AD-R172 386 PRELIMINARY ASSESSMENT OF THE RELATIVE TOXICITY OF 1/1R13-2S837 IN ANIMALS(U) ARMY ENVIRONMENTAL HYGIENEAGENCY ABERDEEN PROVING GROUND NO G J LEACH SEP 6

UNCLASSIFIED USE-75-51- 6528-B6 F/06/2 NL

Eummmomo

4

Sq

4

III -~11111 1.11liii, ~* 11111 ihirn'"* 11111 *se. E315 1J11122

4 II~

H 1e1IH e *~ GII~0hut______ -111111111 liii

2I.,

JI.1's

.1ISI.

S.

4

SI

SI.

~4II

5-5%

SI, 5%-p

SS'.*.

p

* UNITED STATES ARMYV ENVIRONMENTAL HYGIENE* AGENCY

ABERDEEN PROVING GROUND, MD 21010-5422

PRELIMINARY ASSESSMENT OF THE RELATIVE TOXICITY OFA13-20837 IN ANIMALS

STUDY NO. 75-51-0528-86MARCH 1985 - JULY 1986

el.

+:OEEnSEP 3 01986

A

C-D Approved for public release; distribution unlimited.

LiJ

.3,.86 930 54

tL.-

UNCLASSIFIEDSECURITY CLASSIFICATION OF THISPAGE

REPORT DOCUMENTATION PAGE0 a. REPORT SECURITY CLASSIFICATION lb. RESTRICTIVE MARKINGS

UJNCLASSIFFID_______________________2a. SECURITY CLASSIFICATION AUTHORITY 3 DISTRIBUTION /AVAILABILITY OF REPORT

2b. DECLASSIFICATION /DOWNGRADING SCHEDULE Approved for public release; distributionunlimited.4. PERFORMING ORGANIZATION REPORT NUMBER(S) S. MONITORING ORGANIZATION REPORT NUMBER(S)

75-51-0528-86 ________________________6a. NAME OF PERFORMING ORGANIZATION 6b OFFICE SYMBOL 7a. NAME OF MONITORING ORGANIZATIONUS Army Environmental Hygiene O (f appicable)

Agency HSHB-MO-T ___________________

6c. ADDRESS (City, State, and ZIP Code) 7b. ADDRESS (City, State, and ZIP Code)-

Sa. NAME OF FUNDING /SPONSORING B b. OFFICE SYMBOL 9. PROCUREMENT INSTRUMENT IDENTIFICATION NUMBERORGANIZATION j(if aplica ble)

Sc. ADDRESS (City, State, and ZIP Code) 10. SOURCE OF FUNDING NUMBERSPROGRAM IPROJECT ITASK IWORK UNITELEMENT NO. NO. NO. IACCESSION NO.

*11. TITLE (include Security Classification) Preliminaty Assessment of the Relative Toxicity of A13-20837

* In Animals, Study No. 75-51-0528-86

12. PERSONAL AUTHOR(S)* Glenn J. Leach

13a. TYPE OF REPORT 113b. TIME COVERED 1.DAE OF REPORT (Year, Morith,Da)15PG ONStudy FRMMr8 -OJul 861 86 Sep I 12

16. SUPPLEMENTARY NOTATION

17. COSATI CODES 10. SUBJECT TERMS (Continue on reverse if necessary and identify~ by block number)FIELD GROUP SUB-GROUP

19. ABSTRACT (Continue onrevervelfnectuaty and identy by~oc number) o provide preliminary toxicity datafor the candidate cockroach repellent A13-30827. Vhese data are intended to provideguidance in selecting compounds for further entomological and toxicological evaluation. Inaddition, the dAta may be useful in developing preliminary safety guidelines for handling

*this compound~

20. DISTRIBUTION /AVAILABILITY OF ABSTRACT 121. ABSTRACT SECURITY CLASSIFICATIONJUNCLASSIFIEDIUNLIMITED 0 SAME AS RPT. DTIC USERS UNCLASSIFIED

22a. NAME OF RESPONSIBLE INDIVIDUAL 22zb TELEPHONE (Include Area Code) 22.OFISMBOL

DO FORM 1473,84 MAR 83 APR edition may be used unilexhausted. SECURITY CLASSIFICATION OF THIS PAGE' '- .. ~~~All other editions are obsolete UCASFIE tS

~A4

~ * .UNCLASSIFIED

DEPARTMENT OF THE ARMYU.S. ARMY ENVIRONMENTAL HYGIENE AGENCY

ABERDEEN PROVING GROUND. MARYLAND 210104422

ATTENTION OF

HSHB-MO-T 19 September 1986

SUBJECT: Preliminary Assessment of the Relative Toxicity of A13-20837 inAnimals, Study No. 75-51-0528-86, March 1985 - July 1986

Executive DirectorArmed Forces Pest Management BoardForest Glen Section, WRAMCWashington, DC 20307-5001

4 -

EXECUTIVE SUMMARY

*The purpose and a summary of the recommendations of the enclosed reportfollow:

a. Purpose. To provide preliminary toxicity data for the candidatecockroach repellent A13-20837. These data are intended to provide guidancein selecting compounds for further entomological and toxicological evalua-tion. In addition, the data may be useful in developing preliminary safetyguidelines for handling this compound.

b. Recommendations. Based on professional scientific judgment, thefollowing recommendations are offered.

(1) A13-20837 should be considered for more extensive entomolo-gical and toxicological testing.

(2) Personnel handling this compound should avoid contact with theskin and eyes. In case of contact, the area should be flushed with plentyof water.

FOR THE COMMANDER:

Encl K^N. JOE THOMPSONColonel, MCDirector, Occupational and

Environmental Health

CF:HQDA(DASG-PSP) (wo/encl)Comdt, AHS (HSHA-IPM) (w/encl)Dir, Advisory Cen on Tox, NRC (2 cy) (w/encl) - . . __USDA, ARS (Dr. Terrence McGovern) (w/encl) ®ro.USDA, ARS - Southern Region (w/encl) .1;1_.Cdr, USMRDC (SGRD-DPM/COL Reinert) (w/encl) - ...

J.........I

DEPARTMENT OF THE ARMYU.S. ARMY ENVIRONMENTAL HYGIENE AGENCY

ABERDEEN PROVING GROUND. MARYLAND 210104422

AEPL TOATTINTION Of

HSHB-MO-T

PRELIMINARY ASSESSMENT OF THE RELATIVE TOXICITY OFA13-20837 IN ANIMALS

STUDY NO. 75-51-0528-86MARCH 1985 - JULY 1986

1. AUTHORITY.

a. Letter, US Department of Agriculture - Agricultural ResearchService, Southern Region, Insects Affecting Man and Animals ResearchLaboratory, Gainesville, Florida, 5 December 1984.

b. Memorandum of Understanding between the US Army EnvironmentalHygiene Agency; the US Army Health Services Command; the Department of theArmy, Office of The Surgeon General; the Armed Forces Pest Control Boardand the Department of Agriculture, Agricultural Research, Science andEducation Administration; titled, Coordination of Biological andToxicological Testing of Pesticides, effective 23 January 1979.

2. REFERENCES.

a. Guide for the Care and Use of Laboratory Animals, US Department of, Health and Human Services, NIH Pub No. 86-23, revised 1986.

b. Topical Hazard Evaluation Program Procedure Guide, ToxicologyDivision, US Army Environmental Hygiene Agency (USAEHA), October 1985.

c. Standing Operating Procedures, HSHB-OT, Toxicology Division, USAEHA.

d. Final Report, Mutagenicity Evaluation of A13-20837D in the AmesSalmonella/Microsome Reverse Mutation Assay, Hazleton BiotechnologiesCompany, HBC Project No. 20988, July 1986.

* 3. PURPOSE. To provide preliminary toxicity data for the candidatecockroach repellent A13-20837. This report summarizes the toxicologicaldata for USDA candidate cockroach repellent A13-20837. These data areintended to be used in selecting compounds for more extensive entomologicaland toxicological testing. The data may also be used in establishingpreliminary safety guidelines for handling the material.

4. BACKGROUND.

a. General. The preliminary toxicological evaluation of candidatecockroach repellents consists of a series of acute screening tests designedto assess potential hazards from single expesures by various routes of

- administration. The test battery included:-

Use of company names does not imply endorsementby the US Army, but is intended only to assist inidentification of a specific product.

-- I I L - .- - - .. -..--

Z-

Study No. 75-51-0528-86, March 1985 - July 1986

atoral approximate lethal dose (ALD)i

1 Primary Irritation (skin and eye)*

Ck Dermal sensitizati on

Saturated vapor (inhalation hazard)

"< Physiological scree~v

(' Mutagenicity (Ames test).

b. Project Information.

(1) All raw data from this study may be found in project filenumber 75-51-0528-86 or USAEHA Laboratory Notebooks Numbered 101 and 106.

(2) In conducting the studies described in this report, theinvestigators adhered to reference 2a. In addition, these studies wereperformed in animal facilities fully accredited by the American Associationfor the Accreditation of Laboratory Animal Care.

5. PROCEDURES.

a. Test Compound. Two lots (c & d) of AI3-20837 were synthesized andsupplied for use in the toxicological evaluations by Dr. Terrance McGovern,USDA, Beltsville, Maryland. A13-20837 is a clear oily liquid with a sweetodor. It exhibits low solubility in water but is soluble in acetone andother organic solvents. It has a molecular weight of 197 and boils at 83°C at 0.1 mm Hg.

b. Methods.

(1) Acute Toxicity Tests. Detailed descriptions of themethodology for tests (a) through (d) listed below are published inreference 2b. Methodology for test (e) is published in reference 2c.

(a) Rat ALD.

(b) Skin irritancy.

(c) Eye irritancy.

(d) Dermal sensitization (Buehler technique).

(e) Saturated vapor.

(2) Mutagenicity. Mutagenicity testing was performed by HazletonBiotechnologies Company under contract DAADO5-86-M-L723 with USAEHA. Acomplete description of the methodology and results may be found in thefinal report (reference 2d).

'S 2

Study No. 75-51-0528-86, March 1985 - July 1986

(3) Physiological Screening. The physiological screening testswere designed to obtain basic information on the underlying mechanisms ofaction for this compound. Male Sprauge Dawley rats weighing between 270 -380 gms were anesthetized with sodium pentabarbital (30 mg/kg). A hepari-nized cannula (15 cm length of PE5O tubing) was inserted into the leftcarotid artery for blood pressure monitoring. A similar catheter wasinserted into the right external jugular vein for drug injection. AStatham P23-AC fluid filled pressure transducer (Gould Instruments) wasused for blood pressure monitoring. The signals were processed by a BuxcoModel 6 Pulmonary Function Analyzer (Buxco Electronics) and printed on aTexas Instruments Silent 700 terminal. EKG's were monitored from LEAD IIand fed through a pre-amplifier and Buxco EKG analyzer. A digitalrecording of wave heights and intervals was printed on a second TexasInstruments TI terminal. Following a short period of time, usually 10-15minutes, stable physiological recordings were obtained and the animals weretreated with challenge doses, of standard pharmacological drugs includingepinephrine, nor-epinephrine, acetylcholine and histamine. Salineinjections served as a volume control. Preliminary experiments wereperformed in order to find optimum dosage levels. In most cases, thedosage chosen produced a marked change in blood pressure (10-50 mm Hg)lasting less than 5 minutes. Following the initial drug challenges, thetest compound A13-20837 was injected intraperitoneally, and the drugchallenges were repeated 15 minutes post injection. For each drug, themaximum change from baseline condition was recorded and the pre- andpost-dosing values compared. In this way, each animal served as its owncontrol. The data were analyzed using a two-way analysis of variance withrepeated measures program on an IBM PC microcomputer. A least significantrange test was used to compare pre and post-dosing values. A probabilityof less than 0.05 was used as the level of significance.

6. RESULTS.

a. ALD. The rat oral ALD was found to be 1480 mg/kg (Appendix A,Table A-1). This was the lowest dose that produced lethalities. Allanimals treated with A13-20837 (as little as 293 mg/kg) exhibited markedsalivation. Animals receiving the ALD or higher dose died between 10minutes and 18 hours post administration.

b. Skin Irritation. Compound A13-20837 produced a total irritancyscore of 4.75 in the Draize rabbit skin irritancy test. A description ofthe scoring system employed in these tests is provided at Appendix B.Based on this scoring system, A13-20837 would be considered a moderate tosevere skin irritant. Scurf and/or eschar formation was evident at I weekpost application.

c. Eye Irritation. Based on our Draize eye test in rabbits, thiscompound is a mild eye irritant with a total irritancy score of 29. Itproduced injury to both cornea and conjunctiva; however, all but one rabbitwas healed by 7 days post application. Washing the eyes with waterimmediately post application reduced the eye injury.

d. Skin Sensitization. Challenge doses of A13-20837 did not produce areaction in pretreated guinea pigs and, based on these data, it is notconsidered to be a sensitizer.

3

Study No. 75-51-0528-86, March 1985 - July 1986

e. Saturated Vapor. As indicated in Table A-i, exposure to atmospheresof A13-20837 for 8 hours did not produce any mortalities during the exposureor for up to 14 days post exposure. Nominal chamber concentrations, basedon amount of material volatilized, were 4.23 and 12.52 mg/L for the 22 °Cand 100 °C bubblers, respectively. Rats exposed to the higher concentrationexhibited excessive salivation and rapid breathing, suggesting that thecompound is a respiratory irritant. Twenty-four hours post exposure, theseanimals appeared normal. Table A-2 (Appendix A) presents the body weightgain and organ-to-body weight ratios from this experiment. There were nosignificant differences in any of these parameters between the exposuregroups.

f. Physiological Studies. Table A-3 (Appendix A) illustrates thecardiovascular effects of exposure to sublethal intraperitoneal injectionsof A13-20837. The values presented represent the maximum change fromresting or baseline levels in response to injections of the challengedrug. The only statistically significant change was an apparent increasein epinephrine responsiveness in repellent-treated rats. This probablyreflects a reduced heart rate in the baseline tests since in preliminarycontrol experiments using saline as the test compound, rats typicallyexhibited a much higher heart rate in response to epinephrines in both thebaseline and post exposure drug challenge.

g. Mutagenicity. A13-20837 did not exhibit mutagenic activity underthe test conditions employed. It was negative in all test strains used(Salmonella typhimurium strains TA-1535, TA-1537, TA-1538, TA-98 andTA-100) and at dosages ranging from .1 pL to 25 pL per plate bothactivated and nonactivated test systems (reference 2d).

7. CONCLUSIONS. Compound A13-20837 is moderately toxic by the oral routeof exposure. It is a mild eye irritant and a moderate to severe skinirritant. This compound presents no acute inhalation hazard at room-temperatures though at higher temperatures or if the repellent is atomizedas an aerosol, it may cause skin, eye and respiratory irritation. He foundno indication of a sensitization reaction and it was not mutagenic in theAmes test. When administered at approximately 0.5 x the ALD toanesthetized, catherized rats, it did not produce any marked cardiovasculareffects.

8. RECOMMENDATIONS. The following recommendations are based on

professional scientific judgment.

a. A13-20837 should be considered for more extensive entomological andtoxicological testing. Toxicological tests should include a more detailedevaluation of acute toxicity by multiple routes of administration, anassessment of the effects of repeated dosing and a complete evaluation ofmutagenic potential.

b. Personnel handling this compound should avoid contact with the skinand eyes. In case of accidental contact, the area should be flushed withplenty of water.

4

Study No. 75-51-0528-86, March 1985 - July 1986

9. ACKNOWLEDGEMENT. The project personnel shown in Appendix C assisted inthe experiments.

GLENN J. LECHBiologistToxicology Division

APPROVED:

MAURICE&. WEEKSChief, Toxicology Division

* 5

Study No. 75-51-0528, March 1985 - July 1986

APPENDIX A

RESULTS

TABLE A-1. SUMMARY OF TOXICITY DATA CANDIDATE COCKROACH REPELLENT A13-20837

ALD Skin Eye Sensitization Sat Physlo Ames(Mg/Kg) Category Category Vapor

1480 IV C Negative No No CV* Negativedeaths effectsHigh conc-irritant

* Cardiovascular - Parameters monitored included blood pressure, heart rate andelectrocardiogram in anesthetized rats.

.- 1

A-

-'p

Study No. 75-51-0528-86, March 1985 -July 1986

0'

+1 +4 4 -C0o% co % m r-4

0 0 - C~D 400I

.0 0L0-

o ao

4.- 4 +4 +1 w.-COx M m -1- j V

en kn4'I

o too 1 o

0 0 ~~ 0 0 ,

x +1 +4C, .jZ 01

ION ~ % d0)O Oo%010 aO 00 $- =L

coo 00 .jC! . 00 C; C; Q

00CD4 0 0 1004

c., 10 1 V, .C0 &01 Go0 J41 L a

to1. ILC0- +4 H+ +4 W 0

tN 0 D10 r- ...' 00 " NO I0 W0 z 0 100.! 1: 40 (A~0I -~

0 I 0 4 D00 C -3

0-I 0)Ill 0. 41A OI-

00

V. -0 00 -ftJ +n1 +O 1 01

-0r a -e - I 010 o

Im GP U.' ~0

W +1 +4 +4 13

>-0 00 C3,nC C wC

I 4 04.' C4

4C +4U 4 +4 01 4'D 3 C In

w 6 1..0 m w

C-

fe I0I.

A-2

Study No. 75-51-0528-86, March 1985 - July 1986

o Q * 0

c ; I . c

o 0

0 0

o . o 0 0

T 0

44- 44)-44 4

44'

* a -

0'0

;ri

,.°3

Study No. 75-51-0528-86, March 1985 - July 1986

APPENDIX B

DEFINITIONS OF CATEGORIES OF SKIN AND EYE IRRITANTS

1. Skin irritants.

a. Category I - Compounds producing no irritation of intact skin or nogreater than mild primary irritation of the skin surrounding an abrasion.

b. Category II - Compounds producing mild primary irritation of theintact skin and the skin surrounding an abrasion.

c. Category III - Compounds producing moderate primary irritation ofthe intact skin and the skin surrounding an abrasion.

d. Category IV - Compounds producing moderate to severe primaryirritation of the intact skin and of the skin surrounding an abrasion andin addition, producing necrosis, vesiculation, and/or eschars.

e. Category V - Compounds impossible to classify because of stainingof the skin or other masking effects owing to physical properties of thecompound.

2. Eye irritants.

a. Category A - Compounds noninjurious to the eye.

b. Category B - Compounds producing mild Injury to the cornea.

c. Category C - Compounds producing mild injury to the cornea and inaddition some injury to the conjunctiva.

d. Category - Compounds producing moderate injury to the cornea.

e. Category E - Compounds producing moderate injury to the cornea and

in addition, some injury to the conjunctiva.

f. Category F - Compounds producing severe injury to the cornea andto the conjunctiva.

-B

"S

~B-I.

5,

5.

-5

Study No. 75-51-0528-86, March 1985 - July 1986

APPENDIX C

PROJECT PERSONNEL

The experiments described in this report were performed by a multidisci-plinary group under the direction of Glenn Leach. The group included thefollowing:

1. Lynn M. Balczewski, SGT.

2. John G. Harvey, Blo Lab Tech.

3. John T. Houpt, Bio Lab Tech.

4. R. David Russell, CPT, VC.

C-1

4.

/0O

I

.f i .