STUDY SPONSOR: 

Kao Corporation 

Division of Product Safety 

PAGE 1 OF 12 PAGES 

SafePharm 

Laboratories 

EMULGEN B-66: 

ACUTE DERMAL IRRITATION 

IN THE RABBIT 

SPL PROJECT NUMBER: 140/1099 

AUTHOR: 

TEST FACILITY: 

A Sanders 

Safepharm Laboratories Limited 

P.O. Box No. 45 

Safety & Environmental Research Center 

2606,1えkab組 e,Ichikaimachi 

DERBY 

DE12BT 

Haga 

TOCHIGI32ト3497

JAPAN 

140-1099.doclJS 

U.K. 

Telephone: (01332) 792896 

Facsimile: (01332) 799018 
「 ー

SPL PROJECT NUMBER: 140/1099 PAGE2 

QUALITY ASSURANCE REPORT 

百せsstudy type is classedぉ short-term.百les岡 d紅 dtest me也odfor白isstudy type (“General 

Study Plan" in OECD terminology) wお おviewedfor compliance once only on initial production 

Inspection of由eroutine and repetitive procedures也atconstitute the study is carried out as a 

con出 uousprocess designed to encompぉs血emajor phases at or about the time也isstudy was in 

progress. 

This report hぉ beenaudited by Safepharm Quality Assurance U凶t,and is considered to be an 

accurate account of也eda包 generatedand ofthe proced町 esfollowed. 

In each case, the outcome of QA evaluation is repo巾 dto也eStudy Director and Management on 

也eday of evaluation. Audits of study documentation, and process inspections appropriate to白E

type and schedule of也isstudy wereぉ follows:

13 August 2001 Standard Test Method Compliance Audit 

06 February 2002 Test Material Preparation 

15 February 2002 Animal Preparation 

15 February 2002 Dosing 

15 February 2002 Assessment ofResponse 

~ 29 March 2002 Draft Report Audit 

~ DateofQA S明 a加 E Final Report Audit 

~ Evaluation specific to也isstudy 

百三b.1反応三: DATE: 

For Safepharm Quali匂TAssurance U凶t本

• Authorised QA Signatures: 

Head ofDepa血 nent JR Pateman CBiol MIBiol DipRQA FRQA 

JMCrow由erMIScT MRQA 

1 9 APR 2002 

Deputy Head ofDepar回目立Senior Audit S阻ff: JV Johnson BSc MRQA; G Wren ONC MRQA; R Hurst MRQA 
SPL PROJECT NUMBER: 140/1099 PAGE3 

GLP COMPLIANCE STATEMENT 

百lework described wぉ performedin compliance wi也 UKGLP standards (Schedul巴 1,Good 

Laboratory Practice Regulations 1999 (SI 1999β106)). These Regulations紅 'ein accord組 cewi吐1

GLP standards published as OECD Principles on Good Laboratory Practice (revised 1997, 

ENVぶ1C/CHEM(98)17); and紅 ein accord組 .cewith, and凶 plement,出巴 requirementsof 

Directives 87/18AEC (as amended by Directive 1999/11厄C)and 88/320厄EC(ぉ amendedby 

Directive 1999/12AC). 

百leseinternational standards紅 eacceptable to the Regulatory agencies of出efollowing 

coun凶es:Australia, Austria, Belgium, Canada, the Czech Republic, Denmark, Finland, France, 

Germany, Greece, Hungary, Iceland, Ireland, Israel, Italy, Japan, Republic of Korea, 

Luxembourg, Mexico,百leNetherlands, New Zealand, Norway, Poland, Portugal, Slovenia, 

Sp出n,Sweden, Switzerland, Turkey, the United Kingdom, and the United States of America. 

This report fully and accurately reflects the procedures used and da匂 generated.

~...~~. A Sanders 

Study Director 
SPL PROJECT NUMBER: 140/1099 PAGE4 

CONTENTS 

QUALITY ASSURANCE REPORT 

GLP COMPLIANCE STATEMENT 

CONTENTS 

SUMMARY 

1.町TRODUCTION

TESTMATE阻 AL

2.1 Description, Identification and Storage Conditions 

2.2 Preparation ofTest Material 

2.3 Me俗 世'ementofpH

METHODS 

3.1 Animals and Animal Husbandry 

3.2 Procedure 

3.3 Interpretation ofResults 

4. ARCHlVES 

5. RESULTS 

6. CONCLUSION 

2. 

3. 

Table 1 Individual Skin Reactions 

Appendix 1 Statement of GLP Compliance in Accordance wi也 Directive88/3201EEC 

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SPL PROJECT NUMBER: 140/1099 PAGE5 

EMULGEN B-66: 

ACUTE DERMAL IRRITATION IN THE RABBIT 

SUMMARY 

Introducnon.百lestudy was performed to鎚 sess吐le凶 t組 cypotential of the test material to the 

skin ofthe New Zealand White rabbit 刀leme也odwas designed to meet the requirements of the 

f01l0羽ng:

• OECD Guidelines for the Testing of Chemicals No. 404“Acute Dermal IrritationlCoITosion" 

(adopted 17 July 1992) 

• Commission Directive 92/69.厄ECMe白odB4 Acute Toxicity (Skin Irritation) 

Resulぉ. A single 4-hour, semi-occluded application of the test material to the intact skin of也氏e

rabbits produced no evidence of skin iηitation. 

Conc[usion. The test material produced a primary irrita討onindex of 0.0祖 dwas classified as 

non-irritant to rabbit skin according to也eDraize classification scheme. N 0 cOITosive effects 

werenoted. 

刀letest material did not meet也ecriteria for classificationぉ註rit組 tor cOITosive according to也e

EU labe1ling regulations Commission Directive 93/21厄EC.
SPL PROJECT NUMBER: 140/1099 PAGE6 

EMULGEN B-66: 

ACUTE DERMAL IRRITATION IN THE RABBIT 

1.悶TRODUCTION

百lestudy was performed to assess也eirri匂ncypotential of由etest material following a single, 

4・hour,semi-occluded application to也eintact rabbit skin刀lemethod was design吋 tomeet the 

requirements of仕lefollowing: 

• OECD Guidelines for也eT,白血19of Chemicals No. 404“Acute Dermal IrritationlCoITosion" 

(adopted 17 Ju1y 1992) 

Commission Directive 92/69厄ECMethod B4 Acute Toxicity (Skin Irritation) 

刀lealbino rabbit has been shown to be a suitable model for也istype of study and is 

recommended in也etestme血od.百leresu1ts of the study紅巳 believedto be of value in predic出 g

白elikely skin irritancy potential of the test material to man. 

百lestudy was performed between 21 February 2002 and 24 February 2002. 

2. TEST MATERIAL 

2.1 Description, Identification and Storage Conditions 

Sponsor's identification 

Description 

Batch number 

Date received 

Storage conditions 

EMULGEN B-66 

light brown slightly加rbidviscous liquid 

2376 

31 December 2001 

room temperature in也巴 dark

Data relating to the identity, purity and stability of也etest material釘 e白eresponsibility of the 

Sponsor. 

2.2 Preparation of Test Material 

For也ep町 poseof the study the test material wぉ usedas supplied. 

τbe absorption of the test material was not deter百lIned
SPL PROJECT NUMBER: 140/1099 PAGE7 

2.3 Measurement of pH 

The pH of a 90% v/v aqueous preparation of the test material was determined prior to 

commencement ofthe study and found to be approximately 8.5. 

3. METHODS 

3.1 Animals and Animal Husbandry 

Three New Zealand White rabbits were supplied by David Percival Ltd, Moston, Sandbach, 

Cheshire, UK. At the s匂rtof the study吐leanimals were加 theweight range of2.0 to 3.5 kg組 d

were twelve to sixteen weeks old. After組 acclimatisationperiod of at least five days each 

g註malwas given a number unique within the study which was wri抗告nwl仕1a black indelible 

marker-pen on也emner s町 faceof血ee紅 釦don也ecage label. 

百lear廿malswere individually housed in suspended metal cages. Free access to IIl抵ns合i叫cing

water and food (Certified Rabbit Diet (Code 5322) supplied by PMl Nu凶tionIntemational, 

No口ingharn,UK) w俗 allowed耐 oughout血estudy. 百lediet and drinking water were 

considered not to∞nta凶 anycontarninant of a level白紙 mighthave affected也.epurpose or 

integrity of也estudy. 

The tempera旬 reand relative humidity were set to achi巴velimits of 17 to 230C and 30 to 70% 

respectively. Any occasional deviations企'omthese targets wer巴 considerednot to have affected 

也epurpose or integrity of the study. The rate of air exchange was at least fifteen changes per 

hourand也elighting w.ぉ con甘olledby a tInle switch to give twelve hours continuous light (06:00 

to 18:00) and twelve hours darkness. 

The animals were provided with environmental enrichment items which were consider巴dnot to 

co脚 m組 ycontan1Inant of a level白紙 might have affected白epu中 oseor integrity of也estudy. 

3.2 Procedure 

On the day before the test each of a group of吐rreerabbits was clipped free of fur企om血e

dorsal/flank訂 'eausing veterinary clippers. Only animals with a healthy intact epidermis by gross 

observation were selected for the s知 dy.

On the day of the test a suitable test site wぉ selectedon the back of each rabbit. A quantity of 

0.5 ml of也etest material wぉ血仕oducedunder a 2.5 cm x 2.5 cm co柱。ngauze patch and placed 
SPL PROJECT NUMBER: 140/1099 PAGE8 

in position on the shom skin.百1巴 patchwas secured in position with a strip of s町 gicaladhesive 

tape. To prevent也ea由nalsinterfering wi由 也epatches,也巴仕阻止 ofeach rabbit was wrapped in 

組 elasticatedcorset釦 d也ea由nalswere returned to their cages for the duration of the exposure 

period. 

Four hours after application the corset and patches were removed企omeach anintal and any 

residual test material removed by gentle swabbing with cotton wool soaked in distilled water 

Approximately one hour follow泊gthe removal of the patches, and 24, 48 and 72 hours later, the 

test sites were exanined for evidence of primary irritation and scored according to the following 

scale: 

EV ALUATION OF SKIN REACTlONS 

Erythema and Eschar Formation Value 

Noe巧社lema......................,.・H ・.....・H ・.....・H ・..............................・H ・................・.......・H ・-・ 0 

Veηr slight e可証lema(barely perceptible)...-

Well-defined e巧司hema..........・H ・........・H ・-...............................・M ・................................. 2 

Moderate to severe ery也ema.......-.............・H ・.....................・H ・......・H ・....・H ・....... 3

Severe erythema (beet re的 55)to slight eschar formation 何 回esin depth) .............. 4 

Oedema Formation 

Nooedema .........・.......・H ・.......・H ・-...・・・......・H ・....・ ...・・ ・... .........・H ・., ........・H ・..... .......... 0 

Very slight oedema (barely perceptible)...・・

Slight oedema (edges of area well-defmed by defi凶teraising)............ H ・M ・....... 2

Moderate oedema (raised approximately 1 millimeむの................................-...... 3 

Severe oedema (raised more than 1 millimetre and extending beyond也E紅 eaof

exposure)......・H ・..................・H ・........・H ・-........・H ・.......・H ・H ・..・H ・-......・H ・.....................・H ・ 4

Any other skin reactions, if present, were also recorded. 

3.3 Interpretation of Results 

Calculation ofPrimary Irritation Index and Grading oflrritancy Potential Using the Draize 

Scheme 

百lescores for erythema and oedema at血e24 and 72-hour readings weぉ totalledfor the也reetest 

rabbits (12 values)刷 出sto凶 wasdivided by six to give the primary irritation index of批 test

material. 百lete温tmaterial wぉ classifiedaccording to the following scheme devised by 
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SPL PROJECT NUMBER: 14011099 PAGE9 

Draize J H (1959)唱 ermalToxicity" In: Appraisal of the Safety of Chemicals in Foods, Drugs 

and Cosmetics. Association ofFood and Drug Officials ofthe United States, Austin, Texas, p.47: 

Primary Irritation Index 

。

> Ot02 

>2 to 5 

> 5 to 8 

Classification of Irritancy 

Non-irritant 

Mild凶 t如 t

Moderate irritant 

Severe irritant 

If irreversible alteration of吐ledermal tissue is noted in組 yrabbit, as judged by the Study 

Director, which include ulceration and clear necrosis or signs of sc紅 tissue,the test material is 

classified as coπ'Osive to rabbit skin. Classi五cationaccording to Draize may, therefore, not be 

applicable. 

4. ARCHIVES 

Unless instructed otherwise by也eSponsor, all original data副 社lefinal report will be retained 

m吐leSafepharm archives for five ye紅 s,after which ir首位uctionswill be soughtぉ toお 巾er

retention or disposal. 
SPL PROJECT NUMBER: 140/1099 PAGE IO 

5. RESULTS 

百leindividual scores for erythema/eschar組 doedem丸 紅'egiven in Table 1. 

No evidence of skin訂ritationwas noted during the study. 

6. CONCLUSION 

百le test material produced a prim紅 y irritation index of 0.0 飢 d was classified as 

NON-IRRITANT to rabbit skin according to也,eDraize classification scheme. No corrosive 

effects were noted. 

百letest material did not meet the criteria for classification as i凶tantor corrosive according ω也E

EU labelling regulations Commission Dir巴ctive93/21AEC. 
SPL PROJECT NUMBER: 140/1099 PAGE 11 

EMULGEN B-66: ACUTE DERMAL IRRITATION IN THE RABBIT 

Table 1 Individual Skin Reactions 

Individual 8cores -Rabbit Number and 8ex 

8kin Reaction Observation Time Total 

95 Female 98 Female 103 Female 

1 Hour 。 。 。 (0 ) 

Ery白ema/Eschar 24 Hours 。 。 。 。

Formation 48 Hours 。 。 。 (0 ) 

72 Hours 。 。 。 。

1 Hour 。 。 。 (0 ) 

Oedema Formation 

24 Hours 。 。 。 。

48 Hours 。 。 。 (0) 

72 Hours 。 。 。 。

8umof24叩 d72-ho町 Read皿 gs(8) 。

Primary Irri阻lionIndex (8/6) 。/6= 0.0 

Classification NON-IRRITANT 

( ) = Total values not used for calculation ofprimary ir吋阻むonindex
SPL PROJECT NUMBER: 140/1099 PAGE 12 

Appendix 1 Statement of GLP Compliance in Accordance with Directive 88/320lEEC 

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SAFEPHARM LABORATORIES LTD 

EMULGEN B-66: 

ACUTE DERMAL IRRITATION IN THE RABBIT 

SPL PROJECT NUMBER: 140/1099 

1 verifシ也at白isis an exact copy of吐leoriginaI report which is located in the Archives of 

Safepharm Laboratories Ltd., Derby, UK. 

¥ 

A Sanders 

Study Director 

DATE:・......・2.2APR却02・H ・...