# STILLMEADOW I N C 0 R P 0 R A T E D

VOLUME OF OF SUBMISSION 

### Alcohols, C18-22, distn. residues (CAS No: 1160164-88-4)
FINAL REPORT 

### GUINEA PIG MAXIMIZATION TEST FOR TOPICALLYAPPLIED TEST SUBSTANCE OECD 406 & 

### OPPTS 870.2600
AUTHOR- Janice 

### 0. Kuhn, PhD, DABT
STUDY INITIATION DATE: 

### 11 September 2009 STUDY COMPLETION DATE: 23 November 2009
CONDUCTED BY: 

### STILLMEADOW, Inc. 12852 ParkOne Drive SugarLand, TX 77478

LABORATORYSTUDYNUMBER 

### 13296-09
VOLUME_l_ 

### OF _l_ 

### OF STUDY PAGE _l_ 

### OF_lQ


### SUBMITTED TO: 

### Sasol Germany Gmbll

Anckelmannsplatz 1 Hamburg, Germany 20537

12852 Park One 

### Drive • Sugar Land, Texas 77498 • 281 240-8828 • Fax 281 240-8448 www.stillmeadow.com

### 13296-09 Page 2 of 16 GOOD LABORATORYPRACTICE COMPLIANCE STATEMENT


### This study was 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) The provided Certificate ofAnalysis was not accompanied by a GLP compliance statement. Section 792.105 (b) Solubility was not performed. Section 792.113 (a) Mixture analysis was not performed. This study was designed and performed at STILLMEADOW, Inc. and was conducted in compliance with Organization for Economic Cooperation & 

### Development Principles ofGLP, 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 of Analysis was not accompanied by a GLP compliance statement. Section IT, 6.2 (5) Mixture analysis was not performed. This study was designed and performed at STILLMEADOW, Inc. and was conducted in compliance with JapanMinistry ofAgriculture, Forestry & 

### Fisheries, NotificationNo. 11 Nousan 6283, DirectorGeneral ofAgricultural Production Bureau with exception of: Article 3.1 (18), 12.5 and 12.7 The provided Certificate ofAnalysis was not accompanied by a GLP compliance statement. · Article 12.8 Mixture analysis was not performed.

Signature ofAgent ofSponsor
Agent Name Sponsor: Sasol Germany GmbH
Signature ofAgent ofSubmitter
AgentName Submitter: Sasol NA Date

Date

Date

STILLMEADOW, Inc.

### TABLE OF CONTENTS 13296-09 Page 

### 4 of 16

GOOD LABORATORYPRACTICE COMPLIANCE STATEMENT
................................................

### 2 QUALITY ASSURANCE STATEMENT
............................................................................................

### 3 SUMMARY
..........................................................................................................................................5- 

### INTRODUCTION
.................................................................................................................................5 TEST SUBSTANCE
.............................................................................................................................6 ADJUVANT
..........................................................................................................................................6 VEIDCLE AND/OR OTIIERMATERIALS
........................................................................................6 TEST SYSTEM
.....................................................................................................................................6 Experimental Animals
.....................................................................................................:.................6 An:hnal Husbandry
.............................................................................................................................7 POSITIVE CONTROL INFORMATION
.............................................................................................7 Positive Control Material
...................................................................................................................7

- Positive Control Testing
....................................................................................................................7 PROCEDURES
.................................................."
...................................................................................8 Test Substance Preparation
...............................:................................................................................8 Test Substance Application
...............................................................................................................8 Observations and Scoring Methods
...................................................................................................9 RESULTS AND DISCUSSION
............................................................................................................9 CONCLUSION
.....................................................................................................................................9 SIGNATURE
.........................................................................................................................**~** 

### ..............9 STUDY PERSONNEL
..........................................................................................................................9 PRETEST SCREENING
..................................................................................................................... 10 LEGEND TO TABLE 1
...................................................................................................................... 10 TABLE 1 - Skin Reaction Scores and Averages
.................................................................................. 11 TABLE 2.- BodyWeights
................................................................................................................... 13 APPENDIX A- Positive Control Tables
....................................'
...................................... _ .. ;................ 14 APPENDIX B - Certificate ofAnalysis
............................................................................................... 16

STILLMEADOW, Inc.

### QUALITYASSURANCE STATE:MENT

Test Substance: Alcohols, C18-22, distn. residues (CAS No: 1160164-88-4) Study Title: Guinea Pig Maximization TestFor TopicallyApplied Test Substance 13296-09 Page 3 of16

The studyreport and datahave been audited in accordance with GoodLaboratoryPractice Standards and STILLMEADOW, Inc. Standard Operating Procedures (SOPs). The fmal report accuratelyreflects 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 26 Oct 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 were reported to the Study 2 

### DirectorandManagement as follows:
Critical Phase Inspected Date Inspected Reported to Reported to Study Director Management Protocol Review 08 Sep 09 08 Sep 09 08 Sep 09 Dosing 14 Oct 09 14 Oct 09 14 Oct 09 Report/Data Audit 19 Nov09 19 Nov 09 19Nov09



### Richard L. 

### Martin, MS, CPhT Date QualityAssurance, STILLMEADO , Inc.

STILLMEADOW, Inc.

### SUMMARY 13296-09 Page 5 of16

A maximization test for topically applied test substances was conducted on 30 short-haired male and female albino guinea pigs to determine if the test substance, Alcohols, C18-22, distr1. residues (CAS No: 1160164-88-4) produced a sensitizingreaction. Thetest substance was warmedfor use in the intradermal portions ofthe study. Group I animals, the test group (10/sex), eachreceivedthree pairs of intradermal injections (adjuvant, a solution oftest substance in com oil, and a 50:50 mixture ofadjuvant and the test substance solution) followed one week later by a single topical application of the test substance in comoil. Ten additional animals (5/sex) served as a control group (Group II). Control animals were treated atthe sametimeperiods and locations butwiththe vehicle usedinplace of thetest substance solution. Two weeks afterthetopical application, thetest animals were challenged with a secondtopical application ofthetest substance in corn oil at a virgintest site. Control animals were also given a topical application ofthe test substance in com oil. The percentage of animals exhibiting erythema with or without edema afterthe challengetreatmentwas usedto assignthetest substance a sensitizationpotency rating. Sincenone ofthe testanimals exhibited scores greaterthan zero, Alcohols, C18-22, distn. residues (CAS No: 1160164-88-4) was given a sensitization potency rating of non-sensitizing (Grade 0). I 

### INTRODUCTION The objective ofthis study was to determine the sensitizing potential ofthe test substance, using the methods ofMagnusson and Kligman (J. Invest. Dermat52: 268-276 (1969)). This studywas conducted for Sasol Germany GmbH, according to the approved protocol (OECD 406, OPPTS 870.2600 and Canadiannotification) and STILLMEADOW, Inc. SOP's. Therewereno deviations from theprotocolthat affected the quality or outcome ofthe study. All procedures used in this study are in compliance with Animal Welfare ActRegulations. The protocol, raw data, this report and a sample oftest substance are archived at STILLMEADOW, Inc. The study was initiated on 11 Sep 09, and the experimental rangefmdingportionbegan on 04 Oct09. Thetreatment schedulewas as follows, andthe studywasterminated on31 Oct09:
Treatments Intradermal Injections Topical Aoplications Groups Initial Treatment Second Insult Challenge Test and Control 07 Oct 09 14 Oct 09 28 Oct 09

STILLMEADOW, Inc.

### Label Identification: Quantity & 

### Date Received: Physical Description: Storage: Purity: Stability: Concentrations Administered:
Vehicle: 

### TEST SUBSTANCE

·, 

### 13296-09 Page 6 of 16 Alcohols, C18-22, distn. residues (CAS No: 1160164-88-4); Lot: 03585/MA; Spec: 59BlRN2 11 Sep 09; 246.3 g (GW) Tan solid Room temperature See Certificate ofAnalysis Exp: Aug 2013 perprovided information 4% v/v solution oftest substance in vehicle with and without adjuvant (50:50 v/v) for intradermal injections; 10% w/v in vehicle for topical and challenge applications Corn ojl Datageneratedfor characterization and stabilityis theresponsibility of the sponsor. Records pertainingto identity, synthesis methods and location ofdocumentation are the responsibility ofthe sponsor. A copy ofthe Certificate ofAnalysis is included in report Appendix B.

Label: Manufacturer: Physical Description: Storage: Concentration Administered: Purity & 

### Composition: Stability: 

### ADJlNANTFreund's Complete Adjuvant Lot# 093K8932 Sigma Clear liquid Store in a cool, dark place at 2 to 8°C, do not freeze Diluted at 50% v/v in saline Available from manufacturer Exp: Apr 2010 

### VEIDCLE AND/OR OTHER MATERIALS Label: Manufacturer: Expiration Date: Label: Manufacturer: Expiration Date: Label: Manufacturer: Expiration Date:
Experimental Animals Species & 

### Strain: Justification ofSpecies: Source: Quantity & 

### Sex: Quarantine Period: Date Born/Date Received: Animal Identification: WeightWhen Tested: Sodium Chloride Lot # L771147 Baxter Nov 2010 Corn Oil Parade Apr 2010 Petrolatum Walgreen's Jan 2015 

### TEST SYSTEM
GuineaPig; Hartley-Albino The guinea pig is conventionally used in skin sensitization studies to provide information on which human hazard can bejudged. Charles River, Hdq: Wilmington, MA 15/sex (defmitive test) and 3/sex (screen); females nulliparous and non-pregnant 5 days 25 Aug 09 and 29 Aug 09 

### I 28 Sep 09 Ear punch Males (331-403 g); Females (320-380 g) STILLMEADOW, Inc.

### Animal Husbandry Cage Type: Housing: Environmental Controls

Set to Maintain: Actual Temp/Rel. Humidity:
Food: Water: Suspended, wire bottom, stainless steel Housed individually for 3 days (during wrapping) 1-5 animals per cage (males separate from females) 13296-09 Page 7 of16

·Temperature 20°± 3° C
· 12-hour light/dark cycle 19-21 o 

### C 

### I 47-92% ·Relative Humidity30-70%
· 10-12-air changes/hour Protocol deviation: Relative humidity was outside protocol range, but did not affect study outcome. PMIFeeds, Inc.TM Guinea Pig Diet#5025 available ad libitum Municipal water supply analyzed by TCEQ Water Utilities Division; available adlibitum from water bowls or automatic system Animalhusbaf1.dry and housing at STILLMEADOW, Inc. complywith standards outlined inthe "Guide for the Care andUse ofLaboratoryAnimals" (NRC Publ.). No contaminants were expectedto havebeen present in the feed or water that would have interfered with or affected the results ofthe study. 

### POSITIVE CONTROL INFORMATION Positive Control Material Label: Manufacturer: Physical Description: Concentrations Administered: Purity, Composition

& 

### Stability: Positive Control Testing alpha-Hexylcinnamaldehyde 85% Batch 13102MO Batch 03110TA 138630-100G Aldrich Clearyellow liquid 25% v/v solution oftest substance in safflower oil with and without adjuvant (50:50 v/v) for intradermal injections; 100% undiluted test substance for topical and challenge applications. Available from manufacturer

The sensitivity ofguinea pigs to a positive control material is confirmed in this laboratory periodically. Thepositive control animals usedto conductthis study were supplied by Charles River Laboratory, and were tested according to the Magnusson and Kligman 

### (J. Invest. Dermat 52: 268-276 (1969)). STILLMEADOW, Inc. StudyNo. 12290-08 In-life start: 27Aug 08; In-life completed: 20 Sep 08 Results: Datafromthis study are presented inAppendixA. 

### Since all animals of the test group exhibited moderate to intense erythema after the challenge treatment; and none ofthe animals ofthe naive control group exhibited erythema after the challenge treatment, the test substance is considered an extreme sensitizer and confirmed the sensitivity ofguinea pigs to the positive control material.

STILLMEADOW, Inc.

### PROCEDURES 

### Test Substance Preparation 13296-09 Page 8 of16

A pre-test screen with 3 animals/sexwas conductedto determine induction concentrations producing no more than moderate irritation, and a maximum non-irritating challenge concentration. Based on screen results, a 4% v/v solution of test substance in com oil was selected for intradermal injection, and a 10% w/v concentration oftest substance in com oilwas selected for the topical applications (induction and challenge). Healthy, short-haired, albino guinea pigs (males and females) were released from quarantine prior to testing. Five animals per sex were assigned to a control group (Group 

### ll). Ten animals per sex were assigned to the test group (Group I). On the day prior to each treatment, the animals were prepared by clipping the appropriate exposure areas free ofhair. Individual bodyweights were recorded on Days -1 and 24. The animals were treated on Days 0, 7, and 21.
Test Substance Application Induction: Intradermal Injections: 

### The animals were treated on Day 0 by making three pairs of symmetrical intradermal injections on the upper back ofeach animal within a 4 x 6 em exposure area running laterally across the shoulders. Forthe test animals (Group I), the first pair ofinjections (one on each side of the spinal column and approximately 3.5 em apart), consisting of Freund's Complete Adjuvant diluted to 50% v/v in saline, was made at the anterior edge ofthe exposure area. The second pair ofinjections, consisting of4%v/vtest substance in com oil, was made approximately 0.5 embehind the first pair. The third pair ofinjections, consisting ofa 50:50 mixture ofFreund's Complete Adjuvant (dilutedto 50% v/v in saline) and a solution of4%v/vtest substance in comoil was made approximately 0.5 em behind the second pair ofinjections. Group 

### ll control animals received the same injections with thevehicle substituted for thetest substance in the second andthirdpairs ofinjections. Allinjectionswere within a 2 x 4 em area ofthe 4 x 6 em exposure area. A volume of0.1 

### mLwas administered at each site. Induction: Topical Applications: 

### On Day 7, 0.5 

### mL of 10% w/v oftest substance in com oil was appliedto the exposure area ofeachtestgroup animal (Group I) to coverthe intradermalinjection sites. A 25 mm glass fiber filter patch was used to cover the dose site. The patch was then occluded with an adhesive maskingtape and secured inplacewith an elastic adhesive wrap wound aroundthetorso ofthe animal. Control group animals (Group 

### ll)received 0.5 

### mL ofcom oil and a 25 mmglass fiber filterpatch was placed overthe dose sites. The wrappings and patches wereremovedafter48 hours. Test sites 3 and 4 were observed for dermal irritation on Day 10. Challenge: Test and ControlAnimals: 

### On Day20, a 5 x 5 em area was clipped onboth the left and right flanks ofeach test and control animal. Forthe challengetreatment (Day21), 0.5 

### mLof 10% wlv of the test substance in com oil was applied topicallytothe rightflank ofeach animal in a manner identical to theDay 7 treatment.

## A25 

## riim glass fiber filter patchwas usedto coverthe dose site. A dose of0.5 

### mL of50:50 com oil:petrolatum was applied topicallyto the left flank ofeach animal. A 25 mm glass fiber filter patch was placed on the dose site. Patches were secured as above.

STILLMEADOW, Inc.Observations and Scoring Methods 13296-09 Page 9 of16
OnDay22 (24 hours after challenge), the wrappings andpatches wereremoved. OnDay23 (24 hours after unwrapping), the test sites were observed for skin reactions, and again observed on Day24 ( 48 hours after unwrapping). Observations were made ofright and leftflanks ofeach animal in GroupsI andII. The scoring scaleusedfor grading skinreactions afterboth intradermal andtopical exposures is presented inthe Legend to Table 

### 1. After both induction treatments (intradermal injections and topical applications) and the challenge exposure, average skin reaction scores were calculated. These data appear in Table 1. Any Group 

### I animals which exhibited scores greater than 0 forerythema with orwithout edema for the treated right flank after the challenge treatment were considered possibly sensitized. However, the skin reactions ofthe leftflank treatedwithvehicle alone, andthe skinreactions ofthe naive controls were also evaluated. Thetest substancewas graded andrated for sensitizationpotencybaseduponthepercentage of animals sensitized using the scoring scale presented below: 

### % Sensitized 

### 0 1-8 9-28 29-64 65-80 81-100 Grade 0 

### I 

## n ill IV 

## v Rating Non -sensitizer Weak Mild Moderate Strong Extreme 

### RESULTS AND DISCUSSION Skin reaction scores and average skin reaction scores are presented in Table 1. Body weights are presented in Table 2. Bodyweight gain was unaffectedbythe administration ofthetest substance. The challengetreatmentswith eithervehicle alone ortest substance invehicle producedno erythemain any ofthe test and or control group animals. 

### CONCLUSION Sincenone ofthetest animals exhibited scores greaterthanzero, Alcohols,**C18~22,** distn. residues (CAS No: 1160164-88-4) was given a sensitization potencyrating ofnon-sensitizing (Grade 0).
Study Director: J "ce 

### 0. Kuhn, PhD, DABT Date 

# 23 

# hArz! () 

# 1 S n· r Toxicologist, STILLMEADOW, Inc. 

### STUDYPERSONNEL Technical Staff: Carol Morris, BA Robert Preston
Data Services: Jeanne Poorman, BS Report Preparation Paul Siemens, BA Nancy Casajuana, LAT

STILLMEADOW, Inc.**PRETESTSCREE~G** Guinea Pig Maximization Test For Topically Applied Test Substance Intradermal Route

- FirstInsult: Dose amount: 0.1 mL Vehicle' Corn Oil

Animal Body Number Wt (g) 285-M 344 286-F 351 LF 1 1 Observation Time After Initial Dose 24 Hour Score 48 Hour Score

RF LR RR LF RF LR

1 0 0 0.5 0 0 0.5 0 0 0 0 0
External Cutaneous Route

- Second Insult and Challenge: Dose amount: 0.5 g Vehicle' Petrolatum Animal Body Number Wt(g) 281-M 341 282-M 340 283-F 343 284-F 347

Intradermal Site Dose LF 5%v/v RF 4%v/v LR 2%v/v RR 1%v/v 48 Hour Observation LF RF

1 1 0 0 0.5 1 0.5 0

Animals Topical Application treated Site Dose LF 25%w/v 285-M RF 10%w/v LF 5% w/v 286-F RF 1%w/v RR 0 0

Animals treated 281-M 282-F 283-M 284-F
Chosen Dose=4% v/v in Corn Oil Chosen Dose= 10% w/v in corn oil'



### LEGEND TOT ABLE 1 Evaluation ofSkin Reactions 13296-09 Page 10 of 16

Magimsson and Kligman Grading Scale for the Evaluation ofChallenge Patch TestReactions* Erythema Formation No visible change Discrete or patchy erythema Moderate and confluent erythema Intense erythema and swelling Score 0 

### 1 2 

### 3 * - OECD Guidelines for the Testing ofChemicals, Volume 2, Section 4, Number 406, Skin Sensitization, Paragraph 23, page 4/9, Adopted 17 Jul92

STILLMEADOW, Inc.

### TABLE 1- Skin Reaction Scores and Averages Guinea Pig Maximization Test For Topically Applied Test Substance Test Substance: Alcohols, C18-22, distn. residues (CAS No: 1160164-88-4) Group I - Test 13296-09 Page 11 of 16

Intradermal Induction Topical Induction Challenge Animal Scores for each site on Day 6 Day 10 Scores Number 1 

# I 2 

# I 3 4 

## I 5 

# I 6 Site 3 Site 4

11-M 1 

# I 1 0.5 0.5 0 

# I 0 0 0 12-M 1 

# ! 1 0.5 

# I 0.5 

# I 0 0 0 

# I 0 13-M 2 

### I 2 

# I 0 

# I 0 

# I 0 

# I 0 0 

### i 0 ! I 14-M 0.5. 

# I 0.5 0 

# I 0 0 

# I 0 0 

# I 0 15-M 1 

# I 1 0 0 0
- 0 0 0 16-F 0.5 

# I 0.5 

# i 0 0 i 0 0 0 

# I 0 I 17-F 0.5 

# I 0.5 

# I 0 

# I 0 

# i 0 

# I 0 0 

# i 0 18-F 1 

# I 1 

# I 0 0 

# I 0 

### I 0 I 0 

# I 0 19-F 0.5 

# I 0.5 0 0 0 I 0 0 

# I 0 I 

# I I I I 20-F 1 1 0 0 0 0 0 I 0 I I I ! 21-M 2 

# I 2 

# I 0 

# I 0 

# I 0 

# I 0 2 

# i 0.5 22-M 0.5 

# I 0.5 0 0 0 0 1 i 0.5 I 23-M 0.5 

# I 0.5 

# I 0.5 

### I 0.5 

# I 0 

# I 0 0 

# I 0 24-M 1 

# I 1 0 0 0 

## I 0 0 

# I 0 25-M 0.5 

# I 0.5 I 

### I 0 0 

# I 0 i 

### I 0 0 0 26-F 0.5 

# I 0.5 

# I 0 0 

# I 0 

# I 0 0 

# I 0 27-F 1 

# I 1 

# I 0 0 0 0 0 

# I 0 28-F 0.5 

# I 0.5 0 0 0 

# I 0 0 0 

# I I I 

# I i 29-F 0.5 0.5 0 0 I 0 0 0 

### I 0 30-F 0.5 

# I 0.5 

# I 0 

# I 0 

# I 0 

# I 0 0 

# I 0
Mean:

# ~L_o.8 0.1 0.1 0.0 0.0 0.1 0.3 Incidence of Reactions: %Sensitized: 0% Sensitization Potency Rati:Q.g: Non-sensitizer Injection Sites: Sites 1 & 2 - Freund's Complete Adjuvant

- 50% v/v in saline Sites 3 & 4 - Test Substance 4% v/v in com oil Day23 LF 

# I RF 0 0 0 0 0 

# I 0 0 

# I 0 0 0 0 0 0 

# I 0 0 0 0 0 0 

# I 0 0_ 

# I 0 0 0 0 

# I 0 0 0 0 

# I 0 0 I 0 

### I 0 0 0 0 0 

## I 0 0 

# I 0 

# O.?j_ 0.0



### --- 0/20

Sites 5 & 6 -Mixture (50:50) ofFreund's Complete Adjuvant in saline and Test Substance in com oil Induction topical and Challenge Treatments (0.5 rnL): 10% w/v mixture ofTest Substance in com oil LF

- Left Flank; RF

- Right Flank M - Male; F - Female Day24 LF RF 0 0 0 

### i 0 0 0 0 0 0 0 0 

### I 0 0 

# i 0 0 

# I 0 0 0 0 0 0 0 0 

### ! 0 0 

# I 0 0 0 0 0 0 0 0 0 0 I 0 0 0 0 0

# ~~J-~ 0/20

STILLMEADOW, Inc.

### TABLE 1- Skin Reaction Scores and 

### Averages (cont.) Guinea Pig Maximization Test For Topically Applied Test Substance 13296-09 Page 12 of 16

Test Substance: Alcohols, C18-22, distn. residues (CAS No: 1160164-88-4) Group II

- Control Intradermal Induction Animal Scores for each site on Day 6 Number 1 2 3 4 5 6 1-M 1 1 

# I 0 

# I 0 

# I 0 0 2-M I 0 

# I 0 

# I 0 0 1 

## I 1 i 3-M 1 

# I 1 0.5 

## I 0.5 0 

# I 0 I 4-M 1 1 0 

# I 0 

# ! 0 0 5-M 1 

# I 1 

# I 0 

# I 0 

# I 0 

# I 0 6-F 1 

# I 1 0 

# I 0 i 0 0 7-F 

# I 0.5 I 0.5 0 

# I 0 1 1 

# I I 8-F i 0 i 0 0 

# I 0 1 

# I 1 I 9-F 1 

# I 1 

# I 0.5 

# I 0.5 

# I 0 

# I 0 10-F 0.5 0.5 0 0 0 0
Mean: 1.0 1.0 0.2 0.2 0.0 0.0

---- ---- -- 0.4 Injection Sites: Sites I & 2 - Freund's Complete Adjuvant

- 50% v/v in saline Sites 3 & 4- Vehicle (corn oil) 

### Topical Induction Day 10 Scores Site 3 

# I Site 4

I 0.5 

# I 0.5 0.5 0.5 0.5 0.5 0.5 

# I 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 

# I 0.5 0.5 

# I 0.5 0.) Incidence of

Reactions:

Sites 5 & 6 -Mixture (50:50) ofFreund's Complete Adjuvant in saline and vehicle Induction Topical Treatments: 0.5 mL ofvehicle Challenge Treatments (0.5 mL): I0% w/v ofTest Substance in vehicle LF

- Left Flank; RF

- Right Flank M- Male; F- Female 

### Challenge Day23 Day24 LF RF LF 

# I RF 0 0 0 

# I 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 

# I 0 0 0 0 0 0 0 0 

# I 0 0 0 0 

# I 0 0 

## I 0 0 

# I 0 0 0 0 

# I 0

# o~J_i_?_

# /-?!__~~--

0/10 0/10

STILLJylEADOW, Inc.

### TABLE 2- Body Weights Guinea Pig Maximization Test For Topically Applied Test Substance Test Substance: Alcohols, C18-22, distn. residues (CAS No: 1160164-88-4)

Animal Day ofStudy Number Day -1 Day24 Group I- Test 11-M 340 550 12-M 331 520 13-M 403 500 14-M 395 483 15-M 378 445 16-F 327 492 17-F 344 376 18-F 338 386 19-F 320 409 20-F 321 394 21-M 350 457 22-M 384 486 23-M 396 518 24-M 337 506 25-M 332 460 26-F 351 416 27-F 380 450 28-F 365 443 29-F 351 449 30-F 349 409 Group ll- Control 1-M 339 465 2-M 375 484 3-M 368 476 4-M 345 465 5-M 357 481 6-F 374 440**7~F** 362 436 8-F 360 455 9-F 349 441 10-F 321 422 M - Male; F -Female Note: weights are in grams 

### 13296-09 Page 13 of 16 ·

STILLMEADOW, Inc.

### APPENDIXA - Positive Control Tables Guinea Pig Maximization Test For Topically Applied Test Substance Skin Reaction Scores and Averages Positive Control Substance: alpha-He:Xylcinnarnaldehyde 85% StudyNumber: 12290-08 Group I- 

### Test

Animal Number 1

11-M 3 12-M 0 13-M 0 14-M 0 15-M 0 16-F 0 17-F 2 18-F 3 19-F 3 20-F 2
Mean: 1.3

% Sensitized: 100% Injection Sites: 

### Intradermal Induction Scores for each site onDay 6 2 3 4 5 3 3 3 0 0 3 3 0 0 3 3 0 0 3 3 0 0 0 0 0 0 3 3 0 2 3 3 0 3 3 3 0 3 3 3 0 2 2 3 3 1.3 2.6 2.7 0.3 1.4 6 0 0 0 0 0 0 \ 0 0 0 0 0.0
' 

### Topical Induction Day 10 Scores Sitd Site 4

3 3 3 3 3 3 3 3 3 3 

## * * * * * * * * * * 3.0 Incidence of

Reactions:

Sensitization Potency Rating: Extreme Sensitizer
Sites 1 & 2

- Freund's Complete Adjuvant

- 50% v/v in saline Sites 3 & 4

- Test Substance 25% v/v in safflower oil 

### 13296-09 Page 14 of16

Challenge Day23 Day24 LF RF LF RF 3 3 3 3 3 3 2 3



# - - - 

# - 3 3 2 2 3 3 3 3 3 3 3 3 3 3 3 2 3 3 3 2 3 3 2 3 3 3 3 3 3.0 3.0 2.7 2.7 919 9/9

Sites 5 & 6- Mixture (50:50) ofFreund's Complete Adjuvant in saline and Test Substance (25% v/v) Induction Topical and Challenge Treatments: 100% undiluted Test Substance LF

- Left Flank; RF

- Right Flank " M- Male; F -Female; 

## *
- Scores notrecorded Note: Animal 13-M found dead on Day 15

STILLMEADOW, Inc.

### APPENDIXA - Positive Control Tables (cont.) Guinea Pig Maximization Test For TopicallyApplied Test Substance Skin Reaction Scores and Averages Positive Control Substance: alpha-Hexylcinnamaldehyde 85% StudyNumber: 12290-08 Group ll
- 

### Control
Intradermal Induction

Animal Scores for each site on Day 6 

### Topical Induction Day 10 Scores Number 1 2 3 4 5 

### 6 · Site 3 Site 4

1-M 2 3 3 4 0 0 2-M 2 3 3 3 0 0 3-M 2 3 3 3 0 0 4-M 2 2 3 3 0 0 5-M 3 3 3 3 0 0 

### 6-F 2 3 3 3 0 0 

### 7-F 2 3 3 3 0 0 

### 8-F 2 2 3 3 0 0 

### 9-F 2 2 3 3 0 0 10-F 2 2 3 3 0 

### o·
Mean: 2.1 2.6 3.0 3.1 0.0 0.0 

### 1.8
Injection Sites: Sites 1 & 2

- Freund's Complete Adjuvant

- 50% v/v in saline Sites 3 & 4

- Safflower oil 1 2 2 2 2 2 2 2 2 2 3 2 3 2 1 2 2 3 2 3 2.1 Incidence of

Reactions: Day23 

### LF RF 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.0 0.0 0/10 

### 13296-09 Page 15 of 16

Day24 · 

### LF RF 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0.0 0.0 0/10

Sites 5 & 6- Mixture (50:50) ofFreund's Complete Adjuvant in saline and Safflower oil Induction Topical Treatments: 0.5 mL ofSafflower oil Challenge Treatments: 100% undiluted Test Substance LF

- Left Flank; RF

- Right Flank M - Male; F - :f;:male

STILLMEADOW, Inc.

### APPENDIX B - Certificate ofAnalysis

C E R T I F 

### I C A T E OF 13296-09 Page 16 of 16 

# sasoL reaching nsw{rtllltiers 

### Brunsbuettel,**9/09/2~09**

ANALYSIS

Product; Lot No.: 

### Alcohols, ClS-22, distn. 

### Residues 03585/MA (59BlRN2) 

### Manufacturing date: E,xpiry date:

Hydroxyl

- Numl;:ie:r Ester No. Acid. N'o. Water Iodine !lo. 25/08/2009 08/2013

[mg :KOH/g] [mg KOH/g] [mg :KOH/gl [wt.%1 [mg I/lOOmg] 50, 41, l7 0,09 0,02 5,34



### Sasol Germany GmbH P.o. :Sox 

### 1160**D-255~4** Brunsbuettel 

### Phone: 049 (0)4852/392-0 Fax: 049 (0)4852/3285 

### Best regards 

### works inspector M. Sprung

DiE**OllerS~;ndung dte~~~** AnalysEinz.erllfil<<IIs· etiolglltlcllglich :wi b1forma1ion uod sh:-lll keihec BeschalfNlheits- ui1d**Haitbarkei!~­** g;u;;mU!> dar: Ole Dbersenc!ung entt:ilntiefden En'l!xlinger**n~~i':.t.**von**d~r** Durcllfil!mJng elner OJtii1UI1Q!lgemaBen Wal1lfleJr,tmng6prufung, Dics<'f Allal)•ssnzartiiU;at begr\indst keine Af;spi-cicl\e Dnller. an die**~;; W6ii~l\)ereldi1**  wird, lhl**()br~en** ge!ten unseio; Allgenwinr,>o**;~** . .-

Thiil: CE11lfio.ate of**l!l'!lll)'si~** i:; loriniormatlon**oo~·**mddoes not gua• ri!/Jle_e af!y patticu!ar**Pi~dU<;t** propertieS. It dOe!>not fi·aa fh!i recipi· en! ofihaobligaiibn toca•'IYou1 a prodvcl receiving tnsp;.;:::li(ln, This certiik:""" ofanalysis r:Jre:. nm create claimr. o!Ulird j)llrUes In,-.4\lt:h li Is**~.-.;** o!·, Ali lransactkms aro·**subje~t**iri**ot~·** Generi;l Bui>ii1*S D:mdilions .as arnenct>.'d 1mlo 11\.;time**<:oi1C2rn~d.** STILLMEADOW, Inc.