 days. 

## Executive summary 

## The test item Leuco Sulfur Black 1 was investigated for its ready biodegradability in a manometric respirometry test over 28 days according to the EU Commission Directive 92/69 EEC, C.4-D Commission Regulation (EC) No 440/2008, C.4-D and the OECD Guideline forTesting ofChemicals No. 301 F. The biochemical oxygen demand (BOD) ofthe test item Leuco Sulfur Black 1 in the test media was in the normal range found for the inoculum controls. Consequently, Leuco Sulfur Black 1 was not biodegradable underthe test conditions within 28 days. In the toxicity control, containing both Leuco Sulfur Black 1 and the reference item sodium benzoate, Leuco Sulfur Black 1 had no inhibitory effecton the activity ofactivated sludge microorganisms atthe tested concentration of 142 mg/L. In the procedure controls, the reference item sodium benzoate was degraded by an average of 80% by Exposure Day 14, and reached an average biodegradation of82% bythe end ofthe test (Day 28), thus confirming suitability ofthe activated sludge.

# Endpointstudyrecord: Waiving_Simulation testing on ultimate degradation in test water
UUID IUC5-3030b992-103b-4c44-a310-62e187968f4c Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad9732B1 ea Author Date Remarks uensenba I Clariant Produkte (Deutscl11and) GmbH I Sulzbacl1 am Taunus I Germany 2010-05-21 14:47:53 CEST



# Administrative Data Data waiving Justification for 

### data waiving otherjustification Waiving according 

### to "column 2" 

### of Regulation (EC) No 190712006 (CSA doss not indicate need

### for further Investigations). Due

### to the very high molecularweight (>8000 DaKon) 

### the bioavallabilily 

### ofthe substance Is negligible (low potential to cross biological membranes) and no biodegradation could 

### be expec1ed. Therefore addilional testing is notjustified

# 5.2.2 Biodegradation in water and sediment: simulation tests Endpointstudyrecord: Waiving_Biodegradation in waterandsediment: simulation tests UUID IUC5-al01798a-aa7f-4eed-a3b4-185f56ae58f8 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author uensenba I Clariant Produkte (Deutschland) GmbH I Sul-ch am Taunus I Germany Date 2010-05-21 14:48:27 CEST Remarks 

# Administrative Data Data w.Wing otherjustification Justification 

### for Waiving according lo "column 2" 

### of Regulation (EC) No 190712006 (GSA does 

### not indicate need 

### !Of further Investigations). Due lo lhe very high molecularweight 

### data .. 

### lvtng (>8000 Dakon)

### lhe bioavallability of

### lhe substance is nagliglble (low potential tocross biological membranes) and no biodegradation oould 

### be expected. TherafO<e additional testing is notjustified

# 5.2.3 Biodegradation in soil Endpointstudyrecord: Waiving_Biodegradation in soil
UUID IUC5-09b6bac3-5674-4a86-82f7-494edffie4f81 Dossier UUID IUCS-292149d1-a677-4feb-9692-1fad973281 ea Author Date Remarks uensenba I Clarian1 Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-05-21 16:32:10 CEST



# Administrative Data Data waiving Justification for 

### data-lvlng olherjustification Waiving according 1o "column 2" of Regulation (EC) 

### No 190712006 (CSA does not indicale need for lurlher investigations). Dua IX> lhevery high molecularweight (>8000 Dafton) 

### lhe bioavallability of

### lhe subslence Is negligible (low potential to cross biological membranes) and no biodegradation could 

### be expected. The<efore additional testing is 

### not justified.

# 5.3 Bioaccumulation 5.3.1 Bioaccumulation: aquatic 

# Isediment Endpointstudyrecord: Waiving_ Bioaccumulation: aquaticI sediment
UUID IUC5-e9a0ae0f-a42c-47cd-9824-47c1186fcfdf Dossier UUID IUC5-292149d1-a677-4feb-9892-1fad973281ea Author Data Remart<s uensenba 

### I Clarianl Produkte (Deutschland) GmbH I Sulzbach am Taunus 

### I Germany 2010-05-21 14:52:55 CEST



# Administrative Data Dllbl

### waiving otherjustification Justification

### for Waiving according

### to "column 2" of Regulation (EC) No 190712006 (CSA does not indicate need for

### further Investigations). Due

### to the very high molecularweight data waiving (>8000 DaHon) 

### the bioavailabiiHy of

### the substance 

### Is negligible (low potential to cross biological membranes) and no bioaccumulation could 

### be expected. Therefore additional testing 

### Is notjustified

# 5.4 Transport and distribution 5.4.1 Adsorption I 

# desorption Endpointstudyrecord: Waiving_ Adsorption I desorption
UUID IUC5-25c2af70-1 080-4bd3-b58a-5d5ada3ccbf7 Dossier UUID IUC5-292149d1 -a677-4feb-9692-1fad973281 ea Author Date Remarks uensenba I Clariant Produkte (Deutschland) GmbH I Sulzbach am Taunus IGermany 2010-05-21 14:58:55 CEST



# Administrative Data Datawaiving Justlflc:ation for

### data 

### waiving sludy technically notfeasible Due tothe physico-<:hemical properties 

### ofthe UVCB substance no analytical method is available. Aocording to Reach Regulation Annex XI, section 2 the sludy is not required.

# 6 Ecotoxicological Information Endpointsummary: Ecotoxicologicallnformation
UUID IUC5-3ed53458-0d75-4f7 e-bco4-aedd3d11421d Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks uensenba I Clariant Produkte (Deutschland) GmbH I Sulzbach am Taunusl Germany 2010-05-21 14:28:45 CEST



# Administrative Data PNEC aqua (freshwater) Justification for (no} PNEC freshwater derivation Derivation 

### of PNEC is not possible. Justification: The substancewas tested in concentrations in the range ofits water solubility. No

### effects were observed in all test systems. Due to 

### the high molecularweight, bloavailability of

### thetest substance is negligible. Consequently, no LOEC, NOEC, or ECx- value can 

### be calculeted. Since this data is a prerequisite to estimate a PNEC HIs 

### not possible 

### to derive thisvalue. 

# PNEC aqua (marine water) Justification for (no} PNEC marine water derivation Derivetion 

### of PNEC is not

### possible. Justification: The substancewas tested in concentrations in the range ofHs water solubility. No effects were observed 

### in all test systems. Due 

### to the high molecularweight, bloavallablHty of

### thetest substance is negligible. Consequently, no LOEC, NOEC, or ECx- value can 

### be calculated. Since this data is a prerequisite to estimate a PNEC it is not possible 

### to derive this velue. 

# PNEC aqua (intermittent releases) Justification for (no} PNEC Intermittent releases derivation Derivation of PNEC is not possible. Justification: The substance was tested In concentrations in the range of its water solubility. No effects were observed in all test systems. Due to 

### the high molecularwaight, bioavailabllity

### ofthe test substance is negligible. Consequentiy, no LOEC, NOEC, or ECx- value can 

### be calculated. Since this data is a prerequisHe toestimate a PNEC it is 

### not possible 

### to derive this value. 

# PNEC STP

## Justification for (no) PNEC STP derivation Derivation of PNEC is not possible. Justification: The substance was tested in concentrations in the range of hs water solubility. 

### No effectswere observed in aH test systems. 

### Due to the high molecularweight, bloevailability ofthetest substance is negligible. Consequentiy, no LOEC, NOEC, or ECx- value can 

### be calculated. Since this data is a prerequisite

### to estimate a PNEC His not possible to derive this value. 

# PNEC sediment (freshwater) Justification for (no) PNEC sedimentderivation Derivation of PNEC is not possible. JusHfication: The substancewas tested in concentrations in the range ofHs water solubility in aquatic organisms. 

### No effects

### were observed in all test systems. Due

### to the high molecularweight, bioavallabllity ofthe test substance is negligible and effects in otherorganisms could also not

### be expeded. Consequenity, no LOEC, NOEC, or ECx -value can 

### be calculated. Since this data is a prerequisHe to estimate a PNEC it Is not possible

### to derive this value. 

# PNEC sediment (marine water) Justification for (no) PNEC marine sediment derivation Derivation 

### of PNEC is not possible. JusHfication: 

### The substance was tested in ooncantrations in the range of its water solubility in aquaticorganisms. No effects were observed in aH test systems. Due

### to the high molecular weight, bloevailability ofthe testsubstance Is negligible and effects in other organisms could also not

### be expeded. Consequenlty, no LOEC. NOEC. or ECx- value can 

### be calculated. Since this data is a prerequisHe to estimate a PNEC His not possible to derive this value. 

# PNEC soil Justification for (no) PNEC soli derivation Derivation 

### of PNEC is not possible. JustifiC8tion: The substance was tested in concentrations in the range ofitswater solubility in aquatic organisms. Noeffects

### were observed in an test systems. Due 

### to the high molecularweight, bioavallablllty ofthe test substance is negligible and effects in otherorganisms could also not 

### be expected. Consequenlty, no LOEC, NOEC, or ECx- value can 

### be calculated. Since this data is a prerequisHe to estimate a PNEC H

### Is not possible 

### to derive this value. 

# PNEC oral Justificationfor (no) PNEC oral derivation
Derivation of PNEC is not possible. JusHfication: 

### The substance was tested in ooncantrations in the range ofhs water solubility in aquatic organisms. 

### No effects

### were observed in

# an test systems. Due

### to the high molecularweight, bioavailability ofthe test substance is negligible and effects in otherorganisms could also 

### not be expected. Consequenlty, no LOEC, NOEC, or ECx -value can 

### be calculated. Since this data is a prensquisHe to estimate a PNEC it

### Is not possible

### to derive this value. 

# Conclusion on classification Environmental classificationjustification 

## All acute tests showed no effects and the substance has no potential forbioaccumulation. Thereforethe information is conclusive but not sufficient for classification.

# 6.1 Aquatic toxicity Endpointsummary: Aquatic toxicity
UUID IUC5-da01>46d9-4779-455a-607o-89c9c27392b2 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author uensenba I Clariant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany Date 2010-05-11 17:25:39 CEST Remarks 

# Administrative Data Discussion Alga The influence of DIRESUL BLACK ROT LIQUID on the growth ofthe green algae Scenedesmus subspicatus was investigated. The studywas performed according tothe EEC Directive 92/69. The test method was modified to differentiate between a reduced growth ofalgae due to real toxic effects ofthe test substance on the algae cells or dueto an indirect effect. a reduced algae growth by light absorption in coloured test solutions. The test included two experimental parts: Experimental part A: the algae grew in test medium with dissolved dyestuffin Erlenmeyerflasks. Experimental part B: the algae grew in separate containers (testtube) which were surrounded by the black coloured solutions ofthe same concentration as tested in partA. Because ofdifficulties concerning the solubility of DIRESUL BLACK ROT LIQUID in differenttest media, several tests were performed. Finally, in experimental partA the following nominal concentration range was tested: 

# 0.01 mg/1 to 

# 200 mg/1. In experimental part B, the nominal concentration range of

# 0.01 mg/1 to 

# 50 mg/1 was tested.

## The biological results of both experimental parts A and B were almost identical. This was shown by several calculated parameters, the NOEC, LOEC and the EC 50 values: In part 

# Athe NOEC was 1.0 mg/1, and in part B the NOEC was found to be between 1 and 5 mg/1. The LOEC was found to be between 1 and 5 mg/1 in part

# A and itwas about 5 mg/1 in part B. The percentage inhibition of algae biomass and the algae growth rate (IJ) after 72 hours ofexposure were in the same orderof magnitude. The EC 50 values calculated for both growth parameters, showed no significant differences. Experimenta part A EbC 50: Experimental Part B 1.84 mg/1 5.99 mg/1 

# Ere 50: 54.1 mg/1 8.59 mg/1 Thus, the growth inhibition of Scenedesmus subspicatug was in the same magnitude when the algae grew in test medium without test substance but under reduced light intensities bythe filter effect (experimental part B) as in experimental part A, where the algae grew in test solutions without dissolved test substance. In conclusion, this modified algae test clearly demonstrated thatthe observed inhibition effect ofthe substance was caused only bythe indirect effect, the light absorption in the coloured test solutions. Thus, a toxic effect ofthe test substance on the algae cells can be excluded upto a concentration of50 mg/1. 

## Fish, acute The acute toxicity ofthe test item Leuco Sulfur Black 1to zebra fish (Brachydanio rerio) was determined in a 96-hour semistatictestwith a dailytest medium renewal according to the Commission Regulation (EC) No440/2008, Part C.1 and the OECD Guideline for Testing ofChemicals No. 203 (1992). A limit test with a daily test medium renewal was performed in accordance with the test guidelines to demonstrate that the test item has no toxic effecton the test fish up to the loading rate of 1 00 mg/L.

## Daphnia, acute The acute toxicity ofthe test item Leuco Sulfur Black 1to Daphnia magna was determined in a 48-hoursemi-static test according to the EU Commission Directive 92/69/EEC, Part C.2, Commission Regulation (EC) No440/2008, Part C.2 and the OECD Guideline for Testing ofChemicals, No. 202 (2004). A limit testwith a test medium renewal after24 hours was performed in accordance with the test guidelines to demonstrate thatthe test item has notoxic effect on the test organisms up to the solubility limit ofthe test item in test water. Due to the low water solubility ofthe test item, the test medium was prepared priorto the start ofthe test and priortothe test medium renewal as follows: a dispersion ofthe test item with the loading rate of 100 mg/L was ultrasonic treated for 15 minutes and continuously stirred at room temperature in the dark over 96 hours. Then, the dispersion was filtered. The undiluted filtrate ofthe dispersion was used asthe test medium. Additionally, a control was tested in parallel. The preparation ofthe test medium was based on the OECD Guidance Documenton Aquatic Toxicity Testing of Difficult Substa Thences and Mixtures, 2000. No remarkable observations were made concerning the appearance ofthe test medium. It was clearthroughout the entire test duration. Noanalytical work was performed due tothe specific properties ofthe test item (insolubility in water). At the beginning and end ofthe test medium renewal periods, the pH values ofthe test medium were between 7.6 and 8.0. The dissolved oxygen concentrations in the test medium and control were at least 7.9 mgll. The watertemperature was 20 

# 'C during the test. The biological results were based on the loading rate ofthe test item of 100 mg/L.

## In the control and atthe loading rate of 100 mg/L no immobilized daphnids were determined during the test period of48 hours. The biological test results (based on the loading rate) were asfollows: Loading rate (mg/L)

- 24-hour EC50: >100

- 24-hour ECO: :;:::, 100

- 24-hour EC100: >100 -48 -hour EC50: >100

- 48 -hour ECO: :;:::, 100

- 48 -hour NOEC: :;:::, 100 -48 -hour EC100: >100 In conclusion, the test item Leuco Sulfur Black 1 had no acute toxic effects on Daphnia magna up to its solubility limit in the test water underthe present test conditions. Toxicity to bacteria The inhibitory effect ofthe test item Leuco Sulfur Black 1 on the respiration rate ofaerobic wastewater microorganisms of activated sludge was investigated in a 3-hour respiration inhibition test according to the EU Commission Directive 88/302/EEC, Part C.11, Commission Regulation (EC) No. 440/2008, Part C.11 and the OECD Guideline for Testing of

## Chemicals, No. 209. Based on the EU testing guidelines, a limittestwith a nominal test concentration of 1000 mg/Lwas performed. In addition, two controls and three different concentrations ofthe reference item 3,5-dichlorophenol (5, 16, and 50 mg/L) were tested in parallel. The results ofthese treatments confirmed suitability ofthe activated sludge and the method used. At the test item concentration of 1000 mg/L, thetest item was not completely dissolved in the test medium. Thus, the test concentration was clearly above the water solubility limitofLeuco Sulfur Black 1 underthe test conditions. The test item had no inhibitory effect on the respiration rate of activated sludge after the incubation period ofthree hours at a loading rate of 1000 mg/L. The saturation concentration (water solubility limit of Leuco Sulfur Black 1 underthe test conditions) was reached and no inhibitory effectwas determined afterthree hours contact time. Thus, the 3-hour NOEC (EC15) ofthe test item to activated sludge microorganisms was at least 1000 mg/L (i.e. the saturation concentration ofthe test item). This value might even 

## be higher butconcentrations above 1000 mg/L were not tested. The 3-hour EC20, ECSO, and EC80 could not 

## be calculated but were clearly higher than 1000 mg/L. Page

# 6.1.1 Short-term toxicitytofish Endpointstudyrecord: Peither(2009) I Short-term toxicity to fish
UUID IUC5-90518ea2-2e99-4c52-b767-43cb6e3332d9 Dossier UUID IUC5-292149d1-a677-4feb-9692-11ed973281ea Author Date Remarks jregano 

### I Clariantlberlca Producci6n S.A. 

### I El Prat de Llobregat (Barcelona) 

### I Spain 2010-05-20 16:33:12 CEST



# Administrative Data key study**Pu~ellag** Study resulttypa Reliability experimental result Study period 2009-05-0911112009-05-15 1 (reliable without restriction) Rationale for reliability Incl. deficiencies Guideline conform study under GLP without deviations 

# Data source Reference

Reference Author Year

type 

### study report Peither 2009 A 

## Data access data submitter Is data owner 

## Data protection claimed
yes Title
Acute Toxicity 

### of Leuco Sulfur Black 1 to Zebra Fish (Brachydanio rerlo) in a 96-Hour Test



# Materials and methods Test guideline Blbllognphlc 

### aource Testing 

### laboratory
Harlan Laboralories Ltd. ltingan (Switzerland) Report Owner Company Report 

### no. company 

### atudy no. 

# - C31144 Clariant 

### 2009-08- 07Qu
..

### lfler Guideline according EU Method C.1 (Acute Toxicity for Fish) (Convnission Regulation (EC) No44012008

### of30 May 20081aying down test methods pursuantto Regulation (EC) 

### to No 1907/2006 

### ofthe European Parliament and oftha Council on the Registration, Evaluation, Authorisation and Restriction ofChemicals (REACH), C.1 :) according 

### otherguideline: EU Commission Directive 92169/EEC, C.1: Acute Toxicityfor Fish, 1992. 

### to according OECD Guideline 203 (Fish, Acute Toxiclty Test) (OECD Guideline for the Testing ofChemicals, No. 203, Fish, Acute Toxiclty Test, 1992.) 

### to 

### GLP compliance yes (ind. certificate) 

## Test materials Identity oftest material same as for substance defined In section 1 

## (If not read-across)
yes 

### Test material Identity

Identifier Identity CAS number 66241-11-0 ECnumber 266-273-5 

### Details ontest material

- Name 

### of test material : Leuco Sulfur Black 1 -Physical 

### stele: thlxotrophic, black, -tpaste -Composition: Leuco Sulfur Black 1:52.75% (w/w) (calculated by amount

### of by-<:c>mpounds) 2,4-Dinitrochlorobenzane: 

### < 20 ppm 2,4-Dinitrophenole: < 20 ppm SuWate: 970 ppm Chloride: 750 ppm Sodium: 1.3% (w/w) Water: 45.75 % (w/w) by Karl-Fischer titration

- Lot/batch No.: ESA0007595 -Expiration date oftha lot/batch: 2011-07-11

- Stability undertest conditions: Stable under storage conditions.

- Storage condition oftest material: AI room tempsrature (ranga of20 

### ± 5 'C), light protected. 

### Analytical monitoring Devt.tlons 

### no

no

nono 

### Details on sampling Noanalytical 

### work was performed due tothe specific properties ofthe test item (insotubiity in water). 

### Details on analyticalmethods Noanalytical work was performed due tothe specific properties of the test item (insotubiHty in water). 

### Vehicle

no 

### Details on test solutions Due to the low water solubility ofthe test Item. the test medium was prepared priortothe start ofthe test and prior to the test medium renewals asfollows: a dispersion ofthetest item with the loedlng rate 

### of 100 mgilwas continuously stirred at room temperature in the derl< over 96 hours. 

### Then, the dispersion wasfiKered. The undiluted fiKrete 

### with the maximum concentration ofdissolved test**~em** was used as the test medium. The preparation ofthe test medium was besed on the OECD Guidance Document on Aquatic Toxicity Testing of Difficutt Substances and Mixtures, 2000. 

## Test organisms Test organisms (species) Brechydanio rerio (new name: Denio reno) 

### Details on test organisms TEST ORGANISM

- Common name: zebrefish (Brachydanio rerio)

- Source: breeding cuKure at Hartan Laboratories -Length at study inHietion (length definHion, mean, range and SO): 10 fish were measured at the start 

### ofthe 

### test. The mean body length ofthe fish was 2.6 :t 0.05 em (Mean :t SO), -Weight at study initiation (mean and range, SO): 10 fish were measured at the start ofthe test. The mean bodywet weightwas 0.11 :t 0.02 g (Mean :t SO).

- Feeding during test: notfeed during test
ACCLIMATION

- Acclimation period: ona week

- AccHmation condntons (same as test or not): same as test (test water and temperature)

- Type and amount 

### offood, Feeding frequency: During holding and acclimatization until one day before the start 

### ofthe test, the fish were fed with a commercial fish diet (Tetra Min® Hauptlutter, supplied by TETRA-Werl<e, 49304 Melle I Germany)

- Heatth during acciimation (any mortaiHy obsarved):During the last two weeks priorto the test, no fish died in the test fish belch and all fish were heatthy

- No medication was applied during holding and accilmatization.

## Study design Test

## type semi-static 

### Water media

## type freshwater 

### Limit test yes 

### Total exposure duration 

### 96h Post exposure observation period

none 

## Test conditions Hardness
1.25 mmoUL resp. 125 mg/L as CaC03 

### Test temperature The watertemperature was 20-22 

### 'C during the test 

## pH The pH values 

### in the test medium and the control ranged 

### from 7.5to 7.7 

### Dissolvedoxygen The oxygen concentration was always 8.1 mgll or higher, and thus higherthan 

### 60% oxygen saturation. 

### Salinity

see table In section "Details on test condnions" 

### Nominalandmeasuredconcentrations Nominal: 

### 100 mg/L

### Details on 

### test conditions TEST SYSTEM -Testvesset:One glassiestvessel with 51Hers 

### oftest medium was used for each treatment. -Aeration: Thetest vessels were slightly aerated during 

### the test period.

- Renewal rate 

### oftest solution (frequency/flow rate): dally renewal



### - No. oforganisms pervessel: 7

- No. ofvessels 

### peroonoentretion (replicates): 1 -No. 

### ofvessels 

### percontrol (replicates): 1 -Biomass loading rate: 0.16g fish wet weight per Illertest medium
TEST MEDIUM 

### I WATER PARAMETERS

- Source/preparation 

### ofdilution water: reconstituted testwater
Reconstituted Test Water Ingredients 

### I Properties Conosntration mmoVlmg/L CaCI2 • 2H20 1.0 147 MgS04 • 7H20 0.25 61.5 NaHC03 0.38 32.5 KCI 0.038 2.9 Water Hardness 1.25 125 as CaC03 Alkalintly 0.4
-

OTHER TEST CONDITIONS -Photoperiod, Light intensity: A 16-hour light to 8-hourdar!< photoperiod, with a 30-mlnute transition period was used (light intensity during the light period was approximatelywHhin the range of 100to 

### 600 Lux).

EFFECT PARAMETERS MEASURED (with observation intervals 

### ifappliosble): Mortaltly and VIsible AbnormaiHies, at 3, 24, 48. 72 and 

### 96 hours
TEST CONCENTRATIONS

- Spadng fadorfor 

### test conosntrations: limH test 1OOmg/L nominal conoantrallon

- Range finding 

### study: The selection 

### ofthe test conosntration was based on 

### the resuhs ofa range-finding test**(non~LP).** 

### Reference substance (positive control)

no

## Any other Information on materials and methods Incl. tables 

## A semi-static test with dailytest medium renewal was performed to keep the test item concentrations in the test media as constant as possible during the test period of96 hours. During this semi-statictest, the testfish were daily placed into a newtest vessel with freshly prepared test medium. The preparation ofthe test medium was performed as far as possible in the dark to avoid photolytic degradation ofthe test item. At the start ofthe test and before each test medium renewal, the test medium with the loading rate of 100 mg/L was freshly prepared by completely dissolving 499.98, 499.91, 500.1 and 500.0 mg ofthe test item in 5 liter oftest water, respectively using ultrasonic treatment for 15 minutes and stirring for 96 hours at room temperature in the dark. The long stirring period was chosen in consultation with the sponsor. The test medium was freshly preparedjust before start ofthe test and prior to each test medium renewal. 

# Results and discussions Effectconcentrations Dumlon Endpoint Etr.c:tconc. Nominal/Measured Cone. 

### based on 

### Basisforeffect Remarks (e.g. 95% CL) 96h LCO >= 100mgll 

### nominal test mat. mortality 96h LC50 > 100 

### mgll 

### nominal test mat. mortality 96h LC100 > 100 

### mgll nominal test mat. mortaUty 96h NOEC >= 100 

### mgll nominal test mat. mortaUty 96h LOEC > 100 

### mgll 

### nominal test mat. mortaUty 

## Details on results Behavioural abnormalities: In 

### the control and In the test mediumwith 

### the loading rate of 100 mgll no visible abnormalities were observed atthe testfish.

## Results with 

## reference substance (positive control)

no reference substance tested 

## Reportedstatistics anderrorestimates

no statistical analysis 

## Any other lnfonnatlon on results Incl. tables 

## Mort alitvand VisibleAbnormalities Observed atthe Test Loading rate Numberofabnormal and dead fish 

# I number ofdead fish

# Type ofvisible abnormalities Observation time (mgiL) 3 hours 24 hours 48 hours Control 

# 010 0/0 0/0

100 0/0 0/0 0/0 LC50 > 

## 100 mg/L based on the loading rate 

## FishNumber offish tested at the s1ngle concentration and the control: 7 Measurements 

# I Observations Loading Exposuretime rate Oh 24 h 48 h 72 hours 0/0 0/0

new old new 

# old new old Appearance 100 0 0 0 0 0 0 (1) pH (2) Control 7.6 7.7 7.6 7.6 7.6 7.5 100 7.6 7.7 7.6 7.6 7.6 7.5 Oxygen (3) Control 8.6 8.8 8.9 8.7 8.7 8.7 100 8.1 8.8 8.3 8.8 8.5 8.6 Temperature (4) Control 21 21 21 20 21 

# 22 100 21 21 21 20 21 

# 22 (1) Appearance ofthe Freshly Prepared and Old Test Med1um 

## 96 hours 0/0 0/0

72 h new 0 7.6 7.6 8.5 8.6 21 21 96h old 0 7.5 7.6 8.6 8.5 22 21

### (2) pH in the Freshly Prepared and Old Test Medium and in the Control (3) Oxygen Concentrations (mg/L) in the Freshly Prepared and Old Test Medium and in the Control (4)Temperatures 

### ('C) in the Freshly Prepared and 0 ld Test Medium and in the Control 

# Applicant's summary and conclusion Validity criteriafulfilled yes 

### Conclusions In oonclusion, lhe test item had no acute toxic effects on the test fish up to 

### lhe loading rate 

### of 100mg/L. 

### Executive summary 

## The acute toxicity ofthe test item Leuco Sulfur Black 1 to zebra fish (Brachydanio rerio) was determined in a 96-hour semistatictestwith a dailytest medium renewal according to the Commission Regulation (EC) No 440/2008, Part C. 1 and the OECD Guideline forTesting ofChemicals No. 203 (1992). A limit test with a dailytest medium renewal was performed in accordance with the test guidelines to demonstrate that the test item has no toxic effect on the testfish uptothe loading rate of 100 mg/L. Due to the low watersolubility ofthe test item, the test medium was prepared priorto the start ofthe test and priorto the test medium renewals asfollows: a dispersion ofthe test item with the loading rate of 100 mg/L was continuously stirred at room temperature in the darkover 96 hours. Then, the dispersion was filtered. The undiluted filtrate with the maximum concentration ofdissolved test item was used asthe test medium. Additionally, a control was tested in parallel. The preparation ofthe test medium was based on the OECD Guidance Documenton AquaticToxicity Testing of Difficult Substances and Mixtures, 2000. No analytical work was performed due tothe specific properties ofthe test item (insolubility in water). The biological results were based on the loading rate ofthe test item.

## In the control and atthe loading rate ofthe test item of 100 mg/L no mortality orothervisible abnormalities were determined during the test period of96 hours. The pH values in the test medium and the control ranged from 7.5 to 7.7. The oxygen concentration was always 8.1 mg/L or higher, and thus higherthan 60% oxygen saturation. The watertemperature was 20-22 

# 'C during the test. The summary ofthe biological results is as follows: Loading rate (mg/L)

- 96-hour LC50: >100

- 96-hour LCO:**~** 

## 100

- 96-hour LC100: >100

- 96-hour NOEC:**~** 

## 100

- 96-hour LOEC: >100 In conclusion, the test item had no acute toxic effects on the test fish up to the loading rate of 100 mg/L.

# 6.1.2 Long-term toxicitytofish Endpointstudyrecord: Waiving_ Long-term toxicity to fish
UUID IUC5-2398d7e1-69ae-45c7-93e8-1e4440ee91af Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks uensenba I Clariant Produkte (Deutschland) GmbH I Sulzbach em Taunus I Germany 2010-05-21 14:33:09 CEST



# Administrative Data Datew•lvlng**Justlfle~~tlon** 

### for -waiving olherjustification Walving according to "column 2" of Regulation (EC) No 1007/2006 (CSA does not indicale need lorfurther Investigations). Due lo lhe very high molecularweight (>8000 Dafton) lhe bloevailability 

### ofthe subslance is negligible (low polantial tocross biological membranes) and no loxiceffects could 

### be expected. Therefore additional testing 

### Is noljustified

# 6.1.3 Short-term toxicity to aquatic invertebrates Endpointstudyrecord: Peither(2009) I Short-term toxicity to aquatic invertebrates, Daphnia magna UUID IUC5-4b26e921-<:de1-40b6-9968-b814ba3ee187 Dossier UUID IUC5-292149d1-a677-41eb-9692-11ad973281ea Author Da1e Remarks jregano 1Clariantlbenca Producci6n S.A. I El Prat de Llobregat (Barcelona) I Spain 2010-05-20 16:33:36 CEST



# Administrative Data key study 

### Purpoeeflag Study !HUll

### type Reliability experimental result Study

### period 2009-05-08 ti112009-05-14 1 (reliable without restriction) Ra1ionale 

### lor reliability Inc'- deficiencies 

### Guideline conlonn study under GLP without deviations 

# Data source Reference

Reference Author



### type study report Pelther A 

## Data access Yur
2009

data submitter is data owner 

## Data protection claimed yes 

### Title Bibliographic 

### source Acute Toxicity 

### of Leuco SuWur Black 1 to Daphnia magna in a 48-Hour Immobilization Test Testlng laboretory**Ha~an** Labora1ories Ltd. Hingen (Swilzeriand) Report Owner Company Report 

### no. company 

### study

### no. 

# - C31155 Clariant 

### 2009-08- 07

# Materials and methods Test guideline

Qualifier Guideline according EU Method C.2 (Acute Toxicityfor Daphnia) (Commission Regulallon (EC) No44012008 of30 May 20081aying down

### test methods pursuant to Reguiallon 

### to (EC) No 1907/2006

### ofthe European**Pa~iament** and 

### ofthe Council on the Registration, Evaluation, Authorisallon and Restriction ofChemicals (REACH), C.2,) according EU Method C.2 (Acute Toxicity for Daphnia) (EU Commission Directiva 92/69/EEC, C.2, Acute Toxicity for Daphnia, 1992.) 

### to according OECD Guideline 202 (Daphnia sp. Acute immobllisation Test) (DECO Guideline for Testing 

### of Chemicals, No. 202, Daphnia sp., Acute lmmobiUsatlon Test, 

### to 2004.) 

### GLP compliance
yes (Ind. certificate) 

# Test materials Identity oftest material same as for substance defined In section 1 

### (If not read-across)
yes 

### Test material Identity

Identifier 

### Identity CAS number 66241-11-0 EC number 266-273-5 

### Details on test material

- Name 

### oftest meteriai : Lauco Sullur Black 1 -Physical state:thixotrophic, black,

### wet paste • CornposHion: Leuco Sulfur Black 1:52.75% (w/w) (ceicuiated by amount ofby-compounds) 2,4-Dinitrochlorobenzene: 

### < 20 ppm 2,4-0initrophenole: < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3 % (w/w) Water. 45.75% (w/w) by**Ka~-Fischer** titration

- Lot/batch 

### No.: ESA0007595 Deviations 

### no

no

no- Expiration date of

### the lot/batch: 2011-07-11

- StabUity under test conditions: Stable under storage conditions. -Storage condition oftest material: At room temperature (range of20 ± 5 'C), light protected. 

### Analytical monitoring

no 

### Details 

### on sampling No analytical work was perfonnad due to the specific properties ofthe test item (insolubility in water). 

### Details 

### on analyticalmethods No analytical work was performed due to the specific properties ofthe test item (insolubility in water). 

### Vehicle

no 

### Details 

### on testsolutions Due to 

### the low water solubility of

### the test**~em.** the test medium was prepared 

### prior to 

### the start ofthe test and priorto the test medium renewals as folows: a dispersion ofthe test Item**~the** loading rete of 100 mg/L was continuously 

### stinred at room temperature in the dark over 96 hours. Then, the dispersion was**fi~ered.** 

### The undiluted fiHrate**~the** maximum concentration ofdissolved test**~em** was used as the test medium. The preparation ofthe test medium was based on 

### the OECD Guidance Document on Aquatic Toxicity Testing of Dlfficutt Substances and Mixtures, 2000. 

## Test organisms Test organisms (species) Daphnia magna 

### Details 

### on test

### organisms TEST ORGANISM

- Common name: Daphnia magna Tha studywas perfonnad with young daphnlds 

### of the species Daphnia magna Straus. A clone 

### ofthis species (defined bythe supplier as clone 5) was originally supplied by the University ofSheffield 

### I UK in 1992. Since that time, the clone has baen bred at Harlan Laboratories in**reconst~uted** water of

### the quality idantical to 

### the water quality used In the tests (in respectto pH, main ions. and total hardness) and undertemperature and light conditions identical to those of

### the tests. Atthe start ofthe**tes~** the organisms used In the test were 6-24 hours old and were notfirst brood progeny. Tth asphnldswere notfed during 

### the test For evaluation of

### the qualityof

### the daphnia clone and 

### the experimental conditions, potassium dichromate is tested as a positive control twice a year. 

### The resutt ofthe latest positive control test In Februar 2009 (46-hour ECSO: 0.71 mg/L, study C36915) Indicated that

### the sensttivity ofthetest organisms waswithin the internal historical range (48-hour EC50 from 1996 to2009: 0.53-1.1 

### mg/l). 

## Study design Test

## type semi.static 

### Water media

## type freshwater 

### Limittest yes 

### Total exposure duration 48h 

## Postexposure observation

## period

none 

## Test conditions Hardness 2.5 

### mrnoi/L rasp. 250mg/L as CsC03 

### Testtemperature Water temperature: 20'C 

## pH 7.6
- 8.0 (In test medium and control) 

## Dissolvedoxygen 7.9-8.3 

### mg/l (in test medium and control) 

## Salinity

sae table in saction "Details on test conditions•

### Nominalandmeasuredconcenlnltlons nominal 1 OOmg/L 

### Details on testconditions TEST SYSTEM -Testvessel: The test was perfonned In 100-mlglass beakers filled 

### with 50 ml oflest medium. 

### The test vessels

### ware covered 

### with glass plates

### to reduce 

### the loss ofwater by evaporation and to avoid the entry ofdust Into

### the solutions. -Aeration: Thetestwaterwas aerated priorto

### the start 

### of

### the study until oxygen saturation was reached. During 

### the lestperiod,

### thetestwaterwas not aerated.

- Renewal rate oftestsolution (fraquencylflow rate): daily ren-al

- No. 

### oforganisms pervessel: 20 -No. ofvessels 

### perconcentration (replicates):

- No. ofvessels percontrol (replicates): 1
TEST MEDIUM 

### IWATER PARAMETERS

- Source/preparation ofdilution water: reconstitutes testwater Reconstituted TestWater Ingredients 

### I Properties Concentration mmolll mg/L CaCI2 • 2H20 2.0 294 MgS04 • 7H20 0.5 123 NaHC03 0.75 65 KCI 0.075 5.8 Water Hardness 2.5 250 as CaC03 Alkalinity 0.8- 

### The ratio 

### ofCa:Mg and Na:Kwas 4:1 and 10:1, raspectively, 

### basedon molarily OTHER TEST CONDITIONS A 16-hourlightto 8-hourdark cycle with a 3Q.minute transHion period was used. Light Intensityduring 

### the light period was approximately be-n 520 and 680 Lux.
EFFECT PARAMETERS MEASURED 

### (with observation intervals 

### W applicable): Thedaphnids were observed for immobility after 24 and 48 hours 

### ofexposure (daphnids

### not being able 

### to swim within 15seconds aftergentle agHation 

### of

### the test beakerare considered 

### to be Immobilized).
TEST CONCENTRATIONS -Spacing factor

### for test concentrations: limittest, 100mg/L nominal

- Range finding study: The limit test

### was besed on 

### the results 

### ofa rangefinding test (non-GLP). 

### Reference substance (positive control) yes (potassium dichromate, sae •results 

### with reference substance)

## Any other information on materials and methods incl. tables 

## Study Design A semi-static test with test medium renewal after24 hours was performed to keep the concentration ofthe test item in the test medium as constant as possible during the test period of48 hours. After24 hours, the test organisms were placed in clean test vessels with freshly prepared test medium ofthe corresponding concentration.

Due to the lowwater solubility ofthe test item, dispersions with the loading rate of 100 mg/Lwere prepared before start of the test and before the renewal ofthe test medium by dispersing 100.3 and 100.0 mg ofthe test item in 1000 ml oftest water, respectively using ultrasonictreatmentfor 15 minutes and intense stirring. No auxiliary solvent or emulsifier was used. The dispersions were stirred by a magnetic stirrerat room temperature in the dark over96 hours to dissolve a maximum amount ofthe test item in the respective dispersion. The long stirring period was chosen in consultation with the sponsor. Afterthe 96-hour stirring period, the dispersions ofthe test item were filtered through a membranefilter (Schleicher & Schuell, Type NC45, pore size 0.45 IJm). The undiluted filtrates containing the maximum concentration ofdissolved test item were used as test medium atthe start ofthe test 

## (= addition ofdaphnids) and atthe test medium renewal. The preparation ofthe test medium was based on the OECD Guidance Document on Aquatic ToxicityTesting of Difficult Substances and Mixtures, 2000 

# Results and discussions Effectconcentrations Duntlon Endpoint Effec:tconc. Nominal/Measured Cone. based on 

### Baals lor

### effec:t Remarks (e.g. 95% CL) 

### 48 h 

## ECO >= 

### 100 mgll nominal last

### mat. mobility48h ECSO > 100 mg/L 

### nominal test mat. mobility 48h EC100 > 100 mgiL nominal tastmat. mobility 48h NOEC >= 100 mg/L nominal tastmat. mobility 

## Details on results

• Behavioural abnormalities: none • In 

### the control and at the loading rate of 100 mgiL no immobilized daphnids were determined during 

### the last period 

### of 48 hours. 

## Results with reference substance Reference Substance For evaluation of

### the quality ofthe daphnia clone and the experimental conditions, potassium dichromate Is tested as a 

### positive oontrot twice a year. The resuH 

### ofthe latest positive control test In Februar2009 (48-hour ECSO: 0.71 mg/L, study C36915) indicatad thatthe sensHivity of

### the test organisms waswHhin 

### the Internal historical range (48-hour ECSO 

### from 1996to2009: 0.53-1.1 mg/L). 

## Reportedstatistics enderrorestimates

no statistical analysis 

## Any other Information on results Incl. tables 

# Effectof Leuco Sulfur Black 1 onthe Mobilltv ofDaDhnla 

# magna

Loading No. of daphnlds Immobilized daphnlds Immobilized daphnlds

rate tested after24 hours after48 hours (mall) No. % No. % Control 

# 20 0 0 0 0

100 20 0 0 0 0 Measurements 

# I Observations Loading Exposure time rate Oh 

# I 24h 

# I 48h new 

# I old 

# I new 

# I old

## Appearance (1) Control 0 0 0 0 100 0 0 0 0 pH (2) Control 7.8 7.8 8.0 7.9 100 7.6 7.6 8.0 8.0 Oxygen (3) Control 8.3 8.4 7.9 8.0 100 8.0 8.1 8.0 8.1 Temperature (4) Control 20 20 20 20 100 20 20 20 20 (1) Appearance ofthe Freshly Prepared and Old Test Medtum (2) pH in the Freshly Prepared and Old Test Medium and 

## in the Control (3) Oxygen Concentrations (mg/L) in the Freshly Prepared and Old Test Medium and in the Control (4) Temperatures 

### ('C) in the Freshly Prepared and 0 ld Test Medium and in the Control 

# Applicant's summary and conclusion Validity criteria fulfilled
yes (In 

### lhe control no immobilized daphnids were determined during the test period of48 hours; Oxygen content >7.9mg/L.) 

### Conclusions In conclusion, lhe test item Leuco Sulfur Black 1 had noacute toxic effecls on Daphnia magna up

### to Its solubility limll in lhetestwater under

### lhe presenttest conditions (loading rate 

### 1OOmg/L nominal). 

### Executive summary 

## The acute toxicity ofthe test item Leuco Sulfur Black 1 to Daphnia magna was determined in a 48-hoursemi-static test according to the EU Commission Directive 92/69/EEC, Part C.2, Commission Regulation (EC) No 440/2008, Part C.2 and the OECD Guideline for Testing of Chemicals, No. 202 (2004). A limit test with a test medium renewal after24 hours was performed in accordance with the test guidelines to demonstrate that the test item has no toxic effecton the test organisms up to the solubility limitofthe test item in test water. Due to the low water solubility ofthe test item, the test medium was prepared priorto the start ofthe test and priorto the test medium renewal as follows: a dispersion ofthe test item with the loading rate of 100 mg/L was ultrasonic treated for 15

## minutes and continuously stirred at room temperature in the dark over 96 hours. Then, the dispersion was filtered. The undiluted filtrate ofthe dispersion was used as the test medium. Additionally, a control was tested in parallel. The preparation ofthe test medium was based on the OECD Guidance Document on Aquatic Toxicity Testing of Difficult Substa Thences and Mixtures, 2000. No remarkable observations were made concerning the appearance ofthe test medium. It was clearthroughout the entire testduration. No analytical work was performed due tothe specific properties ofthe test item (insolubility in water). At the beginning and end ofthe test medium renewal periods, the pH values ofthe test medium were between 7.6 and 8.0. The dissolved oxygen concentrations in the test medium and control were at least 7.9 mg/L. The watertemperature was 20 

# 'C during the test. The biological results were based on the loading rate ofthe test item of 100 mg/L. In the control and atthe loading rate of 100 mg/L no immobilized daphnids were determined during the test period of48 hours. The biological test results (based on the loading rate) were as follows: Loading rate (mg/L)

- 24-hour EC50: >100

- 24-hour ECO: 

## ? 100

## - 24-hour EC100: >100 -48 -hour EC50: >100 -48-hourECO:**~100** 

## -48 -hour NOEC:**~100** 

## -48 -hour EC100: >100 In conclusion, the test item Leuco Sulfur Black 1 had no acute toxic effects on Daphnia magna up to its solubility limit in the test water underthe present test conditions.

# 6.1.4 Long-term toxicity to aquatic invertebrates Endpointstudyrecord: Waiving_Long-term toxicityto 

# aquatic invertebrates UUID IUC5-fa6ba43d-cc0c-46a4-bdec-3c12f05eafa4 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks uensenba I Clariant Produkte (Deutsdlland) GmbH I Sulzbach am Taunus I Germany 2010-{)5-21 14:33:51 CEST



# Administrative Data Data w•lvlng 

### Justificationfor ut.-lvlng otherjustification Waiving according to "column 2" of Regulation (EC) No 1907/2006 (CSA does not indicale need forfurther investigations). Due to the very high molecularweight 

### (>8000 DeHon) the bloevailabilily ofthe subslence is negligible (low polential tocross biological membranes) and no toxic effects could 

### be expected. Therefore addltionallesting is noljuslified

# 6.1.5 Toxicityto aquatic algae and cyanobacteria Endpointstudyrecord: Key_201_1993_RCC_333977
UUID IUC5-73675ea2-8fc8-4fc9-a942-2603464c8c95 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks uensenba I Clartant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-05-t812:12:38 CEST


# Administrative Data Purpose 

### flag Study 

### result

### type Reliability key study experimental result 1 (reliable without restriction) Rationale

### for reliability Incl. deficiencies Study performed according to OECD-Guideline and GLP 

# Data source Reference

Re,.,..nce Author



### type study report RCC 

## Data access Year
1993

data submitter is data owner 

## Data protection claimed yes. but

### wHIIng to share Title
Acute Toxicity 

### of Diresul Black ROT Liquid to scenedesmus subspicatus



# Materials and methods Test guideline Bibliographic source Testing laboratory
RCC- Umweltchemie AG ltingen Report Owner Company Report 

### no. company study no. 

### elate 333977 Clariant

### Guideline OECD Guideline 201 (Alga, Growth Inhibition Test)

Method was modified 

### to differentiate between a reduced growth ofalgaedue

### to real toxic effects ofthe test substance 

### or due to an indirect effect, a reduced algae growth 

### by light absoprtion in coloured test solutions. 

## GLP compliance
yes (incl. certificate) 

# Test materials Identity oftest material same as for substance defined In 

## section 1 (If

## not read-across)

no 

## Test material identity ldenttfler Identity other. Leuco Sulfur Black 

### 1 formulation 

### see confidential details

### on test material 

## Confidential details on test material
Leuco Sulfur Black 

### 1: 30.2 

### % Sodium hydroxide: 

### 2.7% Sodium sulfide: 

### 3.6 

### % Inorganic sodium salts: 

### 3.9 

### % water. 59.6% 

## Analytical monitoring yes 

## Details 

## on analyticalmethods Apparatus: Perkin-Elmer Lambda 15 UVIVis spectrophotometer Wavelength: 580 nm 

## Vehicle

no

## Test organisms Test organisms (species) Scenedesmus subspicatus (new neme: Desmodesmus subspicatus) 

## Study design Testtype static 

### Water media type freshwater 

### Umlttest

no 

### Total exposure duration 72h 

## Test conditions pH IV. the start 

### ofthe tests, the pH ranged from 8.1 to 9.5. 

### IV. the end 

### ofthetests, 

### the pH ranged from 8.1 

### to 8.9. 

### Nominaland

### measuredconcentrllt/ons 200 mg DIRESUL BLACK ROT LIQUID were dissolved in test medium and filled toa volume 

### of200 mi. From this stock solution further dilutions in test mediumwere prepared and the respective amount ofalgae added. 

### The finel nomine! test concentrations were: 50, 5, 1, 0.5, 0.1 and 0.01 mg/1. DIRESUL BLACK ROT LIQUID wasfound to 

### be stable under the test conditions. The concentration in all lasts ranged from 85.5%to 107.7% ofthe nominal concentration. 

### Reference substance (positive control) yes (potassium dichromate) 

# Results and discussions Effect concentrations72h ECSO 5.99 mg/L nominal test 

### mat. biomass Experimental Part B 72h ECSO 1.84 mg/L nominal test mat. biomass Experimental PartA 72h ECSO 54.1 mg/L nominal test mat. growth rate Experimental PartA 72h ECSO 8.59 mg/L 

### nominal test mat. growth rate Experimental Part B 72 h NOEC 1-Smg/L 

### nominal test mat. 

### biomass Experimental Part B 72 h LOEC ca. 5 mg/L nominal test mat. 

### biomass Experimental Part B 72 h NOEC 1 mg/L 

### nominal test mat. 

### biomass Experimental Part A 72 h LOEC 1-5mgll. 

### nominal test mat. biomass Experimental Part A 

## Details on results The test included two experimental parts: Expertmantal pert A: the algae grew In test medium**w~h** dissolved dyestuff in Erlenmeyer

### flasks. Experimental pert B: 

### the algae grew in separate containers (test tube) which were surrounded by

### the black coloured solutions ofthe same concentration as tested In pert A. 

# Applicant's summary and conclusion Conclusions The growth Inhibition of Scenedesmus subspicatuswas In the same 

### magnitude when 

### the algae grew In test medium**~t** test substance but under reduced light intensities by

### the filter effect (experimental part B) as in experimental partA, where 

### the algae grew intest solutions -•dissolved lest substance. In conduslon, this modified algae test clearly demonstrated that

### the obsarved**lnhi~n** elfed ofthe substence was caused only by 

### the indlrad effect, 

### the lightabsorption in the coloured test solutions. Thus, a toxic effect of

### the

### test substence on the algae ceHs can be exduded up to a concentration of50 

### mg/1. 

## Executive summary 

## The influence of DIRESUL BLACK ROT LIQUID on the growth ofthe green algae Scenedesmus subspicatus was investigated. The study was performed according to the EEC Directive 92/69. The test method was modified to differentiate between a reduced growth ofalgae due to real toxic effects ofthe test substance on the algae cells or due to an indirect effect, a reduced algae growth by light absorption in coloured test solutions. The test included two experimental parts: Experimental part A: the algae grew in test medium with dissolved dyestuffin Erlenmeyerflasks. Experimental part B: the algae grew in separate containers (testtube) which were surrounded by the black coloured solutions ofthe same concentration as tested in partA.

## The biological results of both experimental partsA and B were almost identical. This was shown by several calculated parameters, the NOEC, LOEC and the EC 50 values: In part 

## A the NOECwas 

## 1.0 mg/1, and in part 

## B the NOEC wasfound to be between 

## 1 and 

## 5 mg/1. The LOEC was found to be between 

## 1and 

## 5 mg/1 in part

## A and itwas about

## 5 mg/1 in part 

## B. The percentage inhibition ofalgae biomass and the algae growth rate (IJ) after

# 72 hours of exposure were in the same order of magnitude. The EC 50 values calculated for both growth parameters, showed no significant differences. Experimental Part

# A EbC 50: Experimental Part B 

# 1.84 mg/1 5.99 mg/1 Er C 50: 

# 54.1 mg/1 8.59 mg/1 Thus, the growth inhibition of Scenedesmus subspicatus was in the same magnitude when the algae grew in test medium withouttest substance but under reduced light intensities by the filter effect (experimental part B) as in experimental part A, where the algae grew in test solutions without dissolved test substance. In conclusion, this modified algae test clearly demonstrated thatthe observed inhibition effect ofthe substance was caused only bythe indirecteffect, the light absorption in the coloured test solutions. Thus, a toxic effect ofthe test substance on the algae cells can be excluded upto a concentration of50 mg/1.

# 6.1.7 Toxicity to microorganisms Endpointstudyrecord: Memmert (2009) I Toxicity to microorganisms
UUID IUCS-9961e854-53954e6b-a2b4-316347d06ad2 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks jregano 

### IClariantlbl!rica Producd6n S.A. 

### I El Prat 

### da Llobregat (Barcelona) 

### I Spain 2010-QS-20 16:34:03 CEST



# Administrative Data Purposeflag 

### Study result

### type Reliability key study experimental**resu~** 1 (reliable without restriction) Rationale

### lor reliability Incl. deflclellCiea Guideline conform study under GLP without deviations 

# Data source Reference

Re...,..nce Author Year



### type study report Memmert 2009 

## u Data access data submitter is data owner 

## Data protection claimed yes Title Toxicity ofLeuco Sulfur Black 1

### to Activated Sludge in a Respiration Inhibition Test



# Materials and methods Bibliographic 

### source Teatlnglabonltory Report no. Harlan Laboratories Ltd. C31166 llingen (Switzerland) Owner Company Report company 

### study no. 

### date Clarlant 2009-oS19

### Test guideline OwlIIIIer 

### Guideline according EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test) (Com. Reg. (EC) 

### No 44012008 of30 May2008 laying down test methods to pwsuantto Reg. (EC) No 1907/2008 ofthe European Parliament and 

### ofthe Council on the**Reglstra~on.** Evaluation, Authorisation and Restriction 

### of Chemicals (REACH), Part C.11, Activated S) according EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test) (EU Commission Directive 881302/EEC, Part C.11, Activated Sludga 

### to Respiration Inhibition Tast, 1988) according OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test) (OECD Guideline

### for Testing ofChemicals, No. 209, Activated Sludge, Respiration 

### to Inhibition Test, 1984.) 

### GLP compliance yes (incl. cartificate) 

# Test materials Identity oftest material same as for substance defined In section 1 

### (If not read-.cross) yes 

### Test material Identity

ldentlfter Identity CASnumbar 66241-t1-0 ECnumbar 266-273-5 

### Details on test material

- Name 

### oftest material : Leuco Sulfur Black 1

- Physical state: thlxotrophic, black, wet paste -Composition: Leuco SuWur Black t :52.75% (w/w) (calculated by amount ofby-compounds) 2,4-0initrochlorobanzene: < 20 ppm 2,4-Dinltrophenole: 

### < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3 % (w/w) Water: 45.75% (w/w) by Karl-Fischer titration

- Lot/batch 

### No.: ESA0007595 -Expiration date ofthe lot/batch: 2011-07-11

- Stabiity undertest conditions: Stable under storage conditions. Deviations 

### no

no

no- Storage condition oftest material: AI room lemperature (range of 20 

### ± 5 

### 'C), light protected. 

### Analytical monitoring
not required 

### Details on sampling
For measurement

### ofthe respiration rate a wall-mixed sample 

### of each test medium was poured into a BOO-flask after three hours incubation time, and was notfurther aerated. 

### Details on analyticalmethods
For measurement of

### the respiration rate a well-mixed sample 

### ofeach test medium was poured into a BOD-flask after three hours incubation time, and was notfurther aerated. Then

### the dissolved oxygen concentration was measured 

### with an oxygen electrode

## f"NTW TriOxmatic® 300and an oxygen meterWTW Oxi 539, Wlssenschaftlich-Technische Wer1<staetten WTW, Weilheim/Germany), 

### end was continuously 

### recorded. During measurement, the samples

### wera continuously stirred on a magnetic stirrer. The oxygen consumplion rale (in 

### mg 02 L-1 minute-1)was delennined from the linear part 

### ofthe respiration curve in the range 6.5-2.5

### mg 02/L. In case ofvery rapid oxygen consumption, the range used was belowthe limits indicated above butalways within 

### the linear part ofthe respiration curve. In case of

### low oxygen consumption 

### the rate was detennined over a period 

### ofat least ten minutes. 

### Vehicle

no 

### Details on testsolutions PREPARATION AND APPLICATION OF TEST SOLUTION (especially for dlfficuK test substances) The final nominal concentration tested was 1000 mg/1.. Based on 

### the resuils ofa**~xparimenl** (without GLP), no stock solution could 

### be prapared due to

### the lowwatersolubility 

### ofthetest "em under the pre-test**cond~ns.** Therefore, 500.0 

### mg ofthe test hem was weighed by means ofan analytical balance and transferred to the designated test vessel with 284 mL tapwater. The test Item was mixed intotha tap water by uKrasonictreatmentforfifteen minutes and intense 

### stinring for 24 hours at room temperature in the dari< to dissolve a maximum amount of 

### the lest hem and/ordisperse " as homogeneouslyas possible. No emulsifiers orsolvents were 

### used. Afterstirring, the appearance ofthe lest medium wasevaluated, and the test concentration 

### was dearly above the watersolubility**li~** 

### of Leuco Sulfur Black 1 under the test cond"ions. Then, 16 mL synthetic sewage feed and 200 

### mL activated sludge inoculum were added 

## Test organisms Test organisms (species)
activaled 

### sludge ofa predominantlydomestic sewage 

### Details on Inoculum
The study was performed 

### with aerobic activated sludge from a wastewatertreatment plant (ARAErgotz II, Fullinsdiorf I Switzerland) treating predominantly domesticwastewater. The sludgewas washed twice 

### with tap water by centrifugation and the supernatant liquid phase was decanted. A homogenized aliquot 

### ofthe final sludge suspension wasweighed,thereafter dried and the ratio ofwetto dry weightwas calculated. Based on

### this ratio, an aliquot ofwashed sludgewas suspended in

### tap water

### to obtain a concentration equivalent

### to 3 

### g dry material per**l~er.** During the holding period ofthree days 

### prior

### to use, the sludgewasfed dailywith 

### 50 ml synthetic sewage feed" per literand was keptat 

### room temperature undercontinuous aeration until use. Before use, the dry weightof the activated sludge was measured again In the inoculum usedforthe test. The pH ofthe activated sludge Inoculum was adjusted from 

### 8.7to

### 7.3 with a diluted sulfuric 

### acid solution. • Synthetic sewage

### feed: 16g peptone 11 g meatextract 

### 3 g 

### urea 0.7 

### g NaCI 

### 0.4 

### g CaCI2 • 2H20 0.2 

### g MgS04 • 7H20 2.8 gK2HP04 filled 

### up 

### to a final volume of 1 literwith deionized water 

## Study design Test 

## type static 

## Water media 

## type freshwater 

## Limittest
yes 

## Total exposure duration 3h 

### Post

## exposure observation period

none 

## Test conditions Hardness
notdertermined 

## Test temperature 20'C

## pH

see table "Remarks on results" 

### Dissolvedoxygen
see table *Remarks on results" 

### Salinity synthetic waste water 

### Nominalandmeasuredconcentrations 1000mg/L 

### Details on 

### test conditions TEST SYSTEM

- Test vessel: 2000mt glass beakers

- Aeration: During the Incubation period of 3 hours the single test medium and the controls were rontinuously aarated by Intense**st~ring** on magnetic stirrers to avoid possible foaming and/or stripping ofthe test Hem.

- No. ofvessels per concentration (replicates): 1 -No. ofvessels percontrol (replicates): 2

TEST MEDIUM 

### I WATER PARAMETERS -The study was performed with aerobic activated sludge from a wastewater treatment plant (ARA Ergotz II, FOUinsdorf I Swilz-nd)traating predominantly domesticwastewater. The sludge was washed twice with tap water by centrifugation and the supernatant liquid phese was decanted. A homogenized aliquot ofthe final sludge suspension wasweighed, thereafter dried and the ratio ofwetto dry weightwas calculated.

- Based on this ratio, an aliquot ofwashed sludge was suspended in tap waterto obtain a concantration equivalent

### to 3 g dry material per IHer. During the holding period ofthree days prior

### to use, the sludge was fed daily with 50 ml synthatic sewage feed• per IHer and was kept at room temperature under rontinuous aaratlon until use. Bafore use, the drywelght of the activated sludgewas measured again in the inoculum 

### used forthe test. The pH ofthe activated sludige inoculum wasedjusted

### from 8.7

### to 7.3 with a diluted sulfuric acid solution. • Synthetic sewage 

### feed: 16

### g peptone 11 g meat extract 

### 3 g urea 0.7 g NaCI 0.4 g CaCI2 • 2H20 0.2 g MgS04 • 7H20 2.8 gK2HP04 filled up

### to a final volume of 1 literwith deionized waterEFFECT PARAMETERS MEASURED (with observation intervals Wapplicable): respiration rate
TEST CONCENTRATIONS 1 OOOmg/L nominal 

## Reference 

## substance (positive control)
yes (3,5-<llchlorophenol) 

## Any other lnfonnatlon on materials and methods Incl. tables 

## At the start ofthe test (after the stirring period of24 hours), synthetic sewage feed and activated sludge inoculum were added. The inoculum had a sludge concentration of2.7 g/Ldry weight (corresponding to about 1.1 g dry material per liter test medium). The sludge was added in time intervals of 15 minutes (an arbitrary but convenient interval) first to a control, secondlyto the test solutions ofthe reference item, thirdly to the test solution ofthe test item, and finally to the second control. During the incubation period of3 hours the single test medium and the controls were continuously aerated by intense stirring on magnetic stirrers to avoid possible foaming and/or stripping ofthe test item. The concentration ofdissolved oxygen did not drop below 2.5 mg/L during the incubation period. Just before measurementofthe respiration rates the dissolved oxygen concentrations were at least 8.5 mg/L. The temperature in the test media, measured in one control, was 20 

# 'C atthe start and at the end ofthe incubation period. 

# Results and discussions Effectconcentrations Remarks 

### (e.g. 95% CL)



### M. thetest Item concentration of 1000 mgll, the test ttem was not completely dissolved In 

### the test medium. Thus, the test concentration was clearly above the watersolubility limit 

### of Leuco SuWur Black 1 underthe test conditions. 

## Results with reference substance (positive control)
The 3-hour EC50 ofthe reference**~em** 3,5-<llchlorophenol (positive control) was calculated to be 18 mgll (the 95% confidence limits could not be calculated due to mathematicalreasons). The :Hlour

### EC50 is within 

### the guideline-recommended range of

### 5-30 mgll, confirming suitability ofthe activated sludge used. 

## Reportedstatistics anderrorestimates

no statistical analysis done 

## Any other Information on results Incl. tables 

## Influence of Leuco Sulfur Black 1 (Test Item) and 3,5-Dichlorophenol (Reference Item) on the Oxygen Consumption ofActivated Sludge Vessel Nominal Oxygen Inhibition pH Oxygen

no. concentration consumption values Concentration 

## oftest rate (mg 02/L) chemical

(mg/L) (mg 02 L-1 (%) start* end* start* end* min-1) Control 

# A 0 1.415 7.5 8.4 7.9 8.5 B 0 1.423 7.9 8.3 7.9 8.9 Mean 1.419 Deviation (%) 0.6 Test Item 5 

# I 1000 1.397 1.6 8.1 8.2 7.9 8.7 Reference Item 1 5 1.520 -7.1 7.8 8.2 7.9 8.9 2 16 0.734 48.3 8.0 8.3 8.0 8.8 3 50 0.204 85.6 8.0 8.4 8.2 9.2 • Start and end ofthe 3-hour mcubatlon penod -%inhibition: increased oxygen consumption rate relative to control 

# Applicant's summary and conclusion

## Validity criteria fulfilled yes 

## Conclusions The 3-hour NOEC (EC15) 

### of Leuco Sulfur Black 1 to activated sludge microorganisms was at least 1000 mg/L. This value mighteven 

### be higher butconcentrations 

### abova 1000 mg/L 

### were nottested. 

## Executive summary 

## The inhibitory effect ofthe test item Leuco Sulfur Black 1 on the respiration rate ofaerobic wastewater microorganisms of activated sludge was investigated in a 3-hour respiration inhibition test according to the EU Commission Directive 88/302/EEC, Part C.11, Commission Regulation (EC) No. 440/2008, Part C.11 and the OECD Guideline for Testing of Chemicals, No. 209. Based on the EU testing guidelines, a limittestwith a nominal test concentration of 1000 mg/Lwas performed. In addition, two controls and three different concentrations ofthe reference item 3,5-dichlorophenol (5, 16, and 50 mg/L) were tested in parallel. The results ofthese treatments confirmed suitability ofthe activated sludge and the method used. At the test item concentration of 1000 mgll, the test item was not completely dissolved in the test medium. Thus, the test concentration was clearly above the water solubility limit of Leuco Sulfur Black 1 underthe test conditions. The test item had no inhibitory effect on the respiration rate ofactivated sludge after the incubation period ofthree hours at a loading rate of 1000 mg/L. The saturation concentration (water solubility limitof Leuco Sulfur Black 1 underthe test conditions) was reached and no inhibitory effectwas determined afterthree hours contact time. Thus, the 3-hour NOEC (EC15) ofthe test item to activated sludge microorganisms was at least 1000 mg/L (i.e. the saturation concentration ofthe test item). This value might even 

## be higher butconcentrations above 1000 mg/L were not tested. The 3-hour EC20, EC50, and EC80 could not 

## be calculated but were clearly higher than 1000 mg/L. Page

# 6.2 Sedimenttoxicity Endpointstudyrecord: Waiving_ Long term testing Sediment toxicity
UUID IUC5-2ced649d-4c42-41Of-a7ce-{;14d2506104a Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks uensenba I Clariant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-05-21 16:20:09 CEST



# Administrative Data Data 

### waiving Justification for 

### data waiving otherjustification Waiving according to "column 

### 2" of Regulation (EC) No 190712006 (CSA does not indicate need forfurther Investigations). Due tothe very high molecularweight (>8000 Dation) the bloevallabiiHy ofthe substance is negligible (low potential to cross biological membranes) and no toxic effects could be expected. Therefore additional testing is notjustified

# 6.3 Terrestrial toxicity 6.3.1 Toxicity to soil macroorganisms except arthropods Endpointstudyrecord: Waiving_Short term toxicity to soilmacroorganlsms except arthropods UUID IUC5-aafbb6a9-8c65-416d-a161-35e7ec89e6e1 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author uensenba 

### I Clarianl Produkte (Deutschland) GmbH 

### I Sulzbach am Taunus 

### IGermany Date 2010-05-2116:21:59 CEST Remarks 

# Administrative Data D81a waiving otherjustification Justificationfor Waiving according to "column 2" of Regulation (EC) No 190712006 (CSA 

### does nollndicale need forfurther investigations). Due lothe very high molecularweight data waiving (>6000 DaHon) 

### the bioavailability ofthe substance is negligible (low potential to cross biological membranes) and notoxic effects could 

### be expected. Therefore additional tasting is notjustified

# Endpointstudyrecord: Waiving_Long term toxicity

# to soilmacroorganisms except arthropods
UUID IUC5-19196dal>-9944-4cec-8531-e6234741ab34 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remartc.s uensenba I Clariant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-{)5-21 16:24:01 CEST



# Administrative Data Data 

### waiving Juatlflcatlon for 

### data 

### waiving olherjustification Waiving accon:ling 1o "column 2' 

### of Regulation (EC) No 190712006 (CSA does nol indicate 

### nead for lurlher Investigations). 

### Due lo lhe very high molecularweighl (>8000 DaMon) lhe bioavailabillly ofthe substance is negligible (low potentiallo cross biological membranes)

### and no loxic

### allecls could be expected. Therefore addHional testing

### Is noljustified

# 6.3.3 Toxicitytoterrestrial plants Endpointstudyrecord: Waiving_Short-term toxicity to terrestrialplants UUID IUCS-28691 b63-da93-4d15-a3df-f0fd4fcd43a6 Dossier UUID IUCS-292149d1-a677-4feb-9692-1fad973261ea Author Date Remarks uensenba I Clarlant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-05-21 16:2627 CEST



# Administrative Data Data waiving 

### Justificationfor --lYing otherjustification Waiving according to "column 2" of Regulation (EC) No 190712006 (CSAdoes not indicate need forfurther Investigations). Due 

### to the very high molecularweight (>8000 Dalon)the bloevallability ofthe subBiance Is negligible (low potential tocross biological membranes) 

### end notoxic

### effects could 

### be expected. Therefore additional testing Is noljustifled

# Endpointstudyrecord: Waiving_Long term toxicity to terrestrialplants
UUID IUC5-d4fc96dc-5719-4787-a3db-18d850f1e51a Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remart<s uensenba I Clariant Produkte (Deulschland) GmbH I Sulzbach am Taunus I Germany 2010-05-21 16:27:46 CEST



# Administrative Data Data w•lvlng JustlliCIIUon for 

### data-lvlng otherjustification Waiving acx:ording 

### to "column 

### Z' of Regulation (EC) No 1907/2006 (CSA

### does 

### not indicate need for further investigations). Due 

### to the very high molecularweight (>8000 Dakon) the bioavailability ofthe substance Is negligible (low potential 

### to cross biological membranes) and notoxic effects could be expected. Therefore additionaltesting 

### is 

### not justified

# 6.3.4Toxicity to soil microorganisms Endpointstudyrecord: Waiving_Toxicity 

# to soilmicroorganisms
UUID IUC5-a35b509b-8257-4e91-8b57-Q88e9fcaf22c Dossier UUID IUC5-292149d1-a6n-4feb-9692-1fad973281ea Author Date Remarks uensenba I Clarlant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-05-21 16:28:01 CEST



# Administrative Data Dahl

### waiving 

### Justificationfor -waiving otherjusliflcation Waiving according to "column 2" 

### of Regulation (EC) No 1907/2006 (CSA does nol indicate need forfurther Investigations). Due to the 

### very high molecularweight (>8000 Dalon)the 

### bloavaolability ofthe substance is negliglbla (low potential to cross biological membranes) and notoxic effects could 

### be expected. Therefore additional testing is notjustified

# 6.3.5 Toxicity to birds Endpointstudyrecord: Waiving_ Toxicity to birds
UUID IUC5-149aa32c-5909-4821-b6a2-2f1605c38b2f Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks uensenba I Clariant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-05-21 16:29:42 CEST



# Administrative Data Data 

### waiving Justification

### for data waiving otherjustification Waiving according to "column 

### 2"of Regulation (EC) No 190712006 (GSA doas not indicate need lorfurther investigations). Due to the very high molecularweight (>8000 Dakon)the bloevailabilky ofthe substance is negligible (low potential to 

### cross biological membranes) and notoxic effects could 

### be expected. Therefore additional testing is notjustified

# 7 Toxicological information Endpointsummary: Toxicologicalinformation UUID IUCS-1a26f429-de64-4dfb-b484-e7a7ba13a7e6 Dossier UUID IUC5-292149d1-a6n-4feb-9692-11ad973281 ea Author Date Remarks uensenba I Clarlant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-05-21 15:25:38 CEST



# Administrative Data
Workers Acute 

# Ishort-term exposure - systemic effects Dermal DN(M)EL Justification for (no) DN(M)EL derivation 

## Iapplied assessmentfactors Leuco Sulfur Black 1 is not dassified loracute systemic effects. Thus, the derivation ofthe DNEL is not required (see also "discussion"). 

## Inhalation DN(M)EL Justification for (no) DN(M)EL derivation 

## Iapplied assessmentfactors Leuco Sulfur Black 1 is not dassified lor acute systemic effects. Thus, the derivation ofthe DNEL is not required (see also "discussion"). 

## Acute 

# Ishort-term exposure • local effects Dermal DN(M)EL Justification for (no) DN(M)EL derivation 

## Iapplied assessmentfactors Leuco Sulfur Black 1 is not dassified loracute systemic effects. Thus, the derivation ofthe DNEL Is not required (see also "discussion"). 

## Inhalation DN(M)EL Justification for (no) DN(M)EL derivation 

## Iapplied assessmentfactorsLeuco SuHur Blacl< 1 is not classified for acute systemic effects. Thus, thederivation ofthe DNEL is not required (see also "discussion"). 

# Long-tenn exposure - systemic effects Dermal DN(M)EL

DNEL (Derived No Effect Level) 25 mglkg bwlday 

### Aueument 40 DOH NOAEL Moat repeated dose toxicity 

### bctor descriptor 

### Hnsltlve starting endpoint point 

## Justification for (no) DN(M)EL derivation I 

## applied assessmentfactors


## Inhalation DN(M)EL

DNEL (Derived No Effect Level) 176

### mglm' A

... 

### ssment 10 DOH NOAEC 

### Most repeated dose toxicity 

### factor descriptor 

### Hnslllve starting endpoint point 

## Justification for (no) DN(M)EL derivation 

## Iapplied assessmentfactors

see •discussion" 

# Long-tenn exposure • local effects Dermal DN(M)EL Justificationfor (no) DN(M)EL derivation I 

## applied assessmentfactors Leuco Sulfur Blacl< 1 is not classified for acute systemic effects. Thus, the derivation ofthe DNEL is not required (see also "discussion"). 

## Inhalation DN(M)EL Justification for (no) DN(M)EL derivation/applied assessmentfactors Leuco SuHur Blacl< 1 is not classified for acute systemic effects. Thus, the derivation ofthe DNEL is not required (see also "discussion"). 

## Discussion

## Acute/shortterm exposure - systemic and local effects Leuco Sulfur Black 1 did not induce anytoxic effects in the acute oral and dermal and oral repeated dose toxicity study. It did not induce skin irritation in the acute dermal toxicity study in rats, induced only minimal signs of irritation to the rabbit eye shortly after instillation without any relevance for classification and was not sensitising inthe LLNA. 

# As Leuco Sulfur Black 1 did not show acute systemic noracute local toxicity, the derivation ofacute DNELs for systemic and local effects is not required. This is in accordance with the ECHA guidance document on information requirements and chemical safety assessment, chapter R.8. Long-term exposure - systemic effects In a repeated dose toxicity study with reproduction/developmental toxicity screening according to OECD guideline 422 and GLP Leuco Sulfur Black 1 was administered by gavage to groups of 10 rats per sex and dose at 0, 100, 300 and 1000 mg/kg bw/d. Male rats were treated for 29 days, female rats for 14 days priorto pairing, though the pairing and gestation periods until the F1 generation had reached day4 postpartum. The only effect seen was a dark discolouration offaeces in all high dose animals starting on day3 ofthe study and lasting until termination. Discolouration of organs or urine or substance deposits in organs were not found. This is strongly indicates that systemic absorption did not occur. Based on the results obtained in this study a NOAEL of 1000 mg/kg bw/d was derived. Leuco Sulfur Black 1 is an UVCB substance with a mean molecularweight in the range of 8000 to 9000 Da and a numberaverage molecularweight (Mn) ofabout 3000 Da. It is very poor1y or even notsoluble in common polar and non-polar solvents such as water (solubility of62.4 mg/L), DMSO, methanol, DMF, THF, chloroform and not soluble in acids like sulfuric acid and trifluoroacetic acid. Due to these substance characteristics it is very unlikely that Leuco Sulfur Black 1 can cross any biological membrane. In summary, no evidence oftoxicity was seen in the OECD 422 guideline study. The outcome ofthe study and the substance characteristics show that an absorption oftoxicologically significant amounts of Leuco Sulfur Black 1 subsequent to oral, dermal or inhalation exposure is negligible. Nevertheless, a DNEL for the dermal and inhalation exposure route is derived assuming absorption of5 

# % for all exposure

## routes. Dermal exposure to Leuco Sulfur Black 1 and the formulated Sulfurdye preparation can occur at the workplace. Inhalation exposure to Leuco Sulfur Black 1 is not possible due to pasty consistence and can only occurwhen the substance is formulated in aqueous media. The NOAEL of 1000 mglkg bw/d from the OECD guideline 422 study in rats is used as starting points forthe establishment ofthe DNELs. The oral NOAEL reflects also the dermal NOAEL because no difference in absorption is expected between both routes. Forthe derivation ofthe worker inhalation DNEL (assuming a daily exposure for 8 hours), the oral NOAEL is converted into a repeated inhalation NOAEC according to the following formula: inhalation NOAEC 

# = oral NOAEL 

# * 1/sRV(rat)

# *ABSoral-raUABSinh-human 

# * sRV(human)JwRV(human) with: oral NOAEL 

# = 1000 mg/kg bw/d sRV(rat)

# = standard respiratory volume (rat)

# = 0.38 m3

## /kg (8 h) ABSoral-raUABSinh-human 

# = 1 sRV(human)/wRV(human) 

# = 6.7 m3

## /10m3 

## Thus, the inhalation NOAEC is 1763 mg/m3 

## forthe worker. The following assessmentfactors are used forthe derivation ofthe long-term systemic DNELs: lnterspecies (allometric scaling rat-> human (not to be applied for inhalation exposure): 4 (no additional factor for remaining differences is needed because ofthe substance characteristics (see above)) lntraspecies: 5 (worker) Exposure duration: 2 (subacute to chronic) (the factor of2 instead of6 is used as there was no effect seen in the OECD guideline 422 studyjustifying a higherfactor) Dose-response: 1 Quality ofdatabase: 1

## The resulting overall assessmentfactors and DNELs are: worker (dermal exposure) overall assessmentfactor: 4 x 5 x 2 x 1 x 1 

# =40 DNEL: 1000 mg/kg bw/d /40 

# = 25 mg/kg bw/d worker (inhalation exposure): overall assessment factor: 5 x 2 x 1 x 1 

# = 1 0 DNEL: 1763 mg/kg bw/d /10 

# = 176 mg/kg bw/d Long-term exposure - local effects Leuco Sulfur Black 1 does not induce any local toxic effects. Moreover, no evidence oftoxicity was seen in any ofthe available tests. Thus, the derivation ofa long-term DNEL for local effects is not required. This is in line with the ECHA guidance documenton information requirements and chemical safety assessment, chapter 

# R.B. 

# General population Acute 

# Ishort-term exposure - systemic effects Dermal DN(M)EL Justification for (no) DN(M)EL derivation 

## Iapplied assessmentfactors
Leuco Sulfur Black 1 is not classified loracute systemic effects. Thus. thederivation ofthe DNEL Is notrequired (sse also "discussion"). 

## Inhalation DN(M)EL Justificationfor (no) DN(M)EL derivation 

## Iapplied assessmentfactors Leuco Sulfur Black 1 Is notclassified for acute systemic effects. Thus, thederivation ofthe DNEL Is notrequired (sse also "discussion"). 

## Oral DN(M)EL Justificationfor (no) DN(M)EL derivation 

## Iapplied assessmentfactors Leuco Sulfur Black 1 is 

### not classified for acute systemiceffects. Thus. the derivation ofthe DNEL is notrequired (sse also "discussion"). 

# Acute 

# Ishort-term exposure -local effects

## Dennal DN(M)EL Justificationfor (no) DN(M)EL derivation 

## Iapplied assessmentfactors Leuoo Sulfur Black 1 is not dassified for acute systemic effects. Thus, 

### the derivation ofthe DNEL Is 

### not required (see also "discussion"). 

## Inhalation DN(M)EL Justlflcationfor (no) DN(M)EL derivation I applied assessmentfactors Leuoo Sulfur Black 1 is not dassified for acute systemic effects. Thus, 

### thederivation ofthe DNEL is not required (see also "discussion"). 

# Long-term exposure - systemic effects Dennal DN(M)EL
DNEL (Derived No Effect Level) 12.5 mg/kg bw/day AOHaarnent 80 Dose NOAEL Moat repeated dose toxicity 

### factor descriptor sensitive startlng endpoint point 

## Justification for (no) DN(M)EL derivation 

## Iapplied assessmentfactors

see "discussion" 

## Inhalation DN(M)EL
DNEL (Derived No Effect Level) 43.5 mglm' Assessment 20 Dose NOAEC Moat repeated 

### dosetoxicity 

### factor descriptor sensitive startlng endpoint point 

## Justification for (no) DN(M)EL derivation I applied assessmentfactors
see "discussion" 

## Oral DN(M)EL
DNEL (Derived No Effect Level) 12.5 mglkg bw/day AOHaarnent 80 Dose NOAEL Moat repeated dose toxicity 

### factor descriptor sensitivestarting endpoint point 

## Justificationfor (no) DN(M)EL derivation I 

## applied assessmentfactors

see "discussion" 

# Long-tenn exposure • local effects Dennal DN(M)EL Justificationfor (no) DN(M)EL derivation 

## Iapplied assessmentfactors Leuco Sulfur Black 1 is not dasslfied for acute systemic affects. Thus, the derivation of

### lhe DNEL Is not required (see also 'discussion'). 

## Inhalation DN(M)EL Justificationfor (no) DN(M)EL derivation 

## Iapplied assessmentfactors Leuco Sulfur Black 1 is notdassified for acute systemic effects. Thus, the derivation ofthe DNEL is not required (see also 'discussion'). 

## Discussion 

## Acute/shortterm exposure -systemic and local effects Leuco Sulfur Black 1 did not induce anytoxic effects in the acute oral and dermal and oral repeated dose toxicity study. It did not induce skin irritation in the acute dermal toxicity study in rats, induced only minimal signs of irritation to the rabbit eye shortly after instillation without any relevance for classification and was not sensitising inthe LLNA. As Leuco Sulfur Black 1 did not show acute systemic noracute local toxicity, the derivation ofacute DNELsfor systemic and local effects is not required. This is in accordance with the ECHA guidance document on information requirements and chemical safety assessment, chapter R.8. Long-term exposure - systemic effects No evidence oftoxicity was seen in the OECD 422 guideline study (NOAEL of 1000 mg/kg bw/d). The outcome ofthe study and the substance characteristics show that an absorption oftoxicologically significant amounts of Leuco Sulfur Black 1 subsequentto oral, dermal or inhalation exposure is negligible. (See also discussion under DNEL derivation for the worker.)

## Leucosulfur Black 1 is only used professionally and not included in articles intended to release the substance. Nevertheless, a DNEL for the oral, dermal and inhalation exposure route is derived assuming absorption of5 %forall exposure routes. The NOAEL of 1000 mglkg bw/d from the OECD guideline 422 study in rats is used as starting points forthe establishment ofthe DNELs. The oral NOAEL reflects also the dermal NOAEL because no difference in absorption is expected between both routes. Forthe derivation 

# ofthe consumer inhalation DNEL (assuming a daily exposure for 24 hours), the oral NOAEL is converted into a repeated inhalation NOAEC according to the following formula: inhalation NOAEC = oral NOAEL 

# * 1/sRV(rat) 

# * ABSoral-rat/ABSinh-human with: oral NOAEL = 1 000 mg/kg bw/d sRV(rat) = standard respiratory volume (rat)= 1.15 m3

## /kg (24 h) ABSoral-rat/ABSinh-human = 1 Thus, the inhalation NOAEC is 870 mg/m 3 

## for the consumer. The following assessment factors are used forthe derivation ofthe long-term systemic DNELs: lnterspecies (allometric scaling rat-> human (not to be applied for inhalation exposure): 4 (no additional factorfor remaining differences is needed because ofthe substance characteristics (see above)) lntraspecies: 10 (consumer) Exposure duration: 2 (subacute to chronic) (the factor of2 instead of6 is used as there was no effect seen in the OECD guideline 422 studyjustifying a higherfactor) Dose-response: 1 Quality ofdatabase: 1

## The resulting overall assessmentfactors and DNELs are: consumer (oral exposure) overall assessmentfactor: 4 x 10 x 2 x 1 x 1 

# =80 DNEL: 1000 mg/kg bw/d /80 

# = 12.5 mglkg bw/d
consumer (dermal exposure) overall assessmentfactor: 4 x 10 x 2 x 1 x 1 

# = 80 DNEL: 1000 mg/kg bw/d /80

# = 12.5 mglkg bw/d
consumer (inhalation exposure): overall assessmentfactor: 10 x 2 x 1 x 1 

# = 20 DNEL: 870 mg/kg bw/d 

# I 20 

# = 43.5 mg/m3 

## Long-term exposure - local effects Leuco Sulfur Black 1 does not induce any local toxic effects. Moreover, no evidence oftoxicity was seen in any ofthe available tests. Thus, the derivation ofa long-term DNEL for local effects is not required. This is in line with the ECHA guidance document on information requirements and chemical safety assessment, chapter R.8.

# 7.1 Toxicokinetics. metabolism and distribution Endpointsummary: Toxlcokinetics, metabolism anddistribution
UUID IUC5-e019da60·99cd-4350-99ca-82e9faf3be47 Dossier UUID IUC5-292149d1-asn-4feb-9692-1fad973281ea Author lhr I SCC GmbH I Wendelsheim I Germany Date 2010-05-2617:11:52 CEST Remarl<s 

# Administrative Data Discussion The sulfur dye Leuco Sulfur Black 1 is a black pasty UVCB substance with a mean molecularweight in the range of8000 to 9000 Da and a number-average molecularweight (Mn) of about 3000 Da. It is very poorly oreven not soluble in common polar and non-polar solvents such as water (solubility of62.4 mg/L), DMSO, methanol, DMF, THF, chloroform and not soluble in acids like sulfuric acid and tritluoroacetic acid. The vapour pressure ofthe pure substance, calculated from the boiling point of 

## > 400 

# 'C, is 1.8 x 1OE-5 hPa at 25 

# 'C. Based on these properties it is very unlikely that Leuco Sulfur Black 1 can cross any biological membrane. Thus, absorption oftoxicologically significant amounts of Leuco Sulfur Black 1 following oral, dermal or inhalation exposure can 

# be excluded. This is substantiated by the results obtained in toxicity and ecotoxicity studies, where no substance related local and/or systemic effects indicative oftoxicity were seen in all tests, induding acute animal and acute ecotoxicity studies, sensitization, repeated dosetoxicity and mutagenicity tests. In the acute oral toxicity study black stained faeces was observed for maximally four days starting the day after administration ofthe substance at 2000 mglkg bw. Dark discoloured faeces were also seen in the repeated dose toxicity study (OECD 422) at 1000 mg/kg bw/d, starting on day 3 ofthe study and lasting throughout the administration period until termination. The discolouration offaeces was the only effect observed in both studies and can be attributed to the strong

## staining properties ofthe substance. This indicates that Leuco Sulfur Black 1 is directly excreted and that absorption in the gastrointestinal tract does not occurfollowing oral administration, especially since organs or urine were notdiscoloured and signs ofsubstance deposits were not found in histopathological analysis ofthe organs. Based on the substance properties and as absorption via any route is negligible, metabolism and distribution in the body is also negligible. After oral administration Leuco Sulphur Black 1 is excreted via faeces as demonstrated in the acute oral as well as repeated dose toxicity study (OECD 422).

# 7.2 Acute Toxicity Endpointsummary: Acute Toxicity
UUID IUC5-<:e8d63d8-f0f4-4df6-ab3a-ee2829d2e160 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author lhr 

### I SCC GmbH 

### IWendelsheim 

### I Germany Date 2010-05-26 16:22:28 CEST 

### Remarks 

# Administrative Data Short description of key information

The LD50 oral and dermal in 

### the rat is above 2000 

### mg/kg bw. Leuco SuWur Black 1 did neithercause lethality nor clinical signs indicative 

### oftoxicity in both arute toxicity studies. 

# Discussion The acute toxicity of Leuco Sulfur Black 1 by oral application was investigated according to the OECD Guideline 423 under GLP. Female rats were treated by oral gavage administration at a dosage of2319 mg active ingredient 

# I kg bw (corresponding to 4396 mg test item 

# I kg bw). No deaths occurred during the study. Exceptfor black stained faeces from test day2 up to test day 5 atthe latest, no clinical signs were noted in all animals during the observation time. The body weight ofthe animals was within the range commonly recorded for this strain and age. No macroscopic findings were observed at necropsy. The LD50 is greaterthan 2000 mglkg bw. The acute toxicity by dermal application of Leuco Sulfur Black 1 was investigated according to OECD Guideline 402 under GLP.

## Five male and five female rats were treated at2319 mg active ingredient/kg (corresponding to 4396 mg test item 

# I kg bw) by dermal application. The test item was formulated in purified water and administered at a volume dosage of 6 mUkg. The application period was 24 hours. No deaths occurred during the study. No clinical signs were observed during the course ofthe study. Due to moderate black staining ofthe treated skin afterthe removal ofthe application patch, the assessmentoferythema was first prevented on test day 2 in all animals. From test day 3 to test day 7 at the latest, a slight black staining was still present. Erythema was assessable, but not noted in all animals from test day 3 to the end ofthe study. No oedema was observed throughoutthe whole observation period. Scaling was observed in one male on testday4. The bodyweightofthe animals was within the range commonly recorded for this strain and age. No macroscopicfindings were observed at necropsy. The LD50 is greaterthan 2000 mg/kg bw. 

# An acute inhalation toxicity study has not been performed because exposure of humansvia inhalation is not likely due to the physical state (wet paste) and vapour pressure (calculated as 1.8 x 10E-5 hPa at25 

## 'C) ofthe substance. 

# Justification for classification or non-classification Based on the available data Leuco Sulfur Black 1 is notsubjectto classification and labelling according to Directive 67/548/EEC and Regulation 1272/2008/EC.

# 7.2.1 Acute toxicity: oral Endpointstudyrecord: Simon (2009) I key (class method)
UUID IUC5-cdf11aa3-76c1-4e25-9054-e23a5a96f2o4 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks lhr I SCC GmbH I Wendelsheim I Germany 2010-05-26 16:20:05 CEST


# Administrative Data Purpc>HIIag Study 

### result

### type Reliability key study; robust study summery; used for dasslfication; used for MSDS experimental result Study

### period 2009-01-20

### til 2009-02-12 1 (reliable without restriction) Rationalefor rallablllty Incl. deficiencies Guideline-conform study under GLP wrthout deviations 

# Data source Reference

Rafentnce Author



### type Year 

### Title study report Simon 2009 Acute Oral Toxicity Study 

# c Data access data submitter Is data owner 

## Data protection claimed
yes in Rats



# Materials and methods Test

## type BlbUographlc Testing labonltory source Harlan Laboratories Ltd.• FOHinsdorf I Switzerland Report 

# o..- Compeny study Report 

### no. compeny 

### no. 

# - C31043 Clariant 2009-08- 24acute toxic class method 

## Umlttest
yes 

## Test guideline

Qwollfier Guideline 

### according to OECO Guideline 423 (Acute Oraltoxicity- Acute ToxicClass Method) according to EU Method 8.1 tris (Acute Oral Toxicity -Acute Toxic Class Method) 

## GLP compliance
yes (incl. osrtificate) 

# Test materials Deviation•
no no



## Identity oftest material same as for substance defined In section 1 (If

## not read-across)
yes 

## Test material Identity

Identifier Identity CAS number 66241-11-Q ECnumber 266-273-5 

## Details on test material

- Name 

### of

### test material : Leuco Sulfur Black 1

- Physical state: thixotrophic, black, wet paste

-**Compos~ion:** Leuco Sulfur Black 1:52.75% (w/w) (calculated by amount of by-compounds) 2.4-Dimrochlorobenzene: < 20 ppm 2,4-0initrophenole: < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3 % (w/w) Water: 45.75% (w/w) by Kar1-Fischer titration

- Lot/batch

### No.: ESA0007595 -Expiration date of

### the lot/batch: 2011-Q7-11

-**Stabil~** undertest conditions: Stable understorage conditions. -Storage condition oftest material: At room temperature (range of20 

### ± 5 

### 'C), light protected.

## Test animals Species
rat 

### Strain Wistar 

### Sex female 

### Details on 

### test animals andenvironmental conditions TEST ANIMALS -Source: Harlan Laboratories Ltd., FOIIinsdorf 

### I Switzerland 

## -AGe at study**in~iation:** 11 weeks

- Weightat study Initiation: 175.0 g - 197.7 g -Fasting period before study: lasted for approximately 15to16 hours (access towater was permitted)

- Housing: In groups 

### ofthree In Makrolon 

### type-4 cages**w~** wire mesh tops and standard 

### softwood bedding -Diet (e.g. ad libitum): Pelleted standard Provimi Kllba 3433 rat/mouse maintenance diet, ad libldum -Water (e.g. ed libitum):**Commun~** tap waterfrom FOIIInsdolfad libitum -Acclimation period:**2009~1-20** to**2009~1-26** (females, 2000 mglkg),**2009~1-22** to 2009-01-28 (females, 2000 mglkg)
ENVIRONMENTAL CONDITIONS

- Temperature (t:}: 22 

### ± 3 

## t: -Humidity(%): 30-70%

- I'Jr changes (per hr): 10
- 15 -Photoperiod (hrs dark 

### I hrs light): 12112 

## Administration 

## I exposure Route ofadministration oral:gavege 

### Vehicle water

### Details on oralexposure VEHICLE -Concentration in vehicle: 0.439 g substance/mL vehicle

- Amount ofvehicle 

### (W gavage): 10 mllkg

- Justification for choice ofvehicle: found to 

### be a suitable vehicle

- Purity: purified water
MAXIMUM DOSE VOLUME APPLIED: 10 mllkg bw
DOSAGE PREPARATION (ffunusual): prior to dosing
CLASS METHOD (ffapplicable) -Rationale forthe selection ofthe starting dose: nojustification (no data available) 

### Doses 2319 mg active ingredient/kg bwequals 4396 mg test item I kg bw (Lueco Sulfur Black 1: 52.75% w/w) 

### No. ofanimals persex perdose 3 females at 2319 mglkg bw 3 females at2319 mglkg bw 

### Control animals

no 

### Details on studydesign

- Duration ofobservation period following administration: 14 days -Frequency ofobservations and weighing: Viability I Mortality: Daily during the acclimatization period, during the first 30 minutesand at approximately 1, 2, 3 and 5 hours after administration on test day 1 and twice dallyduring days 2-15. Body Weights: 

### On test days 1 (priorto administration), 

### 8 and 15.

- Necropsy 

### of survivors performed: yes -Other examinations performed: clinical signs (Daily during the acclimatization period, during

### thefirst30 minutes and at approximately 1, 2, 3 and 5 hoursafteradministration on test day 1. depending on the occurrence of clinical signs oftoxicity. Once daily during days 2-15. 

### Statistics Nostatistical analysis was usad.

## Any other information on materials and methods Incl. tables
nment Femalee 

### 3



# Results and discussions Effect levels

No deaths occurred duringthe study. 

## Clinicalsigns Forthe first group of animals treated at 2319 mglkg, no clinical signs were observed in any 

### ofthe animals on test 1. 

### On test day 2, all threefemales were noted with black-stained feces and the 

### cage ofthe animals was changed tofadlitate further assessment. All animals were free ofany signs from test 

### day

### 3 until theend

### ofthe study on test day 15. The second group ofanimals treated at 2319 mglkg were 

### free ofclinical signs on test 

### day 1 as well. On test day 2 up to test day 5 all animals were observed with black-stained feces. On each 

### ofthese 4 days the animals were moved 

### to 

### new ceges. All animals were 

### free ofanyclinical signs from test day

### 6 until the end ofthe study. 

## Body-lght The 

### bodyweight ofthe animalswaswithinthe renge commonly recorded forthis strain and age. 

## Grosspathology Nomacroscopic findings were observed at necropsy. 

## Otherfindings None 

## Any other Information on results Incl. tables Table 2: Numberofanimals dead (and with evidenttoxicity] [and time range within which mortality occurred]

## Dose Mortality Time range ofdeaths lmalkabwl I#deadltotall (hours} 

## 2319 

## 0/3 n.a. 

## 2319 

## 013 n.a. 

# Applicant's summary and conclusion Interpretation 

## ofresults
notclassified 

## Criteria usedfor Interpretation ofresults EU 

## Conclusions Numberwith evident toxlclty(#ltotal) 

## 013 013

The median lethal dose LDSO (tamale rat)of Leuco Sulfur Black 1 after single oral administration

### to female rats. observed over a period of 14 days is greaterthan 2000 mglkg body weight 

## Executive summary 

## The acute toxicity of Leuco Sulfur Black 1 by oral application was investigated according to the OECD Guideline423 under GLP. Two groups, each ofthree female HanRcc:WIST (SPF) rats, were treated by oral gavage administration ata dosage of 2319 mg active ingredient 

# I kg body weight (corresponding to 4396 mg test item 

# I kg bodyweight). The test item was formulated in purified water according to the content ofthe active ingredient at a concentration of0.439 glmland administered at a dosing volume of 10 mllkg. No deaths occurred during the study. Exceptfor black stained feces from test day 2 up totest day 5 atthe latest, no clinical signs were noted in all animals during the observation time. The body weight ofthe animals was within the range commonly recorded forthis strain and age. No macroscopic findings were observed at necropsy. LD50 was greater than 2319 mglkg bw.

# 7.2.2 Acute toxicity: inhalation Endpointstudyrecord: waiving
UUID IUC5-fa34d11O-e335-4b41-be7b-26db45547cd2 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks uensenbe I Clartant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-llS-21 13:21:33 CEST



# Administrative Data Dats waiving otherjustification 

### Justification for In accordance with Regulation (EC) No 190712006 Annex VIII, column 2, an 

### acute inhalation toxicity study

### hasnotto be performed because exposure ofhumans data waiving 

### via inhalation 

### is not likelytaking into accountthe vapour pressure ofthe substance and the possibility 

### of exposure toaerosols, particles ordroplets ofan lnhalable size. The dermal route has

### been studied in addition to the oral route because skin contact In production and/or use 

### is likely.

# 7.2.3 Acute toxicity: dermal Endpointstudyrecord: Arcelin (2009) I acute dermal toxicity
UUID IUC5-986964c8-cbf4-4b59-8183-60f9b94f8559 Dossier UUID IUC5-292149d1-a6n-4feb-9692-1fad973281ea Author Date Remarks thr I SCC GmbH I Wendelsheim I Germany 2010-05-26 16:25:27 CEST



# Administrative Data Purpose 

### Illig Study 

### result

### type Relleblllty key study; robust study summary; usad for dasslfication; usad for MSDS expertmenlal resun Study period 2009-01-20

### tiH 2009-02-10 

### 1 (reliable without reslriction) R.tlon11le for ralleblllty Incl. deficlencleo Guideline-conform study under GLP without deviations 

# Data source Reference

Referance Author 

### Year



### type study report Aroelin 2009 G 

## Data access data submitter Is data owner 

## Data protection claimed
yes Title
Acute Dermal Toxidty Study In Rats



# Materials and methods Test

## type Bibliographic Testing leboralory 

### source Harlan 

### Laboratories Ltd.• FOIIinsdorf 

### I Switzerland 

### Report Owner Company study Report no. company 

### no. date C31054 Clariant 2009-09- 18standard 

### acute melhod 

## Umlttest yes 

## Test guideline

Qualifier 

### Guideline according

### to EU Method B.3 (AcuteToxicity (Dennal)) according

### to OECD Guideline 402 (Acute Dermal Toxicity) 

## GLP compliance
yes (incl. certificate) 

# Test materials Devlotlono
no no



## Identity oftest material same as forsubstance defined In section 1 (If

## not read-across)
yes 

## Test material Identity

Identifier Identity CAS number**66241-11~** ECnumber 266-273-5 

## Details ontest material

• Name oftest material : Lauco Sulfur Black 1 • Physical state: lhixotrophic, black, wet paste • ComposKion: Leuco Sulfur Black 1:52.75% (w/w) (calculated by amountof by-compounds) 2,4-Dinitrochlorobenzene: < 20 ppm 2,4-DinKrophenola: < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3 % (w/w) Water: 45.75% (w/w) by Karl..flschertitration • Lot/batch 

### No.: ESA0007595 ·Expiration date of

### the lot/batch:**2011~7-11** • StabiKy undertest conditions: Stable under storage conditions. • Storage condition oftest material: At room temperature (range of 20 

### ± 5 'C), light protected.

## Test animals Species rat 

## Strain Wistar 

## Sex male/female 

### Details on

## tast animals andenvironmental conditions TEST ANIMALS

- Source: Harlan Laboratories ltd., FOIIinsdorf 

### I Switzerland -Age 

### at study initiation: Males: 9 weeks; females: 11 weeks -Weight at study initiation: 182.9 g- 267.1 g

- Fasting period befora study: 

### no 

### data

- Housing: Individually In Makrolon type-3 cages wtth standard softwood bedding

- Diet (e.g. ad libttum): Pelleted standard Provimi Kliba 3433 rat/mouse maintanance diet, ad libidum -Water (e.g. ad libttum): Community tap water from FOIIinsdorf ed libitum -Acclimation 

### period: 2009.01-20 to 2009-Q1-26
ENVIRONMENTAL CONDITIONS

- Temperature ('C): 22 ±3 'C

- Humidtty (%): 30
- 70%

- Air 

### changes (per hr): 10 • 15



### - Photoperiod (hrs dark 

### I hrs light): 12 112 

## Administration 

# Iexposure Type of coverage semiocclusive 

## Vehicle water 

### Details on dermal

## exposure TEST SITE- Area ofexposure: bad<

- % coverage: 10 % of

### the total body surface -Type ofwrap

# nused: not specified
REMOVAL OF TEST SUBSTANCE -Washing (ifdone): flushed wi1h lukewarm tap waterand dapped offwith disposable papertowels.

- Time afterstertofexposure: 24 hrs
TEST MATERIAL

- Arnount(s) applied (volume or weight with**un~):** 2319mglkg body weight active ingredient (corresponding to 4396 mglkg of

### the test item; based on 

### the amount of 52.75% (wlw) Leuco Sulfur Black 1] or6 mlapplicationvolume/kg body weight -Concentration (ff solution): 0.73 g/ml

- Constant volume or concentration used: 

### yes -For solids, paste formed: 

### yes
VEHICLE

- Purity: purified water 

### Duration ofexposure 24 hrs 

### Doses Males and Females: 2319 mglkg bw active ingredient (corresponding to 4396 mglkg ofthe test**~em:** based on the amount of 52.75% (wlw) Leuco Sulfur Blad< 1)

Nola: Tha dose levels are In terms oftha active ingredient (Leuco Suller Bled< 1: 52.75 % wlw) in the test**~em.** 

### No. ofanimals per sex per dose
5 malas, 5 females 

### Control animals

no 

### Details on studydesign -Duration ofobservation period following edministralion: 14 days

- Frequencyofobservations and weighing: Viability 

### I**Mortal~:** Dailyduring the acclimatization period, during the first 

### 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on testday 1andtwice daHy during days 2-15.Body Weights: On test days1 (priortoadministration), 8 and 15.

- Necropsy

### of survivors performed: yes

- Otherexaminations performed: dinical signs (Daily during the acclimatization period, during the first 

### 30 minutes and at approximately 1, 

### 2, 3 and 5 hours after administration on test day 1. Once daHy during days 

### 2- 15.) Local Dermal Signs: Once daily during days 2 (following dressing removal) through day 15 

## Statistics
No statistical analysis was usad. 

## Any other Information on materials and methods Incl. tables Table 1: Animal assignment

Dose (mglkg Males Females bwl 

## 2319 mo/ko* 5 

# - 2319 mg/kg* 

# - 5 

## *2319 mg/kg active 1ngred1ent (corresponding to 4396 mg/kg ofthe test item; based on the amount of 52.75% 

## (w/w) Leuco Sulfur Black 1) 

# Results and discussions Effect levels

No daalhs

### occurred during the study. 

## Clinicalsigns Noclinical signs were observed during

### thecourse ofthe study. 

## BodyweightThe body weight of

### the animals was within the range oommonly recorded forthis strain and 

### age. 

### Gross pathology No maaosoopicfindings were recorded at necropsy. 

### Otherfindings Local Dermal Signs Due to moderate black staining ofthe

### traaled skin afterthe removal 

### of the application patch, the assessment oferythema wasfirst pravenled on test day 2 in all animals. From test day 3 totestday 7 atthe**lates~** a slight black stainingwas still present. Erythema was assessable, but not noted In all animals from test day 3

### to the end

### of the study. No oedema was observed throughout the whole oberservation period. SceHng was observed In one male on test day 4. 

### Any other Information on results Incl. tables Table 2: Numberofanimals dead [and with evidenttoxicity] [and time range wHhln which mortality occurred]



### Dose Cone. MortaiHy 

## (Idead/total) Time Number

## with evident range of toxlclty(lltotal) (mglkg In vehicle deaths



### bw) 

### (%1* Male Female Combined 

## Ihours

## I Male Female Combined Control Control 

### n.a. n.a. n.a
.. n.a
.. n.a
.. n.a
.. n.a
.. 2319* 0.73g/ml 0/5 

# - n.a. n.a. 0/5 

# - n.a. 2319* 0.73g/ml



# - 0/5 

### n.a. n.a. 

# - 0/5 

### n.a. n.a
.. notapplicable *2319 mg/kg active Ingredient (corresponding 

### to

### 4396 mgJkg ofthe test item; based on the amount 

### of 

### 52.75% (w/w) Leuco Sulfur Black 

### 1) 

## Table 3: Clinical/local signs (Males)

Dose Ani- Signs Testdavs mg/kg mal Sex 1 2 3 4 5 6 

### 7 8 

### 9 10 11 12 13 14 15 

### bw No. 0.5* 1* 2* 3* 5* 231ga 1 

## M No 

# .r .r .r .r .r .r .r .r .r .r .r .r .r .r .r .r .r .r .r clinical

### signs Scaling 

# .r Black 

# 2 1 1 1 1 1 stainina Erythema 

# § § § § § 

# # 0 0 0 0 0 0 0 0 0 0 0 0 0 Oedema 

# § § § § § 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Eschars 

# § § § § § 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 

## M No clinical 

# .r .r .r .r .r .r .r .r .r .r .r .r .r .r .r .r .r .r .r signs Black 

# 2 1 1 1 staining Erythema 

# § § § § § 

# # 0 0 0 0 0 0 0 0 0 0 0 0 0 Oedema 

# § § § § § 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Eschars 

# § § § § § 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 M No clinical 

# .r .r .r .r .r .r .r 

# .r .r .r .r 

# .r .r .r .r 

# .r .r .r 

# .r signs Black 

# 2 1 1 1 1 1 staining Erythema 

# § § § § § 

# # 0 0 0 0 0 0 0 0 0 0 0 0 0 Oedema 

# § § § § § 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Eschars 

# § § § § § 0 0 0 0 0 0 0 0 0 0 0 0 0 0 

# 4 M No 

# .r .r .r .r .r .r 

# .r .r .r .r .r .r .r .r .r .r .r .r .r clinical

## sians Black 

# 2 1 1 1 1 1 stainina Erythema 

# § § § § § 

# # 0 0 0 0 0 0 Oedema 

# § § § § § 0 0 0 0 0 0 0 Eschars 

# § § § § § 0 0 0 0 0 0 0 

# 5 M No clinical 

# .r .r .r .r .r .r .r .r .r .r .r .r signs Black 

# 2 1 1 1 1 1 staining Erythema 

# § § § § § 

# # 0 0 0 0 0 0 Oedema 

# § § § § § 0 0 0 0 0 0 0 Eschars 

# § § § § § 0 0 0 0 0 0 0 Key: 0 nofindings, 1 slight, 2 moderate,/"noted 

## * Examinations were performed within the first 30 minutes and 1, 2, 3 and 5 hours aftertreatment. §Assessment ofdermal findings was not possible underneath the application patch #Assessment oferythema was not possible due to black staining ofthe skin 

# 0 0 0 

# .r

0 0 0

82319 mglkg active Ingredient (corresponding 

### to 4396 mglkg 

### ofthe test Kem; 

### baaed on the amount

### of 52.75% (wlw) Leuco SuWur Black 1) no dlnlcal signs were evident in any animal during the acdimatization period. 

# Overall remarks, attachments Overall remarks 

## Table 4: Clinical/local signs (Females) 

# 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 

# .r .r .r .r .r .r

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

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

### bw 

### No. 0.5* 1* 2* 3* 5* 231ga 

## 6 F No clinical 

# .r .r .r .r 

# .r .r .r .r .r .r .r .r .r .r .r .r .r .r 

# .r signs Black 

# 2 1 1 1 1 1 staining Erythema 

# § § § § § 

# # 0 0 0 0 0 0 0 0 0 0 0 0 0 Oedema 

# § § § § § 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Eschars 

# 0 § § § § 0 0 0 0 0 0 0 0 0 0 0 0 0 0 7 F No clinical 

# .r .r .r .r .r 

# .r .r .r .r .r .r .r .r .r .r .r .r 

# .r .r signs Black 

# 2 1 1 1 staining Erythema 

# § § § § § 

# # 0 0 0 0 0 0 0 0 0 0 0 0 0 Oedema 

# § § § § § 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Eschars 

# § § § § § 0 0 0 0 0 0 0 0 0 0 0 0 0 0 No 

## 8 F clinical 

# .r .r .r 

# .r .r .r 

# .r .r 

# .r .r .r .r 

# .r .r .r .r .r .r 

# .r signs Black 

# 2 1 1 1 staining Erythema 

# § § § § § 

# # 0 0 0 0 0 0 0 0 0 0 0 0 0 Oedema 

# § § § § § 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Eschars 

# § § § § § 0 0 0 0 0 0 0 0 0 0 0 0 0 0 9 F 

# .r .r .r .r 

# .r .r .r .r .r .r .r .r .r 

# .r .r .r .r .r .r No clinical

## sians Black 

# 2 1 1 1 1 1 staining Erythema 

# § § § § § 

# # 0 0 0 0 0 0 Oedema 

# § § § § § 0 0 0 0 0 0 0 Eschars 

# § § § § § 0 0 0 0 0 0 0 10 F No clinical 

# .r .r .r .r .r .r .r .r .r .r .r .r signs Black 

# 2 1 1 1 1 1 staining Erythema 

# § § § § § 

# # 0 0 0 0 0 0 Oedema 

# § § § § § 0 0 0 0 0 0 0 Eschars 

# § § § § § 0 0 0 0 0 0 0 Key: 0 no findings, 1 slight, 2 moderate, noted • Examinations were performed within the first 30 minutes and 1, 2, 3 and 5 hours after treatment. §Assessment ofdermal findings was not possible underneath the application patch ..Assessment oferythema was not possible due to black staining ofthe skin 

# 0 0 0 

# .r

0 0 0

82319 mglkg active ingr>ldient (corresponding to 4396 mglkg ofthe

### test ttem; based on the amount of 52.75% (wlw) Leuco Sulfur Black 1) 

### no clintcal signs were evident in any animal during the acdlmatization period. 

# Applicant's summary and conclusion Interpretation ofresults notdasslfled 

## Criteria usedfor Interpretation ofresults 

## EU Conclusions 

# 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 

# .r .r .r .r .r .r

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

### The median lethal 

### dose L050 (rat) ofLeuco Sulfur Black 1 aftersingle dermal adminislration to rats ofboth sexes, observed overa period of 14 days is greaterthan 2000 mglkg body weight 

## Executive summary 

## The acute toxicity by dermal application of LEuco Sulfur Black 1. was investigated according to OECD Guideline 402 under GLP. Five male and five female HanRcc:WIST (SPF) rats were treated with Leuco Sulfur Black 1 at 2319 mg active ingredienUkg (corresponding to 4396 mg test item/kg) by dermal application. The test item was formulated in purified water according to the content ofthe active ingredient at a concentration of0.73 g/mL and administered at a volume dosage of 6 mL/kg. The application period was 24 hours. No deaths occurred during the study. No dinical signs were observed during the course ofthe study. Due to moderate black staining ofthe treated skin afterthe removal ofthe application patch, the assessment oferythema was first prevented on testday2 in all animals. From test day3 to test day 7 at the latest, a slight black staining was still present. Erythema was assessable, but not noted in all animals from test day 3 to the end ofthe study. No oedema was observed throughout the whole oberservation period. Scaling was observed in one male on test day4. The body weight ofthe animals was within the range commonly recorded forthis strain and age. No macroscopic findings were observed at necropsy.

# 7.3 Irritation 

# I corrosion Endpointsummary: Irritation 

# I corrosion UUID IUC5-50fa682b-20ac-4b90-87a3-d06b3088e470 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author thr I SCC GmbH I Wendelsheim I Germany Date 2010-05-26 16:52:29 CEST Remarks 

# Administrative Data Short description of key information
Leuco Sulfur Black 1 ceused none to very mild signs 

### of skin oreye irritation In the rat or rabbh, respectively, without any relevance for dasslficetion and labeHing. 

# Key value for chemical safety assessment Skin Irritation 

## I corrosion
not Irritating 

## Eye Irritation not irritating 

## Respiratory Irritation
not irritating 

# Discussion The skin irritation potential of Leuco Sulphur Black 1was assessed in the acute dermal toxicity study according to OECD Guideline 402 under GLP.

## Five male and five female rats were treated at2319 mg active ingredient/kg (corresponding to 4396 mg test item 

# I kg bw) by dermal application. The test item was formulated in purified water and administered at a volume dosage of6 mUkg. The application period was 24 hours. No deaths occurred during the study. No clinical signs were observed during the course ofthe study. Due to moderate black staining ofthe treated skin afterthe removal ofthe application patch, the assessmentoferythema was first prevented on test day2 in all animals. From test day 3 to test day 7 atthe latest, a slight black staining was still present. Erythema was assessable, but not noted in all animals from test day 3 to the end ofthe study. No oedema was observed throughout the whole observation period. Scaling was observed in one male on test day 4. The primary eye irritation potential of Leuco Sulphur Black 1 was investigated according to OECD test guideline No. 405 and GLP. The test item was applied by instillation of0.1 g into the lefteye ofeach ofthree young adult NewZealand White rabbits. Scoring ofirritation effectswas performed approximately 1, 24, 48 and 72 hours aftertest item instillation. The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation)for each animal for corneal opacity, iris, redness and chemosis ofthe conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for all three animals for the conjunctival reddening and swelling were 0.00 and 0.00, respectively. The instillation of Leuco Sulfur Black 1 into the eye resulted in mild, early-onset and transient ocularchanges, such as reddening and swelling ofthe conjunctivae, redness ofthe sclera as well as discharge. These effects were reversible and were no longer evident24 hours aftertreatment in all animals. Black test item remnants were observed in the eye or conjunctival sac ofall three animals at the 1-hour reading. No abnormal findings were observed in the cornea or iris ofany animal at any ofthe examinations. Thus, the test item did not induce significant or irreversible damageto the rabbit eye. Based on the lacking skin and eye irritation potential, the high boiling point and consequently very lowvapro pressure, a respiratory irritation can be excluded. 

# Justification for classification or non-classification Based on the available data Leuco Sulfur Black 1 is not subjectto classification and labelling according to Directive 67/548/EEC and Regulation 1272/2008/EC.

# 7.3.1 Skin irritation 

# I corrosion Endpointstudyrecord: waiving_Skin irritation I corrosion
UUID IUC5-49607fb8-499b-48f2-94ed-debdl2ee647b Dossier UUID IUC5-292149d1-a6IT-4feb-9692-1fad973281ea Author Date Remarks uensenba I Clariant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-QS-21 13:27:28 CEST



# Administrative Data Data w•lvlng otherjustifocalion 

### Justlllcallonfor data In accordance with Regulation (EC) No 190712006 Annex VIII, column 2, a skin irritation 

### study does not

### need to 

### be conducted because 

### the acule toxicity study 

### w.lvtng by

### the dermal route did not Indicate skin irritation ata dose level even higherthan 

### the limit

### dose of 2 000 mglkg bw.

# 7.3.2 Eye irritation Endpointstudyrecord: Arcelin (2009) Eye irritation (in vivo)
UUID IUCS-341a0a6c-347c-4c5a-95e7-ba9a6fa01e2c Dossier UUID IUC5-292149d1-a6n-4feb-9692·1fad973281ea Author Date Remarks lhr I SCC GmbH IWendelsheim I Germany 2010-05-26 16:27:16 CEST



# Administrative Data key sludy Purposeft.lg Study reault

### type Reliability experimental resun Study period 2009-02-18 tin 2009-03-02 1 (reUeble wilhout restriction) Rationalefor reliability Incl. deficiencies Guideline conform study under GLP wilhout deviations 

# Data source Reference

Reference Author

type Year 

### Title sludy report Arcelin 2009 Primary Eye Irritation Sludy G 

### Data access data submitter is data owner 

### Data protection claimed
yes 

### in Rabbits



# Materials and methods Type ofmethod Bibliographic Testing laboratory 

### source Hanan Laboralories Ltd. FOIIinsdorf (Swilzeriand) Report Owner Company study Report no. company 

### no. data C31065 Clarianl 2009-08- 20

### in vivo 

### Test guideline

Qualifier Guideline 

### Deviationa according lo EU Method B.5 (Acule Toxicily: Eye IITitation I Corrosion) 

### no according lo OECD Guideline 405 (Acule Eye IITitation IConosion) 

### no GLP compliance
yes (ind. certificate) 

### Identity oftest material same as for substance defined in section 1 

### (If not read-ecross)
yes 

## Test materials Test material Identity

Identifier Identity CAS number 66241-11-0 EC number 266-273-5 

### Details on test material

- Name oflest material : Leuco Sulfur Black 1 -Physical stale: lhixotrophlc, black, wet paste

- ComposHion: Leuco Sulfur Black 1:52.75% (w/w) (calculated by amount of by-rompounds) 2,4-Dinitrochlorobenzene: < 20 ppm 2.4-Dinitrophenole: < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3 % (w/w} Water: 45.75% (w/w) by Karl-Fischer titration -Lot/batch 

### No.: ESA0007595 -Expiration dateof

### the lot/batch: 2011-07-11

- StebiiHy undertest conditions: Stable under storage condHions. -Storage condition oftest material: At room temperature (range of20 ± 5 'C), light protected. 

## Test animals

### Species rabbit 

### Strain New Zealand Whhe 

### Details on 

### test animals andenvironmentalconditions TEST ANIMALS: Young AduH Naw Zaaland WhKe RabbK, SPF -Source:**Ha~an** Laboratories B.V. 5960AD Horst I The**Nethe~ands** -Age at study initiation: 15 weeks (male), 15- 16 weaks (females) -Weight at study initiation: 2608 g (male), 2729-2892 g (females)

- Housing: Individually In stainless steel cages equipped with feed hoppars and drinking water bowls. Wood blocks and haysticks for gnawing. -Diet (e.g. ad libKum): Pelleted standard Provimi Kllba 3418 rabbit maintenance diet ad libKum -Water (e.g. ad libitum): Communitytapwaterfrom FOIIinsdorf ed libitum

- Acclimation :Under laboratory oonditions after health examination. Only animals without any visual signs of illness were used 

### for the study -Randomization: Selected by hand at time ofdelivery. No computer generatad randomization program. ENVIRONMENTAL CONDITIONS -Temperature 

### (q: 17-23 

### 'C -Humidity(%): between 30-70% (values above 70% during daaning process possible) -Air changes (per hr): Air-mnditioned 

### with 10-15airchanges per hour -Photoperiod (hrs dart< I hrs**~ght):** automaticaUy controlled light cyde of 12 hours lightand 12 hours dart<, music during the daytime light period.



# Test system Vehicle unchanged (no vehide) 

### Amount/concentration applied TEST MATERIAL

- Amount(s) applied (volume orweightwith**un~):** 0.1 

### gl animal (undiluted)VEHICLE: none 

## Duration oftreatment I 

## exposure The eyes ofall animals were rinsed with sterile physiological saline (B. Braun, Emmenbrllci<e. Swltzet1and; Lot-No. 7443A191. expiry date 30-Sep-2009) afterthe 1-hourobservation because black test Item remnants (partides) were observed atthis observation time point. 

## Observation period
72h 

## Numberof

## animals NumberofAnimals

### per Test: 3 (Animals ofboth sexeswere used) 

## Control animals other: lefteye 

### ofeach animal treated, right eye served for reference control 

## Details on studydesign REMOVAL OF TEST SUBSTANCE The eyes

### ofall animals were rinsed with sterile physiological saline (B. Braun, Emmenbroci<e, Swltzet1and; Lot-No. 7443A191. expiry date 3()-Sep-2009) afterthe 1-hour observation because blaci< test**~em** remnants (particles) were observed atthis observation time point.
SCORING SYSTEM: The eye reectlons were assessed according to the numerical scoring system listed In the Commission Regulation (EC)

### No. 440/2008, B.5, at approximately 1, 24, 48 and 72 hours after administration. Sderel reddening and oculardischarge were also assessed. TOOL USEDTO ASSESS SCORE: Eye examinations were madewith a Varte Cliptrix diagnostic-jamp 

## Any other Information on materials and methods Incl. tables 

## The pH ofthe test item was measured before the start ofthe study in a 1% dilution in purified water and was found to be 6. The eyes ofthe animals were examined one day prior to test item administration. Animals with overt signs ofocular injury or irritation which may have interfered with the interpretation ofthe results were not used in the test. On the day oftreatment, 0.1 g of Leuco Sulfur Black 1 was placed in the conjunctival sac ofthe left eye ofeach animal after gently pulling the lower lid away from the eyeball. The lids were then gently held togetherfor aboutone second to

## prevent loss oftest item. The right eye remained untreated and served as the reference control. 

# As an irritant effect seemed to be unlikely, butcould not be fully excluded, a single animal (one female) was treated first. 

# As neither a corrosive effect nor a severe irritant effect was observed afterthe 1- and 24-hourexaminations, the testwas completed using the two remaining animals The eyes ofall animals were rinsed with strile physiological saline (B. Braun, EmmenbrOcke, Switzerland; Lot-No. 7443A191, expiry date 30-Sep-2009) afterthe 1-hourobservation because black test item remnants (particles) were observed atthis observation time point. Observations Viability 

# I Mortality: Daily from acclimatization ofthe animals to the termination oftest. Clinical Signs (systemic): Dailyfrom acclimatization ofthe animals to the termination oftest. Body Weights: At start of acclimatization, on the day ofapplication and at termination ofobservation. Necropsy No necropsy was performed on the animals sacrificed at termination ofobservation. The animals were killed by intravenous injection of pentobarbitone into the earvein at a dose ofat least 1 mUkg body weight (162 mg sodium pentobarbitone/kg body weight) and discarded. 

# Results and discussions Overall Irritation 

## Icorrosion results

lrrlhlllon p•rameter overall irritation scoreoverall irritation seo<e mean 48hours overall irritation seo<e mean 72 hours 

## Irritant/corrosive response data

no irritation ellects obseJVed 

## Othereffects Viability I Mortality: 

### No mortality occurred. 

### 0 0 0 0

Clinical 

### Signs: No clinical signs were obseiVed during the course 

### ofthe study. BodyWeights: The body weights 

### ofall rabbits were considered to

### be within the nonnal range ofvariability. Pathology, Macroscopic Findings: No necropsy was performed attha end ofthe study. 

## Any other information on results incl. tables 

## The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis ofthe conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for all three animals for the conjunctival reddening and swelling were 0.00 and 0.00, respectively. No abnormal findings were observed in the cornea or iris of any animal at any ofthe measurement intervals. Slight conjunctival reddening and swelling, slight scleral reddening, as well as slight ocular discharge was noted in all three animals atthe 1-hour reading. No abnormal findings were observed in the treated eye ofany animal 72 hours after treatment, the end ofthe observation period for all animals. Coloration: No staining produced bythe test item ofthe treated eye was observed. Test Item Remnants: Black test item remnants were evident in the eye or conjunctival sac of all three animals at the 1- hour reading.

## Corrosion: No corrosion ofthe cornea was observed at any ofthe reading times. 

# E I

. S :ye mtat1on cores

Animal 

## Sex Evaluation Corneal Area ofCorneal 

### Iris Conjunctivae Number Interval* Ooacitv Ooacitv Redness Chemosis Sclera



# 19 M 

# 0 0 0 1 1 1 20 

# F 1 hour 

# 0 0 0 1 1 1 21 

# F 0 0 0 1 1 1 19 M 

# 0 0 0 0 0 0 20 

# F 24 hours 

# 0 0 0 0 0 0 21 

# F 0 0 0 0 0 0 19 M 

# 0 0 0 0 0 0 20 

# F 48 hours 

# 0 0 0 0 0 0 21 

# F 0 0 0 0 0 0 19 M 

# 0 0 0 0 0 0 20 

# F 72 hours 

# 0 0 0 0 0 0 21 

# F 0 0 0 0 0 0



# .ExaminatiOns were performed at the specified times after InstillatiOn ofthe test item

. 

# EY'e Irritation Scores
- Mean Values after24 48 and 72 Hours Animal Sex 

# Corneal N Iris N Conjunctivae Number Opacity Redness 

# N Chemosis 

# N 19 M 

## 0.00 

## 3 0.00 

## 3 0.00 

## 3 0.00 

## 3 20 

# F 0.00 

## 3 0.00 

## 3 0.00 

## 3 0.00 

## 3 21 

# F 0.00 

## 3 0.00 

## 3 0.00 

## 3 0.00 

## 3

Evaluated Intervals Corneal Opacity Con unctivae

## 24hours 48 hours Not Irritating 72 hours 

# Applicant's summary and conclusion Interpretation ofresults
not Irritating 

## Criteria used for Interpretation ofresults EU 

## Conclusions Redness 

## Chemosis
Not Irritating Not Irritating Not Irritating

Based upon the referred classification (Commission Directive 2001159/EC ofAugust 06, 2001), Leuco Sulfur Black 1 is considered

### to be 'not irritating'

### to the rabbit eye. 

## Executive summary 

## The primary eye irritation potential of Leuco Sulfur Black 1 was investigated according to OECD test guideline No. 405 and Commission Regulation (EC) No. 440/2008, B.5. The test item was applied by instillation of0.1 g into the left eye ofeach ofthree young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours aftertest item instillation. The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis ofthe conjunctivae, separately. The individual mean scores for corneal opacity and iris were 0.00 for all three animals. The individual mean scores for all three animals for the conjunctival reddening and swelling were 0.00 and 0.00, respectively. The instillation of Leuco Sulfur Black 1 into the eye resulted in mild, early-onset and transient ocular changes, such as reddening and swelling ofthe conjunctivae, redness ofthe sclera as well as discharge. These effects were reversible and were no longer evident 24 hours aftertreatment in all animals. Blacktest item remnants were observed in the eye or conjunctival sac ofall three animals at the 1-hour reading. No abnormal findings were observed in the cornea or iris ofany

## animal at any ofthe examinations. No corrosion was observed at any ofthe measuring intervals. No clinical signs were observed. Thus, the test item did not induce significant or irreversible damageto the rabbit eye. Based upon the referred classification criteria (Commission Directive 2001/59/EC ofAugust 06, 2001 ), Leuco Sulfur Black 1 is considered to be "not irritating" to the rabbit eye.

# 7.4 Sensitisation Endpointsummary: Sensitisation
UUID IUC5-5039afad-2a7e-4143-1!483-350bl9bc60cd Dossier UUID IUC5-292149d1-a6n-4feb-9692-1fad973281ea Author thr I SCC GmbH I Wendelsheim I Germany Date 2010-05-26 16:47:38 CEST Remarks 

# Administrative Data Skin sensitisation Short description ofkey information
Leuco Sulfur Black 1did

### not cause skin sensitisetion 

### in the LLNA In mice. 

# Key value for chemical safety assessment Skin sensitisation
not sensitising 

# Discussion The skin sensitisation potential of Leuco Sulfur Black 1 was investigated in the LLNA according to OECD test guideline No. 429 and GLP. Three groups each offour female mice were treated daily with the test item at concentrations of5, 10and 25% (w/w) in dimethylformamide bytopical application to the dorsum ofeach ear lobe (left and right) for three consecutive days. A control group offour mice was treated with the vehicle (dimethylformamide) only. Five days afterthe first topical application the mice were injected 3H-methyl thymidine. Approximately five hours later, the mice were sacrificed, the draining auricular lymph nodes excised and pooled per group. Single cell suspensions of lymph node cells were prepared from pooled lymph nodes and the proliferative capacity of pooled lymph node cells was determined by the incorporation of

## 3H-methyl thymidine measured in a scintillation counter. All treated animals survived the scheduled study period and no signs oftoxicitywere observed. A test item is regarded as a sensitizer in the LLNA ifthe exposure to one or more test concentration resulted in 3-fold or greater increase in incorporation of3HTdR compared with concurrent controls, as indicated bythe Stimulation Index (S.I.). The estimated concentration oftest item required to produce a S.l. of3 is referred to asthe EC3 value. In this study Stimulation Indices of 1.42, 1.12 and 1.61 were determined with the test item at concentrations of5, 10 and 25% in dimethylforrnamide. The EC3 value could not be calculated, since none ofthe tested concentrations induced a S.l. greaterthan 3. Thus, the test item was not a skin sensitizer. 

# Respiratory sensitisation Discussion At presentthere is no accepted experimental test system available to evaluate this endpoint Based on the substance properties (mean MW >8000 Da and very poor solubility in water), the fact that no toxicity was seen in any ofthe available studies and thatthere are no human data available indicative ofa respiratory sensitisation potential, it is concluded that Leuco Sulfur Black 1 does not raise any concern regarding respiratory sensitisation. 

# Justification for classification or non-classification Based on the available data Leuco Sulfur Black 1 is not subjectto classification and labelling according to Directive 67/548/EECand Regulation 1272/2008/EC.

# 7.4.1 Skin sensitisation Endpointstudyrecord: LLNA
UUID IUC5-e412fb21 -5b5c-4bbe-9b09-b92dclb419a6 Dossier UUID IUCS-292149d1 -a677-4feb-9692-1lad973281 ea Author Dale Remarks lhr I SCC GmbH I Wendelsheim I Germany 2010-{)5-26 16:33:34 CEST


# Administrative Data Purpose 

### flag key study; robust study summary; used for dassification; used for MSDS Study result

### type expertrnental resuH Reliability 1 (reliable without restriction) 

# Data source Reference

Reference 

### Author Year



### type study report Vogel 2009 

### J 

## Data access data submitter is data owner 

## Data protection claimed
yes 

### Tille
Local Lymph 

### Node Assay (LLNA) in Mice



# Materials and methods Type ofmethod tnvivo Bibliographic Testing 18boralory 

### source Hanan CCR GmbH. Rossdorf (Grmany) Report no. 

### Owner Company

### study Report company 

### no. dele 1234901 Clariant 2009-{)9. (C31100) 

### 17

### Type ofstudy Mouse locallymphnode assay (LLNA) 

### Test guideline Guideline OECD Guideline 429 (Sidn Sensttisation: Local Lymph Node Assay)

yes (ind. cet11ficate) 

## Test materials Identity oftest material same as for substance defined in section 1 

## (if not read-across) yes 

### Details on test material Identification: Leuco Sulfur Black 1 CAS 

### No.: 66241-11-0 Description: Thlxolhrophic, black, wet paste 

### Batch No.: ESA0007595 Composition: Leuco Sulfur Black 

### 1: 52.75% (calculated by amount ofby-compounds) 2,4-Dinttrochlorobenzene: 

### < 20 ppm 2,4-Dinttrophenole: < 20 ppm Sulfate: 970 

### ppm Chloride: 750 ppm Sodium: 1.3 

### % (wlw) Water: 

### 45.75 

### %(wlw) by**Ka~-Fischar** titration Stebittty of

### the test rtern Stable under s1orage condttions Stability in solvent: 

### > 4 h in water Storage: 

### />J room temperature (range 20 

### ± 5 

### 'C), light protected Expira1ion Date: July 11, 2011 

## Test animals Species
mouse 

### StrainCBA 

### Sex female 

### Details on 

### test animals andenvironmentalconditions TEST ANIMALS • Source: Harlan Laboratories B.V., Postbus 6174, 5960 AD Horst 

### I The Netherlands •Age atstudy initiation: 8 • 12 weeks • Weight at study initiation: 15 • 25 g • Housing:single caging 

### • Diet (e.g. ad libi1um): pelleted standard dial (Harlan Laboratories GmbH, 33178 Borchen), ad Hbidum • Water (e.g. 

### ad libitum): tap water, (Gemeindewerl<e, 64380 Rossdorf), ad libitum • Acctlmatlon period: At least 5 days prior tothe start 

### ofdosing undertest condHions after heaHh examination. Only animalswithout anyvisible signs 

### of Illness will 

### be used for

### the study ENVIRONMENTAL CONDITIONS • Temperature ('C): 22 + 2'C • HumidHy (%): 45-65% • Photoperiod (hrs dar!< I hrs light): artificial light 6:00 a.m. • 6:00 p.m. 

## Test system LLNA Vehicle dirnethytformamida 

### Concentration 5, 10, and 25% 

### No. ofanimals per dose 4 

### Details on studydesign (LLNA) To determine the highest non-irritant test concentration. a pre-testwas performed in two animals. Two mice were treated with concentrations 

### of 10 and 25% each on three consecutive days. In

### the pr&-tast clinical signs were recorded within 1 hourand 24 ± 5 hours after each application as well as on day 7. At the tasted concentrations lheanimats did

### not show any signs of 

### irritatlon or systemictoxicHy. The 1ast Hem in the main studywas assayed at 5, 10 and 25%. Thetop dose is 

### the highest technically achievable concentration whilst avoiding systemictoxicity and excessive local imtatlon. No severe imtant effects were tolerated choosing 

### the test concentrations.

## Positive controlsubstance(s) hexyl dnnamic aldehyde (CAS No**101~)** 

### (In acetone:olive oil (4+1) using CBAICeOtaHsd mice in June 2009) 

## Statistics The mean values and standard deviations were calculated in the body weight tables. 

## Any other lnfonnatlon on materials and methods Incl. tables 

# Interpretation of Raw Data The proliferative response of lymph node cells 

### is expressed as the number of radioactive disintegrations per minute per lymph 

### node (DPM/node) and as the ratio oA-iTdR incorporated into lymph 

### node cells oftest lymph nodes relative to that recorded for oontrollymph nodes (Stimulation Index; S.l.). Befora 

### DPM/node values were determined, mean scinUIIetionbeckground DPM was subtracted from 

### test and control raw data. Atest Item is regarded as a sensltiser In 

### the LLNA 

### ffthe following criteria are fuffllted: First, that exposure to at least one concentration ofthe test Item resuned In an incorporation oFHTdR at least 3-fotd orgreaterthan that recorded in oontrol mice, as indicated by

### the stimulation Index. 

### Second,that 

### thedata are compatible with a oonventional dose response, anhough allowance must be made (especially at high topical concantrations) for enher local toxicity or Immunological suppression. 

# Observations In**adld~ion** 

### to the sensHising reactions the following observations and data were recorded during the test end observation period: MortaiHy I ViebiiHy:once daily (week day) from experimental start to necropsy. Body weights:priorto tha first application and priortotreatment wlthltiTdR.
CHnical signs (local/systemic): 

## In the pre-test clinical signs were recorded within 1 hour and 24±4hours after each application as well as on day 7. In the main experiment clinical signs were recorded within 1 hour and 24±4 hours afterthe first and second application (days 1 and 2) as well as on the day of preparation (day 6).EspeciallylhetreatrnentsHeswereobservedcaretully. 

# Results and discussion Positive controlresults Experiment performend in January 2009, 5, 10, and 25% alpha-Hexylcinnamaldehyde yielded a S.l. of 1.49, 4.17, and 4.90, respectively. The EC3 value calculated was 7.8% 

## LLNA Disintegrations per minute (DPM)

see table below

## Stimulation Index
1.4 (at 5 %). 1.1 (at 10 %), 1.6 (at 25 %) 

## Any other Information on results Incl. tables 

## Test item Calculation Result concentration Group Measurement number of DPM per 

## % (w/w) DPM DPM-BGa) lymph nodes lymph nodeb) 

# S.l.



# -- BGI 25 

# -- --- 

# -- -

-- BGII 39 

# -- --- 

# -- -- 0 1 2735 2703 8 337.9 

# 1.00 5 2 3869 3837 8 479.6 

# 1.42 10 3 3067 3035 8 379.4 

# 1.12 25 4 4386 4354 8 544.3 

# 1.61



# - 

# - - BG = Background (1 ml 5% trichloroacetic acid) In duplicate 1-Control Group 2-4-Test Group S.I.-Stlmulation Index •l=The mean value was taken from thefigures BG I and BG ubl=Since 

### the lymph nodes ofthe animals of a doss groupwere pooled, DPM/node was determined by dividing the measuredvalue by

### the number

### of lymph nodes pooled 

## The EC3 value can not be calculated, since all S.l.'s are below 3. 

# Viability 

# I Mortality No deaths occurred during the study period. 

# Clinical Signs No symptoms of local toxicity atthe ears ofthe animals and no systemic findings were observed during the study period. 

# Body Weights

# The body weight ofthe animals, recordedprior to the first application and priorto treatment with3

# HTdR, was within the range commonly recorded for animals ofthis strain and age. 

# Applicant's summary and conclusion Interpretation of

## results
not sensitising 

## Conclusions The test item Leuco Sulfur Black 1

## was not a skin sensiliser under

### the described conditions. 

## Executive summary The present study (Vogel2009) analyses 

### the sensilislng potential ofleuco Sulfur Black 1
Three groups each offourfemale mice ware treated daily 

### with the lest Hem atconcentrations of 5, 10and 25% (w/w) In dimethylformamlde by topical applicetion totha dorsum ofeach ear lobe (left and right) 

### lorthree consecutive days. A control groupoffour mice

## was treated with the vehicle (dimathylformamlde) only. Five days after

### the first lopical application the mioa were injected intravenously inlo alallvein with redio-labetled thymidine eH-methyi thymidine). Approximately five hours after intravenous injection, 

### the mice were sacrificed, the draining auricular lymph nodes excised and pooled pergroup. Single cell suspensions of lymph node oalls were prepared from pooled lymph nodes. which were subsequenUy washed and incubated with trichloroacetic acid overnight. The proliferative capacity ofpooled lymph node oalls was determined by the incorporation offi-methyl thymidine measured in absclntHiation counter. All treated animalssurvived 

### the scheduled studyperiod and no signs oftoxicity were observed. A test Hem is

### regarded as a sensHiser in the LLNA ifthe exposure 1o one ormore test concentration resuHed in 3-fold orgreater increase in incorporation ofiHTdR compared 

### with concurrent controls, as indiceled bythe Stimulation Index (S.I.). The estimated concentration oftest item required

### to produce a S.l. of3 is refemed lo as

### the EC3 value. In this study Stimulation Indices of 1.42, 1.12 and 1.61 were delermlned 

### with the test item at concentrations of5, 10and 25% in dlmethylformamide. The EC3 value could 

### not be calculated, since none ofthatested concentrations induced a S.l. greater than 3. The test Hem Leuco Sulfur Black 1 was not a skin sensiliserunderthe described condHions.

# 7.5 Repeated dosetoxicity Endpointsummary: Repeateddose toxicity UUID IUC5-074041c6-f34c-4c11-9b71-6382281125b6 Dossier UUID IUC5-292149d1-a6n -4feb-9692-11ad973281ea Author thr I SCC GmbH I Wendelsheim I Germany Date 2010-05-26 17:13:00 CEST Remarks 

# Administrative Data Short description of key information
Leuco Sulfur Black 1did not cause any toxic affects in 

### the repeated 

### dosetoxicity study with reproduction/developmental toxicity screening in rats (OECD 422). 

### The limK

### dose of 1000 rng/kg bwld represents the NOAEL. 

# Key value for chemical safety assessment Repeated dose toxicity: oral
Effect NOAEL 1000 mg/kg bwlday level Tnt subacute 

### type Species rat 

# Discussion In a repeated dose toxicity study according to OECD guideline 422 and GLP Leuco Sulfur Black 1 was administered by gavage to groups of 10 rats per sex and dose at 0, 100, 300 and 1000 mg/kg bw/d. Male rats were treated for 29 days, female rats for 14 days prior to pairing, though the pairing and gestation periods until the F1 generation had reached day4

## postpartum. The test item was administered in water as vehicle at a dose volume of 10 mllkg bw/d. Control animals were doses with the vehicle alone. Observation and examinations consisted of mortality, clinical signs, detailed clinical observations, food consumption, body weight, functional observation battery, clinical biochemistry, haematology, urinalysis, macroscopical examination at necropsy, organ weights and histopathological examination oforgans. The only effect seen was a dark discolouration of faeces in all high dose animals starting on day3 ofthe study and lasting until termination. Based on this finding a NOAEL of 1000 mg/kg bw/d was derived. The discolouration offaeces was the only effect observed in this study and also in the acute oral toxicity study. Organsor urine were not discoloured and signs ofsubstance deposits were notfound in histopathological analysis ofthe organs. This indicates that Leuco Sulfur Black 1 is directly excreted and that absorption in the gastrointestinal tract does not occur following oral administration. The mean molecularweight and number-average molecularweight (Mn) of Leuco Sulfur Black 1 is in the range of8000- 9000 Da and about 3000 Da, respectively.lt is very poorly oreven notsoluble in common polar and non-polarsolvents such as water (solubility of62.4 mg/L), DMSO, methanol, DMF, THF, chloroform and notsoluble in acid likes sulfuric acid and trifluoroacetic acid. The vapour pressure ofthe pure substance, calculated from the boiling point of>400 

# 'C, is 1.8x 1 OE-5 hPa at25 

# 'C. These substance properties supported by the results oforal toxicity studies demonstrate thatthe absorption of Leuco Sulfur Black 1 via the oral, dermal or inhalation route is negligible and, consequently, that systemic bioavailability is also negligible. Based on the substance properties and the fact that no toxicity was seen in any ofthe available toxicity studies it is concluded that exposure to Leuco Sulfur Black 1 does not raise any health concern. In line with Regulation (EC) No 1907/2006Annex XI and taking into accountthe REACH paradigm that newtests on vertebrates shall only be conducted or proposed as a last resort when all other data sources have been exhausted, further animals studies to examine the repeated dose toxicity of Leuco Sulfur Black 1 are not necessary and should not be performed. 

# Justification for classification or non-classification Based on the available data Leuco Sulfur Black 1 is not subjectto classification and labelling according to Directive 67/548/EECand Regulation 1272/2008/EC.

# 7.5.1 Repeated dose toxicity: oral Endpointstudyrecord: Repeateddose toxicity: ora/.001 OECD 422 Leuco SulfurBlack 1 C31087
UUID IUC5-d08b5!2b-3ffd-40b5-8bde-37609a519786 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks thr I SCC GmbH I Wendelsheim I Germany 2010-05-26 16:34:35 CEST



# Administrative Data Purpoae flag Study result

### type key study; robust study summary experimental resutt Reliability 1 (reliable without restriction) Study 

### period

Rationale

### for reliability This study has been 

### performed in compliance wllh the: incl. deflcienclea Between 20 May 2009 and 23 March 2010

Swiss Ordinance relating 

### to Good laboratory Practice adopted May 18th, 2005 (SR 813.112.1}. This Ordinance is based 

### on the OECD Principles of

### Good Laboratory Practice, as revised In 1997 and adopted on November26th, 1997 by decision ofthe OECD Council [C (97)1861Final}.



# Data source Reference

Reference Author



### type study report Ceccatelli R These principles are compatible with Good laboratory Practice regulations specified by regulatory authorities throughout 

### the European Convnunity. the Unned States (EPA and FDA). and Japan (MHLW, MAFF and METI). There were no circumstances thet may have affected the qualny or Integrity of the data.

Year Title Blbllogr8phlc Tntlng labo1'8tory Report Owner Compeny 

### Report source 

### no. cornpeny study no. 

### date 2010 Combined Repeated Dose Toxicity Study 

### with the Harten laboratories Ud., C31087 Ciariant 201(). Reproduction/Developmental Toxicity Screening WOiferstrasse 4 4414 03-23 Test In the Han Wlstar Rat Filillnsdort, Switzertand

## Data access data submitter is data owner 

## Cross-referenceto same study Toxicity

### to reproduction Tocicity

### to reproduction Toxicity

### to reproduction.001 OECD 422 in Rat Main Study 

# Materials and methods Testtype combined repeated doss and reproduction 

### Idevelopmantal screening 

## Limittest
yes 

## Test guideline

Qualifier 

### Guideline according to OECD Guideline 422 (Combined Repsated DossToxicity Study

### wi1h the Reproduction I Developmental Toxicity Screening Test) according 

### to other guideline: EPA OPPTS 670.3650 (Combined Repeetad Dose Toxicity Studywith the Reproduction/Developmental Toxidty Screening Test) 

## GLP compliance
yes (ind. certificate) 

# Test materials Identity oftest material same asforsubstance defined In section 1 (If

## not**read;~~cross)** yes 

## Test material Identity
Identifier 

### Identity 

### Common name Leuco Sulfur Black 1 CAS number 66241-11-0 

## Details ontest material Devlatlone no noDesaiplion: Thixolrophic, black, wet paste Batch Number: ESA0007595 Composition: Leuco Sulfur Black 1: 52.75% (calculated by amount ofby-compounds) 2,4-0initrochlorobenzene: < 20 ppm 2,4-0initrophenole: < 

### 20 ppm Sulfate: 970 ppm Chloride: 750 ppm Sodium: 1.3% (w/w) Water. 45.75% (w/w) by Kari Fischertitration S1abiityofTest Item: Stable under storage conditions StabiityofTest Item In Vehide: Seven days Expiry Date (Retest Date): 11-Jul-2011 Storage Conditions: At room temperature (20 ± 5 'C) , light proteded Sefety Precautions: Routine hygienic procedures (gloves, goggles, face mask). 

## Test animals Species
rat 

### Strain Wistar 

### Sex male/female 

### Details on testanimals andenvironmentalconditions Animals: Rat, HanRcc: WIST(SPF) Rationale: Recognized by inlemationel guidelines as a recommended testsystem. Breeder. Harian Laboratories Ltd., LaboratoryAnimal Services, WOifenrtrasse 4, 4414 FOIIinsdorf, Swltzeriand NumberofAnimals: 40 males (10 pergroup), 40females (10 pergroup) Age (at Start ofTnsatment): 11 weel<s Body Weight Range (at StartofTnsatment): Males: 290 to 324 g, Females: 284 

### to 212 g ldentiflcalion of Pnsntal Animals: Cege card and individual animal number (eartattoo). Identification of Pups: On day 1 postpartum, pups were individually tattooed with Indian ink. Randomization: Computer-generated random algorithm. In addition body weights (recorded on lhe day ofallocation)were taken into consideration in orderto ensuns similar mean body weights In ali groups. Acclimatization: Undertestconditions after health examination. Only animals without any visible signs of illness wens used forthe study. Conditions: Standard laboratory conditions. Air-<:anditioned with 10
- 15 air changes per hour, continuouslymonitonsd environmental conditions (temp. range: 22 ± 3 'C; relative humidity range: 37.5- 80.9%). Thens was 12 hourftuorescentlighl/12-hourdarl< cyde wilh music during the light period. 

## Administration 

## I exposure

### Route ofadministration
oral: gavage 

### Vehicle water 

### Details on oralexposure Dose Formulations The doseformulations were prepared weekly using the test Item as supplied bythe Sponsor. Leuco Sulfur Black 1 was weighed into a glass beakeron a tared precision balance and approximately 80%

### ofthe vehicle was added (w/v). Using a magnetic stirrer, a homogeneous suspanslon was prepared. Having obtained a homogeneous mixture, the remaining vehicle was added. Separate formulations ware prepared lor each concentration. Homogeneity ofthe test Hem in the vehicle was maintained during the dally administration period using a magneticstirrer. Storageof Dose Formulations 

### Dose formulations were stored at room temperature (20 

### ± 5 

## q in brown glass beakers. Based upon the results ofstabiHty analyses performed within the Harian Laboratories study no. C31076, dose formulations were stable lor at least one week. Treatment Method: Oral, by gavage Rationale lor Method: Administration by gavage Is a common and accepted route ofexposure for studies ofthis 

### type. Target 

### Dose Levels: Group 1: 0 mg/l<g bwlday (control group) Group 2: 100 mglkg bw/day Group 3: 300 mg/l<g bwlday Group 4: 1000 mg/l<g bw/day Rationale for Dose Level Selection: The dose levels were selectad based on a previous dose range finding toxicity study in Han Wister

### Rats, Harian Laboratories Study C31076, using dose levels of 116, 348 and 1180 mglkgldayadjustad for 52.75% Leuco Sulfur Black 1 dye stuff in

### the product and corresponding to 220, 680 and 2200 mglkg bwlday

### ofproduct. The NOAEL ofthis study resultad to be 1160

### mg dye stuff/kg bw/day or2200 

### mg product/kg bwlday. Dose Volume: 10 mL/kg body weight 

### Analytical verification ofdoses or concentrations
yes 

### Details on analytical verification ofdoses orconcentrations Analysis of

### Dose Formulations On the first treatment day duplicate samples

### from the control group aswell as three duplicate samples (top, middle and bottom) ofabout 2 g ofeach concentration ware taken priorto dosing for analysis of concentration and homogeneity. Samples

### ofabout 2 g 

### ofeach concentration were taken from the middle onlyto confirm stabHity (4 

### hrs and 7 days). During the lastweek ofthe treatment. samples ware taken 

### from the middle to confll'm concentration. The allquots 

### for analysis 

### ofdosa formulations were trensferred Into tightly closed vessels 

### protected againstoxygen. One set

### of samples was delivered at ambienttemperature to 

### Dr. K. Morgenthel (Harlan Laboratories Ltd., ltingen 

### I Switzerland)and stored thera

### at mom temperature (20

### ±5 'C) until analysis. The remaining setof sa mpleswas stored at the facUlty at momtemperature (20

### ±5 'C) in cese 

### of repeated analysis. Analysis

### of homogeneityand concentration was performed gravimetrically by weighing the residue

### attar drying by lyophilization. Stabllllyofdoss formulations was confinned by IRspectroscopy. The application formulations investigated during the studywere found to comprise Leuco Sulfur Black 1 in the range of81.7% to 102.5%. The homogeneous distribution 

### of Lauco 

### Sulfur Black 1 in the preparations was approved beceuse Individual recoveries found 

### did not deviate""""than 14.2% (<15%)

### from the corresponding mean. In addition, the

### test Rem was found to

### be stable in applicetion formulations when kept for4 hours and for 7 days under storage condRions due to slrrilar IR spectra compered tothe reference spectrum obtained 

### from the test Hem. Thus, identityofthe test itam in applicetion fonnulations was confinned. In conclusion, the resuHs indicate the accurate use

### of the tast item Leuco Sulfur Black 1 and highly purified water as vehicle during this study. 

### Duration oftreatment I 

### exposure Males: 29 days Females: Approximately 7 weeks 

### Frequency oftreatment Daily 

### Doses/concentrations 100 mglkg bw/day Baals actual Ingested 300 mglkg bw/day Baals actual ingested 1000 mglkg bw/day S.als actual ingested 

### No. ofanimals per sex per dose
10 

### Control animals
yes. concurrent vehicle 

### Detlllls on studydesign STUDY SCHEDULE MalesAcclimatization: 7 days FirstTest Hem Administration : Day 1ofpre-pairing Pre-Pairing: 14 days Pairing: 14 days maximum Treabnent Ends: On day before sacrifice Necropsy: Aftertreatment ofat least28 days, when no longer

### needed for assessment of repro-ductive effects Females Acclimatization: 7 days FirstTest Hem Administration: Day 1 ofpre-pairing Pre-Pairing: 14 days Pairing: 14 days maximum Gestation: Approximately 21 days Treabnent Ends: On day 4 post pariUm Necropsy:On day 5 postpartum (pups on day 4 post partum) 

## Examinations Observations andexaminationsperformedandfrequency
VIABILITY I MORTALITY Twloe daily CLINICAL SIGNS Dally cage-side clinical observations (once daily, during acclimatization and uptodayofnecropsy). Additionally females were observed for signs ofdiflicuH or prolonged parturition, and behavioral abnormalitias In nesting and nursing. FOOD CONSUMPTION Males: Weekly during pre-pairing and aHer pairing periods Females: Pre-pairing period days 1 -8 and 8- 14; gestation days 0-7, 7- 14 and 14- 21 post coitum, and days 1 -4 postpartum. No food consumption was recorded during the pairing period. BODY WEIGHTS Recorded dailyfrom treatment start to day ofnecropsy. DETAILED CLINICAL OBSERVATIONS Once priortothe first administration ofthe test item and weekly thereafter, detailed clinical observationswere performed outside the horne cage. Animals were observed for the following: changes in skin, fur, eyes, mucous membranes, occurrence ofsecretions and excretions, and autonomicactivity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern). Changes in gail, posture and response 

### to handling as well as the presence ofclonic ortonic movements, stereotypies or bizarre behaviorwere also reported. FUNCTIONAL OBSERVATION BATERY Atone time during the study (males shortly before 

### the scheduled sacrifice and females on day 3 or4 post partum) relevant parameters were performed wHh five P generation males and five P generation femalesfrom each group. This FOB assessment was conducted following 

### the daily dose administration. Animals were observed for the following:

- Cege-side observations: unusual body movements (e.g. tremors, convulsions), abnormal behavior (e.g. circling, stereotypy) and posture as well as resistance

### to removal -Hand-held observations: palpebral closure, pinna raftex, lacrimation, pupH siza, pupil reactivity, salivation, muscle tone, extensorthrust response, righting reflex and reaction tohandling. -Open field observations: level of ambulatory activity including rearing (one minute evaluation), responsiveness to sharp noise, paw pinch,**ga~** evaluation, quantity of urine and fecal 

### pelletsvoided. -Categorical observations (can be made anytime during 

### the FOB): haircoat, behavior, respiration, muscle movements, eyes, hearing**abil~** (Preyar's reflex), urine orfeces, soiling, ganerel abnormalities, posture. -Measurements 1Counts: hind limb I fore limb grip strength, landing foot splay, ractal temperature. Any ebnonnal findings were recorded and, where appropriate, graded in severity. Additionally, locomotor**activ~** was measured quantitatively for the same animals. Activitywas measured with an Activity MonitorAMS-0151 (FMI, Germany).**Activ~** of

### the anlmals (besed on beam count)was recorded for 6-minute intervals over a period of 30 minutes. These data and the**totals~** over 30 minutes were reporied. CUNICAL LABORATORY INVESTIGATIONS Blood and urine sampleswere obtained on the day ofthe scheduled necropsyfrom 5 males from each group. Males were kept in metabolism cage and fasted overnight (but allowed acoassto waterad Nbitum) for approximately 18 hours. Blood samples from 5 lactating females from each group were obtained on day 5 post partum. The females ware fasted for approximately 18-20 hours before blood sampling but allowed 

### acoass to waterad libitum. Blood samples wera drawn sublingually from all animals under light isoflurane anesthesia. The samples were collected eariy in the working dayto reduca biological variation caused by circadian rhythms. HEMATOLOGY The following hematology parameters were determined: Complete Blood 

## een Count Erythrocyte count Reticutocyte Count Hemoglobin Hematoait Mean corpuscularvolume Red cell volume distribution width Mean corpuscular hemoglobin Mean corpuscular hemoglobin concentration Hemoglobin concentration distribution width Leukocyte count, total Differential leukocyte count Platelet count Coagulation: Prothrombin time (= Thromboplastin time) Activated partial Thromboplastin time CUNICAL BIOCHEMISTRY The following clinical biochemistry parameters were determined: Glucose Urea Creatinine Bilirubin, total

### Cholesterol, total Triglycerides Aspartate aminolransferase Alanine aminotransferase 

### Alkaline phosphatase Gamma-glutamy14ranslerase Bile acids 

### Sodium Potasalum Chloride Calcium Phosphorus Protein, total Albumin Globulin Albumin/Globulin ratio URINE ANALYSIS 

### The following urine parameters were detennined: 

### Relative density colour Appearance pH Protein 

### Glucose NKrile Ketones Urobilinogen Bilirubin Erytrocytes Leukocytes Volume (18 

### hours) 

## Sacrificeandpathology TERMINATION OF THE STUDY Pups 

### were sacrificed on day 

### 4 postpartum. NECROPSY All animals wera 

### saaificed by an injection 

### ofsodium pentobarbital. 

### Dead pups, except those excessively cannibalized, were examined macroscopically. All pups were examined macroscopically for any structural changes, eKher atthe scheduled necropsy or during the 

### study

# n death occurred. ORGAN WEIGHTS The testes and epididymides of aU parantal males were weighed as pairs. In addition, from 

### 5 malas and females killed atthe end 

### ofthe 

### study which 

### were selected from 

### each group, the following organs were trimmed from 

### any adherent tissue, es appropriate,and lhelrwetweighttaken. Adrenal glands (weighed as pairs) Brain Heart Kidneys (weighed as pairs) Liver Thymus Spleen TISSUE PRESERVATION 

### Tha following tissues from aN parental males were preserved in neutral phosphate buffered 4% formaldehyde solution: Prostate Seminal 

### vesicles with coagulating gland Testes (in Bouin'sfixative) Epididymides (in Bouin's fixative) 

### Tha following tissues from an parental females were preserved in neutral phosphate buffered 4% formaldehyde solution: Ovaries In addition, 

### from thefive males and females pargroup selected for organ weights and from all animalsfound 

### dead or 

### kHied in extremis, 

### the following tissues were preserved In neutral phosphate buffered 4% formaldehyde solution: 

### Gross lesions 

### Brain Spinal 

### chord SmaH and large intestines (incl. Payer's patches) 

### Stomach Liver Kidneys Adrenals Spleen Heart Thymus Thyroids, and parathyroids 

### ff possible Trachea and lungs (preserved by inflation with fixative and then immersion) Uterus (with vagina) Urinary bladder Lymph nodes (mesenterial, mandibular) Peripheral nerve (sciatic) 

### Bone marrow HISTOTECHNIQUE All organ and tissue samples to

### be examined by the principal investigatorwere processed, embedlded and 

### cut at an approximate thickness of2 - 4 micrometers and stained with hematoxylin and eosin. Additionally, the testis was stained by PAS-hematoxylin. HISTOPATHOLOGYSlides 

### ofall organs and tissues collected atterminal sacrifice from 

### the animals 

### ofthe control 

### and higiHiose groups were examined by the principal investigator. The same applied

### to all 

### occurring gross lesions. Special emphasis was made on the stages ofspermatogenesis and histopathologyof interstitial cell sbuclu111. Histological examination ofovaries was canied 

### out on females that did notgive birth. 

### Statistics The following statistical methods were usedto analyze 

### food consumption, 

### bodyweights, organ weight, locomotoractivity and macroscopical findings and reproduction data: 

### • Means end standard deviations 

### ofvarious data were calculated. • The Dunnell-test (many to one 1-test) based on a pooled variance estimate was applied 

### Hthe variables could 

### be assumed 

### to follow a normal distribution for

### the comparison oflhe treated groups and 

### the control groups for each sex. ·The Steel-test (many-ooe renk test) was applied Instead ofthe Dunnell·taslwhen the data could 

### not be assumed to follow a normal distribution. • Fishers exact-testwas applied 

### Hthe variables could 

### be dichotomized wllhoutloss ofinformation. 

# Results and discussions Effect levels Endpoint Effect 

### Bued 

### Sex lev., on Basis 

### foreffect

### level/Remark•
NOAEL >= 

### 1000 test male/female With the exception for a dark discoloration 

### offeces observed In all animals treated wtth the testttem at

### the dose level of 

### 1000 mglkg 

### body 

### mat. 

# Results ofexaminations Clinicalsigns andmortality weight/day, no othertest tterJHelaled effects were observed during the study. The discoloration offeces was considered to

### be due tothe staining property of

### the test ttem and

### not to

### be adverse.

yes (dark dlsoololl!tion 

### offeces at

### the dose level of 

### 1000 mg/kg bw/day) 

### Body -lghtand -lghtgain

no

### effects 

## Food consumption andcompoundIntake (If

### feeding study)

no effects 

## Food efficiency 

### not examined 

### Waterconsumption andcompoundIntake (Ifdrinking waterstudy)

### not examined 

### Ophthalmoscopic

## examination
not examined 

### Haematology

no effects 

### Clinicalchemistry

no effects 

### Urinalysis

no effaciS 

### Neurobehaviour

no

### effects 

## Organ weights

no 

### effects 

### Gross pathology

no effects 

### Histopathology: neoplastic

no eflects 

### Details on results CLINICAL SIGNS AND MORTALITY BODY WEIGHTANDWEIGHT GAIN FOOD CONSUMPTION AND COMPOUND INTAKE (K feeding study) FOOD EFFICIENCY WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study) OPHTHALMOSCOPIC EXAMINATIONHAEMATOLOGY CLINICAL CHEMISTRY URINALYSIS NEUROBEHAVIOUR ORGAN WEIGHTS GROSS PATHOLOGY HISTOPATHOLOGY: NON-NEOPLASTIC HISTOPATHOLOGY: NEOPLASTIC (ifapplicable) HISTORICAL CONTROL DATA (if applicable) OTHER FINDINGS 

### Any other Information on results Incl. tables IN LIVE OBSERVATIONS Detailed Clinical Observations (Dally) All animals survived until the scheduled necropsy. In group 4, dark discoloration offeces was observed in males and females starting on day 3 ofthe pre-pairing period onwards. This was likely due to the staining property ofthe test item at higher concentration. In groups 2 and 3, no clinical signs were observed for the entire duration ofthe study. (See attached Tables on pp. 1to 3) Detailed Clinical Observations (Weekly) No findings were observed during the weekly examination. Functional ObservatlonaiBattery

### None 

### ofthe parameters under investigation during the functional observational battery gave an indication ofa test item-related effect. Thefrequency of rearing numberand incidence ofurine puddles during the open field observation did not give indication ofany test item-related effect. Observational as spontaneous vocalization when the rat was removed from the cage was considered to be 

### of incidental nature. Mean values ofgrip strength (fore- and hind paws) and landing foot splay gave no indication ofanytest item-related effect. Bodytemperature in males was statistically significantly lower in group 4 compared to the control group (37.9 

### 'C compared to 38.7 

### 'C in the control group). However, this value was within the range ofthe historical control data and therefore was notconsidered to be a test item-related effect. (See attached Tables on pp. 4 to 7 and attached Historical Control Data on pp. 3 to 14) LocomotorActivity Locomotor activity was assessed quantitatively in terms of low beam counts in activity monitor and did not give any indication ofa test item-related effect. 

### Food Consumption - Males - Pre-pairing and After Pairing Periods Meanfood consumption was not considered to be affected by the treatment with the test item. During the pre-pairing period, the lowerfood consumption observed in groups 2 and 3 (-6.2 and -4.6% compared to the control group)was considered to be incidental sincethis reduction did notfollow a dosEKiependent pattern and in group 4 mean food consumption was similar to the control (-0.4% compared tothe control). During the after pairing period, mean 

### food consumption compared to the control was: +2.2, -1.1 and +1.5%, in groups 2, 3 and 4, respectively (See attached Tables on pp. 8 to 11) Body Weights- Males
- Pre-pairing, Pairing and After Pairing Periods Mean body weight and mean body weight gain were notaffected by the treatment with the test item forthe 

### whole duration ofthe study. The statistically significantly lower body weight gain observed in group2 between day 4 and 7 was considered to be incidental since there was no dosedependency. In order ofascending dose level, body weight gain was: during the pre-pairing period +13.0%, +10.9%, +11.5 and +11.9%, during the pairing period +4.9%, +5.7%, +4.4% and +4.7%, and during the after pairing period +4.1%, +5.6%, +4.8% and +4.5%, respectively. (See attached Tables on pp. 12 to 20)

### Food Consumption
- Females
- Pre-pairing, Gestation and Lactation Periods Notest item-related effects were observed in mean food consumption for

## the entire duration ofthe study. Mean food consumption in groups 2, 3 and 4, compared to the control group, was: duringthe pre-pairing period +5.0%, +2.2% and +2.2%, during the gestation period +0.4%, -2.1% and -0.4%, and during the lactation period +11.3%, -3.1% and -3.7%, respectively. (See attached Tables on pp. 21 to 26) BodyWeights
- Females -Pre-pairing, Pairing, Gestation and Lactation Periods Notest item-related effects were noted in mean bodyweight and mean body weight gain offemales during 

## the whole study atany dose level. In order of ascending dose level, body weight gain was: during the pre-pairing period +7.5%, +8.3%, +9.3% and +8.5, during the gestation period +63.5%, +63.3%, +60.2% and +60.5%, and during the lactation +3.1%, +6.2%, +4.6 and +2.7%, respectively. (See attached tables on pp. 27 to 39) CLINICAL LABORATORY INVESTIGATION Hematology The assessmentof

## the hematology data did not reveal any test item-related effects in males and females. Clinical Biochemistry Males: In group 4, the concentration of potassium was statistically significantly higher (+17.8% compared 

## to the control). Since the mean value was within the range ofthe historical control data, this was not considered to be a test item-related effect. In groups 2 and 3 no changes were noted. Females: The assessment ofthe dinical biochenistry data did not reveal any test item-related effects in females. (See 

### attached Tables on pp. 40 to 41 and attached Historical Control Data on pp. 1

## to 2) Urine analysis The assessment of

## the urine analysis data did not reveal any test item-related effects in males and females.

## TERMINAL FINDINGS Organ Weights Notest item-related effects were noted in absolute and relative weight oforgans in males and females. All 

## the statistically significant changes were within the range of

## the historical control data ordid not follow a dose-dependent pattern. (See attached Tables on pp. 44 to 49 and attached Historical Control Data on pp. 39to 62) 

## Macroscopical Findings The findings noted during the macroscopical examination did not give indication ofany test item-related effect. Males: In group 4, spleen with a constriction was noted in male no. 34. No otherfindings were noted. Females: In group 4, female no. 72 was noted to have the liver caudal lobe adherent to the lateral lobe. In groups2 and 3 nofindings were noted. In group 1, female no. 43 was noted to have tan or dark red or red brown discolored nodules on the 

## left uterine hom. Same female had only two implantation sites and did notdeliverany pups. 

## Histopathology Findings In group 4, in the lungs ofanimal no. 75 some alveolar macrophages (moderate severity degree)were found

## to contain dark pigment. This finding was possibly related to accidental aspiration ofdose solution. All further lesions recorded during the microscopic investigation were within the range of background alterationsthat may be recorded in this 

### type ofstudy, and in rats ofthis strain and age. In group 1, in one infertilefemale (number43) a deciduoma was noted. This lesion was deemed to have contributed

## to infertility. Qualitative staging ofthePAS-stained testes was conducted and there were no abnormal lesions encountered during sperm staging regarding completeness 

### ofstages and maturation ofcell populations. Individual lesions recorded were within 

## the range 

### ofbackground alterations that may be recorded in this 

### type ofstudy, in rats ofthis strain and age. 

# Applicant's summary and conclusion

## Conclusions
This study is a valid investigation 

### ofthe toxicological effects resuHing 

### from repeated oral-gavage administration 

### ofthe test item Leuco Sulfur Black 1 (CAS#66241-11-0) to rats over approximately28 days. Leuco Sulfur Black 1 was admlnisterad in highly purified wateras vehicle atdosages

### of 100,300,

### end 1000

### mglkg bodyweight/day, and controls received 

### the vehicle only. Leuco Sulfur Black 1 was administered to male rats for 29 days and to female ratslor 14 days priorto pairing, through 

### the pairing andgestation periods unlit 

### the F1 generation reached day4 post partum. Administrations at 1000 mglkg body weight/day caused a dark discoloration of

### feces In all animals after

### the onsatof

### thetreatment until

### the end ofsllldy. This was Nkely due tothe staining property 

### ofthe test 

### item and not considered to be adversa. Food consumption and body weight ware

### not affected forthe entire duration ofthe sllldy. The evaluation 

### ofthe functional batteryobservations and 

### the assassmentofclinical laboratory Investigations parametersdid not reveal any test Hem-related effect. All 

### the changes notad in the organ weights were within the bacl<ground ofthe biological variation. Noadversafindings were noted during macroscopical and histopathology examinations. Based on thesa resuhs a general NOAEL (No Observed Adverse Effect Level) was established at 1000 mglkg body weight/day. 

## Executive summary The purpose 

### ofthis sllldy was to generate pretimlnary information on the possible health hazards likelyto arise 

### from repaatad exposure overa relatively limited pariod oftime. In addHion Hprovided information on possible eflecls on male and female reproductive performance such as gonadal function, mating behavior, conception, developmentof

### the conceptus andparlllrltion.

This sllldy should provide information to assess the need to conduct furlher investigations and may provide guidance in the design of subsequent studies. 

### Dose levelswere seleclad in agreement

### wHh the Sponsor, based on the resuhs ofa dose range-finding sllldy (Harlan Laboratories Sllldy C31076). Leuco Sulfur Black 1 was administered to male rats lor29 days and tofemale rats for 14 days 

### prior to pairing, through

### the pairing and gestation parlods until the F1 generation reachad day4 post parlllm. The following 

### doss levels were applied: Group 1: Group2: Group3: Group4: 0 mglkg bodyweight/day (control group) 100 mglkg bodyweight/day 300 mglkg bodyweight/day 1000 mglkg body weight/day A standard dosevolume of 10 mUkg body weight with a dailyadjustmentto the actual body weightwas

### used. Control animalswere dosed with the vehicle alone (highly purified water). The following results were obtained:Gene,.lT olerllblllty All the animals survived until the scheduled necropsy. AI t000 mglkg bodyweightiday, a dati< discoloration 

### offeces 

### was obsarved in all animals starting on day 3 

### ofthe pra-pairing period onwards. 

### Functional Observetlon.ol 8ettery Notest item-related effects ware noted. Food Consumption Food consumption was not affected bythe treatmentwith the test**~em** in males and females. Body Weights Notest Item-related effects ware noted in body weight and body weight gain in males and females for the entire duration ofthe study. Clinical 

### Laboratory Investigations The assessment ofthe clinical laboratory investigations parameters did not give indication ofany test ijem-related effect. Reproduction and BIHdlng 

### Oats Mean precoital time,**fartll~** and gestation indaxand conception rate were not affected bythe treatment with thetest Ham. Implantation rate and post implatation loss ware also

### not alfected by the treatment with the test item. 

### Orvan Weights Weightoforgans was notalfected by the treatment with the test ijem. All the alterations noted ware within the range of historical control 

### data. Macroac:oplclll Findings and Hlatopathologlcal Eumln.otlons No adverseeffects 

### ofthe test Hem were noted during the histopathological examination. Conclusion Basad on thesa resulte a general NOAEL (No Obsarved Adverse Effect Level)was established at 1000 mglkg bodyweight/day.

# Endpointstudyrecord: waiving_Repeated dose toxicity
UUID IUC5-e66359e9-ad84-4e86-a489-5011b522b4bc Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea 

### Author Date 

### Remarks uensenba I Clariant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-05-21 13:43:48 CEST



# Administrative Data Datllwalvlng Justification

### forwaiving study scientifically unjustified In accordance with Regulation (EC) No 190712006 Annex XI, a subchronic toxicity study 

### is scientificaHy notjustified and, thus, should not be conducted because ofthe following: Leuco Sulfur Black 1 has a mean molecularweight in the range of8000 to 9000 Da and a number-average molecularweight (Mn) ofabout3000 Da. It is very poorly oreven no1 soluble in common polar and non-polarsolvents such as water (solubility of62.4 mg/L), DMSO, methanol, DMF, THF, chloroform and no1 soluble In acids Hke sulfuric acid and trlftuoroacelic acid. The vapour pressure ofthe pure substance, calculated 

### from the boiling point of>400 

### 'C, 

### is 1.8x 10E-5 hPa at25 

### 'C. Based on these properties IIis very unlikelythat Leuco Sulfur Black 1 can cross any biological membrane. Thus, an absorption oftoxicologically significant amounts of Leuco Sulfur Black 1following oral, dermal orinhalation exposuns can be excluded. This Is substantiated by

### the results obtained in toxicity and ecotoxicity studies, whans no substance related local and/orsystemic effects indicative oftoxicity wens seen in all tests, including acute animal and 

### acute ecotoxicity studies, sensKization, repeated dosetoxicity and mutegenlcKytests. In 

### the

### acute oral toxicity study black stained faeces was observed for maximally four days starling 

### thedayafteradministration of

### the substance al2000 

### mgl1<g bw. Dart< discoloured faeces wens also seen in the repeated 

### dose toxicKy study (OECD 422) at1000 

### mgl1<g bwld, starling on day 3 ofthe studyand lasting throughout the administration period until termination. The dlscolouration offaeces wasthaonly 

### effect observed in both studies and can beattributed to 

### the strong staining properties of

### the substance. This indicatesthat Leuco Sulfur Black 1 is directlyexcreted and that absorption in tha gastrointestinal tract does notoccur following oral administration, especially since organs orurinewens notdiscoloured and signs 

### of substance deposits were noffound in 

### the histopathological analysis 

### of the organs. In conclusion, 

### the availabletoxicity and ecotoxicity data togetherwith the intrinsic substance properties are sufficienttodemonstrate that exposure to Leuco Sulfur Black 1 does not cause any haaRh concern.

# 7.5.2 Repeated dose toxicity: dermal Endpointstudyrecord: waiving_Repeateddose toxicity: derma/.001
UUID IUC5-6e0385ef-4a64-48c4-9680-a0774b47064c Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author thr I SCC GmbH IWendelsheim I Germany Date 2010-05-26 16:35:53 CEST Remarks 

# Administrative Data Data wslvlng siUdy scientifically unjustified Justification fordata wslvlng see dela waiving under repealed dose loxicity: orel (CSR 

### section 5.6.1.1)

# 7.5.3 Repeated dose toxicity: inhalation Endpointstudyrecord: waiving_Repeateddose toxicity: inhalation.001 UUID IUC5-7e173fd7-29cf-4711-ba10-1e736b54465d Dossier UUID IUC5-292149d1-a6n-4feb-9692-1fad973281ea Author lhr 

### I SCC GmbH 

### I Wendelsheim 

### I Germany Date 2010-05-26 16:36:42 CEST Remarks 

# Administrative Data Data waiving sludy scientifically unjuslifoed Justlllcatlon 

### fordata waiving see dala waiving under repeated dose toxicity: oral (CSR 

### section 5.6.1.1)

# 7.6 Genetic toxicity Endpointsummary: Genetic toxicity
UUID IUC5-455d61b4-0863-40ee-bcd0-03c1ae1075b7 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author lhr 

### I SCC GmbH 

### I Wendelsheim 

### I Germany Date 2010-05-26 16:46:32 CEST Remarks 

# Administrative Data Short description of key information

Leuco Sulfur Black 1 was neither mutagenic In 

### lhe Ames lest nor in the HPRT assay on V79 calls and was non-dastogenic In the in vitro chromosome aberration assay using V79 cells. All assays were performed with and without metabolicactivation sys1am. 

# Key value for chemical safety assessment Genetictoxicity negative 

# Discussion An aqueous Leuco Sulfur Black 1 formulation containing less than typical sulfurdye concentration (30.2% instead of52.75 %) was tested in the bacterial reverse mutation assay with Salmonella typhimurium strains TA1535, TA1537, TA1538, TA98 and TA100 using the plate incorporation method atfive dose levels, in triplicate, both with and without a metabolic activation system (59 mix from induced rat liver). Two independent experiments were conducted. The assay was performed according to OECD test guideline No. 471 and GLP. The dose range was 312.5 to 5000**~g/plate.** 

## Distilled water was used as solvent

## The concurrent positive controls gave increases in revertants within the expected ranges. The test item did not cause toxicity. Precipitations occurred at and above 6251Jg test item/plate. Nosignificant increase in the numberof revertants was recorded for any ofthe bacterial strains with any dose ofthe test item eitherwith orwithout metabolic activation. In this assay Leuco Sulfur Black 1 precipitated at and above test item concentrations of625 IJg/plate, demonstrating that the highest possible Leuco Sulfur Black 1 concentrations which can be studied in theAmestestwere achieved. Thus, the assay is fully valid for the assessment ofthe mutagenic potential of Leuco Sulfur Black 1. Leuco Sulfur Black 1was assessed for its potential to induce gene mutations at the HPRT locus using V79 cells ofthe Chinese hamster according to OECD test guideline No. 476 and GLP. The study was performed in two independent experiments. In the first experimentthe treatment period was 4 hours with and without metabolic activation. The second experimentwas performed with a treatmenttime of4 hours with and 24 hours without metabolic activation. Precipitation ofthe test item was observed at 1251Jg/ml and above with and at250 IJg/ml and above without metabolic activation. Relevant cytotoxic effects defined as a relative cloning efficiency I below 50% in both parallel cultures occurred at500 IJg/mL in both experiments without metabolic activation. In the presence of metabolic activation cytotoxic effects as described above were noted at 125 IJg/ml and above in the first experiment. The positive controls ethylmethanesulfonate and DMBA showed a distinct increase in induced mutant colonies. No relevant and reproducible increase in mutant colony numbers/10E6cells was observed in the main experiments up to the maximum concentration. All mutant frequencies remained well within the historical range of solvent controls. No significant dose dependenttrend ofthe mutation frequency indicated by a probability value of<0.05 was determined in any ofthe experimental groups. In conclusion, Leuco Sulfur Blackwas not mutagenic in theHPRTassay with and without metabolic activation. Leuco Sulfur Black 1, suspended in deionised water, was assessed for its potential to induce structural chromosome aberrations in V79 cells ofthe Chinese hamster in vitro in two independent experiments according to OECDtest guideline No. 473 and GLP. In each experimental group two parallel cultures were set up. 100 metaphases perculture were evaluated for structural chromosome aberrations. Cells were exposed to the test item for4 hours with and without S9 in experiment I and again for 4 hours with S9 and continuouslyfor 18 hours in experiment II. Dose selection forthe cytogenetic experiments was

## performed considering the toxicity data and the occurrence of precipitation. The chosen treatment concentrations were between 2.3 and 300**~g/ml** 

## (with and without S9). The evaluated concentrations were 19.5, 39.1 and 78.1**~g/ml** 

## in experiment I and 37.5, 75 and 150**~g/ml** 

## in experiment II. In the absence and presence ofS9 mix no cytotoxicity was observed up to the highest evaluable concentration. Dueto strong test item precipitation concentrations higherthan those examined could not be evaluated for cytogenetic damage. No clastogenicitywas observed at the concentrations evaluated eitherwith orwithout metabolic activation. However, in experiment II in the presence ofS9 mix a single statistically significant increase in the number ofaberrant cells excluding gaps (3.0 %) was observed at the highest evaluable concentration (150**~g/ml).** 

## This value was dearly in the laboratory's historical solvent control data range (0.0- 4.0 

# % aberrant cells excluding gaps) and is therefore regarded as biologically irrelevant. No evidence of an increase in polyploid metaphases was noticed aftertreatment with the test item as compared to the control cultures. Appropriate mutagens were used as positive controls. They induced statistically significant increases (p < 

## 0.05) in cells with structural chromosome aberrations. In conclusion, the test item was not clastogenic in this in vitro chromosome aberration assay. 

# Justification for classification or non-classification The available data demonstrate that Leuco Sulfur Black 1 has no genotoxic potential. Thus, it is not subjectto classification and labelling according to Directive 67/548/EECand Regulation 1272/2008/EC.

# 7.6.1 Genetic toxicity in vitro Endpointstudyrecord: key_471_Safepharm_1991 UUID IUC5-ac176bd8-fc03-4e62-a18e-3a2266ab21eb Oosaler UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks thr 

### I SCC GmbH 

### IWendelsheim 

### I Germany 2010-05-26 16:44:32 CEST



# Administrative Data Purposeflag key study Study F8sult

### type experlmentel resun Reliability 1 (reiable without restriction) 

# Data source Reference

Reference Author Veer Title



### type study report Jenkinson 1991 Leuco sulphur black 1: reverese mutation 

### PC assay "Ames test"' using salmonella 

### typhimurium 

### Data access data submrtter is data owner 

# Materials and methods Type ofgenotoxicity gene mutation 

### Type ofstudy Bibliographic 

### oource Teeling laboratory R_.t Owner Company Report 

### no. company study no. 

### data Safepharm Laboratories 4711657 Clarlant 1991-02- Ltd. Derby, DE1 2BT, UK 14bacterial reverse mutation assay (e.g. 

### Ames test) 

## Test guideline Guideline DECO Guideline 471 (Bacterial Reverse Mutation

### Assay)

yes (Ind. certificate) 

# Test materials Identity oftest material same as forsubstance defined in section 1 (ifnot read-across)

no 

## Test material identity Identifier other: Leuco Sulfur Black 1formulation 

### see confidential details on - material 

## Confidential details on test material Leuco Sulfur Black 1: 30.2 

### % Sodium hydroxide: 2.7% Sodium sulfide: 3.6 

### % Inorganic sodium salts: 3.9 

### % water: 59.6% 

# Method Species/strain Species/strain other: TA1535, TA1537, TA1538, TA98, TA100 Additional not applicable atreln ch•racterlatlca Met.obollc with and**w~hout** 

### IIC!Iv•Uon Met.lbollc S9 mix derived form Aroclor 1254 inducad rat livers 

### •ctlvatlon ayatem

### Test concentrations 0, 8, 40, 200, 1000 and 5000**~g/plate** (experiment I, 

### with and without S9 mix) 0, 312.5, 625, 1250, 2500 and 5000**~glplate** (experiment II, 

### with and without S9 mix) 

### Vehicle sterile distilled water 

### Controls Negative 

### no controls 

### Solvent I yes (distilled water) 

### vehicle controls 

### True no negative controls 

### Positive yes controls 

### Positive other. -S9 mix: ENNG (TAtOO, TA1535), 9-AA (TA1537), 4-NOPD (TA1538), 4-NQO ( TA98): +S9 mix: 2-AA (TA1535), BP (al otherstrains) control substance 

### Details 

### on testsystem andconditions Five concentrations ofthe test Hem

### ware assayed 

### in triplicate against 

### each tester strain, using the direct plate incorporation method. 0.1 

### mL ofthe diluted test Hem or negative control solutionwas added to 2.0 ml oftrace supplemented top agarat 45'1:: In sterile tubes. A 0.1 mlaliquot ofbacterial suspension and 0.5 ml ofthe S9 liver microsome mixorbufferwas added. The mix was overlaid onto sterile plates ofVogei-Brunner agar. After approximately48 hours 

### at 37"Cthe plates ware scored for revertant colonies and thinning ofbad<ground lawn 

### Evaluation criteria For a substance considered to be positive, Hshould have induced a dose-related and statistically significsnl increase In the mutation rate in one ormore strains. To be considered nagative the numbrof reveriatns compared to spontaneous reveriants should be less than twofold ateach dose level. 

### Statistics All data are statistically analysed using the methods recommendad by UKEMS (Kirkland OJ (ed) Statistical evaluation ofmutagenicHy test

### data. Reprot- part Ill (1989)- Cambridge universHy press) 

# Results and discussions

## Test results Species/strain other. TA1535, TA1537, TA1538, TA98, TA100 

### Metabolk: with and without IICIIvatlon

Test al strains/cell types tested 

### system Genotoxlclty negative 

### Cyt-xlclty no, buttested upto precipitating concentrations Vehicle yes 

### controls valid Negatlve not applicable 

### controls valid Positive yes 

### controls vaHd 

## Add/tiona/Information on results Notoxicitywasexhibited. Precipitation was observed with doses at and above 625**~glplate.** this did not interfere with the scoring 

### of revertant 

### colonies. 

### No significant increases in the numbers 

### of revertant colonies were recorded for any ofthe strains 

### ofSelmonelly used at any dose level etther wtth orwithout S9. 

# Applicant's summary and conclusion Executive summary 

# An aqueous Leuco Sulfur Black 1 formulation containing less than typical sulfur dye concentration (30.2% instead of52.75 %) was tested in the bacterial reverse mutation assay with Salmonella typhimurium strains TA1535, TA1537, TA1538, TA98 and TA100 using the plate incorporation method atfive dose levels, in triplicate, both with and without a metabolic activation system (S9 mix from induced rat liver). Two independent experiments were conducted. The assaywas performed according to OECD test guideline No. 471 and GLP. The dose range was 312.5 to 5000 IJg/plate. Distilled water was used as solvent. The concurrent positive controls gave increases in revertants within the expected ranges.

## The test item did not cause toxicity. Precipitations occurred at and above 625 j..lg test item/plate. No significant increase in the numberof revertants was recorded for any ofthe bacterial strains with any dose ofthe test item eitherwith orwithout metabolic activation. In this assay Leuco Sulfur Black 1 precipitated at and above test item concentrations of625 1Jg/plate, demonstrating that the highest possible Leuco Sulfur Black 1 concentrations which can be studied in the Ames test were achieved. Thus, the assay isfully valid for the assessment ofthe mutagenic potential of Leuco Sulfur Black 1.

# Endpointstudyrecord: 1234903_HPRT
UUID IUC5-7f0c301e-409c-4381>-a151-edcd5b837397 Dossier UUID IUC5-292149c1-a677-4feb-9692-1fad973281ea Author Date Remarks thr I SCC GmbH I Wendelsheim I Germany 2010-05-26 16:43:42 CEST



# Administrative Data Purposeflag Study

### resulttype Reliability key study; robust study summary experimental resuH 1 (reliable without restriction) Rationale for reliability Incl. deflclencleo according to OECD 

### 476 

# Data source Reference

Reference Author Year



### type study report Wollny 2009 H-E 

## Data access data submitter is data owner 

## Data protection claimed yes. butwUiing to share 

### Title
Gene Mutation Assay In Chinese Hemster 

### V79 Cells in vitro 

### ty79I HPRT)



# Materials and methods Type ofgenotoxlclty gene mutation BlbHognphlc 

### T•tlng laboratory Report Owner Compeny Report 

### source 

### no. compeny study no. 

### date Her1an Cytolest Cell C31133 Clarient 1234903 

### 2009-00- Research GmbH 07

### Type ofstudy mammaUan cell 

### gene mutation assay 

### Test guideline Guideline OECD Guideline 476 (In vitro 

### Mammalian 

### Cell Gene Mutation Test)

yes (Ind. certificate) 

## Test materials Identity oftest material same as for substance defined In section 1 

### (If not read..across) yes 

### Details on test material ldenlification: Leuco Sulfur Black 1 CAS No.: 66241-11-0 Description: Thixotrophlc, blacl<, wet paste Belch 

### No.: ESA0007595 Composition: Leuco Sulfur Blacl< 1: 52.75 % (calculated by amount 

### ofby-compounds) 2,4-Dinitrochlorobenzene: < 20 ppm 

### 2,4-0ini1rophenole: < 

### 20 ppm Sulfale: 970 ppm Chloride: 

### 750 ppm 

### Sodium: 1.3%(w/w) Water: 45.75% (w/w) by Kar1-Fischer titration MolecularWeight: 

### Not applicable, because test Kern 

### is an UVCB substance StablityofTest nem: Stable understorage oondKions Storage: At room temperatura (range 20 +1- 5"C), Iight protected 

### Expiry 

### Date: July 11, 2011 On the day 

### ofthe experiment (immediately before treatment), 

### thetest Kern was suspended in deionised water. The final concentration of daionised water in the culture medium was 10 

### %vlv. 

## Method Target gene 

## HPRT

## Species/strain Species/strain Chinese hamster lung fibroblasts (V79) Details -Type and identity of media: MEM

on - Periodically 

### checked 

### for Mycoplasma contamination: yes rnamrnallen
- Periodically 

### checked 

### for karyotype stability: yas

cell
- Periodically "cleansed"against high spontaneous background: 

### yes lines

(If -llceble) 

### Metabolic 

### activation with and whhout 

### -bollc Rat liver S9 

### activation sysam 

## Test concentrations Experiment 

### 1: wfthoul S9 mix: 31.3; 62.5; 125.0; 250.0; and 500.0**~g/ml** wfth S9 mix: 31.3; 62.5; 125.0; 250.0; and 500.0**~g/ml** Experiment II: without S9 mix: 31.3; 62.5; 125.0; 250.0; and 500.0**~g/ml** 

### with S9 mix: 31.3; 62.5; 125.0; 250.0; and 500.0**~g/ml** 

## Vehicle

- Vehide(s)lsolvent(s) used: water -Justification 

### for choice ofsolvent/vehicle: solub!Rty properties 

## Controls Negative no controls Solvent/ yes vehicle controls True no 

### negllllve controls Poeltlve yes controlsPositive ethytmethanesulphonate 

### control oubslllnce Remarko without metabolic activation N-tlve no 

### controll Solvent I 

### yas vehicle 

### controls True no negative 

### controll Poolllve yes 

### controls Positive 

### 7,12-dimelhytbenzanthracene 

### control oubslllnce Rem.orko with metabolic activation 

## Details

## on testsystem andconditions METHOD OF APPLICATION: in medium
DURATION -Exposure duration: experiment 

### 1: 4 hours with and without metabolic activation experiment 

### II: 24 hours without metabolic activation, 4 hours with metabolic activation -Expression time 

### (cells in growth medium): 

### 72 hours

- Selection time (if incubation with a saleclion agent): 1

### 0 days SELECTION AGENT (mutation assays): Thioguanine NUMBER OF REPLICATIONS: 2 NUMBER OF CELLS EVALUATED: >1.5x10exp.6 DETERMINATION OF CYTOTOXICITY -Method: cloning efficiency

## Evaluation criteria
A test ttem producing neither 

### a concentration-related Increase 

### ofthe mutantfrequency 

### nor a reproducible positive response atany

### ofthe test points Is considered to 

### be non-mutagenic in this 

### system. 

### Amutagenic response is described as follows: The test ttem Is classified 

### as mutagenic

# ntt Induces reproducibly with one of

### the concen"'lnltlons a mutation frequency that Is three times higher than the spontaneous mutation fre""quency in

### the experiment. The test ttem Is desslfied as mutagenic If

### there is a reproducible concentration-related Increase 

### ofthe mutation frequency. Such evaluation may

### be considenad also in tha case lhet a threefold increase

### ofthe mutant frequency Is not observed. In a case bycase evaluation this decision depends on the level 

### ofthe correspon"'Cling 

### solvent control data. 

## Stet/sties
Linear

### regression analysis (least squares)

. 

# Results and discussions Test results Speclealatraln Chinese hamster lung fibroblasts (V79) Metabolic with and without actiYIIIlon Genotoxiclly negative Cytotoxicity yes 

### at 500**~g/ml** (-59, both experiments), and>= 125**~g/ml** (+S9, 1st experiment) Vehicle yes controls valid Positive yes controls valid 

## Additionallnfonnation on results TEST-SPECIFIC CONFOUNDING FACTORS

- Effects of pH: 

### not effected

- Effects 

### ofosmolaltty: not Increased

- Watersolubility: soluble -Precipitation: precipttatlon at 250**~g/ml** and above after 4 hours treatmentwithout metabolic 

### activation. 

### at 125**~g/ml** and above after 4 hours treatment with metabolic activation. at 125**~g/ml** and above after24 hours treatmentwithout metabolic activation. -Otherconfounding effects: noneRANGE-FINDING/SCREENING STUDIES: 

### The concentration range of

### the pre-lestwas**llm~ed** by 

### the**solubil~** of

### the test item in waterand aqueous media. Test ttem concentrations between 3.9 and 500**~glmL** were 

### used to evaluate toxicity In the presence (4 h treatment) and absence (4 hand24 h treat-ment) ofmetabolic activation. Relevant cytotoxic effects were notedat250 and 500**~glmL** without metabolic activation. In the presence ofmetabolic activation no**cytotoxic~** oc-rurred upto 

### the rneximum concentration. 

### The last medium of

### the pre-experimentwas checked lorprecipitation at

### the end ofeach treatment period (4 or 24 hours)just priorto removal ofthe test**~em.** Precipitation ofthetest ttem visible to the unakled eye

### occurred in at 125**~glmL** and above lolowing 4 hourtreatment- and -outmetabolic activation. Following continuous traatrnent 

### lor24 hours wtlhout metabolic activation precipitation occurred at250**~glmL** and above.
COMPARISON WITH HISTORICAL CONTROL DATA: controls -in historical range
ADDITIONAL INFORMATION ON CYTOTOXICITY: Relevant cytotoxic effects defined as a relative doning efficiency I below 50% In both parallel cultures occurred at 500**~glmL** in both experiments wtthout metabolicactivation. In the presence of metabolic activation cytotoxic effeds as described abovewere noted at 125**~glmL** and above in

### the first experiment. 

## Any other Information on results Incl. tables Summary Table

relative relative mutant relative relative mutant
cone.**~g** 59 doning doning colonies/ 

### induction doning doning colonies/ 

### induction permL 

### mix efflciency I efficiency II 1o"ce11s lador efficiency I effidency II 106cells factor 

### % % 

### % % 

### column 3 

### 4 5 

### 6 8 9 10 Experiment I, 4 h treatment culture I culture II Solvent control with water 100.0 100.0 25.8 1.0 100.0 100.0 9.4 1.0 Positive control with EMS 150.0 116.4 85.6 133.3 5.2 102.3 79.9 110.0 11.7 Test Item 15.6 97.1 cuhure was not continued' 84.6 culture was not continued" Test item 31.3 71.3 105.4 14.5 0.6 90.1 82.9 15.8 1.7 Testttem 62.5 59.3 105.4 12.0 0.5 98.2 85.8 27.7 3.0 Testttem 125.0 59.4 103.2 11.9 0.5 70.8 76.4 28.7 3.1**Test~em** 250(p) 44.2 102.1 15.7 0.6 84.2 84.5 24.2 2.6TestKem SOO(p) 29.3 81.3 18.4 0.7 37.0 60.3 23.9 2.5

Solvent control wilh water 100.0 100.0 29.1 1.0 100.0 100.0 18.1 1.0 Positive control wilh DMBA 1.1 45.9 92.6 891.7 30.6 40.6 71.2 1421.6 78.6 Test**~em** 15.6 67.0 cunure was not

### contin~ 76.3 culture was not

### contin~ Test**~em** 31.3 70.8 85.1 13.3 0.5 67.6 90.6 17.8 1.0 Test**~em** 62.5 50.5 83.7 19.5 0.7 39.0 90.3 18.5 1.0 Test**~em** 125 (p) 44.0 95.1 13.5 0.5 36.2 88.9 10.6 0.6 Test**~em** 250 (p) 32.8 83.3 22.6 0.8 24.1 93.9 21.9 1.2**Test~em** 500(p) 39.9 107.4 23.3 0.8 24.8 72.5 18.5 1.0 Experiment II, 24 h treatment culture I cunurell Solvent control wilh water 100.0 100.0 17.8 1.0 100.0 100.0 15.8 1.0 Positive control wKh EMS 75.0 107.3 79.4 255.7 14.4 98.0 86.7 383.9 24.2 Test Hem 15.6 109.5 cunure was not

### contin~ 105.4 rutture was not contlnued11 Test Ham 31.3 109.4 83.2 22.9 1.3 111.6 85.9 20.1 1.3 Test item 62.5 94.9 62.9 19.1 1.1 96.3 71.5 35.7 2.3 TestKem 125.0 102.3 73.8 23.1 1.3 100.5 91.5 38.1 2.4 Tes!Kam 250 (p) 62.2 77.6 18.4 1.0 76.8 88.3 32.5 2.1 TestKam SOO(p) 16.9 83.2 12.4 0.7 

### 9.4 70.1 19.0 1.2 Experiment II, 4 h treatment Solvent control wilh water 100.0 100.0 15.2 1.0 100.0 100.0 11.1 1.0 PO&Kive control wilh DMBA 1.1 36.5 88.1 744.4 

### 48.9 24.2 91.0 841.6 75.6 Test**~em** 15.6 106.3 cunure was not

### contin~ 82.8 cullure was not**continued~** Test Hem 31.3 + 94.5 90.1 20.7 1.4 78.9 119.6 12.2 1.1 TestKem 62.5 + 82.2 85.1 15.3 1.0 77.1 100.3 18.0 1.6 Test Hem 125 (p) 78.5 92.8 12.6 0.8 

### 68.2 79.7 23.3 2.1Test Kern 

### 250 (p) 

### + 

### 79.9 89.8 16.9 1.1 54.2 

### 61.8 Test Kern 500 (p) 74.4 95.0 21.4 1.4 54.9 88.4 

## # culture was not continued since a minimum ofonly four analysable concentrations is required P

# = precipitation 

# Applicant's summaryand conclusion Conclusions Conclusion: 

### 22.3 12.1 

### 2.0 1.1

In conclusion itcan 

### be stated that underthe**ex~perimental** conditions reported thetest nem did not Induce gene mutations at the HPRT locus In V79 a>lls. Therefore, Leuco**Su~ur** Blacl< 1 

### is considered 

### to be non-mutagenic in this HPRT assay. 

## Executive summary 

## The test item Leuco Sulfur Black 1 was assessed for its potential to induce gene mutations atthe HPRT locus using V79 cells ofthe Chinese hamster. The study was performed in two independent experiments, using identical experimental procedures. In the first experiment the treatment period was 4 hours with and without metabolic activation. The second experimentwas performed with a treatment time of4 hours with and 24 hours without metabolic activation. Precipitation ofthe test item was observed at 125 J.lg/ml and above with and at 250 J.lg/ml and above without metabolic activation. Relevant cytotoxic effects defined as a relative cloning efficiency I below 50% in both parallel cultures occurred at 500 J.lg/ml in both experiments without metabolic activation. In the presence of metabolic activation cytotoxic effects as described above were noted at 125 J.lg/ml and above in the first experiment. No relevant and reproducible increase in mutant colony numbers/106

## cells was observed in the main experiments up to the maximum concentration. All mutantfrequencies remained well within the historical range ofsolvent controls. The induction factor was equal to or exceeded slightlythe threshold ofthree times the corresponding solvent control in the second culture ofexperiment I without metabolic activation at 62.5 and 125.0 J.lg/ml (induction factor of3.0 and 3.1, respectively). This marginal effectwasjudged as biologically irrelevant since it was based upon the rather low solvent control of9.4 mutant colonies/1

# o 6

## cells. Furthermore, the increase was not reproduced in the parallel culture under identical conditions and not dose dependent as indicated by a lacking statistical significance

## A linear regression analysis (least squares) was performed to assess a possible dose dependent increase ofmutant frequencies using SYSTATastatistics software. No significant dose dependent trend ofthe mutation frequency indicated by a probability value of<0.05 was determined in any ofthe experimental groups. In both experiments ofthis study (with and without S9 mix) the range ofthe solvent controls was from 9.4 up to 29.1 mutant colonies per 1

# o 6cells; the range ofthe groups treated with the test item was from 11.9 up to 38.1 mutants per 106cells. Ethylmethanesulfonate (0.15 mg/ml in experiment I and 0.075 mg/ml in experiment II) and DMBA (1.1 IJg/mL) were used as positive controls and showed a distinct increase in induced mutant colonies.

# Endpointstudyrecord: 1234902_CA79
UUID IUC5-8c54462c-9cd7-434c-be03-8d6abcd1da8a Dossier UUID IUC5-292149d1-a677-4feb-9692-11ad973281ea Author Date Remart<a thr I SCC GmbH IWendelsheim I Germany 2010-05-26 16:45:11 CEST


# Administrative Data Purpose flag Study result

### type key study; robust study summary 

### experimental resutt RellabiiHy 1 (reliable without restriction) Rllllonalefor reliability Incl. deficiencies According to OECD guideline 473 

# Data source Reference

Referenca Author 

### Year



### type study report HaiiC 2009


### Data access data submitter is data owner 

## Data protection claimed yes. butwilling to share Title
In Vitro Chromosome Aberration Test in Chinese HamsterV79 Cells 

### with Leuco Sulfur Black 1



# Materials and methods Type ofgenotoxicity Bibliographic Testing laboratory Report Owner Company Report 

### sourca 

### no. company study no. 

### date Harlan Cytotest Cell Research C31122 Clariant 1234902 20CMHl7- GmbH. Rossdorf (Garmany) 07chromosomeaberration 

## Type of

## study in vitro mammalian chromosome aberration test 

## Test guideline
Guideline OECD Guideline 473 (In llilro Mammalian Chromosome Aberration Test)

yes (inct. certificate) 

# Test materials Identity oftest material same as forsubstance defined In section 1 (If not read-across)
yes 

# Method Target gene Chromosome aberration 

## Species/strain Species/strain Chinese hamster lung fibroblasts (V79) 

### Details



### on mammalian



### cell lines

(If -llcloble) 

### Met8bollc actiYatlon Thawed stock cukures were propagated at 37 

### 'C in 

### 8 0 

### em' plastic flasks (Greiner, 72632 Fricl<enhausen, Germany). About 5 

### x 105cells 

### perflask were seeded into 15 

### mL ofMEM (Minimal Essential Medium; Invitrogen Gibco, 76131**Ka~sruhe,** Germany) supplemented 

### with 10%fetal 

### calfserum (FCS; Invitrogen Glbco, 76131**Ka~ruhe,** Germany). Additionally, the medium was supplemented 

### with 1 % 100x Penicilin/ Streptomycin solution (10.000 Unlls/ml PenicHiin, 10

### mg/ml Streptomycin; PAA Laboretories GmbH, 35091 COibe, Germany) and 1 %Amphotericin B-solution (250**~g/ml,** PAA Laboretories GmbH, 35091 COibe, Germany). 

### The cells were sub-cultured twice weekly. The cell cullureswere incubated at 37 

### 'C In a humidified atmosphere 

### with 1 .5 %carbon dioxide (98.5% air). -Periodically checl<ed for Mycoplasma contamination: yes/no

- Periodically checked for karyotype stability: yes/no 

### with and without 

### -bollc rat liver S9 

### 8CIIY811on system 

## TestconcentrationsWithout S9 mix: 19.5, 39.1, 78.1, 156.3, 312.5, 625, 1250,2500and 5000**~glml** 2.3, 4.7, 9.4, 18.8, 37.5, 75, 150 and 300**~glml** WothS9mlx: 19.5, 39.1, 78.1, 156.3, 312.5, 625, 1250, 2500and 5000**~glml** 2.3, 4.7, 9.4, 18.8, 37.5, 75, 150 and 300**~glml** 

### Vehicle

- Vehicle(s)/solvent(s) used: deionlsed water -Justification

### for choice ofsolwmt/vehicle: solubi6ty (OECD Guideline 473) 

### Controls**~no** 

### controls 

### Solvent I 

### yes (deionised water) 

### vehicle controls 

### True no 

### negative controls Positive yes controls Positive ethylmethanesulphonate (cylophosphamide) 

### control subelllnce 

### RIII1Uirks in tha absence orpresence of S9 

### Details on testsystem andconditions Two independentexperiments were performed. In Experiment I

### the exposure period was 4 hours with and without metabolic activation. In Experiment II 

### the exposure period was 4 hourswith S9 mixand 18 hours without S9 mix. Thechromosomes

### were prepared 18 hours afterstart oftreatmentwith the 

### test Kern. METHOD OF APPLICATION; in minimal essentila medium DURATION

- Exposure duration: 4 and 18 hours -Expression time (cells 

### in growth medium): 18 hours -Fixation time (start ofexposure up to fixation or harvestofcells): 18 hours
SELECTION AGENT (mutation assays): no selection agentSPINDLE INHIBITOR (cytogeneticassays): colcemid STAIN (for cytogenetic assays): Giemsa

NUMBER OF REPLICATIONS: 2 parallel cuKures in each experimental group
NUMBER OF CELLS EVALUATED: 100/cuKure
DETERMINATION OF CYTOTOXICITY -Method: mHotic index; cloning efficiency; relative total growth
OTHER EXAMINATIONS:

- Determination ofpolyploidy: no evidence ofan increase in the rate ofpolyploid cells

- Detennination ofendorepUcation: no evidence in the rate ofendomitotic cells

OTHER: none 

## Evaluation criteria Evaluation ofthe cultures was performed (according to standard protocol 

### ofthe "ArbeHsgruppa der lndustrie, Cytogenetik") using NIKON miaoscopas with 1OOx objectives. Breaks, fragments, deletions, exchanges, and chromosome disintegrations were recorded as structural chromosome aberrations. 

### Gaps- recorded as -II but not inciuded 

### in the calculation 

### ofthe aberration rates. 100 -•spread metaphases parcuKure -rescoredfor cytogenetic demage on

### coded slides. Only metaphases with characteristic chromosome numbers of 22 

### ± 1 were induded in the analysis. To describe a cytoloxic eflect

### the mHotic index (% cells in mitosis)was determined. In addition, the number ofpolyploid cells In 500 metaphases par cuKure was determined (% polyploid metaphases; in 

### the 

### casedthis aneuploid cell line polyploid means a near tetraploid karyotype). 

## Statistics Statistical significance was confirmed by means ofthe Fisher'sexact test (p < 0.05). 

## Any other Information on materials and methods Incl. tables 

## Phenobarbitallb-Naphthoflavone induced rat liver S9 was used as the metabolic activation system. The S9 was prepared from 8- 12 weeks old maleWistar rats (HsdCpb:WU, Harlan Laboratories GmbH, 33178 Borchen, Germany)weight approx. 220
- 320 g induced by intraperitoneal applications of 80 mg/kg b.w. Phenobarbital (Desitin; 22335 Hamburg, Germany) and by peroral administrations of80 mglkgb.w.b-Naphthoflavone(Sigma-Aidrich Chemie GmbH, 82024 Taufkirchen, Germany) each, on three consecutivedays. The livers were prepared 24 hours afterthe last treatment. The S9 fractions

## were produced 

## by dilution ofthe liver homogenate with a KCI solution (1 :3 parts)followed 

## by centrifugation at 9000 g. Aliquots ofthe supernatant were frozen and stored in ampoules at--80 

# 'C. Small numbers ofthe ampouleswere kept at20 

# 'C for up to one week. The protein concentration was 35.0 mg/ml (Lot no. 211108). 

# Results and discussions Test results SpeciH/atrllln Chinese hamster lung fibroblasts (V79) Metabolic with and without IICitvllllon

T•t 

### system Genotoxtclty**C~xlclty** Vehicle 

### controls valid an strains/cell types tested negative no. but tested up

### to precipitating concentrations yes



### Positive 

### yes controls 

### valid 

## AdditionalInformation on results Thetest item Leuoo Sulfur Black 1, suspended in deionised water, was assessed for

### its potential

### to induce structural 

### duomosome aberrations in V79 cells ofthe Chinese hamster in vitro in

### the absence and presence 

### of metabolic activation by 59 mix. Two independent experiments were performed. In Experiment I, the exposure period was 4 hours with and without metabolicactivation. In Experiment II the exposure period was 4 hourswith 59 mix and 18 hourswithout 59 mix. The chromosomes were prepared 18 hours afterstart oftreatment 

### with the- item. In 

### each experimental group two perellel cukureswere set up. 100 metaphases per cukure were evaluated for structure! chromosome aberrations. In both experiments no relevant influence of

### the test item on pH value orosmolarky was observed (Exp. 1: solvent control286 mOsm, pH 7.4 versus 285 mOsm and pH 7.4 at 5000**~g/ml:** Exp. II: solvent control284 mOsm, pH 7.4 versus 290 mOsm and pH 7.4 at 300**~glml).** Precipitation 

### ofthe test item in cukure medium was observed aftertreatmentwith 78.1**~g/ml** and above Inthe absence and presence of59 mix 

### in Experiment I and with 150**~g/ml** and above In 

### the absence and presence of59 mix in Experiment II. Nocleartoxic effacts indicated by reduced mHotic Indices or reduced cell numbers

### were observed aftertreatmentwith 

### thetest Hem (see Table 2, 3 and 6, pages 25 

### to 27 and 30). Due to strong test kern precipitation concentr&tions higherthan those examined could not be evaluated 

### for cytogenetic damage. In both experiments in 

### the absence and presence of59 mix no biologlcaUy relevant increase In 

### the number

### of cells carrying structural chromosome aberrations was observed (see Table 4, 5, 7 and 8, pages 28, 29, 31 and 32). The aberration rates ofthe cells aftertreatment with 

### the- Hem (0.0• 3.0 %aberrantcells. excluding geps) were dose

### to the range 

### ofthe solvent control values (0.5- 2.0 % aberrant ceHs. excluding gaps) and within the range 

### ofthe laboratOf}''s historical solvent control data (0.0- 4.0 %aberrant cells, excluding gaps). A single significant inaeasewas observed In Experiment II 

### in the prasence 

### of59 mix after treatment with 150**~g/ml.** Although this increase 

### of 3.0 % aberrant cells excluding gapswas statistically significant compared 

### to the low response (0.5% aberrant cells) in the solvent control data, the response is within 

### the historical control 

### data range (0.0- 4.0% aberrant cells, excluding gaps). Therefore, this observation has

### to be regarded as biologlcaly Irrelevant. No evidence ofan increase in polyploid melaphaseswas noticed aftertreatment with the test item as compared to the controls. In both experiments, either EMS (400 or 1000**~g/ml)** or CPA (1.4**~glmL)** were used as positive controls and showed distinct inaeases in the number

### ofcells with structural chromosome aberrations. In conclusion, Hcan be stated that underthe experimental 

### condHions reported, the test item Leuco Sulfur Black 1did not induce structural chromosome aberrations in V79 cells (Chinese hamstereel line) when tested up to the highestevaluable concentrations. 

## Any other Information on results lncl- tables 

## Summary of results ofthe chromosome aberration studywith Leuco Sulfur Black 1 Exp. Preparation Test item Cell numbers Mitotic indices Aberrant celts interval concentration in% in% 

### in% in !Jg/ml ofcontrol ofcontrol incl. gaps• exd. gaps• with excha es exposure 

### period 4 hrs without 59 mix 

### 18 hrs Solvent control1 

### 100.0 100.0 1.5 1.0 0.0 Positive control2 n.t. 

### 89.5 14.0 14.05 

### 4.0 19.5 85.0 87.5 1.5 1.5 1.0 39.1 110.8 95.0 2.5 2.5 0.5 78.1p 100.2 107.0 2.0 2.0 0.0 Exposure 

### period 18 hrs without 59 mix 

### 18 hrs Solvent control1 

### 100.0 100.0 1.0 1.0 0.0 Positive control3 n.t. 

### 70.5 13.0 11.55 

### 3.0 37.5 97.5 96.9 0.0 0.0 0.0 75.0 82.2 90.3 2.5 2.0 0.0 15o.oP 77.0 112.8 1.5 1.5 0.5 Exposure 

### period 4 hrswith 59 mix 

### 18 hrs Solventcontrol 1 

### 100.0 100.0 2.0 2.0 0.0

### Positive controt4 n.t. 89.2 10.5 

### 10.01 

### 19.5 79.4 113.5 2.0 1.0 39.1 113.8 120.0 0.0 0.0 78.1p 90.3 93.0 2.5 2.0 18hrs 

### Solvent control1 100.0 100.0 0.5 0.5 

### Positive control4 n.t. 87.0 15.5 

### 13.55 37.5 96.4 104.0 2.5 2.5 75.0 93.1 106.0 0.5 0.5 1so.oP 84.1 113.0 3.5 

### 3.01 

### Including cells carrying exchanges
n.t. Nottested p Precipitation occurred


# s Aberration frequency statistically significant higherthan corresponding control values
Deionised water 10.0% (v/v)

2 

### EMS1000.0 

### IJg/ml
EMS400.0 

### IJg/mL

4 

### CPA 1.4 j.Jg/ml 

# Applicant's summary and conclusion Conclusions 2.5 0.0 0.0 0.0 0.0 2.5 1.0 0.0 0.5In conclusion,**~can** be stated that underthe experimental conditions reported, thetest item did not induce structural chromosome aberrations in 

### V79 eels (Chinese hamstercell Hne) in vitro. Theralore, Leuco Sulfur Black 1 is considerad to be non-dastogenic in this chromosome aberration test in the absence and presence ofmetabolicactivatiOn, when -ed upto the highestevaluable concentrations. 

### Executive summary The test item Leuco Sulfur Black 1, suspended in deionised water, was assessed for its potential to induce structural chromosome aberrations inV79cells ofthe Chinese hamsterin vitroin two independent experiments. In each experimental group two paraUel cullunss

## were set up. 100 metaphases per cuKure wens evaluated lorstructural chromosome aberrations. 

### The highest applied concentration in lhe pre-teston toxicity (5000**~g/ml)** wes chosen with respeclto lhe curnsnl OECD Guideline473. Dose seleclion

### for the cytogenetic experiments was performed considering the toxicity dataand lhe occurnsnce ofprecipitation. 

### The chosen traatment concentrations

### were between 2.3 and 300**~g/ml** (with and without S9). In lhe absence and pnssence ofS9 mix no cytotoxicity wasobserved upto lhe highest evaluable concentration. Due to strong last**~ern** pnscip- concentrations higher lhan lhose examined could not be evaluated 

### for cytogenetic damege. Noclastogenicily was observed atlhe concentrations evaluated eKher with orwKhoul metabolic activation. However, in Experiment II in the presenceofS9 mix a singlestatisticaHy significant increase in the numberofaberrant cells excluding gaps (3.0 %) was observed atlhe highest evaluable concentration (150**~ghnl).** Thisvalua was clearly in lhe laboratory's historical solvent control data range 

### (0.0- 4.0% aberrant cells excluding gaps)and lslhensfore 

### nsgarded as biologically Irrelevant. No evidence ofan increase in polyploid metaphases was**~** aftertraatrnenlwith the test Kem as comparad tothe control cullunss. Appropriate mutagens were used as positive controls. They induced statisticaly significant increases (p < 0.05) in cells with structural chromosome aberrations.

# 7.7 Carcinogenicity Endpointsummary: Carcinogenicity UUID IUC5-9303f711-1e0d-4db9-9899-c0cb658f3d71 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author thr 

### I SCC GmbH 

### IWendelsheim 

### I Germany Date 2010-05-26 16:48:23 CEST Remarks 

# Administrative Data Discussion There is no carcinogenicity study available. However, there is no concernregarding carcinogenicity because Leuco Sulfur Black I is not genotoxic and the mean molecularweight of8000

- 9000 Da does notindicate significantabsorption. 

# Justification for classification or non-classification Based on the available information Leuco Sulfur Black 1 is notsubject to classification and labelling according to Directive 67/548/EECand Regulation 127212008/EC.

# 7.8 Toxicityto reproduction Endpointsummary: Toxicity to reproduction
UUID IUC5-d90b0564-9095-4680-af3b-faed4el68e8e Dossier UUID IUCS-292149d1-a6n -4feb-9692-1fad973281ea Author lhr

### I SCC 

### GmbH 

### I Wendelsheim 

### I Germany Da1e 2010-05-26 16:49:10 CEST Remarks 

# Administrative Data Effects on fertility Short description ofkey information
Leuao Sulfur Black 1did not cause any toxic effects in 

### lhe repeated dose toxicity study wilh reproduction/developmental toxicity saeening in rats (OECD 

### 422). The llmH dose 

### of 1000 IJ19/kg bwld was derived as NOAEL for general parental toxicity and reproductive/developmental toxicity. 

# Key value for chemical safety assessment
Effect NOAEL 1000 mg/kg bwlday level



### lor fertility 

### (01"111 exposurs) 

# Discussion In a repeated dose toxicity study with reproduction/developmental toxicity screening according to OECD guideline 422 and GLP Leuco Sulfur Black 1 was administered by gavage to groups of 10 rats per sex and dose at 0, 100, 300 and 1000 mg/kg bw/d. Male rats were treated for 29 days, female rats for 14 days priorto pairing, though the pairing and gestation periods until the F1 generation had reached day 4 postpartum. The test item was administered in water asvehicle at a

## dose volume of 10 ml/kg bw/d. Control animals were doses with the vehicle alone. Observation and examinations consisted of mortality, clinical signs, detailed clinical observations, food consumption, body weight, functional observation battery, clinical biochemistry, haematology, urinalysis, macroscopical examination at necropsy, organ weights and histopathological examination oforgans. Reproductive parameters examined were examination of reproductive organs with special emphasis on the stages of spermatogenesis and histopathology of interstitial cell structure, mating performance and fertility, duration ofgestation, implantation sites, pre- and postimplantation loss, corpora lutea count. Parameters relevant for developmental toxicity consisted oflitter size, numberoflive and dead pups, sex ratio, pup weight atday4 post partum, macroscopical examination of pups attermination. The only effect seen was a dark discolouration offaeces in all high dose animals starting on day 3 ofthe study and lasting until termination. Based on this finding a NOAEL of 1000 rng/kg bw/d was derived for general toxicity aswell asfor reproductive and developmental toxicity. The discolouration offaeces was the only effect observed in this study and also in the acute oral toxicity study. Organs or urine were not discoloured and signs ofsubstance deposits were notfound in histopathological analysis ofthe organs. This indicates that Leuco Sulfur Black 1 is directly excreted and that absorption in the gastrointestinal tract does not occur following oral administration. The mean molecularweight and number-average molecularweight (Mn) of Leuco Sulfur Black 1 is in the range of8000- 9000 Da and about 3000 Da, respectively. It is very poorly or even notsoluble in common polar and non-polar solvents such as water (solubility of62.4 mg/L), DMSO, methanol, DMF, THF, chloroform and notsoluble in acid likes sulfuric acid and trifluoroacetic acid. Thevapour pressure ofthe pure substance, calculated from the boiling point of

# >400 

# 'C, is 1.8 x 10E-5 hPa at25 

# 'C. These substance properties supported bythe results oforal toxicity studies demonstrate thatthe absorption of Leuco Sulfur Black 1 via the oral, dermal or inhalation route and, consequently, systemic bioavailability is negligible. 

# Based on the substance properties and the fact that no toxicity was seen in any ofthe tests it is concludedthat exposure to Leuco SulfurBlack I does not raise any health concern. In line with Regulation (EC) No 1907/2006Annex XI and taking into account the REACH paradigm thatnew tests on vertebrates shall onlybe conducted orproposed as a lastresortwhen all otherdata sources have been exhausted, further animals studies to examine the reproductive anddevelopmental toxicity ofLeuco Sulfur Black 1are not

## necessary and should notbe performed. 

# Developmental toxicity 

# Iteratogeniticy Short description of key information
Leuco Sulfur Black 1 did not cause any toxic effects in 

### the repeated 

### dose toxicity studywith reproduction/developmental toxicity saeening In rets (OECD 422). The limH 

### doseof 1000 rnglkg bw/d was derived as NOAEL for general perental toxicity and reproductive/developmental toxidty. 

## Discussion
see "effects on fertility" 

# Justification for classification or non-classification Based on the available data Leuco Sulfur Black 1 is not subjectto classification and labelling according to Directive 67/548/EECand Regulation 1272/2008/EC.

# 7.8.1 Toxicity to reproduction Endpointstudyrecord: Toxicityto 

# reproduction.001 OECD 422 Leuco SulfurBlack 1 C31087
UUID IUC5-82c86a1a-8126-46be-b0df-<lc7ab37c29c5 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks uensenba I Clariant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Garmany 2010-05-21 14:10:47 CEST



# Administrative Data Purpose ftag Study resulttype RetlabiiHy key study; robust study summary experimental result 1 (relable without restriction) Study period

Rationale for rallabiiHy This study has been performed in compliance with 

### the: Incl. deficiencies Between 20 May 2009 and 23 March 2010

Swiss Ordinance relating to 

### Good Laboratory Practice adopted May 18th, 2005 (SR 813.112.1). This Ordinance is based on the OECD Principles ofGood Laboralory Practice, as revised in 1997 and adopted on November26th, 1997 bydecision oftha OECD Councii(C (97)186/Final].



# Data source Reference

Rafentnce Author



### type study report Ceccatalli R 

### Data access These principles are compatible with Good Laboratory Practice regulations specified by regulatory authorities throughout

### the European Community, the United States (EPA and FDA), and Japan (MHLW, MAFF and METI). There were no circumstances 

### the! may have affected the quality or integrity ofthe data.

Year Title Bibliographic Testing laboratory Report OWner Company Report source 

### no. company study no. date 2010 Combined Repeatad Dose Toxicity Studywith 

### the Harlan Laboratories ltd., C31087 Clarlant 2011).. Reproduction/Developmental Toxicity Screening WOI!erstrasse 4 4414 03-22 Test in 

### the Han Wistar

### Ret FOIIinsdorf, Switzerianddata submitter is data owner 

# Materials and methods Testtype saeening 

## Limittest

no 

## Test guideline

Qualifier 

### Guideline according 

### to OECD Guideline 422 (Combined Repeated Dose Toxicity Study with tha Reproduction 

### I Developmental Toxicity Screening Test) according

### to otharguideline: EPA OPPTS 870.3650 (Combined Repeated 

### Dose Toxicity Studywith tha Reproduction/Developmental Toxicity Screening Test) 

## GLP compliance yes (Ind. certificate) 

# Test materials Identity oftest material same as for substance defined in section 1 (if

## not read-across)
yes 

## Test material identity
Identifier Identity Common name Leuco Sulfur Black 1 CAS number 66241-11-0 

## Deteils ontest material Description: Thixotrophic, black, wet peste Batch Number: ESA0007585 Composition: Leuco SuWur Black 1: 52.75% (calculated by amount ofby-<:ompounds) 2,4-Dinitrochlorobenzene: 

### <20 ppm 2,4-Dinitrophenole: < 20 ppm Sulfate: 970 ppm Chloride: 750 ppm 

### Devlmlons
no 

### noSodium: 1.3% (wlw) Water. 45.75% (wlw) by Karl Fischertitration Stability ofTest Hem: Stable under

### storage conditions Stability 

### of Test Item in Vehide: Seven days 

### Expiry Date (Retest Date): 11-Jul-2011 Storage Conditions: At room temperature (20 :t 5 'C)•light proteded Safety Preeautions: Routine hygienic procedures (gloves, goggles, face mask). 

## Test animals Species
rat 

### Strain Wistar 

### Sex male/female 

### Details on test animals andenvironmentalconditions Animals: Rat, HanRcc: WIST(SPF) Rationale: Recognized 

### by internationalguidallnes as a recommended test system. Breeder. Harlan Laboratories Ltd., LaboratoryAnimal Services, WOiferstrassa 4, 4414 FOIIInsdorf, Switzerland Number

### ofAnimals: 40 males (10 pergroup), 40females (10 pergroup) Age (at Start 

### ofTreatment): 11 weeks Body Weight Range (at Start 

### of Treatment): Males: 290 to 324 g. Females: 284 to 212 g Identification of Prental Animals: Cage card and individual animal number (eartattoo). Identification 

### of Pups: On day 1 post partum. pupswere Individually tattooed with Indian ink. Randomization: Computer-generated random algorithm. In addition body weights (recorded on the day

### ofallocation)weretaken into consideration in ordertoensure similar mean body weights In all groups. Acclimatization: Undertest condftions after health examination. Onlyanimals without any visible signs

### of Illness were used for

### the study. Conditions: Standard laboratory conditions. Air-conditioned 

### with 10- 15 air changes per hour. continuously monitored environmental conditions (temp. range: 22 :t 3 

### 'C; relative humidfty range: 37.5- 60.9%). Therewas 12 hourfluorescent light 112-hourdark cyde

### with music durlng 

### the lightperiod. Accommodation: Individually in Makrolon type-3 cages 

### with wire 

### mesh 

### tops and stenllzed standard softwood bedding ('Lignocei'J. Rettenmaier & SOhne GmbH & CoKG, 73494 Rosanberg 

### I Ge111111ny, Imported by Provimi Klibe SA, 4303 Kalseraugst 

### I SWitzerland). During the pre-pairing period, cageswith males were interspersed amongstthose holding females to promote the davelopment

### of regular eslrus cydes. Oiet: PeHetad standard Kiiba Nafeg 3433

### rodant maintenancediet (Provimi Kllba SA. 4303 Kaiseraugst I Switzerland) was available ad libftum (batch no. 15109). Water: Communfty tap-waterfrom FOIIinsdorfwas available ad iibftum In water bottles. 

## Administration 

# Iexposure Route ofadministrationoral: gavage 

### Vehicle
water 

### Details on exposure Dose Fonnulations The 

### dose formulatlons were prepared weekly using 

### the test item as supplied bythe Sponsor. Leuco Sulfur Black 1 was weighed Intoa glass beaker on a tered precision balance 

### and approximately 80% 

### ofthe vehicle was 

### added (w/v). Using a magnetic stirrer, a 

### homogeneous suspension was prepared. Having obtained a homogeneous mixture, 

### the remaining vehicle was 

### added. Separateformulations were prepared 

### for each concentration. Homogeneity ofthe test item in 

### the vehicle was maintained during the daily administration period using a magneticstirrer. Storaga of Dose Fonnulations Dose formulations were stored at room temperature (20 

### t 5 

### 'C) 

### in brown glass beakers. Based upon 

### the results 

### ofstablfity analysas performad within the Harlan Laboratories study no. C31076, dose formulations were stable 

### for at least one week. Treatment Method: Oral, by gavage Rationale for Method: Administration by gavaga is a common and accepted route ofexposure for studies ofthis type. Target

### Dose Levels: Group 1: 0 rngJl<g bwlday (control group) Group 2: 100 mglkg bwlday Group 3: 300 mglkg bw/day Group 4: 1000 mg/kg bwlday Rationale for Dose 

### lave! Selection: The dose levels were selected based on a previous dose renga finding toxicity study in Han Wistar Rats, Harlan Laboratories Study C31076, using dose levels of 116, 348 and 1160 rngJl<glday adjusted for 52.75% Leuco Sulfur Black 1 dye stuffin the product and corresponding to 220, 680 and 2200 mglkg bwlday 

### of product. 

### The NOAEL ofthis study resulted to ba 1160

### mg dye stuff/kg bw/day or 2200 

### mg product/kg bwlday. Dose Voluma: 10 mllkg body weight


### Details on mating procedure During 

### the pairing period, females were housed with sexually mature males (1 :1) in special automatic mating cages i.e. with synchronized timing to initiate the nightly mating period, until evidence ofcopulation was observed. This system reduced the variation in 

### the copulation times of

### the differentfemales. The females were removed and housed individually 

### W: -the daily vaginal smaarwas sperm positive, or -a copulation plug was 

### observed. The day

### of mating was designated day 0 post coitum. All damswera allowed togive birth and reartheir Hlters (F1 pups) up to day 4 postpartum. Day 0 was designated as the day on which a female hed delivered all her pups.

### Analytical verification ofdoses or concentrations yes 

## Details on analytical verification ofdoses 

## orconcentrations Analysis of

### Dose Formulations On lhefirst treatment day duplicate samples

### from lhe control group aswell as three duplicate samples (top, middle and bottom) ofaboul2 g ofeach concentration

### were taken prior

### to dosingfor analysis ofconcentration and homogeneity. Samplesofabout2 g ofeach concentration weretakenfrom lhe middle only

### to conflnn stability (4

## tvs and 7 days). During lhe last week of lhetreatment, samples weretaken

### from lhe middle to confirm concentration. 

### The aliquots for analysis ofdose formulations were transferred intotightly closed vessels protected againstoxygen. One set ofsamples was 

### delivered atambienttemperature to 

### Dr. K. Morgenlhel (Harten Laboratories Ltd., ltingen 

### I Switzerland) and slorad lhere 

### at roomtemperature (20

### ±5 'C) until analysis. The remaining 

### sat ofsa mples was stored atlhe facility at room temperature (20

### ±5 'C) in case ofrepeated analysis. Analysis ofhomogeneity and concentration was performed gravimetrically byweighing lhe residue after drying by lyophilization. Stability ofdosa formulations was confinned by IRspectroscopy. The application formulations investigated during lha studywere found 

### to comprise Leuco Sulfur Black 1 In lhe range of81.7% 

### to 102.5%. The homogeneous distribution of Leuco Sulfur Black 1 in lhe preparations was approved because individual recoveries found did 

### not daviate more lhan 14.2% (<15%) from lha corresponding mean. In 

### addilion,lhe

### test item was found to 

### be stable in epplicetion formulations when kepifor 4 hoursandfor 7 days understorage condKions due to similar IR spectra compared to lhe reference spedrum

### obtained 

### from lhe test Hem. Thus, idantity oflhe test Kem In application formulations was confinned. In conclusion, lhe results indicate lhe accurate use oflha test item Leuco Sulfur Black 1 end highly purified water as vehicle during lhis study. 

### Duration oftreatment 

## Iexposure Males: 29days Females: Approximately 7 weeks 

### Frequency oftreatment DeUy 

### Details on study schedule Males Acclimatization: 7 days First Test Item Administration : Day 1 of pre-pairing Pre-Pairing: 14 days Pairing: 14 days maximum Treatment Ends: On day before sacrifice Necropsy: Aftertreatment of at least 28 days, when no longer needed

### for assessment of repro-ductive effects Females Acclimatization: 7 days FirstTest Item Administration: Day 1 ofpre-pairing Pre-Pairing: 14 daysPairing: 14 days maximum Gestation: Approximately 21 days Treatment Ends: On day 4 postpartum Necropsy:On day 5 postpartum (pups on day4 post par1um) 

### Doses I concentrations 100 rng/kg bw/day Baals actual ingested 300 mg/kg bw/day Baals actual ingested 1000 mg/kg bw/day Baals actual ingested 

### No. ofanimals persex perdose 10 

### Control animals yes, concurrent vehide 

### Furtherdetails on studydesign -Dose salectlon rationale: -Rationale for animal assignment (if not random): 

### -Other: 

### Positive control Not requirad. 

## Examinations Parentalanimals: Observations andexaminations VIABILITY I

### MORTALITY Twice daily CLINICAL SIGNS Dally cage-side dlnical observations (once daily, during acclimatization and up to day ofnecropsy). Additionally females were obsarved for signs 

### ofdifficutt or prolonged parturition, and behavioral abnormalities in nesting and nursing.FOOD CONSUMPTION Males: Weekly during pre-pairing and after pairing periods Females: Pre-pairing period days 1-8and 8 -14; gestation days 0-7, 7- 14 and 14- 21 post coilum, and days 1-4 post parium. Nofood consumption was recorded during the pairing period. BODY WEIGHTS Recorded daily from treatment start to day ofnecropsy. DETAILED CLINICAL OBSERVATIONS Once prior to the first administration ofthe test Item andweekly thereafter, detailed clinical observations were performed 

### outside the home cage. Animals were observed for the following: changes in skin, fur, eyes, mucous membranes, ocrurrence ofsecretions and excretions, and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory panem). Changes in gall, posture and response to handling as well as the prasence 

### of clonic ortonic movements, stereotypies or bizarre behavior were also reported. FUNCTIONAL OBSERVATION BATERY Atana time during the study (males shortly before the scheduled 

### sacrifice and females on day 3 or4 postpartum) relevant parameters were performed wllh five P generation males and five P generation females fmm each group. This FOB assessment was conducted following the daily dose administretion. Animals were observed for the following: -Cage-side observations: unusual body movements (e.g. tremors, convulsions), abnonnai behavior (e.g. drciing, stereotypy) and posture as well as resistance to removal -Hand-held observations: palpebral dosure, pinna reflex, lacrimation, pupil 

### size, pupil reactivity, salivation, muscletons, extensor thrust response, righting reflex and reaction to handling. -Open field observations: level 

### of ambulatory activlly induding rearing (one minute evaluation), responsiveness to sharp noise, paw pinch, gait evaluation, quantity of urine and fecal pellets 

### voided.

- Categorical observations (can be made any time during 

### the FOB): hair

### coat, behavior, respiration, musde movements, eyes, hearing abilily (Preyer's reflex), urine or

### feces, soling, generei abnormalities, posture. -Measurements 

### 1Counts: hind limb

### Ifore limb grip strength, landingfoot splay, redal temperature. Any abnormal findings were 

### recorded and, where appropriate, graded in severity. Addllionaily, locomotor activilywas measured quantitativelyforthe same animals. Activity was measured with an Activlly MonitorAMS-0151 (FMI, Germany). Activily ofthe animals (based on beam count) was recorded for 6-minute intervals over a period 

### of 30 minutes. These data and the total activity over 

### 30 minutes were reported. CLINICAL LABORATORY INVESTIGATIONS Blood and urine samples were obtained on the day of the scheduled necropsyfrom 5 males fmm each group. Males were kept in metabolism cage and fasted overnight (but allowed access to 

### water ad libllum) for approximately 18 hours. Blood samples from 51actatlng females fmm each groupwere obtained on day 5post parium. The females were tested for approximately 18-20 hours before blood sampling but allowed 

## access to water ad libitum. Blood samples were drawn sublingually from all animals under Nght isoflurane anesthesia. The samples were colieded early in 

### the working dayto reduce biological variation caused by circadian rhythms. HEMATOLOGY 

### The following hematology parameters were determined: Complete Blood Cell Count: Erythrocyte count Reticulocyte Count Hemoglobin Hematocrit 

### Mean corpuscular volume

### Red cell volume distribution width 

### Mean corpuscular hemoglobin Mean corpuscular hemoglobin concentration Hamoglobin 

### concentration dislribution wldlh Leukocyte count, 

### lolal Differential leukocyte count 

### Plalelet count Coagulation: Prothrombin time(=

### Thromboplastin time) Activaled partial Thromboplastin time CLINICAL BIOCHEMISTRY 

### The following dinical biochemistry paramelers were determined: Glucose Urea Creatinine Bilirubin, tolal Cholesterol, total Triglycerides Aspartate aminotransferase 

### Alanine aminotransferase Alkaline phosphalase 

### Gamma-glulamyl-transferase Bile 

### acids Sodium Potassium 

### Chloride calcium 

### Phosphorus Protein, tolal Albumin 

### Globulin Albumin/Globulin ratio URINE ANALYSIS 

### The following urine parameters were determined: Relative density colour Appearance pH Protein 

### Glucose Nitrite KetonesUrobilinogen Bilirubin Erytrocytes Leukocytes Volume (18 hours) 

### Estrous cyclicity(Parentalanimals) Not reported. 

### Spermparameters (Parentalanimals)
tes1es weight, epididymes weight, stages ofspermatogenesis 

### Litterobservations The litterswere examined for littersize, live births, sbll births and any gross anomalies. The sex ratio ofthe pups was recorded. Pups wereweighed individually (without identificatiion) 

### on days

### 0 (n possible), 1 and 

### 4 postpartum. 

### Postmortem examinations (Parentalanimals) TERMINATION OF THE STUDY Males were 

### sacrificed aftertreatment of

### 29 days, when no longer needed forthe assessmen1 of reproductive effects. Dams were sacrificed 

### on day

### 5 postpartum. 

### When birth did 

### not 

### occur on the expected date (day 21 post coltum), the dam was 

### secrificed on day26 post collum. NECROPSY All animals were 

### sacrificed by an injecbon ofsodium pentobarbital. All P generabon animals were exsanguinated. All parent animals were examined macroscopicallyfor any structural changes, efther atthe scheduled necropsy or during the study

# n death occurred. Forthe parent animals, special attentionwas directed at the organs ofthe reproducbve system. The numberofimplantation sftes and 

### corpora lutea was recorded for all damswith litters. 

### The uteri of non-pregnantfemales were placed In a solution ofammonium sulfide to visualize possible hemorrhagicareas ofimplantation sites. ORGAN WEIGHTS The testes and epididymides ofaM parental males were weighed as pairs. In addftion, from 

### 5 mates and females killed 

### at the end ofthe studywhich were seiected from each group, the following organs were trimmed from any adherenttissue, as appropriate, andtheirwetweight taken. Adrenal glands (weighed as pairs) 

### Brain Heart Kidneys (weighed as pairs) Liver Thymus Spleen TISSUE PRESERVATION

### Thefollowing tissues from 

### al parental males were preserved in neutral phosphate buffered 4% 

### formaldehyde solution: Prostate 

### Seminal 

### vesicles with coagulating gland Testes (in Bouin'sfixative) Epididymides (in 

### Bouin'sfixative) 

### Thefollowing tissues from 

### all parental females were preserved in neutral phosphate buffered 4% 

### formaldehyde 

### solution: Ovaries In addition, from 

### the 

### five males and females per group selected for organ weights and 

### from all animals fOL<Id 

### deed or killed in extremis, the following tissues were preserved in neutral 

### phosphate buffered 4% 

### formaldehyde solution: 

### Gross lesions Brain 

### Spinal chonl SmaN and large 

### intestines (incl. Payer's patches) Stomach 

### Liver Kidneys Adrenals 

### Spleen Heart Thymus Thyroids, and parathyroids W possible Trachea and lungs (preserved by inflation with fixative and then immersion) Uterus 

### (with vagina) Urinary bladder 

### Lymph nodes (mesentarial, mandibular) Paripheral nerve (sciatic) 

### Bone marrow HISTOTECHNIQUE All 

### 0f11811 andtissue samples to 

### beexamined by the principal investigatorwere processed, embedded and 

### cut at an approximate thickness of2 - 4 micrometers and stained with hematoxylin and eosin. Addltlonaly, the testis was stained by 

### PAS-hematoxylin. HISTOPATHOLOGY Slides ofall organs and tissues 

### cotlectad at terminal sacrifice 

### from tha animals 

### ofthe control 

### and high-dose groups were examined by

### the principal investigator. The same applied to all 

### occurring gross lesions. Special emphasis was made 

### on tha stages of spermatogenesis and histopathology of Interstitial cell structure. Histological examination 

### ofovaries was carried 

### out on females that did not give birth.



### Postmortem examinations (Offspring)TERMINATION OF THE STUDY Pups were 

### sacrificed on day 4 post pariUm. NECROPSY All animals were 

### seafficed by an Injection ofsodium pentobarbital. 

### Dead pups, except lhose excessively cannibalized, -reexamined macroscopically. All pups were examined macroscopically for any structural changes, either at

### lhe scheduled necropsy orduring the study 

### W death occurred. 

### Statistics The following statistical melhods -reusedto analyze 

### food consumption, body -lghts, organ -ight,locomotoractivity and macroscopical findings and reproduction data: -Means and standard deviations ofvarious data were calculated. 

### -The Dunnell-test (many to 

### one t-test) based on a pooled variance estimate was applied**~** lhe variables could ba assumed 

### to follow a normal distribution for lhe comparison of

### lhe treated groups and 

### lhe control groups 

### for each sex. 

### -The Steel-test (many-one rank test) was applied instead oflhe Dunnell-testwhen the data could not 

### be assumed to follow a no""aldistribution.

- Fisher's exact-test was applied**~** lhevariables could ba dichotomized without loss of information. 

### Reproductive Indices From 

### lhe**on~ine** recorded reproduction data, the following parameters were calculated: fertility indices, mean precoltal time, post-implantation losses, mean litter size, pup sex ratios. 

### Offspring viabilityIndices From lhe on4ine recorded reproduction data. the following parameters were calculated: deed I ive pups at first litter check, postnatal loss (up 

### to day 

### 4 post parium). 

# Results and discussions Effect levels Endpoint Generation Sex Effect level


### NOAEL p male/female >= 

### 1000 mg/kg bw/day Based 

### Baals for effect level/ Remarks on With the exception for a darl< discoloration offeces obse!Ved 

### In all animals treated with 

### lhetest Hem atthe dose level 

### of 

### 1000 mglkg body -ightlday, no olhertest 

### item-related 

### ellects -reobserved during lhe study. 

### The discoloration of 

### feces was considered 

### to ba duetothe staining property 

### oflhe test Hem and not to 

### beadverse. 

# Results ofexam.nations: parental animals Clinicalsigns (parental animals) yes 

### Bodyweightand

## food consumption (parentalanimals)
no-cts

### Reproductive function: estrous cycle (parental animals)
not examined 

### Reproductive function: sperm measures (parental animals) not examined 

### Reproductiveperformance (parental animals)

no 

### elfecls 

### Organ weights (parentalanimals)

no

### elfecls 

### Gross pathology(parentalanimals)

no effects 

### Histopathology(parentalanimals)

no

### elfecls 

### Details 

### on results (parentalanimals)
PARENT ANIMALS Delalled Clinical Observations (Dally) 

### All animals survived until 

### the scheduled necropsy. In group

### 4, dar1< discoloration offeces was observed in males and females slarting on day 

### 3 of

### the pre-pairing period onwards. 

### This was likely due

### to the slainlng property ofthetest llem al higher concentration. In groups

### 2 and 

### 3, 

### no clinical signs were observed forthe entire duration ofthe study. (See attached Tables on 

### pp. t to3) Delalled Clinical Observations (Weekly) Nofindings were obsarved during the weekly examination. Functional Observational Battery 

### None of

### the parameters under investigation during the functional observational battery gave an indication ofa last ileiTHBiated effect. 

### The frequency of rearing numberand incidence of urine puddles during the opan fiald observation did notgive indicalion ofany test llern-related effect. Observational as spontaneous vocalization when the ratwas removed 

### from the cage was considered to be ofincidenlal nature.Mean values ofgrip strength (fore- and hind paws)and landing foot splay gave no indication 

### of any test Kern-related effect. Bodytemperature in maleswas statistically significantly lower in group4 compared to the control group (37.9 

### 'C compared to 38.7 

### 'C in the control grou p). However, this value was within the range 

### ofthe historical control data and therefore was

### not considered to be a test Ham-related effect. (See attached Tableson pp. 4 to 7 and attached Historical Control Data on pp. 3 to 14) LooomotorActivity Looomotor activtly wasassessed quantitatively in terms of low beam counts in activtly monitorand did

### not give any indication ofa test item-related effect. Food Consumption
- Males
- Pre-pairing and After Pairing Periods Mean food consumption was not considered to be affected bythe treatment with thetest Kern. During the pre-pairing period, the lowerfood consumption observed in groups 2 and 3 (-6.2 and -4.6% compared to the control group) was considered to be incidental sincethis reduction did notfollow a dOSIHlependent pattam and in group4 mean 

### food consumption was similarto the control (-0.4% compared to the control). During the after pairing period, mean food consumption compared tothe control was: +2.2, -1.1 and +1.5%, in groups 2, 3 and 4, respectively (See attached Tabies on pp. 8 to 11) Body Weights
- Males- Pre-pairing, Pairing and After Pairing Periods Mean body weight and mean body weight gain were noteffected bythe treatment with thetest Hem for the whole duration 

### of the study. The stetistically significantly lowerbody weight gain observed in group 2 between day 4 and 7 was considered to be incidental sincethere was no dose-dependency. In order ofascending dose level, bodyweight gain was: during the pre-pairing period +13.0%, +10.9%, +11.5 and +11.9%, during the pairing period +4.9%, +5.7%, +4.4% and +4.7%, and during the after pairing period +4.1%, +5.6%, +4.8% and +4.5%, respectively. (See attached Tables on pp. 12 to20) Food Consumption -Females- Pre-pairing, Gestation and Lactation Pariods Notest Hem-related effects were observed In mean food consumption for the entire duration ofthe study. Mean

### food consumption In groups 2, 3 and 4, compared to the control group, was: during the pre-pairing period +5.0%, +2.2% end +2.2%, during the gestation pariod +0.4%, -2.1% and -0.4%, and during the lactation period +11.3%, -3.1% and -3.7%, respectively. (See attached Tableson pp. 21 

### to 26) BodyWeights- Females -Pre-pairing, Pairing, Gestation and Lactation Periods No test item-related effects were noted In mean body weight and mean bodyweight gain

### of females during the whole study at any dose level. in order ofascending dose level, body weight gain was: during the pre-pairing pariod +7.5%, +8.3%, +9.3% and +8.5, during the gestation period +63.5%, +63.3%, +60.2% and +60.5%, and during the lectation +3.1%, 

### +62%, +4.6 and +2.7%, respectively. (See attached tables on pp. 27 to 39)CLINICAL LABORATORY INVESTIGATION Hematology The assessmentofthe hematology data did not reveal any test Item-related effects in males and females. CMnical Biochemistry Males: In group 4,the concentration ofpotassium was statistically significantly higher (+17.8% compared tothe control). Since the mean value 

### was within the range ofthe historical control data, this was not considered 

### to be a test Hem-relatad effect. In groups

### 2 and 3 no changes

### were noted. Females: The assessment ofthe clinical biochenlstry data did not reveal any test ltem-relatad affects In females. (See attached Tables on 

### pp. 40 to 41 and attached Historical Control Data on 

### pp. 1 to 2) Urine analysis 

### The assessment ofthe urine analysis data did not reveal any test item-related effects in males and females. REPRODUCTIONANDBREEDINGDATA- PARENTANIMALS Please 

### see section "Any otherinformation on results including tables• for the table "Summary of Performance - P Animals Breeding for F1 LHters"

Mating Performance and Fertility All females 

### malad within the first pairing period. 

### The median and mean precoltal times were unaffectad by treatment with 

### the test item. Mean precoital times were 2.6, 3.1, 2.8 and 3.7 days ingroups 1, 2. 3 and 4, respectively. The median precoital time was 3, 4, 3 and 4 days in orderofascending dose level. Due

### to onefemale (no. 58) in group2 which was not pregnant, the fertRily Index and conception rate were 100% in groups 1, 3 and 4, and 90% in group 2. In group 1, one female (no. 43) did not deliverany pup and had only two Implantation sites. Therefore, thegestation index was 

### 90% in group 1, and 100% in groups 2, 3 and 4. Duration ofGestation The mean duration ofgestation was unaffected by the exposure to the test item. Mean duration ofgestation was 21.4, 21.8, 21.5 and 21.4 days, in orderofascending dose level. 

### Corpora Lutea Count Maan number ofcorpora lutea per dam (determined at necropsy) was similar In all groups (16.9, 18.3, 17.2 and 16.4 in orderofascending dose level) and gave no indication ofa test ltem-relatad 

### effect. Implantation Rate and Post-implantation Loss 

### The mean number of implantations perdam was notaffected by 

### the treatment with the test Item.In groups 3 and4, tolal**post~mplantation** loss was statistically significantly higher but within the range ofthe historical control data, therefore was not considered 

### to betest item-related. 

### The mean numbers

### ofimplantations per litterwere 15.0, 16.1, 14.3 and 14.6 in orderof ascending dose level. The mean incidence of post-implantation loss as a percentageoftolal implantations was 1.5, 4.8, 8.4 end 6.2%, in orderofascending dose level. (See attached Tables on pp. 42 to43 and attached Historical Control Data on pp. 15to 38) Utter Size at First Litter Check At first litter

### check, mean litter size was not effected bythe treatmentwith the lest**~em.** Due to the higher number**post~mplantation** loss which occurred in groups 3 and 4, the birth index was statisticeHy significantly lower but wtthin the range 

### ofthe historical control data. Birth index was 98.5, 95.2, 91.6 and 93.8% and mean litter size atfirst littercheckwas 14.8, 15.3, 13.1 and 13.7, In orderofascending dose level. Postnatal Loss Days 0
- 4 Post Parium Postnatal loss up 

### to day 4 p.p. was not affected by the treatment wtth the test Item atanydose level. Mean postnatal loss was 0.3, 0.0, 0.0 and 0.1, corresponding 

### to a**viabll~** index of97.7, 100.0, 100.0 and 99.3% in groups 1, 2, 3 and 4, respectively. TERMINAL FINDINGS Organ Weights Notest Item-related effects were noted in absolute and relative weight oforgans in males and females. All the statistically significant changes were**w~in** the range 

### ofthe historical control data ordid notfollow a dose-dependent psttem. (See attached Tables on pp. 44 to 49 and attached Historical Control Data on pp. 39 

### to 62) Maaosoopical Findings 

### The findings noted during the macroscopical examination did not give indication 

### ofanytest ltem-<etated effect. Males: In group4,**spleen~** a constriction was noted in male no. 34. No otherfindings were noted. Females: In group 4, female no. 72 was noted to have the liver caudal lobe adherentto the lateral 

### lobe. In groups 2 and 3 nofindings were noted. In group 1, female no. 43 was noted to have tan or dark red or red brown discolored noduleson the left uterine horn. Same female had only two Implantation sites and did notdeliver any pups. Histopsthology Findings In group 4, In the lungs ofanimal no. 75 some alveolar macrophages (moderate**sev~** degree) were found to contain dark pigment. This finding was possibly related to accidental aspiration ofdose solution. All further lesions recorded during the microscopic investigation were**~in** the range 

### ofbackground alterationsthat may be recorded 

### in this type 

### ofstudy, and in rats ofthis strain and age.In group 1, in

### one Infertile female (number43)a deciduoma was noted. This lesion was deemed to have contribu1ed to infertitty. Qualitative staging

### ofthePAS-stained testes was conducted and there were no abnormal lesions encountered during sperm staging regarding completeness 

### ofstages and maturation

### of cell populations. Individual lesions recorded were within the 

### range of background aHerations that may be

### recorded In 

### this

### type 

### ofstudy, 

### in 

### rats ofthis strain and ega. 

## Results ofexaminations: offspring VIability(offspring)

no effects 

### Clinicalsigns (offspring)

noeffects 

### Body weight (offspring)

no

### effects 

### Saxualmaturation (offspring) not examined 

### Organ weights (offspring)
not examined 

### Gross pathology(offspring)

noeffects 

### Histopathology(offspring)
not examined 

### Details 

## on results (offspring) LIITER DATA- F1 PUPS External Examination at First Utter Check and during Lactation No abnormalfindings were noted atfirst littercheck orduring the first 4 days postpartum. In group 1, two male pups from sama litter and one male pup from another litterwere noted to have no milk in tha stomach at

### first litter

### check. Sex Retios Sex ratios atfirst litter

### check and on day4 postpartum were unaffected by exposureto the test item.

### The proportion ofmales on day4 post parium was 48, 49, 44 and 50%, in order

### of ascending 

### doss level. Body Weights

### to Day4 Post Parium Mean

### pupweights on day 1 post parium were unaffected by treatmentwith thetest Hem. On day 1 post parium mean 

### pup weightswere6.0,6.1, 6.4and6.0 gin

### order ofascending dose level. During the lactation period (to day 4 post parium), mean pup weight gain was not affected 

### by the treatmentwHh lhe test 

### Hem (+41.7, +39.3, +39.1 and 43.3%, in order

### of ascending 

### doss level). Mean pupweights on day4 post parium were 8.5, 8.5, 8.9 and 8.6 g in order

### of ascending

### doss level. Macroscopical Findings Noabnormal findings ware noted during lhe macroscopical examination 

### of pups. 

## Anyother lnfonnatlon on results incl. tables Summary of Perfonnance • PAnimals Breeding for F1 Litters

Group (mglkg{dayl Female numbers Number offemales paired Number offemales mated Number of non pregnant females (A) Numbers offemales, which did not deliver any pups (B) Number offemales which reared their pups until day 4 post partum (A) Female No. 58 was not pregnant. (0) 

## 41-50

10 10 0



## 9 (B) Female No. 43 had only two implantation sites. 

# Overall remarks, attachments Overall remarks 

## 2 (100) 

## 51-60

10 10 

## 1
0 9 

## 3 

## 4 (300) (1000) 

## 61-70 71-80

10 10 10 10 0 0
0 0 10 10



## Relevant reproduction parameters (number ofcorpora lutea, implantation sites, pre- and post implantation losses, number of pups) were not affected by the treatmentwith the test item. The litter size atfirst littercheck and on day4 postpartum was also not affected.

# Applicant's summary and conclusion Conclusions This study is a valid investigation of

### the loxicological effecls resulting from repealed oral-gavage admlnislralion ofthe test item leuco Sulfur Black 1 (CAS#66241-11-0)to 

### rats over approximately 28 days. Leuco Sulfur Black 1 wasadministered In highly purified waleres vehicle 

### at dosages of 100, 300, and 1000 mg/kgbody weight/day, and controls received 

### the vehicle only. Leuco SUlfur Black 1 was administered 1o male rets for 

### 29 days and tofemale rats for 14 days prior lo pairing, through 

### the pairing and geslelion periods until

### the 

### Fl generation reached day

### 4 post par1um. Administrations at 1000 mglkg body weight/dey caused a dark discoloration of

### feces in all animals afterthe onset of

### the treatment untH 

### theand ofstudy. Thiswas likely due to

### the staining property of

### the test Kern and not considered lo be adverse. Food consumption and bodyweightwere notaffected for

### the entire duration ofthe study. The evaluation of

### the functional 

### battery observations and

### the 

### assessment ofclinical laboratory investigations parametersdid not reveal any test Kern-related effect. Relevant reproduction parameters (numberofcorpora lulea, implantation sKes, pre- and post Implantation losses, numberof pups) were not affected by 

### the treatment with the lest Kern. 

### The Iiiier size atfirst litter

### check and on dey

### 4 postpartumwas also notaffected. All 

### the changes noted in 

### the organweights were within the background ofthe biological variation. No adverse findings were noted during macroscopical and histopathologyexaminations. Based on these results a general NOAEL (No Observed Adverse Effect Level)was established at 1000 mglkg body weight/day. The NOEL (No Observed Effect Level)for reproduction/developmentalloxicKywas considered to be 1000 mgJkg/dey. 

### Executive summary The purpose ofthis studywasto generate pretiminary information on the possible heaKh hazards likely to arise from repeated exposure overa relatively limKed period oftime. In addition Kprovided information on possible effects on male 

### and female reproductive performance such as gonadal function, mating behavior, conception, development of

### the conceptus and parturition.

This study should provide information 1o assess the need lo conduct furlhar investigations and may provide guidance in the design of subsequent studies. 

### Dose levels were selected in agreementwith 

### the Sponsor, based on the results ofa 

### dose range-finding study (Harlan Laboratories Study C31076). Leuco Sulfur Black 1 was administered to male 

### rats for29 days and to female rats for 14 days prior lo pairing, through the pairing and gestation periods until the Ft generation reached day

### 4 post partum. The following dose levels were applied: Group!: Group2: 

### 0 mglkg body weight/day (control group) 100 mglkg body weight/day

### Group3: Group4: 300 mg/kg body weighUday 1000 mg/kg body weighUday A standard dosevolume of 10 mL/kg bodyweightwith a dailyadjustment to the actual bodyweightwas 

### used. Control animals were 

### dosad wHh lhavehicle alone (highly purified water). 

### The following resuHs were obtained: PARENTANIMALS 

### Genenol T-.blllty Alllha animals survived untillha schaduled necropsy. 

### AI. 1000 mglkg bodywelghUday, a darkdiscoloration 

### offeces was observed in all animals starting on day 3 of lhapre-pairing period onwards. FunctloiUII Ob•rvatlon81 

### Battery 

### No test item-related effects were noted. 

### Food Consumption Food consumption was not afleded by the treatmentwHh the test Hem in males and females. 

### Body Weights Notest Hem-related effects were noted 

### in body weight and body weight gain in males and females forlha entire duration 

### of the study. Clinical Laboratory lnvestlgatlions 

### The assessment

### of the clinical laboratory investigations parameters did not give indication ofany test Item-related efled. 

### Reproductlion and a..-lng 

### Data Mean pracoital time, fertility 

### end gestation index and concaplion rate were not afleded by tha treatment 

### with thetest Hem. Implantation rate and post implatation loss were also not affected by lha treatment with lha test Hem. Organ Weights Weight oforgans was 

### not affeded by the treatmentwith lhatest Hem. Ail lha alterations notedwere within lha range 

### of historical control data. Macroscopical Findings and Histopathological Examinations Noadverse effects 

### ofthe test item were noted during the histopathological examination. 

### UTTER DATA- 

### F1 PUPS Findings at Firat 

### Uttar Check and during Lactation The mean number

### of pups at firstiHter check and on day4 post parium were not affected bythe treatment

### with lhatest item. The sex ratiowas also not efleded. No abnormal pupwas noted at any dose level. Pup Weights to 

### Day 4 Post Parium Mean pupweight gain to dey

### 4 post parium was not aflectd by the treatment with the lest item. Macroscopical Findings 

### AI. necropsy 

### of pups, lhare were no abnormal findings.

### Conclusion Based on these results a general NOAEL (No Observed Adverse Effect Level)was established al1000 mg/kg body weight/day. The NOEL (No Observed Effect Level)for raproductionldevelopmental toxicity was considerad to

### be 1000 mg/kg body weight/day.

# Endpointstudyrecord: waiving_Toxicity to reproduction.001
UUID IUC5-89c0c88b-a9a4-4963-9b56-b0b1Oc476117 Doss

### Ia< UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea 

### Author Date Remarl<s uensenba I Clarlant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2011J-05-2114:11:40 CEST


# Administrative Data Date w.Jvlng 

### JustlllcMion 

### fordata -lvlng study scientifiCally unjustified In accondance with Regulation (EC}

### No 1907/2006Annex XI, a one or

### two generation reproductive toxicity study Is scientifically notjustified and, thus, should not be conducted because of

### thefollowing: Leuco Sulfur Blacl< 1 hasa mean molecularweight in the range of8000 to 9000 Da and a number-average molecularweight (Mn) ofabout 3000 Da. hisvery poorly oreven not soluble in common polarand non-polarsolvents sucheswater (solubillly of62.4 mgll), DMSO, methanol, DMF, THF, chloroform and notsoluble In acids tike suWuric acid and trifluoroacetic acid. The vapour prassure ofthe pure substance, calculatedfrom the bolllng point of

### > 400 

### 't:, Is 1.8x 1 OE-5 hPa at 25 'C. Basad on these properties it Is 

### very unlikelythat Leuco Sulfur Black 1 can cross any biological membrane. Thus, an absorption oftoxicologically significant amounts of Leuco Sulfur Blacl< 1following orel, dermal or Inhalation exposure can beexdudad. This is substantiatad by

### the results obtalnad In toxicity and ecoloxicity studies, where nosubstance reiatad local and/or systemic effects Indicative oftoxicity were seen in all tests, including acute animal and acute ecoloxicity studies, sensitization, genoloxicllytests and a repaatad dose toxicity studywtth reproductiveldevolpmental toxicity screening. In 

### the acute oral toxicity study black stainad faeces was observed for maximallyfour days starting the deyafter administration ofthe substance at 2000 mglkg bw. Dart< discoloured faeces were also seen in the repaatad dose toxiclly study

### with reproductive/devolpmental toxicity screening (OECD422) at 1000 mglkg bwld, starting on day 3 

### ofthestudy and lasting throughout 

### the administration pariod untiltermination. The discolouration offaeces was the onlyeffectobservad and can be attributad to the strong staining properties ofthe substance. This indicates that Leuco Sulfur Blacl< 1 isdirectly excreted and thatabsorption in

### the gastrointestinal tract 

### does not occurfollowing oral administration, espacially since organs or urine wera not discoloured and signs ofsubstance depositswere nolfound In the histopathological analysis of

### the orgens. In conciuslon, the availabletoxicity and ecotoxiclly data togetherwith the intrinsic substance properties are sufficient to demonstrate that exposure to Leuco Sulfur 

### Blacl< 1 does nol cause any heahhconcern.

# 7.8.2 Developmental toxicity 

# Iteratogenicity Endpointstudyrecord: waiving_Developmental toxicityI teratogenicity.001
UUID IUC5-cd2b3712-<>4<:6-41b4-b396-7ce7bdcaa2d5 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks uensenba I Clariant Produkte (Deutschland) GmbH I Sulzbach am Taunus I Germany 2010-05-2114:12:26 CEST



# Administrative Data Date w•lvlng 

### Justification for

### date -lvlng study scientifically unjustified In accordance 

### with Regulation (EC) No 1907/2006Annex XI, a teratogenicily study is scientifically notjustified and, thus, should not be conducted because ofthe following: Leuco Sulfur Black 1 has a mean molecular weight in the range of8000 to 9000 Da and a number-average molecularweight (Mn) 

### ofabout 3000 Da. It isvery poorly oreven not soluble in common polar and non-polar solvents such as water (solubility 

### of62.4 mg/L), DMSO, methanol, DMF, THF, chloroform and 

### not soluble In acids**~ke** sulfuricacid 

### and 

### 1riftuoroacetic acid. The vapour pressure of

### the pure substance, calculated from 

### the bolllng pointof> 400 

### 'C, is 1.8 x 1OE-5 hPa at 25 

### 'C. Based on

### these**properties~** 

### Is very unlikelythat Leuco Sulfur Black 1 can cross any biologk:el membrane. Thus, absorption oftoxicologically significant amounts of Leuco Sulfur Black 1following oral, dermal or Inhalationexposure Is negHgible. This is substantiated by

### the resuhs obtained intoxicity and ecotoxicily studies, where no substance related local and/orsystemic effects indicative oftoxicity were -n In all tests, including acute animal and acute ecotoxlcily studies, sensitlzatlon, genotoxicity tests and a repeated 

### dose toxicity study

### with reprodudlveldevolpmental toxicity screening. In the acute 

### oral toxlcily study black stained faeces was observed lor maximally four days starting the day alferadministration of

### the substance 

### at 2000 mg/kg bw. Dark discoloured faeces

### were also seen in 

### the repeated dose toxicity study 

### with reproductlveldevolpmental toxlcily screening (OECD 422) at 1000 mglkg bw/d, starting on day 3 ofthe 

### study and lasting throughout

### the administration period until termination. The discolouration offaeces was 

### the only 

### ailed observed and can 

### be attributed to thestrong staining properties ofthe substance. This indicates that Leuco Sulfur Black 1 Is dlrectiyexcreted and that absorption In 

### the gastrointestinal tract does notoccurfollowing 

### oral administration, especially since organs or urine were not discoloured and signs ofsubstance depositswere notfound in 

### the histopathological analysis 

### oftha organs. In conclusion, the available toxicity and ecotoxicity data togetherwith 

### the intrinsic substance properties are sufficient to demonstrete that exposure to Leuco Sulfur Black 1 does 

### not cause any health concern.

# 7.9 Specific investigations 7.9.1 Neurotoxicity Endpointsummary: Neurotoxicity
UUID IUC5-a04392ee-dbfd-4d39-95ec-aef5eb8bf917 Dossier UUID IUC5-292149d1 -a677-4feb-9692-1fad973281 ea Author thr 

### I SCC GmbH 

### IWendelsheim 

### I Germany Date 2010-05-26 16:49:43 CEST Remarks 

# Administrative Data Discussion There are no studies available specifically addressing this endpoint However, based on the substance properties (mean MW >8000 Da and very poor solubility in water) and the fact that no toxicity was seen in any ofthe available, it is concluded that Leuco Sulfur Black 1does not raise any concern regarding this endpoint 

# Justification for classification or non-classification Based on the available data Leuco Sulfur Black 1 is not subject to classification and labelling_

# 7.9.2 lmmunotoxicity Endpointsummary: lmmunotoxicity
UUID IUC5-351fad61-1d5a4814-a2fb-b9ec0c7209e6 Dossier UUID IUC5-292149d1-a6n-4feb-9692-1fad973281ea Author lhr I SCC GmbH I Wendelsheim I Germany Date 2010-05-26 16:51:40 CEST Remarks 

# Administrative Data Discussion There are no studies available specifically addressing this endpoint. However, based on the substance properties (mean MW >8000 Da and very poor solubility in water) and the fact that no toxicity was seen in any ofthe available, it is concluded that Leuco Sulfur Black 1 does not raise any concern regarding this endpoint. 

# Justification for classification or non-classification Based on the available data Leuco Sulfur Black 1 is not subject to classification and labelling.

# 11 Guidance on safe use Additionalinformation: Guidance on safe·use.001
UUID IUCS-be2cb819-599a-4105-b877-c0ea861c5267 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks pbasto I Clariant Quimicos (Portugal) Lda /lei"' do Balio I Portugal 2010-06-11 10:40:39 CEST



# Administrative Data

Flrst..,ld measures Ifsomeone exposad

### to the productfeels unwell seek medical assistance In case ofcontact with skin wash immediaUy with water 

### Fire-fighting measures Suitable exlinguishing media water sprayjel, dry powder , carbon dioxide 

### Acddent.l release measures Take up mechanically

. Do not allow to entryto drains, water courses orsoil. Dispose 

### the absorbed material in accordance with local ragulalions 

### Handling and storege Handling: Observe the usual precautions for handling chemicals keep awayfrom 

### food. Wash hand before breaks and afterwork. Storage Stable under normal conditions Keep containertighUy closed, dry in a cool areaAvoid impact , friction and accumulationofeleclm5tacticcharges Keep 

### awyfrom ignition sources. Shlbtlltyand 

### ""'ctlvlty Stable under usual conditions Disposal considerations Dispose according to local regulations for chemicel produdS

# 13Assessment Reports Additionalinformation: CSR
UUID IUC5-0d168ec0-7fc5-4eeb-bd1a-177f823bbf5d Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea 

### Author Date Remarks jregano 

### I Clariant lbertca Producci6n SA 

### I El Prat de Llobregat (Barcelona) 

### I Spein 2010-06-23 08:51:01 CEST



# Administrative Data Type ofreport REACH Chemical safely 

### report (CSR) 

### Document LSBI<1_CSR_23 06 2010.pdf/284.52 KB (application/pdf)

# Reference substance: sodium sulphate UUIO ECB5-<>db493fc-4080-4e80-8761-<>c151 506fddf Dossier UUIO IUC5-292149d1 -a677-4feb-9692-1fad973281ea Author Date Remarks mkuehn I Clariant Produkte (Deulschland) GmbH I Sulzbach am Taunus I Germany 2010-06-Q1 15:53:58 CEST 

# General information Reference substance n•m• sodium sulphate 

# EC inventory EC number 23H!20-9 CAS number 7757-82-8 EC n•me sodium sulphate Molecularfonmula H204S.2Na 

# Reference substance information CAS information CAS number 7757-82-8 CAS name Sulfuric acid sodium salt (1 :2) 

# IUPAC name disodium sulfate 

# Synonyms Name Sulfuric acid disodium satt Name Sulfuric acid disodium satt 

# Group 

# I category information

DSL Category: lnorganics 

# Molecular and structural information Molecularfonmul• H204S.2Na

### Molecul•r-lght range 142.0421 SMILES nolatlon [Na+].[Na+].[O.]S(=O)(=OMO-J InChi lnChi=1/2Na.H204S/c;;1-5(2,3)4/h;;(H2,1,2,3,4)/q:z-+1 ;/p-2 Slructuralfonnula

# Reference substance: 2.4-dinitrophenol UUID ECB5-16d37b76-0b88-40cb-8cc5-07c7d66002ef Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973261ea 

### Author Date XML Transformation V1.0 Plug-In 2007-05-10 11 :00:00 CEST Remarks Successfully migrated lo IUCLID 5.2 formal. 

# General information Reter.nce oubagnce name 2,4-dinltrophenol 

# EC inventory ECnumber EC name 200-067-7 CAS number 51-26-5 2,4-dinitrophenol -cularformula C6H4N205 

# Reference substance information CAS Information CAS number 51-26-5 

# IUPAC name 2,4-dinrtrophenol 

# Synonyms Name Phenol, 2,4-dinrtroName Phenol, 2,4-dlnitro

# Group 

# I category Information

DSL Category: Organics 

# Molecular and structural information -cularformula C6H4N205 Molecular-lght range 184.1064

### SMILES 

### notation InChi 

### SlrUclur•l formula Oc1ccc(cc1[N+}(=O)(O-})[N+](=O)[O-} lnChi=1/C6H4N205/c9-6-2-1-4(7(10)11 )3-5(6)8(12)131h1-3,9H



# OH 

# NQ

# Reference substance: water UUID ECB5-910811d3-a683-4371-b479-777b663a18c5 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks XML Transformation V1.0 Plug-In 2010-04-{19 16:35:06 CEST Successfully migrated to IUCLID 5.2 format 

# General information Reference •ubstance name water 

# EC inventory
EC number EC name 231-791-2 CAS number 7732-18-5 water 

### Molecularformula H20 

# Reference substance information EU:REACH 

# CAS Information CAS number 7732-18-5 CAS name Water 

# IUPAC name water 

# Synonyms Name Water Name Water 

# Group I 

# category information

DSL 

### Category: lnorganics 

# Molecular and structural information

### EU:REACH Molecularformula H20 Molecular-lght range 18.0153 SMILES notation 0 lnChl lnChi=1/H20/h1H2 Structural formula

# Reference substance: sodium chloride UUID ECB5-fdal2f4f-2a29-4181-ae78-5b5622d4912b Dossier UUID IUC5-292149d1-a6n-4feb-9692-1fad973281 ea Author Date Remarks XML Transformation V1.0 Plug-In 2010-02-23 13:59:30 CET Successfully migrated to IUCLID 5.2 format. 

# General information R.,.,..nce substance name sodium chloride 

# EC inventory EC numbsr EC name 231-598-3 CAS numbsr 7647-14-5 sodium chloride Molecularformula GINa 

# Reference substance information CAS Information CAS number 7647-14-5 CAS name Sodium chloride 

# IUPAC name sodium chloride 

# Synonyms Name Sodium chloride (NaCI) Name Sodium chloride (NaCI) 

# Group 

# I category Information

DSL Category: lnorganics 

# Molecular and structural information Molecularformula GINa

### Molecul•r-lghtf8nge 58.4428 SMILES no18tlon (Na+].[CI-) InChI**lnChi=1/CIH.Nalh1H~q;+1/p-1** 

### Structuf81 formula



# Cl

# Reference substance: 1-chloro-2.4-dinitrobenzene UUID ECB5-f3f3978e-cfca-442d-bd31-9e04ac3b4354 Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks XML Transformation V1.0 Plug-In 2007-05-1011:00:00 CEST Successfully migrated lo IUCLID 5.2 format. 

# General information Reference substance name 1-chloro-2,4-dlnnrobenzene 

# EC inventory EC number EC name 202-551-4 CAS number 97-00-7 1-chloro-2,4-dinilrobenzene Molecularformula C6H3CIN204 

# Reference substance information CAS Information CAS number 97-00-7 

# IUPAC name 1-chloro-2,4-dlnitrobenzene 

# Synonyms N"""' 1-chloro-2,4-dinnrobenzene Name Benzene, 1-chloro-2,4-dinitroName Benzene, 1-chloro-2,4-diMro

# Group 

# I category information

DSL Celegory: Organics USEPA Category: Neutral Organics;Polynilroaromatics 

# Molecular and structural information

### Molecularformula C6H3CIN204 

### Molecular-lght range 202.552 

### SMILES 

### notation [O-)[N+K=O)c1ccc(CI)c(c1MN•](=OMO-] 

### InChI InChI=1/C6H3CIN204/c7 -5-2-1-4(8(10)11**)~(5)9(12)13/h**1-3H 

### Structural formula



# Cl


# NO?

# Reference substance: phenol.2.4-dinitro-,sulfurized, leuco derivatives UUID**IUC5~a653db5-12844**7db-9384-05f22219el4d Dossier UUID IUC5-292149d1-a677-4feb-9692-1fad973281ea Author Date Remarks XML Transfonnation V1.0 Plug-In 2010-01-15 08:28:39 CET Successfully migrated to IUCLID 5.2 format. 

# General information Reference substance name phenol,2,4-<linitro-,sulfurized, leuco derivatives 

# EC inventory EC number 266-273-5 

### CAS number 66241-11-0 

### EC name phenol, 2.4-<linitro-, sulfurized, leuco derivatives 

# Reference substance information CAS 

# information CAS number 66241-11-0 

# Molecular and structural information Molecularweight renge > 

### 8000-- < 

### 9000 Remarks It is a UVCB ,**~does** not have a defined molecularformula because**~was** not possible to establish a defined chemical structure

# ISO 

# ON SNIVlNOO