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NATIONAL BUREAU OF STANDAR DSM CPOCOPY RESOLUTION TEST CHART

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TOPICAL HAZARD EVALUATION_____

OF CMDIDATE INSECT REPELLENT A13—36562~~~~

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4-~~~THyL— 1-[ (2-t€THy L~CyCt~Oi.m XyL) CARBONYL ] PIPE RAZ INE_____ STUDY NO. 51—0 824—77_____

OCTOBE R 1975 — MARCH 1977

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Approved for public release; distribution unlimited .

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ENVIRONMENTAL HYGIENE AGENCY

~~ ABERD EEN PROVING GROUND, MD 21010~:

NJLA SSIFIED ___________

$ !C URITY CLA $S IF ICAT% ~~~~ØP Tp~ S (Wb~~ GM. E,il.f. ~~

DED?~or f~~sf i u~~kr rAY IV~II D AI READ INSTRUCT ION Sr.. r~ j i~ I,’~ ~~~~~~~~ ‘S~~S flJr~ SEFORE COMPLETING FORM

I. REPO RT ~uuStA -

~ 2. GOVT ACCEUION NO. 3. R E C I P I E NT S C A T A L O G NUMB ER

_________ ____________________________— __________________________

4. T~ F S. TYPE OF REPORT A PERIOD COVERED

Toj~~ca1 ~~~‘ ~r~1 Evaluation of Candidate Insect Final - Oct 75 — Mar 77Rep el1er~ ~d3-36562 ______________________________

A . PERFORMING ORG. K€PORT NUMBER51—0824—77

AIJ T~ 4OR(~ ) 0 A. CONTRACT OR GRANT NUMBER(.)

MAURICE H . WEEKS HRENDA J. DeSENA

A . PERFORMI NG O R G A N I Z A T I O N NAM E AND AD DRESS 10. PROGRAM ELEMENT. PROJECT , T A S KAR E A A WORK UNIT NUMBERS

Conunande rUS Army Environn~ental Hygiene AgencyAberdeen Proving Ground , l.~ 21010

II. CONTR OLLING OFFICE NAME AND ADDRESS 12. REPORT DATE

Commander Oct 75 - Mar 77US Army Health Services Command 13 . N U N B E R O F P A G E S

Fort Sam Houston , TX 78234 7T4 MONITORI NG AGE NCY NAME & ADDRESS(1I dIll.r ,f f,ea~ Coøtrolllng OllIc.) IS. SECURITY CLASS. ( Of ehI. ,.pr~rn

Unclass i f ied

5.. DEC L*SSIFICAT ION ” DOW NG RAOINGSCHEDULE

IA. DISTRIBUTION STATEMENT (of thu R.port)

Approved for public release; distribution unlimited.

IT. DISTRIBUTION STATEMENT (of A . ab~f,.ct ~if.,.d In Block 20, If dlfI.r~ 1I ko., R.pofl)

IA. SUPPLEMEI4’fARY NOTES

19. KEY WORDS (Contirn,. on ,•c.,a. .Id. If n.c..w .,. wd Id.nVIfy by block n m,b.,)

4-methyl-i- ( (2-methylcyclohexyl) carbonyl)piperazineA13-36562 eye irritationTopical Hazard Evaluation sensitizationcandidate repellent oral toxicityskin irritation Photochemical Skin Irritation

20. AB STRACT (Conlfflu. ~ , ,.. ~~.. •I~~ If n.cs.~~~~ a~d ld..nllfy by block .n b.r)

‘A hazard evaluation of A13-36562 was conducted using New Zealand White rabbitsfor skin and eye studies; Hartley guinea pigs for a skin sensitization study;and Sprague-Dawley, Wistar—derived rats for acute oral toxicity. AI3-36562caused mild skin and eye irritation reactions in rabbits and should be usedwith caution around the eyes and mucosa. It was recommended that A13-36562 beapproved for ftirther testing as a candidate insect repellent...

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,. ‘ DEPARTMENT OF THE ARM Y

• U. S. ARMY £NV~RONMENTAL J IYGIENE AGENCYA B E R D E E N PR O~~ NG ~~~ Q~~~P~~~~A RYLAND 21010

HSE-LT/WP ,“ - . (17 1 7 JUN ~77- . . . . . TOPICAL HAZARD EVALUATION

OF CM~DIDATE INSECT REPELLENT A 13-365624-METHYL-1-~ (2-METHYLCYCLOHEXYL) CARBONYI3 PIPERAZINE-

STUDY NO. 51-0824-77OCTOBE R 1975 - MA RCII.J.977.--— ‘ ‘

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~~~~~ ~‘ J ’ ~ ,, .- --~~- .. ,•-ABSTRACT

A hazard evaluation of A13—36562 was conducted using New Zealand Whiterabbits for skin and eye studies ; Hartley guinea pigs for a skinsensitization study; and Sprague—Dawley, Wistar—derived rats for acute oraltoxicity. A13—36562 caused mild skin and eye irritation reactions in rabbitsand should be used with caution around the eyes and mucosa. It wasrecommended that A13—36562 be approved for further testing as a candidateinsect repellent

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DEPARTMENT OF THE ARMYU. S. ARMY ENVIRONM ENTAL HYGIENE AGENCY) ABE RDEEN PROVING GROUND . MARYLAND 21010

HSE-LT/WP

TOPICAL HAZARD EVALUATIONOF CAND IDATE INSECT REPELLENT AI3-36562

4 —M E T H Y L - l-E (2-METHYLCYCL OHEXYL) CARBONYL] PIPERAZINESTUDY NO. 51—0824—77

OCTOBER 1975 - MARCH 1977

1. AUTHORITY .

a. Letter, US Department of Agricul ture - Agricultural Research Service ,- - Southern Region , Insects Affecting Man Research Laboratory, Gainesville , —

Florida, 17 October 1975.

b. Memorandum of understanding between the US Army Environmental HygieneAgency ; the US Army Health Services Command ; the US Departmsnt of the Army ,Of f i ce of The Surgeon Genera); the Armed Forces Pest Control Board ; and theUS Department of Agriculture , effective Decembe r 1970 with Amendment No. 1,e ffec tive August 1974.

2. REFERENCE. Toxicology Division Procedural Guide , U SAEHA , 1972 .

3. PURPOSE. The purpose of this prv-~gr axn is to proviae guidance for fur therentomological testing of the candidate insect repellent A13—36562.

4 . SUMMARY OF FINDINGS . A hazard evaluation of the candidate repellentA13—36562, 4—methyi—l— [(2—tnethylcyclohexyl)carbonyl] piperazine was conductedby this Agency using New Zealand White rabbits for skin and eye studies ,Hartley guinea pigs fo~ a skin sensitization study arid Sprague-Dawley,Wistar-derjved rats for determination of oral toxicity. A tabularpresentation of animal toxicity data developed in this Agency follows :*t

* In conducting the studies described in this report, the investigatorsadhered to the “Guide for the Care and Use of Laboratory Animals,” USDepartment of Health , Education and Welfare Publication No. (N IH ) 74—23 ,revised 1972 — second printing 1974.t The experiments reported herein were performed in animal facilities , fu llyaccredited by the American Association for Accreditation of Laboratory AnimalCare.

Approved for public release; distribution unlimited. J0’

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Study No. 51— 0824—77, Oct 75 — Mar 77

_TEST — - RESULTS

- INTEPP•RETATION

SKIN IRRITATION STUDIES

Rabbits

Single 24—hour application Compound A13—36562 USAEHA Categoryto intact and abraded skin produced mild II ~ref Appendix).of New Zealand White irritation torabbits, the intact skin

and of the skin0 .5 ml technical grade surrounding ancompound to each of abrasion .six rabbits.

EYE IRRITATION STUD IES

Rabbits

Single 24—hour application Compound A13—36562 USAEHA Categoryof 0.1 nil technical produced mild C (ref Appendix),grade compound to one injury to the corneaeye of each of six and some injuryNew Zea.larid White to the conjunctivarabbits, in six of six rabbits

lasting up to 3 days .No signs at 7 days.

APPROXIMATE LETHAL DOSE (ALD)

ORAL

Rats (Male ) — no diluent ALP 3300 mg/kg Presents littlelethal hazardfrom acuteaccidentalinqestion

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Study No. 51—0824—77, Oct 75 — Mar 77

TEST RESULTS INTE RP RETATION

PHOTOCHEMICAL SKIN IRRITATIONSTUDIES

Rabbits

A single application (0.05 A 25 percent solution Compound A 13— 36562 didml) of a 25 percent (w/v) of A13—36562 in not cause a photochemicalsolution of the compound ethanol did not cause irritation reaction under(A13—36562) and of a 10 a photochemical irrita— test conditions and is notpercent (w/v) oil of tion reaction under expected to cause a photo—Bergamot solution test conditions, chemical irritation in(positive control) in However, this humans.95 percent ethyl alcohol , solution caused awere applied to the intact very mild irritationskin of six rabbits. Five reaction for up tominutes after application, 3 days afterthe rabbits were exposed to application ,on bothUV light (365 run) for 30 irradiated and unirradi-minutes at a distance of ated skin areas.10— 15 cm.

Control

Following UV exposure of Positive controlthe rabbits 0.05 ml of test application aridcompound, positive control irradiation causedarid diluent were applied greater irritantto additional skin areas effects than into serve as unirradiated un irradiated skincontrol sites. Applica— areas.tion areas we re checkedfor skin irritationreactions at 24 , 48 arid72 hours.

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Study No. 51—0824—77, Oct 75 — Mar 77

— TEST RESULTS INTERP RETAT ION

SENSITIZATION STUDIES

Guinea Pigs (Mal e)

Intradermal injections of0,1 ml o~ a 0.1 percentsuspension (w/v) ofAI3—36562 or ofdinitrochlorobenzene(DNcB) * in a mixturecontaining 1. volumeof propylene glycoland 29 volumes ofsaline.

Ten test guinea pigs challenge dose of Compound A13-36562 didreceived and challenged test compound (last not produce a sensitizationwith a 0.1 percent intradermal injection) reaction under these testssolution of A13-36562, did not produce a conditions and is not expected

sensitization to produce a sensitizationreaction, reaction in man.

Ten positive control Positive control (DNCB)guinea pigs received produced a markedand challenged sensitization reactionwith 0 .1 percent in ten out of tensuspension of DNCB. guinea pigs .

Ten cage control guinea pigs Cage control guineapigs showed no greater

Five receiving challenge reaction to test compounddose of test compound and DNCB th an were seenwithout prior in original test groups.sensitizing doses.

Five recieving challengedose of I~JCB withoutprior sensitizing doses.

* A known skin sensitizer.

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Study No. 51—0824—77, Oct 75 — Mar 77

5 • CONCLUSION • T.chrd ca]. grade and 25 percent ethanol solutions ofA13—36562 cause mild sk in irritation reactions in rabbits. Eye irritationinjury was mild and reversible after 7 days.

6. RECOMJ~~NDATION . Unde r the provisions of the Memorandum of Understanding(reference paragraph ib) it is recommended that A13—36562,4-Tnethyl—l-[ (2—methylcyclohexyl)carbonyl] piperazine , be approved for furthertesting as a candidate insect repellent. It is to be used with cautionaround the eye. and mucosa.

MAURICE H. WEEKSChief , Toxicity Evaluation Bran chToxicology Divisicrn

BRENDA J . DeSENAPFCVeterinary SpecialistToxicology Division

APPROVED:

Mc~~~~~~ , Ph.D,Chie f , Toxicology Division

BRENDON E. JOYCE, .D.LTC , MSCDirector, Laboratory Services

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Study No. 51—0824 ~77 , Oct 75 - Mar 77

APPE)t)IX

TOPICAL )~~ZARD rVAWATIO.1 P~ )GRAZ4DD INITIONS OF CATEGORIES O~ CONPOWIDS BEING

CONSIDERED FOR ACUTE SKIN APPLICATIONS

CATEGORY I - Compounds producing no primary irritation of the intact skin orno greater than mild primary irritation of the skin surrounding an abrasion.(INTERPRETAT ION : No restriction for acute application to the human skin.)

CATEGORY II — Compounds producing mild primary irri tation of the intac t sk inand the skin surrounding an abrasion. (INTERPRETATION : Should be used onlyon human skin found by examination to have no abrasions or may be used as aclothing itnpregnant.)

CATEGORY III - Compounds producing moderate primary irritation of the intactskin and the skin surrounding an abrasion. (INTERP RETATION: Should not beused directly on the skin without a prophetic patch test having beenconducted on humans to determine irritation potential to human skin . May beused without patch testing, with extreme caution, as clothing ixnpregnants.Compound should be resubmitted in the form and at the intended useconcentration so that its irritation potential can be reexamined using othertest techniques on animals , prior to human testing.

CATEGORY IV - Compounds producing moderate to severe primary irritation ofthe intact skin and of the skin surrounding an abrasion and, in addition,producing necrosis , vesiculation and/or eschars. (INTERPRETAT ION: Should beresubmitted for testing in the form arid at the intended use concentration,Upon resubmission, its irritation potential will be reexamined using ohtsrtest techniques on animals , prior to possible prophetic patch testing inhumans , at concentrations which have been shown not to produce primaryirritation in animals.)

CATEGORY V - Compounds impossible to classify because of staining of the skinor other masking effects owing to physical properties of the compound.(INTERPRETATION: Not suitable for use on humans.)

EYE CATEGORIES:

A. Compounds noninjurious to the eye. INTERPRETATIONt Irritation ofhuman eyes is not expected if the compound should accidentally get into theeyes , provided it is washed out as soon as possible.

B. Compounds producing mild injury to the cornea. INTERPRETATION:Should be used with caution around the eyes .

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Study No. 51—0824 77, Oct 75 — Mar 77

C. Compounds p~roducing mild injury to the cornea, and in addition someinjury to the conjuxtctjva. INTERPRETATIONt ~~ould be used with cautionaround the eyes and mucosa.

D . Compounds producing moderate injury to the cornea. INTERPRETATION SShould be used with extreme caution around the eyes .

E . Compounds producing moderate inj ury to th. cornea, and in additionproducing some injury to the conjunctiva. INTERPR ATION: Should be usedwith extreme caution around the eyes and miacosa.

F. C~~pounds p~roducing severe injury to the cornea and to theconjunctiva. INTERPRETATION: Should be used with extreme caution. It isrecommended that use be restricted to areas other than the face ,

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