A

iE FILE -COPY

INSTITUTE REPORr NO. 224

00

, , IMARY' DERMAL IRRITATION POTENTIAL OF TRIMETHYLOLETHANET INITRATE (TMETN) IN RABBITS

II

EARL W. MORGAN, DVM, MAJ VC

and

DON W. KORTE, JR., PhD, MAJ MSC

TOXICOLOGY BRANCH,

DIVISION OF COMPARATIVE MEDICINEAND TOXICOLOGY

DTICELECTESEP 1.9 1988 D

OCTOBER 1986 Toxicology Series 114

LETTERMAN ARMY INSTITUTE OF, RESEARCHPRESIDIO OF SAN FRANCISCOp CALIFORNIA 94129

'DfSTUUTION MTAWMET

Approved for public rW*Mra;jM r in-t ,o U ,,o I -" 2

I . . .. .. . . .,

Primary Dermal Irritation Potential of TrimethylolethaneTrinitrate (TMETN), (Toxicology Series 114)--Mbrgan and Korte

Reproduction of this document in whoie or in part is prohibited cxccptwith the permission of theCommander, Lcttcrman Army Institute of Rcsearch, Presidio ofSza-Francisco, California 94129.Itowever, tie Defense Tcchnical Information Center is authorized to reproduce the document forUnited Sttes Government purposes.

Destroy this report when it is no longer needed. Do not return it to the originator.

Citation of tradr names it thLs report does not constitute in official endorsement or approval of theuse of such Imnes.

This material has been reviewed by Lc:tcrman Army Instituteof Research and there is no objectiun to iO presentazion and/or publication. The opinions or assertions contained hereinare the private views of rhe author(s) and are not to be con-strucd as official or as reflecting the views of the Dcpartmcntof~the Army or-the Department of Defense. (AR 3605)

(Signature nd dat)

'This documenthas been approved for public release and s-Jc; its distribution is unlimited.

UNCLASSIFIED

-CCURITY CLA,-IFICATIOtR OF THIS PAOG (Whn Da.a Krt.rs4l)

IR5AD II4STRUCTiONS•_REPORT DOCUMENTATION PAGE BRE^ NRCOMPLETINGORMREPORT NUMUCRT C2. GOVT ACCCSSION HO. 3. RECIPICHT*S CATALOG NUMBER

LAIR Institute Report No. 224

. Tt .'LC fond Sobtblui S YYPC OF IMPORT & PERIOO COVCREOPrimary Dermai Irritation Potential of FinalTrimethylolethane Trinitratce (THETN) in Rabbits 24 Oct - 13 Nov 84

6. PEcFORMiNG ORG. REPORT NUMBIIR

7. AUTBOR() U. CONTRACT Olt GANT HUMULACD)

Earl W. Morgan, DVH, KV VCDon W. Korte Jr., PhD, MAJ MSC

9. PERFORMING OG1ANIZATION NAMC AND ADDRESS 10. PnOGRAM CLCMENT. PROJECT. TASIK

Toxicology Branch, Div of Camp Med and Tox AREA 6 WORK UNIT NUMUCRS

Letterman Army Institute of Research 3EI62720A835Presidio of San Francisco, CA 94129-6800 WU 180; APC TL09

I1 CONTRtLING OFFICE NAME AND ADDRESS IZ. REPORT OATO

US Army Medical Research and Development Command October 1980Fort Detrick, MD 21701-5010 5 NUmUenOFPAGES

1.714. MONITORING AGENCY NAME A AOORESS'It dlltt i tIeam Controllni Oic..j It$ SECURITY CLASS, (oti fhol pot)j

US Army Medical Bioengineering Research and I UNCLASSIFIEDDevelopment Laboratory IFort Detrick, RD 21701-5010 flCELA-SSIFICATIO*OOWHGHAOING

I. OISTRIOUTION STATEMENT (fo this RePOUJ

TIllS IDCUtIENT 11AS BEEN CLEARED FOR PUBLIC RELEASE AN SALE: ITS DISTRIBUTIONIS UNLINITEUI

RUUllON .T&TCM% .NT to# Ith* ab. ef anterod In Mufoc hO, f dilfeent tm N foI)

WS. SUPPLEMENTARY NOTES

19. KEY WORDS (C.wntinue on tevr.,a aide It n#c*Jauy a id Identity by block numbi t)

Primary Dermal Irritation, Trimethylolethane Trinitrate, TMETN,Mammaliar. Toxicology, Rabbit

20. A!STRACT (Co lnss m% ,reveres &ld. I I ei. *ary and Idenllr by bltock 114mbaJ

The primary dermal irritation potential of trimethylolethane trinitrate (TRETN)was determined ii male and female New Zealand White rabbits using a modifiedDraize method. The test compound was non-irritating. Very slight erythema wasobserved in 2 rabbits by 1/2 hour after dosing and in 4 rabbits at 24 hoursafter dosing. All rabbits had returned to normal by 48 hours after dosing.Neither edema nor any other recognizable skin reaction was detected at any timeduring the14-day observation period.

DD I Fo n 1473 EDITION or INOV 6sIS OOSOLETN.;Ajt 73UNCLASSIFIED

SECURITY CLASSIFICATIONf OF THIS PAGE (When Data Entecod)

. . . . . . . . . . . . . . .. . . . . I

ABSTRACT

Tite primary dermal Irritation potential of trimethyl.oltthanetrLnitrare (TMI*TN) was determined in nLie and feMaie Neu Zealand WhIterabbies using a modified Draize maethiod. The Lest compound w-ks non-irritacting. Very slight erythema tins ooserved in 2 rabbits by 1/2hour , cr dosing and in 4 rabbits at 24 hours nEater dosing. Allrabbits had returned to normal by 48 hours after dosing. Neitheredema nor any other recognizable skin reaction was detected at tinytime during the 14 day observation period,

Key words: Primary Dermal Irritation, TrimethylolethaneTrinitrate, ThETN, Mammalian Toxicology, Rabbit

a

Acoession For

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PREFACE

TYPE REPORT: Primary Dermal lvritacton GLP Study Report

TESTING FACILtY: US Army Medicatl Research and Development CommandLetterman Army Institute of ResearchPresidio of $an Frnncisco, CA 94129-6800

SPONSOR: US Army Medical Rescarch and Development CommandUS Army Medical Bioengineering Researchand Development Laboratory

Fort Detrick, Maryland 21701-50OProject Officer: Gunda Reddy, PhD

WORK UNIT: 3EI62720A835

Toxicity Testing of PropellantsWU 180; \PC TL09

GLP STUDY NUNBER: 84041

';TUDY uIRECTOR: '-A.I Don Wt. Kor~e JIr., PhD, ',413C

PIWNCIPAL INVEST -(ATOR: CP Earl W. Merrin, IVi, VCDi ,Iomace of Ameriecan College ofVeterinary Preventive ,edictne

REPORT AND DATA .AGE% I1r'&T: A copy of the final, report, studyprotocol, retired SOPs, raw data,analytical, stbilicy, and purity data ofthe test compound, ind an aliquot of thetest compound will be retained in theLAIR Archives.

TEST SUBSTANCE: Trimechyloletitne Trinitrnre

INCLUSIVE STUDY DATES: 24 October - 13 November 1,984

OBJECTIVE: Thn objective of this study was to determine the primarydermal trritation potential of TrimethylolethaneTrinitrate in mile and female New Zealand White rabbits.

iiL

-S I

ACKNOWLEDGMENT

SGT Steven K. Sano, B.S and Yvonne C. Johnson, SS, assisted in theresearch; SP4 James J. Fisher, SP4 Scott L. Schwebe, and CharlotteSpeckman provided care for the animals; and Callie B. Crosby, MA, andBrenda V. Goce, provided secretarial assistance.

iv

SIGNATURES OF PRINCIPAL SCIENTISTS AND KANAGERS NVOLVED IN TIlE STUDY

We, the undersigned, declare thjt GLP Study 84043, was performedunder our supervision, according to the procedures deccribed herein,and that this report is an accurate record of the results obtained.

DON W. KOTh JR., (VhD ^DATEMAJ, MSStudy Divector

#/

EARL W. ORGAN, DViW/ DATCPT, VC */Principal Investigntor

CONRAD R. WHEELER, PhD / DATEDACAnnlyticnl Chemist

V

% DEPARTMENT OF THE ARMYLETTERMAN ARMY INSTITUTE OF RESEARCH

PRESIDIO O. ' SAN rHANCISCO, CAUrORNIA 4,41" "M

SGR)-U .QKA 14 Januiy 198r

ZM0RANN 1 M'R RWXORD

SuJFCT: Reort oC CLP Ccttipliance

1. t hereby certify tl1.ft in relation to FAIR GLP Study 84043 tho following

insp..ections were mide:

6 Novc 19R4

2. The report and raw dati for this study were amlitc d on 10 October M..

3. Routine inspxctions with no adverse findings are reported qunrterly,thus these inspections nrm also included in the 21. janury 1985 repoct toMarkigement and the Study Direc'tor.

-GARY ' IIE.RSqGj, LA

Quaility Assurmncc Unit

vI

TABLE OF IONMENTS

Abst~r&CL ....................... . . . . . . . . . . . . .. . . . . .Pref~tac .. . . . . . . . . . .. . . . . . . . . ... . • 9 55 5 * * * *SS*SSSSSS**S**S*S*S**S9 t* • •. ......... SiL

Aeknow.edsm4.nt s .......................... .................... . v

Sib iaLtures of Priacipa ;i SC,lrlLt V .............. . ....... ... . . V

ReporL of quAlity Avuurance Un . . . . . . . . . . . . . . . . . . . . . ...... . . . vi

Table of Con.ent ................ .. . . ...... . ....... ......... . vil

BODY OF REPOKT

Objectivye of Study*...................*........ . . . . . . .

Z4AThRIALS

TeL SubStaI : ............................................... IAnimt~al Wlhla ........... o.......s................................ 2

IIuh1i ndry ....................................................

HI:fLIIOi)S

Group As! lll.WnL/.CC I l ol ................................ 2Test Procedures .................................. .... .. .2Observatiois ... ...... ...................................... 3Duration of Study ... ...................... .. ....... 4Clnges/Deviations ............................ .............. 4Raw Data and Final Report Storat ........................... 4

RESULTS ............. *.......................*...................... 4

DISCUSSION ......................... o .......... ........ ...... 4

CONCLUSION ............................... • ....................... 5

REFERENCES ......................... ................

vii

Table of Contents (continued)

APPE# DICES

Appendix A. Chemical t)ata ................................Appe".ix $. Animal Uata .................................... ,,11Appendix U. iIstortcul Lis:nj; of tudy Evens ................ 13Appendix D. Tabular Scoring Uata ............................

O F 'CIAL DISTRIBUTION lAST.....................................17

viii

I

Primary Deurmal Irrition Potentila) of Triwur~yloleitane Trinitrate ithale ant! Female' Robbits--horgon at 31i

Tile Uepartwnmi of Uense is consilserinij theQ use OCdiethyleneglycol dinicrate (DEGDN), triethyleneglycol dinitratu(TEUNfl, or tC1%eihyjlOlethwnO trinitrate (ThICTN) its u replact-ownt fornitroglycerin In mounition formulations. A "health eiffects" reviewconducted for the US Army ?Iedical Bioengineering Research andDevelopment Laboratory (USAXBKDL) identified numerous Scaps in thetoxicology database of those coraipounds (1). Conequently, USAWRUI4hans tusked the Toxicology Brstnch, LAIR, to conduct an initial. healtheffects evaluation of UIUN, THMrN, TIELiN, Mo~d tWO D1EUUN-binedpropellaiits, JA-2 and ll.-U This illiLtial -V3i1llALion intcludes tuAlIS MuUICiLy 48811y, IiCULO o1ral oXie'ity LvtjtS In rcits and mice,acute derrml toxicity tests in rabbits, dermal aind ocular Irritationstudies in rabbiLS, anid dermal sensiti~ation studies in guinea pigs.This report containus the results of a sc-.idy to assess the primaryderwal, 1 1rr tcat.ion I'1.nl.IIU l of' TiI1'N, fit vtbbiu;.

7The objCCtLie 0f thli$ SLudy W.18 LO d~'eri-1 14110thL* priuunry dertmillirritationi Potential or vNEV4T in macle' and femaile New Zealand Whiterabbits.

HIATERALS

Test Substanice

Chemical name: Trirnethyloletivine Tritnitrat

Chemical structure:

CHzONO2

CHI- C-CH 2ONO2

CH20NO2

HoLecular form~uLa: C 5 1 N 3()

Other test substance information is presented in Appendix A.

?Irgaii-- 2

Animal )aca

Four uale and four female New Zealand White rabbits (ElkhornRabbitry, Watsonville, CA), Identified individisally with ear tattooes,numbered 84059b to SFW597, 8AF99 to 84Fb03, itclusive, and 84FOllwere assigned to the uiLdy. The antimal weights on dosing day ('10 Oct)ranUed from 3. to 3.5) kg. AdiWIl0o1al ahilwil dauiL, appliAir i AIIdH.

Husbandry

The rabblts were housed individually in stainless steel, screen-bottomed, battery-type cages with automatically flushing dumptanks.The diet consisted of I5U g per day of Certified Ilurina , Chov Ulut 5322(Ralston Purina Company, St Louis, MO); water was provided bycontinuous drip from a cent'al line. The animal room temperature wasmaintained at 20.0 to 22.2C with a relative humidity range of 48 to6b percent with short spikes up to 82 percoxat associated with roomcleaning. The photoperiod was 12 hours of light per day.

KiETIIOUS

Group Assignment/Acclimation

Study animals were occlimated for 6 days to the sLudy roomfollowing a 14-day quarantine by the Animal Resources Group. IVringthis period they were observed daily for signs of illness. They weretreated once prophylactically for ear mites with Canex* and mineraloil in the eara.

Test Procedures

This study was conducted in accordance with EPA guidell-,,,s (2) andLAIR SOP-OP-STX-34 (3).

The backs of 8 rabbits were close-clipped 24 hours before theactual dosing. The clipped area was divided into 4 quadrantsdesignated I-V (4, 5). Sites I and IV were shani patches. Sites I1 andIII were test compound sites. Since the TMETN is a liquid, a staidardidose of 0.5 ml of the test compound was placed on 1-inch (2.5 cMisquare gauze patches which were taped to Sites 11 and III. Bleuderr*,(Hedical Products Division of 3h, Saint Paul, HN), a seuiimpervious,hypoallergenic surgical tape, was used to hold the patches in place.Vet WrapV (Animal Care Products Division of 3H, Saint Paul, MN) wasthen wrapped securely around the animal. The test compound was left;in contact with the skin for 4 hours. At the end of the exposure'period the wrapping and patches were removed, and the areas wereIscored 1/2 hour later.

Morgan--3

Observations

Tit grading and scoring for dermal reactions were performedaccording to Table 1. Observations were made daily from 30 October to13 November L4. Scoring and grading were perLorwd at 1/2, 24, 48,and 72 hours after removal of the pmtch.

TABLE IEVALUATION OF SKIN REACTIONS

Erythema and Eschear Frormation

No erytLheumitVery slight erythema (barely perceptible) IWell-defined erythema 2Moderate-to-severe erythema 3Severe erythema (beet redness to slight

eschar formation (injurious In depth) 4Possible total erythema score: 4*

Edema Formation

No edema 0Very slight edena (barely perceptible) 1Slight edema (edges of area well-defited

by definie raising) 2Modi.taLe e.denla (ieIge.'n ra lIed approxilltaily 1 111h,) 3Severe edlema (rlaJsed .01-C L1.n1 1 mm atdexLendin, buyond area oL exlposare)Possible total edema score 4*

Possible total score for primary irritation 8

*Any skin reaction more serious than severe edema, vesiculation,ulceration, or necrosis places the chemical in Category V.

Morgan--4

Duration of Study

Appendix C iS a complete listing of historical events.

Chan&es/Deviations

Rabbit number 84F601 had the test compound and the sham patchsites inadvertently reversed. All other aspects of this study wereconducted in accordance with all applicable SOPs and addenda.

Raw Data and Final Report Storage

A copy of the final report, study proLocoAh, raw data, retiredSOPs, and an aliquot ot the test compound were retained in the LAIRArchives.

RESULTS

Animals were scored for erythea and edema at each patch site.iFive rabbits exhibited very slight erythems (score 1) at test compounosites. Rabbits 84F597 and 84F602 were observed to have very slightierythema at 1/2 hour after dosing. Rabbits 84F596, 84F597, 84F600,and 84F611 exhibited very slight erythema 24 hours after dosing. Allrabbits had returned to normal by 48 hours after dosing. Neither,edema nor any other recognizablq skin rectlon was detected at anytime during the 14-day observation period. The sham patch sites were

inormal throughout the study. Results of scoring the dermal irritation1potential in each rabbit were tabulated (Appendix D).I$DISCUSSION

THETN produced very slight erythema at the test patch sites infive of eight rabbits after a 4-hour dermal patch test. Neither edema

nor any other recognizable skin reaction was detected at any timeduring the 14-day observation period.

The primary irritation index adapted from McCreesh and 3teinberg(5) was used as a basis for categorization. Non-irritating compounds(Category 1) have peak net mean scores of 0.0-0.5. Mild irritants(Category II) have peak net mean scores from 0.51 to 2.0. CategoryIII Compounds are moderately irritating with indices between 2.1 and5.0. Chemicals are considered severe irritants (Category IV) if theyhave indices between 5.1 and 7.9 and they produce necrosis,vesiculation, ulceration, and/or eschars. Compounds which areimpossible to classify because of staining or masking of effects dueto physical properties are placed in Category V. The peak net meanscore from the test compound was 0.5. Therefore, TMETN was classifiedas a non-irritating chemical (Category I).

Morgan--5

Jones et al (6) studied the toxicity of Propylene Glycol 1, 2-dinitrate (PUiN) a nitrate eWter structurally similar to TKETN. Theirstudies showed that PG!N was absorbed perctmaitcously but produced noprimary dermal irritation at either 24 or 72 hours after exposure tovapor. The revults of this study with TNh~V4 are consistent with thenon-Arritancy reported for ?N;ON,

CONCLUSION

The test compound, TNETN, Is a non-irritant under conditions ofthis assay.

Morgan--6

REFERENCES

1. Holleman JW, Ross RH, Carroll JW. Problen definition study on thehealth effects of diethyleneglycol dinitrate, triethyleneglycoldinitrate, and trimethylolethane trinitrate and their respectivacombustion products. Frederick, Maryland: US Army Medicallioengineering Research and Development Laboratory, 1983, IDTIC NoADA 127846.

2. Environmental Protection Agency. Office ot Pesticide and ToxicSubstances, Office of Toxic Substances (TS-792). Primary DerowlIrritation. In: Health effects test guidelines. Washinptou, DC:Environmental Protection Agtncy, August 1982; EPA 560/6-82-OUl.

3. Primary Dermal Irritation Study. LAIR Standard OperatingProcedure OP-STX-34, Letterman Army Institute of Research,Presidio of San Francisco, CA. I August 1984.

'4. Draize JII, Woodard G, Calvary 110. Methods for Lhe study ofirritation and toxicity of substances applied topically to theskin and mucous membranes. J Phaarmcol Lxp Ther 1944; 8:377-390.

15. McCreesh AH, Steinberg H. Skin irritation testing in animals.In: arzulli FN-, Haibach III, eds. Dermato-toxicology and

I pharmacology (Advances in modern toxicology, vol 4). Wtshington:Ilemisphere Publishing Corp. 1977: 193-210.

16. Jones RA, Strickland JA, Siegel J. Toxicity of Propylene Glycol1,2-Dinitrate in experimental animals. Tox Appl Pharmacel 1972;22: 128-137.

Appendix B. Animal Data**#..**........ ....

Appendix C. Historical Listing of Study Events ....... #..1I3

ApedxD aulrSoigDt ........

Morgan--9

CHEMICAL DATA

Chemical Nmie: 1,3-Propinediol, 2-sethyl-2 [(nitrooxy)mothyl]-dinitrate (estaer)

Other Listed Names: 1,3-Propane~iol-2-(hydroxymcthyl)-2.-icthyl-,trinitrate; 1,1 1-Trimethylolethane trinitrate(THETi), Netriol trinitrate (MTN);Nitro pntaglycerin

CAS: 3032-55-1

Structural formula: CHONO 2I

CH 3- C-CHON0 2I

CH 2ONO 2

Molecular foromlA: C5 19N3 09

Molecular weight: 255.15

Physical state: Light brown oil

Melting Point: -3"*t

Compound Density: 1.47 g/cm *t

Lot No: 53-84A

Sourco: Naval Ordance StationIndian Head, 1D

Chlaical Annlysis

Tillt; L: r utontil ti:on:

!lltra-violet (UV) spectra were obtatned usinr a itachl 110-A Spectro-photometer (itachi Irnstruments, Inc., Mountain View, CA), infra-redspectra (IR) were obtained with a Perkin-Elmer odel 457 Infra-redSpectrophotometer (Perkin-Elmer, Norwalk, CT) and nuclear magneticresonance (NMR) spectra were recorded on a Varian FT-80 NMR (Varian,Palo Alto, CA) using tetramethylsilane as an internal standard.Chromatographic analysis was performed using a 1090B IIPLC with diodearray detector (Hewlett-Packard, Santa Clara, CA) and a Brownlee RP-18Spheri-5 Column, 4.6 x 250 mm (Brownlee Labs, Inc., Santa Clara, CA).The following conditions were employed for the IPLC assay: solventsystem, 70% methanol, 30% water; flow rate 0.9 ml/min; detectorwavelength, 215 na; oven temperature, 50"C.

* Holleman JW, Ross RI1, Carroll JW. Problem definition study onhealth health effects of diethylenegycol dinitrate, triethyleneglycoldinitrate and trimethylolethane trinitrate and their respectivecombustion products. Frederick, Maryland: U.S. Army MedicalBioengineering Research and Development Laboratory, 1983; DTIC No. ADAL27846, p17.

t Lindner V. Properties of ex-losive aliphatic nitrate esters. Table5. In: Grayson H., exe. ed. K2.rk-Othmer encyclopedia of chemicaltechnology. Volume 9. 3rd ed. New York: John Wiley and Sons, Inc.,1980:573.

Appendix A-1

Results:

UV Spectrum: For UV analysis TXETN was dissolved in acetonitrile. UVabsorbance begins at approximately 240 nm and increases with decreasingvavelength.* No absorption peak was observed. IR (K~r windows): 2900,

1645 (asymetrie stretch of NO group) 1470, 137, 1280 (symetric stretch of

NO2 group), 990, 860, and 755 co .t MKR (CDCI , 80MHZ): 61.22

(S, 3H, CN3), A.44 (S, 611, -CH2 -).t TWTN subjected to HPLC analysiseluted as two peaks with reteRtion times of 8 and 12.5 min.5 Based on

integration of peak areas the first peak represented 98Z of the sample.

The second peak was not identified. No decomposition of TK):TN was detectedby HPLG after storage of ThWT8 (neat or in ethanol) for a period ofnine weeks.l

*Wheeler, CR. Nitrocellulose-Nitroguanidine Projects. Laboratory

Notebook 184-05-010, p 51. Letterman Army Institute of Research, Presidioof San Francisco, CA.

tWheeler, CR. Nitrocellulose-Nitrogunnidine Projects. LaboratoryNotebook #84-05-010.2, p 67. Letterman Army Institute of Research, Presidioof San Francisco, CA.

TIbid., p 68.

SWheeler, CR. Nitrocellulose-Nitroguanidine Projects. LaboratoryNotebook #84-05-010, p 72-75. Letterman Army Institute of Research,Presidio of San Francisco, CA.

IWheeler, CR. Nitrocellulose-Nitroguanidine Projects. LaboratoryNotebook #84-05-010.1, p 34. Letterman Army Institute of Research,Presidio of San Francisco, CA.

Appendilx A-2

Morgon--l1

A.HIKNL DATA

Species: Oryctolagus cuniculus

Strain: New Zealand White (albino)

Source: Elkhorn Rabbitry5265 Starr WayWatsonville, CA 95076

Sex: Male and Female

Age: Young Adults

Animals in each group: 4 males and 4 femaloe

Condition of animals at start of study: Normal

Ilody wetItL raotr;e at. duadnp,: 3.0 - 3.5 K

ldentifictLIon procedures: Ear t08, ceg numbers 84F596-84-597;84Fb99-84VF63, and 84tF6l inclusive.

Pretest conditioning:

1. quarantine from 11 October - 29 Uctobdr 1984

2. Animal were close-clipped and examined *,Vo hours before dosing.

Justific.iton: Laboratory rabbits are ta proven sensitive animal modelfor d'irmal irritation.

Appendix B

Morgan--13

HISTORICAL LISTING OF STUDY IENTS

Date EvoIL

11 Oct 84 Rabbits arrived LAIR.

12 Oct 84 They were tattooed, weighed, examined for illness,and placed under a two week quarantine.

11-23 Occ 84 Animals were checked daily by Animal ResourcesGroup (ARG) personnel.

24 Oct 84 All rabbits were Lr oLtd wlih Gzntexw and mineraloil in their cars. Rabbits were removed fromquarantine after being certified healthy by ARGStuff Veterinarian. The animals were weighed.

25-29 Oct 84 Animals were checked daily.

29 Oct 84 Animals were close clipped and areas marked.

30 Oct 84 Animals were weighed. Test substance was nppl!edfor four hours. Patches were removed and sitesscored within 60 rinthies.

31 uct-13 Nov 4 Animals were observed dully.

31 Oct- 2 Nov 84 Areas were scored at 24, 4tI and 72 hours afterexposure.

5 Nov 84 Reclipped animsals.

0 Nov 4 Animals were welhed.

13 Nov 84 Animals were weightd and sacrificed.

Appendix C

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Morgan--i7

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