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 

TOPICALLY APPLIED 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 Park One Drive 

Sugar Land, 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 LABORATORY PRACTICE 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 of Analysis 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 Japan Ministry of Agriculture, Forestry & Fisheries, Notification No. 11 Nousan 6283, DirectorGeneral of Agricultural Production Bureau with exception of: 

Article 3.1 (18), 12.5 and 12.7 The provided Certificate of Analysis was not accompanied by a GLP 

compliance statement. · 

Article 12.8 Mixture analysis was not performed. 

Signature of Agent of Sponsor 

Agent Name 

Sponsor: Sasol Germany GmbH 

Signature of Agent of Submitter 

Agent Name 

Submitter: Sasol NA 

Date 

Date 

Date 

STILLMEADOW, Inc. 
TABLE OF CONTENTS 

13296-09 

Page 4 of 16 

GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT ................................................ 2 

QUALITY ASSURANCE STATEMENT ............................................................................................ 3 

SUMMARY .......................................................................................................................................... 5-

INTRODUCTION ................................................................................................................................. 5 

TEST SUBSTANCE ............................................................................................................................. 6 

ADJUVANT .......................................................................................................................................... 6 

VEIDCLE AND/OR OTIIER MATERIALS ........................................................................................ 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 ..................................................................................................................... 1 0 

LEGEND TO TABLE 1 ...................................................................................................................... 10 

TABLE 1 - Skin Reaction Scores and Averages .................................................................................. 11 

TABLE 2.- Body Weights ................................................................................................................... 13 

APPENDIX A- Positive Control Tables ................................... .' ...................................... _ ..; ................ 14 

APPENDIX B - Certificate of Analysis ............................................................................................... 16 

STILLMEADOW, Inc. 
QUALITY ASSURANCE STATE:MENT 

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

Study Title: Guinea Pig Maximization Test For Topically Applied Test Substance 

13296-09 

Page 3 of16 

The study report and data have been audited in accordance with Good Laboratory Practice Standards and 

STILLMEADOW, Inc. Standard Operating Procedures (SOPs). The fmal report accurately reflects the 

study data. The Quality Assurance Unit has not been involved in the actual conduct of this 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. 

The fmdings from any study inspections and audits were reported to the Study2 

Director and Management 

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 

Quality Assurance, 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: 1160164-8 8-4) produced a sensitizing reaction. The test substance was warmed for use in the 

intradermal portions of the study. Group I animals, the test group (10/sex), each received three pairs of 

intradermal injections (adjuvant, a solution oftest substance in com oil, and a 50:50 mixture of adjuvant 

and the test substance solution) followed one week later by a single topical application of the test 

substance in com oil. Ten additional animals (5/sex) served as a control group (Group II). Control animals 

were treated at the same time periods and locations but with the vehicle used in place of the test substance 

solution. Two weeks after the topical application, the test animals were challenged with a second topical 

application of the test substance in corn oil at a virgin test site. Control animals were also given a topical 

application of the test substance in com oil. The percentage of animals exhibiting erythema with or 

without edema after the challenge treatment was used to assign the test substance a sensitization potency 

rating. Since none of the testanimals exhibited scores greater than 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 of this study was to determine the sensitizing potential of the test substance, using the 

methods of Magnusson and Kligman (J. Invest. Dermat 52: 268-276 (1969)). This study was conducted 

for Sasol Germany GmbH, according to the approved protocol (OECD 406, OPPTS 870.2600 and 

Canadian notification) and STILLMEADOW, Inc. SOP's. There were no deviations from the protocol that 

affected the quality or outcome of the study. All procedures used in this study are in compliance with 

Animal Welfare Act Regulations. 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 rangefmding portion began on 04 Oct 09. The treatment schedule was as follows, and the study was terminated 

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 of Analysis 

Exp: Aug 2013 per provided 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 

Data generated for characterization and stability is the responsibility of the sponsor. Records pertaining to 

identity, synthesis methods and location of documentation are the responsibility of the sponsor. A copy 

of the Certificate of Analysis 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 of Species: 

Source: 

Quantity & Sex: 

Quarantine Period: 

Date Born/Date Received: 

Animal Identification: 

Weight When Tested: 

Sodium Chloride Lot # L 77114 7 

Baxter 

Nov 2010 

Corn Oil 

Parade 

Apr 2010 

Petrolatum 

Walgreen's 

Jan 2015 

TEST SYSTEM 

Guinea Pig; Hartley-Albino 

The guinea pig is conventionally used in skin sensitization studies to 

provide information on which human hazard can be judged. 

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% 

· 1 0-12-air changes/hour 

Protocol deviation: Relative humidity was outside protocol range, 

but did not affect study outcome. 

PMI Feeds, Inc.TM Guinea Pig Diet #5025 available ad libitum 

Municipal water supply analyzed by TCEQ Water Utilities Division; 

available ad libitum from water bowls or automatic system 

Animal husbaf1.dry and housing at STILLMEADOW, Inc. comply with standards outlined in the "Guide 

for the Care and Use ofLaboratory Animals" (NRC Publ.). No contaminants were expected to have been 

present in the feed or water that would have interfered with or affected the results of the study. 

POSITIVE CONTROL INFORMATION 

Positive Control Material 

Label: 

Manufacturer: 

Physical Description: 

Concentrations Administered: 

Purity, Composition 

& Stability: 

Positive Control Testing 

alpha-Hexylcinnamaldehyde 85% Batch 131 02MO 

Batch 03110TA 138630-100G 

Aldrich 

Clear yellow liquid 

25% v/v solution of test 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 of guinea pigs to a positive control material is confirmed in this laboratory periodically. 

The positive control animals used to conduct this 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. Study No. 12290-08 

In-life start: 27 Aug 08; In-life completed: 20 Sep 08 

Results: Data from this study are presented in Appendix A. Since all animals of the test group exhibited 

moderate to intense erythema after the challenge treatment; and none of the animals of the 

naive control group exhibited erythema after the challenge treatment, the test substance is 

considered an extreme sensitizer and confirmed the sensitivity of guinea pigs to the positive 

control material. 

STILLMEADOW, Inc. 
PROCEDURES 

Test Substance Preparation 

13296-09 

Page 8 of16 

A pre-test screen with 3 animals/sex was conducted to 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 of test substance in com oil was 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 of hair. Individual body weights 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 of each animal within a 4 x 6 em exposure area 

running laterally across the shoulders. For the test animals (Group I), the first pair of injections (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 of the exposure area. The second 

pair of injections, consisting of 4% v/v test substance in com oil, was made approximately 0.5 em behind 

the first pair. The third pair of injections, consisting of a 50:50 mixture of Freund's Complete Adjuvant 

(diluted to 50% v/v in saline) and a solution of4% v/vtest substance in com oil was made approximately 

0.5 em behind the second pair of injections. Group ll control animals received the same injections with 

the vehicle substituted for the test substance in the second and third pairs of injections. All injections were 

within a 2 x 4 em area of the 4 x 6 em exposure area. A volume of0.1 mL was administered at each site. 

Induction: Topical Applications: On Day 7, 0.5 mL of 10% w/v of test substance in com oil was 

applied to the exposure area of each test group animal (Group I) to cover the intradermal injection sites. A 

25 mm glass fiber filter patch was used to cover the dose site. The patch was then occluded with an 

adhesive masking tape and secured in place with an elastic adhesive wrap wound around the torso of the 

animal. Control group animals (Group ll) received 0.5 mL of com oil and a 25 mm glass fiber filter patch 

was placed over the dose sites. The wrappings and patches were removed after 48 hours. Test sites 3 and 

4 were observed for dermal irritation on Day 10. 

Challenge: Test and Control Animals: On Day 20, a 5 x 5 em area was clipped on both the left and 

right flanks of each test and control animal. For the challenge treatment (Day21), 0.5 mL of 10% wlv of 

the test substance in com oil was applied topically to the right flank of each animal in a manner identical 

to the Day 7 treatment. A 25 riim glass fiber filter patch was used to cover the dose site. A dose of0.5 mL 

of 50:50 com oil:petrolatum was applied topically to the left flank of each 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 

On Day 22 (24 hours after challenge), the wrappings and patches were removed. On Day 23 (24 hours after 

unwrapping), the test sites were observed for skin reactions, and again observed on Day 24 ( 48 hours after 

unwrapping). Observations were made of right and left flanks of each animal in Groups I and II. The scoring 

scale used for grading skin reactions after both intradermal and topical exposures is presented in the 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 for erythema with or without edema for the 

treated right flank after the challenge treatment were considered possibly sensitized. However, the skin 

reactions of the left flank treated with vehicle alone, and the skin reactions of the naive controls were also 

evaluated. The test substance was graded and rated for sensitization potency based upon the percentage 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. Body weight gain was unaffected by the administration of the test substance. The 

challenge treatments with either vehicle alone or test substance in vehicle produced no erythema in any 

of the test and or control group animals. 

CONCLUSION 

Since none of the test animals exhibited scores greater than zero, Alcohols, C18~22, distn. residues (CAS 

No: 1160 164-88-4) was given a sensitization potency rating of non-sensitizing (Grade 0). 

Study Director: 

J "ce 0. Kuhn, PhD, DABT Date 

23 hArz! () 1 

S n· r Toxicologist, STILLMEADOW, Inc. 

STUDY PERSONNEL 

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 - First Insult: 

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 of Skin Reactions 

13296-09 

Page 10 of 16 

Magimsson and Kligman Grading Scale for the Evaluation ofChallenge Patch Test Reactions* 

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 of Chemicals, 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: 1160 164-88-4) 

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 I I 0 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) of Freund's Complete Adjuvant in saline and Test Substance in com oil 

Induction topical and Challenge Treatments (0.5 rnL): 10% w/v mixture of Test 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 1 0 0 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 1 0 1 I I 

8-F i 0 i 0 0 I 1 0 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 1 0 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) of Freund's Complete Adjuvant in saline and vehicle 

Induction Topical Treatments: 0.5 mL of vehicle 

Challenge Treatments (0.5 mL): I 0% w/v of Test 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 of Study 

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. 
APPENDIX A - Positive Control Tables 

Guinea Pig Maximization Test For Topically Applied Test Substance 

Skin Reaction Scores and Averages 

Positive Control Substance: alpha-He:Xylcinnarnaldehyde 85% 

Study Number: 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 on Day 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) of Freund'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 not recorded 

Note: Animal 13-M found dead on Day 15 

STILLMEADOW, Inc. 
APPENDIX A - Positive Control Tables (cont.) 

Guinea Pig Maximization Test For TopicallyApplied Test Substance 

Skin Reaction Scores and Averages 

Positive Control Substance: alpha-Hexylcinnamaldehyde 85% 

Study Number: 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 of Safflower oil 

Challenge Treatments: 100% undiluted Test Substance 

LF - Left Flank; RF - Right Flank 

M - Male; F - :f;:male 

STILLMEADOW, Inc. 
APPENDIX B - Certificate of Analysis 

C E R T I F I C A T E OF 

13296-09 

Page 16 of 16 

sa soL 

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:ilntief den 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:; lor iniormatlon oo~· md does not gua• 

ri!/Jle_e af!y patticu!ar Pi~dU<;t propertieS. It dOe!> not fi·aa fh!i recipi· 

en! ofiha obligaiibn to ca•'IY ou1 a prodvcl receiving tnsp;.;:::li(ln, This 

certiik:""" of analysis 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.