# STILLMEADOW I NCO R P 0 RAT E D

VOLUME OF OF SUBMISSION Alcohols, C18-22, distn. residues (CAS No: 1160164-88-4)
FINAL REPORT ACUTE EYE IRRITATION STUDY IN RABBITS OECDNO.405

AUTIIOR: Janice O. Kuhn, 

### PhD, DABT
STUDY INITIATION DATE: 11 September 2009 STUDY COMPLETION DATE: 06 October 2009
CONDUCTED BY: STll.,LMEADOW, Inc. 12852 Park OneDrive SugarLand, TX 77478
LABORATORY STUDY NUMBER: 

### 13294-09
VOLUME.l OF .l OF STUDY PAGE.l OF 

### --.l2.
SUBMITTED TO: Sasol Germany GmbH

Anckelmannsplatz 1 Hamburg, Germany 20537

12852 Park One Drive. Sugar Land, Texas 77498 • 281 240-8828 • Fax 281 240-8448 www.stillmeadow.com13294-09 Page 2 of 15 

### GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT This studywas designed and performed at STILLMEADOW, Inc. and was conducted in compliance with United States Environmental Protection Agency TSCA 40 CFR 792 with exception of: 

### Section 792.31 (d) and 792.105 (a)(b )(e) Theprovided Certificate of Analysis was not accompanied bya GLP compliance statement. . This studywas designed and performed at STILLMEADOW, Inc. and was conducted in compliance with Organization for Economic Cooperation & Development Principles of GLP, Annex 

### 2, C(98)17 with exception of: 

### Section IT, 1.1 (2)(p), 6.1 (1) and 6.2 (2)(4) The provided Certificate ofAnalysis was not accompanied by a GLP compliance statement. This study was designed and performed at STILLMEADOW, Inc. and was conducted in compliancewith Japan Ministry of Agriculture, Forestry & Fisheries, NotificationNo. 11 Nousan 6283, Director-General ofAgricultural Production Bureau with exception of: 

### 11 Nousan 6283 Article 3.1 (18), ·12.5 and 12.7 The provided Certificate of Analysis was not accompanied by a GLP compliance statement.
uhn, PhD, DABT ector, STILLMEADOW, Inc.
Signature ofAgent ofSponsor AgentName Sponsor: Sasol Germany GmbH
Signature ofAgent ofSubmitter AgentName Submitter: Sasol NA Date

Date

Date

STILLMEADOW, Inc.

### QUALITY ASSURANCE STATEMENT
Test Substance: Alcohols, C18-22, distn. residues (CAS No: 1160164-88-4) Study Title: Acute Eye Irritation Study in Rabbits 13294-09 Page 3 of15

The studyreport and datahavebeen audited in accordance with Good LaboratoryPractice Standards and STILLMEADOW, mc. Standard Operating Procedures (SOPs). The fmal report accurately reflects the study data. The Quality Assurance Unit has not been involved 

### in the actual conduct ofthis study. The Quality Assurance Unit performed a recent facility inspection on 10 Jul 09. All fmdings were reported to Management, and the report and responses are kept in the Quality Assurance files. Thefmdings from any studyinspections and audits werereported to the StudyDirector and Management as follows:Critical Phase Inspected Date Inspected Reported to Reported to Study Director Management Protocol Review 8 Sep 09 8 Sep 09 8 Sep 09 Dosing 21 Sep 09 21 Sep 09 21 Sep 09 Report/Data Audit 01 Gct 09 01 Gct 09 01 Gct 09

Richard r:.Martin, MS, CPhT QualityAssurance, STILLMEADO! ,mc.

STILLMEADOW, Inc.

### TABLE OF CONTENTS 13294-09 Page 4 of15
Page GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT.
.............................................2 QUALITY ASSURANCE STATEMENT
..........................................................................................3 SUMMARy
........................................................................................................................................5 INTRODUCTION
............................................................................................................................... 5 TEST SUBSTANCE
........................................................................................................................... 5

, TEST SySTEM
................................................................................................................................... 6

PROCEDURES
................................................................................................................................... 6 Test Substance Administration
......................'"
............................................................................... 6 Observations
....................................................................................................................................7 Irritation Scoring Method
................................................................................................................7 RESULTS AND DISCUSSION
..........................................................................................................7 Evaluation
........................................................................................................................................7 CONCLUSION
...................................................................................................................................7 SIGNATURE
......................................................................................................................................7 STUDY PERSONNEL
........................................................................................................................ 8 LEGEND TO TABLE 1
......................................................................................................................9 LEGEND TO TABLE 2
.................................................................................................................... 11 TABLE 1 - Ocular Reactions
............................................................................................................. 12 TABLE 2 - Scores and Score Summary
............................................................................................. 1 4 APPENDIX A - Certificate ofAnalysis
................................................................:............................ 15

STILLMEADOW, Inc.

### SUMMARY 13294-09 Page 5 of15
An acute eye irritation studywas conducted on three albino rabbits usingtest substance Alcohols, C18-22, distn. residues (CAS No: 1160164-88-4). The test substance, 0.1 mL byvolume (33.1 mg), was placed into the conjunctival sac ofthe right eye ofeach animal selected for testing. All treated eyes were washed with room temperature deionized water for one minute immediately after recording the 24-hour observation. The number ofanimals testing "positive" for each parameter (accordingto theLegendto Table 1) overthe number ofanimals tested is presented below. Time After Treatment Hours _1 24 48 72 Cornea

Opacity 0/3 0/3 0/3 0/3 Iritis 0/3 0/3 0/3 0/3 Conjunctivae

Redness 0/3 0/3 0/3 0/3 Chemosis 0/3 0/3 0/3 0/3 There were no positive effects exhibited in any eyes at any time after treatment. Non-positive irritation was clear by 72 hours, and the test substance is rated minimally irritating. 

### INTRODUCTION The objective ofthis studywas to assess the relative level ofeye irritation following a single exposure of the test substance to rabbits in accordance with OECD 405; OPPTS 870.2400; and Canadian notification. This study was conducted for Sasol Germany GmbH, according to the approved protocol and STILLMEADOW, Inc. SOPs. There were no deviations from the protocol that affected the quality or outcome of the study. All procedures in this study are in compliance with Animal Welfare Act Regulations. The protocol, raw data, this report and a sample of test substance are archived at STILLMEADOW, Inc. The studywas initiated on 11 Sep 09, the pre-dose experimental portionbegan on 20 Sep 09, and the animals were treated with the test substance between 1125 and 1126 on 21 Sep 09. The in-life portion ofthe study was terminated on 24 Sep 09.

Label Identification: Date & Quantity Received: Physical Description: Storage: Purity: Stability: 

### TEST SUBSTANCE Alcohols, CI8-22, distn. residues (CAS No: 1160164-88-4) Lot: 

### 03585iMA Spec: 59BIRN2 11 Sep 09; 246.3 g (GW) Tan solid Room temperature Refer to Certificate ofAnalysis Exp: Aug 2013 per provided information Data generated for characterization and stabilityis the responsibility of the sponsor. Records pertainingto identity, synthesis methods and location ofdocumentation are the responsibility of the sponsor. A copy of the Certificate ofAnalysis.is included as report Appendi-x 

### A.
STILLMEADOW, Inc.Experimental Animals Species & Strain: Justification ofSpecies: Source: Date BornlDate Received: Quarantine Period: Quantity & Sex: Animal/Group Identification: Initial BodyWeights: Final BodyWeights: Animal Husbandry Cage Type: Housing: 

### TEST SYSTEM
Albino rabbit; NewZealand White 13294-09 Page 6 of15

The rabbit is preferred by the various regulatory agencies for use in eye irritation testing. Nichols Rabbitry Inc., Lumberton, TX 31 May & 21 Jun 09 / 19 Aug 

### & 9 Sep 09 5 days 3 female (nulliparous and non-pregnant) Eartag / Cage cards 2.700-3.150 kg 2.750-3.300 kg Suspended, wire bottom, stainless steel 1 per cage Environmental Controls

Set to Maintain:
. Temperature 200± 3° C

. 12-hour light/dark cycle Actual TemplRel. Humidity: 17-22° C / 59-97% .Relative Humidity 30-70%

. 10-12 air changes/hour

Food: Water: Protocol deviation: humidity outside protocol range but did not affect study outcome. PM! Feeds, InC.TM Lab Rabbit Diet#5321,**~8** oz. daily Municipal water supply analyzed by TCEQ Water Utilities Division; available 

### ad libitum from automatic water system Animalhusbandry and housing at STILLMEADOW, Inc. comply with standards outlined inthe "Guide for the Care and Use ofLaboratory Animals" (NRC Publ.). No contaminants were expectedto havebeen present in the feed or water that would have interfered with or affected the results ofthe study. 

### PROCEDURES Test Substance Administration Healthy albino rabbits were released from quarantine. Both eyes ofeach animal were carefully examined within 24 hours priorto treatmentwith a fluorescein sodium ophthalmic solution and cobalt-filtered 

### light. Both eyes of each animal were again carefully examined just prior to treatment, but without the fluorescein sodium ophthalmic solution. Onlythose animals withouteye defects or irritationwere selected for testing. On Day 0, a dose of 0.1 

### mL by volume (33.1 mg) ofthe undiluted test substance was placed into the conjunctival sac ofthe right eye ofeach aniinal by gently pullingthe lowerlid away from the eyeball to form a cup into which the test substance was dropped. The lids were gentlyheldtogether for one second to prevent loss ofmaterial. The untreated left eyes served as comparative controls.

STILLMEADOW, Inc.Observations 13294-09 Page 7 of 15
Thetreated eyes ofall animals were examined without magnification under white room lightingprovided bydaylight-type fluorescent ceilingfixtures, and (ifneeded) an additional source ofwhite light affixed to the examination table or using a handheld flashlight. The grades ofocularreactionwere recorded at 1, 24, 48 and 72 hours after treatment. The corneas of all treated eyes were examined immediately after the 24-hour observation with a fluorescein sodium ophthalmic solution. A Finoffoculartransilluminatorwith cobalt blue filter (Welch Allyn, Skaneateles Falls, NY) was utilized to enhance visualization of fluorescein staining. Any ofthe corneas which exhibited fluorescein staining at the 24-hour observation were re-examinedwith the fluorescein sodium ophthalmic solution at each consecutive observationuntil fluorescein staining of the cornea no longer occurred. All treated eyes were washed with room temperature deionized water for one minute immediately after recording the 24-hour observation. Irritation Scoring Method mdividual irritation scores for each animal at each scheduled observation were determined using the grading scale given inthe Legendto Table 1. 

### An average irritation score for each scheduled observation was then determined, based on the number ofanimals tested. A maximum average irritation score was derived from the observation yieldingthehighest average irritation score. Themaximum average irritation score was usedto rate the test substance accordingtothe ratings presented inthe Legendto Table 2. Any corneal involvement or iridic irritation with a score of 1 or more is consideredpositive. Any conjunctival irritation (redness or chemosis) with a score of2 or more is considered positive. 

### RESULTS AND DISCUSSION Evaluation The number of animals with "positive" findings, if any, at each observation period is presented in the summary section ofthis report. Ocularreactions are presented inTable 1. A summaryofirritation scores is presented in Table 2. The maximum average irritation score of2.0, obtained 1hour aftertreatment, "'[as used to rate A1cohols, C 18-22, distn. residues (CAS No: 1160164-88-4) minimally irritating. Fluorescein staining didnot occur in any ofthe eyes. 

### CONCLUSION Based on the maximum average irritation score of2.0, the test substance Alcohols, CI8-22, distn. residues (CAS No: 1160164-88-4) is rated minimally irritating. There were no positive effects during the study, and non-positive irritation was clear in all.eyes at 72 hours. Study Director: 

### J nice
. Kuhn, PhD, DABT Toxicologist, STILLMEADOW, mc.

STILLMEADOW, Inc.

### Technical Staff: Carol Morris, BA 

### Paul Siemens, BA Robert Preston STUDY PERSONNEL

Nancy Casajuana, LAT Data Services: Connie Pavatte, Report Preparation 13294-09 Page 80f15

STILLMEADOW, Inc.13294-09 Page 9 of15 

### LEGEND TO TABLE 1 ACUTE EYE IRRITATION STUDY 

### IN RABBITS Grading Scale



### 1. Cornea A. Opacity

- degree (area most dense taken for reading) No opacity.......
........................................................................................................... 0 Slight dulling ofnormalluster
........... ................ ....... ............................................... ... 

### + Scattered or diffuse areas ofopacity (other than slight dulling ofnormal

luster), details ofiris clearly visible.....................
..................................................... 

### 1

## * Easily discernible translucent area, details ofiris slightly obscured
......... .................. 2* Nacreous area, no details ofiris visible, size ofpupil barely discernible.................... 3* Opaque cornea, iris not discernible through the opacity
............................................. 

### 4

## * B. Area ofcornea involved One quarter (or less), but not zero.............................................................................. 

### 1 Greater than one quarter, but less than halL.............................................................. 

### 2 Greater than half, but less than three quarters............................................................. 3 Greater than three quarters, up to whole area............................................................. 

### 4 C. Fluorescein staining

- appearance ofyellow-green staining ofcornea Cornea not examined with fluorescein
....................................................................... - No fluorescein staining............................................................................................... 0 Positive fluorescein staining.......................................................................................
. P

Area ofcornea involved

One quarter (or less), but not zero.......................................................................... A Greater than one quarter, but less than halL.........
................................................. B Greater than half, but less than three quarters......................................................... C Greater than three quarters, up to whole area ......................................................... D D. Stippling

- appearance ofpinpointroughening No stippling
................................................................................................................ 0 Presence ofstippling................................................................................................... S

Area ofcornea involved

One quarter (or less), but not zero .......................................................................... A Greater than one quarter, but less than halL
..................:....................................... B Greater than half, but less than thre€quarters......................................................... C Greater than three quarters, up to whole area ......................................................... D

AXBX5 Total Maximum 

### = 80 

# * - Reaction indicates a positive effect. Reference: Draize, John H., Woodard, Geoffrey, and Calvery, Herbert 

### 0., Journal ofPharmacol. Exp. Ther., 82,377-390 (1944).

STILLMEADOW, Inc.13294-09 Page 10 of15 

### LEGEND TO TABLE 1 (cont.) ACUTE EYE IRRITATION STUDY IN RABBITS Grading Scale


## n. Iris 

### A. Grades Normal.
....................................'.................................................................................... 0 Markedly deepened rugae, congestion, swelling, moderate circumcorneal

hyperemia or injection (any ofthese or combination thereof), iris still reacting to light (sluggish reaction is positive)
......................................................... 

### 1

## * No reaction to light, hemorrhage, gross destruction (any or all ofthese).................... 2*

AX5 Total Maximum = 10 

## m. Conjunctivae 

### A. Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris) Blood vessels normal.................................................................................................. 0 Some blood vessels defmitely hyperemic (injected)
................................................... 

### 1 Diffuse, crimson color, individual vessels not easily discernible................................ 2* Diffuse beefy red........................................................
................................................ 

### 3

## * B. Chemosis: lids andlor nictitating membrane No swelling
.................... :........................................................................................... 0 Any swelling above normal (includes nictitating membrane)
............................ :........ 

### 1 Obvious swelling with partial eversion oflids............................................................ 2* Swelling with lids about halfclosed........................................................................... 

### 3

# * Swelling with lids more than**h~lf** closed.................................................................... 4

## * C. Discharge No discharge..........................
............................................................. ........... ...... ........ 0 Any amount different from normal (does not include small amounts .

observed in inner canthus ofnormal animals)
.......................................................... 

### 1 Discharge with moistening ofthe lids and hairsjust adjacent to lids
........ ................. 

### 2 Discharge with moistening ofthe lids and hairs, and considerable area

. around the eye.......................................................................................................... 

### 3 D. Necrosis or Ulceration ofthe palpebral and bulbar conjunctivae or nictitating membrane No necrosis or ulceration............................................................................................ 0 Presence ofnecrosis or ulceration
..................................:........................................... N (A+B+C)X2 Total Maximum

## =20 The total score for the eye is the sum ofall scores obtained for the cornea, iris, and conjunctivae withthe possible maximum total score for the eye being equal to 110. 

## * -Reaction indicates a positive effect.

STILLMEADOW, Inc.LEGEND TO TABLE 2 13294-09 Page 11 of 15 ACUTE EYE IRRITATION STUDY 

### IN RABBITS Rating ofTest Substance Based on Eye Irritationl

Rating Non-irritating Practically non-irritating Minimally irritating Mildly irritating Moderately irritating

Severely irritating

Extremely irritating Maximum Average Score 0.0-0.5
>0.5-2.5
>2.5-15.0
>15.0-25.0
>25.0-50.0

>50.0-80.0

>80.0-110.0 Defmition To maintain this rating, all scores at the 24-hourreading must be zero; otherwise, increase rating one level. To maintain this rating, all scores at the 24-hour reading must be zero; otherwise, increase rating one level. To maintain this rating, all scores at the 72-hourreading must be zero; otherwise, increase rating one level. To maintain this rating, scores at the 7 day reading must be zero; otherwise, increase rating one level. To maintain this rating, scores at the 7 day reading must be less than or equal to 10 for 60% or more ofthe animals; also, the 7 day mean score must be less than or equal to 20. lfthe 7 day mean score is less than or equal to 20, but less than 60% of the animals show scores less than or equal to 10, then no animal with a score greater than 10 can exceed a score of30 if rating is to be maintained; otherwise, increase rating one level. To maintain this rating, scores at the 7 day reading must be less than or equal to 30 for 60% or more ofthe animals; also, the 7 day mean score must be less than or equal to 40. Ifthe 7 daymean**~core** is less than or equalto 40, butless than 60% of the animals show scores less than or equal to 30, then no animal with a score greater than 30 can exceed a score of60 if rating is to be maintained; otherwise, increase rating one level.

NOTE: The rating ofthe test substance is not to be increased more than one level above its maximum average score.

ISlightly modified from Kay, J.H. and Calandra, J.C. (1962) Interpretation ofEye Irritation Tests. J. Soc. Cosmetic Chemists 13 :281-289 STILLMEADOW, Inc.TABLE 1 - Ocular Reactions ACUTE EYE IRRITATION STUDY INRABBITS Test Substance: Alcohols, CI8-22, distn. residues (CAS No: 1160164-88-4)

I. Cornea

A. Opacity B. Area C. Fluorescein Staining D. Stippling SCORE 

### n. Iris A. Grade SCORE Ill. Conjunctivae A. Redness B. Chemosis C. Discharge D. Necrosis or Ulceration SCORE TOTAL SCORE

1. Cornea A. Opacity B. Area C. Fluorescein Staining D. Stippling SCORE 

## n. Iris

A. Grade SCORE Ill. Conjunctivae A. Redness B. Chemosis C. Discharge D. Necrosis or Ulceration SCORE TOTAL SCORE

M - Male; F - Female Duration ofStudy: 

## n Hours 1 0 0



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

1 0 0



### - 0 0 0 0 1 0 0 0 2 2 Rabbit No. 4217-F Hrs. After Treatment 1 Days After Treatment 

### I 24 

### I 48 

### In 14 17 

### I .10 

### I 14 

### I 17

0 0 0 0 0 0 0 

## - - 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 2 2 0 2 2 0 \ \

Rabbit No. 4219-F Hrs. After Treatment 

### I Days After Treatment 124 1 48 

### In 14 17 -' 10 

### I 14 

### I 17

0 0 0 0 0 0 0 

### - 

## - 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 

### I 21



### I 21 13294-09 Page 12 of 15

STILLMEADOW, Inc.

### TABLE 1 - Ocular Reactions (cant.) ACUTE EYE IRRlTATION STUDY IN RABBITS Test Substance: Alcohols, C18-22, distrl. residues (CAS No: 1160164-88-4)

I. Cornea

A. Opacity B. Area C. Fluorescein Staining D. Stippling SCORE IT. Iris

A. Grade SCORE 

## m. Conjunctivae

A. Redness B. Chemosis C. Discharge D. Necrosis or Ulceration SCORE TOTAL SCORE

M - Male; F - Female Duration ofStudy: 72 Hours 1 0 0



### - 0 0 0 0 1 0 0 0 2 2 Rabbit No. 4249-F Hrs. After Treatment / Days After Treatment / 24 / 48 /72 14 17 1 10 / 14 1 17

0 0 0 0 0 0 0 

### - - 0 0 0 0 0 0 0 0 0 0 0 0 ( 1 1 0 0 0 0 0 0 0 0 0 0 2 2 0 2 2 0 / 21 13294-09 Page 13 of15

STILLMEADOW, Inc.

### TABLE 

### 2 - Scores and 

### Score Summary ACUTE EYE IRRITATION STUDY IN RABBITS Test Substance: Alcohols, 

### CI8-22, distn. residues (CAS No: 1160164-88-4)
Time After Rabbit Number Treatment 4217-F 4219-F 4249-F Average Score i , Hour 1 2 2 2 2.0 Hour 24 2 0 2 1.3 Hour 48 2 0 2 1.3 Hour 72 0 0 0 0.0

Day 4 Day 7 Day 10 Day 14 

# I Day 17 Day 21 

# I Maximum Average Irritation Score:

# I 2.0 IMinimally irritating M - Male; F - Female Duration ofStudy: 72 Hrs 13294-09 Page 14 of 15

STILLMEADOW, Inc.

### APPENDIX A - Certificate ofAnalysis



### C E R TIP I 

### CAT E 

### o F 13294-09 Page 15 of 15 

# saSDL
9/09/2008

ANALYSIS

Product: Lot No. t 

### 'ManU£acturing date: 

### Alcoholsl' C1.8-22 t 

### diatn. Residuea 0.3S8S/MA (S9B1RN2) 25/08/2009 Exp:lry date: 

### oa/2\ll.3

Unit Result Teata:**_~=:_.**

# __**~~=**••**~~=~~~~~~;~~~~.g==~~=:~~~~w--~~~~~~==.a_-~~~~~~~~=~~~=~;=;~~~BDk~~.~.=~:=; H~Ox.yl**  

# - Nwribex: [mg KCH/g] SO,**I!:st~** No. Img XOH!g] 41,1'1 Aci." No. [mg KOR/gl**()~O-9** water rwt.. \] 0,02 Iodl,ne No. [mg l/100mq] 5,34



### Sasol Germany GmbH F.O. Box l16{) 

### D-25534 :exunsbuettel Phone: 049 (0) 4852/392 -0 Fax: 049 (0)4852/3.285 Best regards 

### works inspector 

### M. Sprung

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STILLMEADOW, Inc.