UNCLASSIFIED AD NUMBER AD880200 NEW LIMITATION CHANGE TO Approved for public release, distribution unlimited FROM Distribution authorized to DoD only; Test and Evaluation; DEC 1970. Other requests shall be referred to Army Envirionmental Hygiene Agency, Edgewood Arsenal, MD 21010. AUTHORITY usaeha ltr, 10 jun 1971 THIS PAGE IS UNCLASSIFIED
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UNCLASSIFIED
AD NUMBER
AD880200
NEW LIMITATION CHANGE
TOApproved for public release, distributionunlimited
FROMDistribution authorized to DoD only; Testand Evaluation; DEC 1970. Other requestsshall be referred to Army EnvirionmentalHygiene Agency, Edgewood Arsenal, MD21010.
AUTHORITY
usaeha ltr, 10 jun 1971
THIS PAGE IS UNCLASSIFIED
RELATIVE TOXICITY OF CANDIDATE MOTHPROOFINGUNIFORM IMPREGN1ANT HEXADECYLPYRIDINIUM CHLORIDE
SPECIAL STUDY NO. 33-4-68/71NOVEMBER 1967-DECEMBER 1969
' N i
SENTINEL HEALTH
9TA T R T #4 L 4 SSIFI D
Each tra:i- ' ' . tjm-: r' t et i.-c t:o Dcp teimft ofDefense ---------..-.............
US ARMYENVIRONMENTAL HYGIENE AGENCY
EDGEWOOD ARSENAL, MD. 21010
DISCLAIMER NOTICE
THIS DOCUMENT IS BEST QUALITYPRACTICABLE. THE.COPY FURNISHEDTO DTIC CONTAINED A SIGNIFICANTNUMBER OF PAGES WHICH DO NOT
'..REPRODUCE LEGIBLY.
USAEILA-MT November 1967-December 1969
CONTENTS
A b s tra c t . . . . . .References ****,• * 0 o, o
Purpose . * * . • * * * o , o o *• 1Background 2 . . . . , * * * * o o • , * 2Summary of Findings * o , ** *e * ,4, 40 2Figure 1 * * e @ a 12
APPENDIX A - Infrared Spectrum of Hexadecylpyridinium Chloride , 19
APPENDIX B - Glossary of Recurring Definitions, Abbreviationsand Clinical Chemistry Reporting Units . . . . . . . . . . . . 20
APPENDIX C - Numerical Scoring of Primary Irritation Evaluationof Skin Reactions oo , o 24
APPENDIX D - Scale for Numerical Scoring of Injury to RabbitSkin when Determining Nonirritating Concentration . . . o . . 25
APPENDIX E - Tabulation of Toxicity Doses . * * @ . . . . . . . 26APPENDIX F - 90-Day Feeding Study Hexadecylpyridinium Chloride •
Table 1 - Summary of Diet Consumption and Dosage Data ofMale Rats * a s a # a # s * # a e s # o * * * o * , , a 27
Table 2 - Summary of Organ-to-Body Weight Ratios of MaleRats . s s e s s . . o o a * # s a * * .o .o o & . . , 28
Table 3 - Summary of Diet Consumption and Dosage Data ofFemale Rats a . * * . . o . , . . o . , . . . . . . o a * 29
Table 4 - Summary of Organ-to-Body Weight Ratios of FemaleRats . , . 30
APPENDIX G - Formulation of Artificial (Eccrine) Sweat . . . . 31APPENDIX H - 90-Day Application ofImpregnated Cloth to Skin
of Rabbits a s s # e o a ' a & # I * * * . * * * * * Table 1 - Summary of Organ-to-Body Weight Ratios a . . . o . . 32Table 2- Summary of Body Weights # .a . . . . 34
APPENDIX I - 90-Day Application of Impregnated Cloth to Skinof Rabbits, Summary of Clinical Chemistry Results a * . . & #Table 1 - Serum Lactic Dehydrogenase . . . . . o 36Table 2 -Blood Glucose #. ... . . .a. s . . 38Table 3 - Serum Glutamic Oxaloacetic Transaminase .a. .# 40Table 4 -Blood Urea Nitrogen s * s o .,.. , o 42
Table 5 -Total Serum Protein a a .a a * * s*. a s o 44Table 6 - Serum Amylase . s . . . .. . .o . . . . • 46Table 7 - Brain Cholinesterase Activity . . . . . o . . 48
USAEIIA-M1T November 1967-December 1969
CONTENTS (Cont)
APPENDIX J - 90-Day Application of Impregnated Cloth to Skin ofRabbits, Summary of Hematology Results . o . . . . . . . a . *
APPIENDIX K - 90-Day Application of Impregnated Cloth to Skin ofRabbits, Sumnary of Skin Irritation Scores . . a a a . a a a a . a 55
DEPARTMENT OF THE ARMYU.S. ARMY ENVIRONMENTAL HYGIENE AGENCY
EDGEWOOD ARSENAL, MARYLAND 21010
USALILA-MT
RELATIVE TOXICITY OF CANDIDATE MOT1IPROOFINGUNIFORM IMRGN HXADCYLPYRIVINIUM CHtLORIDESPECIAL STUDY NO, 33-4-68/71NOVEMBER 1967-DECEBER 1969
The relative toxicity of a candidate uniform mothproofing impregnant
haxadecylpyridinium chloride (10PC) was investigated using mice, rats, guineapigag rabbits and humans with the following findines:
a. The technical grade compound and aqueous solutions of 1.0 percentconcentration (w/v) or greater caused skin irritation in rabbits,
b. Eye irritation in rabbits resulted from single 24-hour applicationof a 0.1 percent (w/v) aqueous solution but no irritation resulted froma 0.01 percent (w/v) solution.
c. The skin irritating properties of hexadecylpyridinium chloridein guinea pigs prevented the determination of the sensitization potentialof this compound in comparison with dinitrochlorobenzene,
do Chronic ingestion ad libitum of a diet containing 300 ppmhexadecylpyridinim chloride for 90 days caused minimal changes in rats,
as Cloth impregnated with U.5 percent (w/w) hexadecylpyridinium
chloride and applied continuously to the shaved skin of rabbits for 90days produced no remarkable degree of skin irritation or evidence ofpotential toxic hazard.
f, A prophetic patch test on humans of wool fabric impregnatedwith hexadecylpyridinium chloride at a concentration of 0.5 percent byweight produced no evidence of skin irritation or sensitization,
g. Under the conditions of impregnation specified, IIPC shouldnot present a hazard when used as a mothproofing compound in wool fabricat a deposition of 0.5 percent (w/w). Since the weight of wool fabricto be used has not been specified it is impossible to estimate the degreeof safety.
DEPARTMENT OF THE ARMYU.S. ARMY ENVIRONMENTAL HYGIENE AGENCY
EDGEWOOD ARSENAL.
MARYLAND 21010
USAEIA-M.T
RELATIVE TOXICITY OF CANDIDATE MOT11PROOFINGUNIFORM IMPREGNANT IEXADECYLPYRIDINIUM CHLORIDE
SPECIAL STUDY NO. 33-4-68/71NOVEMBER 1967-DECEMBER 1969
1. REFERENCES.
at Letter, Stored-Product Insects Research and Development Laboratory,US Department of Agriculture, Savannah, Georgia, dated 7 August 1967,to this Agency.
b. Letter, Harris Reseerch Laboratories, Washington, D. C., dated18 September 1967, to this Agency.
c. Procedural Guide for the Toxicology Division, US Army EnvironmentalHygiene Agency, 1968 (revised 1969).
d. SOP for Patch Testing, US Army Environmental Hygiene Agency,12 October 1962.
2, PURPOSE. The purpose of this study was to acquire informationconcerning t.he toxicity of haxadecylpyridinium chloride (HPC) in man andanimals. This information provides a basis for advising on possiblehazards associated with the use of this compound as a mothproofing agentat a concentration of 0.5 percent by weight in the wool fabric of uniforms.The weight of the woolen fabrics in which the candidate compound isultimately to be used is not stated in the request for toxicologicclearance (ref para la).
USAEHA-MT Re Toxicity of Candidate Motliproofing Uniform Impregnantg
Study No. 33-4-68/71, Nov 67-Dec 69
3. BACKGROUND. The proposed mothproofing chemical IIPC submitted wasa white crystalline solids produced by Eastman Organic Chemicals,Catalog No. P 5361. Samples of the wool fabric impregnated with thiscompound at a deposition of 0.5 percent by weight were supplied for
testing by 1Uarris Research Laboratories, 4ashington, D. C., a contractor
for the US Department of Agriculture, Savannah, Georgia (ref para lb).
Harris Research Laboratories also provided untreated wool fabric and
fabric which had been treated with a sodium tetraborate (borax) solution.
The treatment of the fabric with HPC was carried out by soaking wool
fabric in aqueous solutions (pH 9.2) containing the mothproofing chemicaland 0,01M sodium tetraborate.
4. SUMSARY OF FINDINGS. The relative toxicity of IIPC was investigatedby this Agency using mice rats, guinea pigs, rabbits and humans. No
data were found which indicated a potential toxicologic hazard from wearingof uniforms impregnated with 0.5 percent (w/w) HPC. An infrared spectrumof the technical grade compound evaluated by USAEHA is found in Appendix A.Definitions, abbriviations, oymbols and clinical chemistry reportingunits used in this report are found in Appendix B. Numerical data foundin the Appendices are expressed in terms of the mean plus or minus onestandard deviation (X + S ), Statistical significance selected in thisreport is at the .01 level. All regression lines were fitted by themethod of least squares. A tabular presentation of toxicity data follows:
2
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USAEHA-MT Rol Toxicity of Candidate Mothproofing Uniform Impregnant,
Study No. 33-4-68/71, Nov 67-Dec 69
5. DISCUSSION.
a. The technical grade compound and aqueous solutions of lO percent
or greater of 11PC were shown to cause primary skin irritation in rabbits.These solutions and the technical grade compound should be handled withcare waring oye and skin protective equipment. Since there was no
difference between the skin irritation in the control animals and therabbits wearing cloth impregnated with HPC and treated with artificialeccrine sweat, it would appear that simulated sweat does not leach the11PC from the cloth in a form that causes topical effects.
b. The relative hazard from accidental chronic ingestion of HPCdoes not appear to be great although some biologic action was evident infemale rats fed moderately low dietary levels of 300 ppm and in male rats
fed 800 ppm. These levels produced increases in caecum-to-body weightratios and a slight decrease in total number of caecal microorganisms.Female rats seem to be slightly more sensitive to dietary levels of HPC
than male rats. A positive correlation of 0.8 between levels of HPC and
increase in caecui-to-body weight ratios was found both in females andmales (Figure 1) which indicates a similar dose response relationship inboth sexes to the compound.
c. The animal laboratory toxicity data and the results from a prophetic
patch test in man indicate 111'C impregnated in wool cloth at 0. 5 percent(w/w) would present little potential hazard to man under the conditions ofevaluation.
6. CONCLUSION. Under the conditions of impregnation specified, IPC should
not present a hazard when used as a mothproofing compound in wool fabric ata deposition of 0.5 percent (w/w). Since the weight of wool fabric to beused has not been specified it is impossible to estimate the degree of safety.
Skin and eye contact with the technical grade compound or with aqueous
solutions of 1.0 percent or greater should be avoided. In the event solutionof HPC or the technical grade compound should come in contact with the skinor eyes it should be washed off with copious amounts of water.
17
USAEHA-MT Rol Toxicity of Candidate Mothproofing Uniform ImpregnantsStudy No. 33-4-68/71, t'ov b7-Dec 69
ACTION OFFICERS:
' AURICE WEEKSPharmacologist
Toxicology Division
op/ .t =47/(O ER E. OLDTO Ph.D.Biochemist
Toxicology Division
STEPNE ROWE .
CPT, VC
Toxicology Division
MARSHALL erINj ERG
LTC, MSC
APPROVED: Chief, Toxicology Division
As ). J, SOMMER
LTCt MCDirector, Medical Services
18
USAEHA-MT Rel Toxicity of Candidate MothprStudy No 33-4-68/71, Nov 6
APPENDIX AINFRARED SPECTRUM OF HEXA
WAVELENGIH IN MICRON(
S
1* ri I JI,11 T'I
-........ ... _.
7) I.
00 3800 3600 3400 3200 3000 2800 2800 2400 2200 2000 1900 1800 1700 1600 1 5OWAVENUMBER CM I
SAMPLE: HEXADECYLPYRIDINIIKBr DISC, AIR I
SLIT: ROUTINE
SPEED: 200CM'/ MIN
GAIN: 3%
PEROID: 2
ORDINATE: 0-100%T
19
T Rel Toxicity of Candidate Mothproofing Uniform Impregnant,
I - ------ i . -.. .. ...... - ...... .. .... ... ...- .
.... .... L
SAMPLE: HEXADECYLPYRIDINIUM CHLORIDE
KBr DISC, AIR REFERENCE
SLIT- ROUTINE
SPEED: 200CM'1/MIN
GAIN: 3%
PEROID: 2
ORDINATE: 0-100%T
* 19
USAEHA-MT Rel Toxicity of Candidate Mothproofing Uniform Impregnant,Study No. 33-4-68/71, Nov 67-Dec 69
APPENDIX B
GLOSSARY OF RECURRING DEFINITIONS, ABBREVIATIONS, SYMBOLS AND CLINICALCHEMISTRY REPORTING UNITS USED BY THE TOXICOLOGY DIVISION, USAEHA
Definitions of medical terms and abbreviations used in this report arein agreement with the New Gould Medical Dictionary, Second Edition,published by the Blakiston Division of McGraw-Hill Book Company, Inc.Statistical terms and abbreviations are in agreement with those foundin J. Maxwell Little's, An Introduction to the Experimental Method, 1961,Burgess Publishing Company, Minneapolis, Minn. The following terms andabbreviations are either not found in the above references or have beenmodified to fit the special purposes of this report. Some of the itemshave been included below for special emphasis.
DEFINITIONS
WORD DEFINITION
Acute Exposure or Application One exposure to exogenous test materialfor no longer than 8 hours. Animalsare normally observed for 14 daysafter exposure.
Approximate Lethal Dose In range finding the first dose of thelowest series of three ascending doses(each being 50% higher in concentrationthan the previous) all of which producefatalities.
Caraway Unit That amount of amylase activity whichcauses the hydrolysis of 10 mg of starchin 30 minutes at 37C to a stage at whichthere is no absorbance at 660 my afterreaction with iodine.
Chronic Exposure Repeated daily or constant exposure toa test material for 60 or more days.Post exposure observation period willvary,
20
mi
USAEHA-MT Rel Toxicity of Candidate Mothproofing Uniform Impregnant,Study No. 33-4-68/71, Nov 67-Dec 69
WORD DEFINITION
Garry & Routh Unit That amount of cholinesterase activitywhich causes the liberation of onemicromole of SH groups from acetylthio-choline in 3 minutes at 37*C per milli-literof serum, plasma or packed redcells or per gram tissue.
Hazard Evaluation A study performed to estimate thedegree of danger associated with theuse of a material under specified
conditions of use.
Karmen Unit That amount of transaminase activitywhich causes at 25"C and 340 my adecrease in optical density of 0.001per minute per milliliter of serum.
Primaty Irritation A local inflammatory reaction of theskin, produced by a compound, whichdoes not produce destruction or irre-versible change at the site of contact.
Skin Sensitizer A compound which produces an allergicdermatitis under the conditions of test.
Subacute Exposure Repeated daily or constant exposure toa test material for no longer than 59days or less than 2 days. Post obser-vation period will vary.
Technical Grade Compound As produced by the manufacturers fortheir commerical compound; definitiondependent upon manufacurers' criteria.
Wacker Unit That amount of dehydrogenase activitywhich causes an increase of opticaldensity at 340 mi of 0.001 per minuteper ml of serum at 25°C.
21
USAEHA-MT Rel Toxicity of Candidate Mothproofing Uniform Impregnant,
Study No. 33-4-68/71, Nov 67-Dec 69
ABBREVIATIONS AND SYMBOLS
ABBREVIATION MEANING
ACh acetylcholine
AChE acetylcholinesterase
ALD approximate lethal dose
antiChE anticholinesterase
BUN blood urea nitrogen
ChE cholines te rase
CNS central nervous system
df degree of freedom
DNCB dinitrochlorobenzene
Hb hemoglobin
ia intra-arterial
im intramuscular
ip intraperitoneal
IR infrared
iv intravenous
LDs median lethal dose
LDII lactic dehydrogenase
meq milliequivalent
p .01 The probability of the change fromnormal or control being due to chance
alone is less than I out of 100.
22
_ _ _._ I
USAEHA-MT Rel Toxicity of Candidate Mothproofing Uniform Impregnant,Study No. 33-4-68/71, Nov 67-Dec 69
ABBREVIATION MEANING
PC percutaneous
ppm parts per million
P.E.G. 200 polyethylene glycol having a modalmolecular weight of 200
SGOT serum glutamic oxaloacetic transaminase
SGPT serum glutamic pyruvic transaminase
sc subcutaneous
SD or (S-) standard deviation
SE or (SEn) standard error
v/v volume-to-volume ratio
w/v weight-to-volume ratio
w/w weight-to-weight ratio
CLINICAL CHEMISTRY TEST REPORTING UNITS
Serum amylase Caraway Units
Acetylcholinesterase Garry and Routh Units
Blood urea nitrogen Milligrams per 100 cc's of serum
Blood glucose Milligrams per 100 cc's of serum
Cholinesterase Garry and Routh Units
Lactic dehydrogenase Wacker Units
Serum glutamic oxaloaeetictransaminase Karmen Units
Serum glutamic pyruvictransaminase Karmen Units
Serum protein Grams per 100 cc's of serum
23
USAEHA-MT Rel Toxicity of Candidate Mothproofing Uniform Impregnant,Study No. 33-4-68/71, Nov 67-Dec 69
APPENDIX C
EVALUATION OF SKIN REACTIONS
Erythema and Eschar Formation
No erythema 0
Very slight erythema (barely perceptible) 1
Well defined erythema 2
Moderate-to-severe erythema 3
Severe erythema (beet redness to slighteschar formation) 4
Edema Formation
No edema 0
Very slight (barely perceptible) 1
Slight edema (edges of area well definedby definite raising) 2
Moderate edema (edges raised approximatelyI m) 3
Severe edema (raised more than 1 mm and
extending beyond area of exposure) 4
An individual irritation score is equal to the sum of the scores foredema formation and erythema and eschar formation.
24
USAEHA4 IT Rel Toxicity of Candidate Mothproofing Uniforn Impregnant,Study No. 33-4-68/71, Nov 67-Dec 69
APPENDIX D
SCALE FOR NUMERICAL SCORING CF INJURY TO RABBIT SKIN WRENDETERMINING NON-IRRITATING CONCENTRATION
SKIN INJURY
ORSERVATION SCORE
No reaction 0
Slight capillary injection 1
Strong capillary injection 2
Slight erythema 3
Strong erythema 4
Slight edema 5
Strong edema 6
Slight necrosis 8
Strong necrosis 10
2Ir25
USAEHA-Mr Rel Toxicity of Candidate Motbproofing Uniform Impregnant,Study No. 33-4-68/71, Nov 67-Dec 69
APPENDIX E
TABULATION OF TOXICITY DOSES*
VARIOUS ROUTES OF ADMINISTRATIONInhalation 4-Hr Vapor
A REPORT DATE 70. TOTAL NO OP PAGES 15. NO. oPl R61PS
I. DEC I 5OI 45CONTRACT opt GRANT NO S.ORIGINA TOR'S REPORT NUKflERI21
6. P4oJEC T No 3J-4-ui/71 33-4-683/71
c 06~~S. 0OTHER NREPORT 7 OM~ (Any other nuishote hat fey be wootipeOhie repert)
NiA
to 046141OUION STATEMINT
hach transmittal of tis document Outside the~ Departmont of Defense must have priorapproval of tho US Army mivironoauntal Hlygiense Aguncy, Ldgavood Arsenal, 'Id. 210lu
rtue relatiVe toXIcitLY Of a uandhidate unifor,, naotliproofinp Lmpregnaint hiezadecylpyridintlaiciklortase was investigated using -tcs, rate, gssinea pine, rabbits and tumans Iitt. tile
a. 11a tecthinical grade iiaiwounbj and ae1ueous solutions of 1.4 percent concentration(w'vi or Swater csusa skin trr1tatsnr in rabbits.
"Ye ICrItAtiOai W.- V446itl resulted frOM Oinale i4-60O41' application Of 8 A.1t Oaercent-0 Aqueous #0otia4 Liii no IrrlistWii resuitot; Ft" a -fo "aekent o(v) cut.
Las cxii. iritatii irapsvttes of t*sadetyipvrldtnioa §.torjI~e in guifls ?i~s
PleesnteA tiet* r~i~i~ of L-a sai.sitiaton pul*Iexttak - tiUs kLsii.6ni to4mmparisca with dinitrachloroberoaene.
lao.i.igestion x4 litbitv- ut a jL*L %.oiittnlng p;.- ~xdxiyit~ii- 4Qrica f.r 0 j dys Xue 12d I .A'*$s til rats.
e..~it~.u..iyto tale sli46ou sktil'it tA.tktt wr lk' Ja ays ir:e-gCe,41 I lsarusi .ie*pro&* V itt iA of OVi,91014a ..t WL6.4W~a t.-xl& -44At.
1- .~rte at A .UQ146tratiw ' Of J,~ .' fit eL -1 .i''. -
ID L . ....... .. W s-.ijm os8ve
USAEdA-MT Rol Toxicity of Candidate Mothproofing Uniform Impregnant,Study No, 33-4-bb/71, Nov 67-Dec 69
Para 13, UD Form 1473 cont
go Under the conditions of impregnation specified, IIPC shouldnot present a hazard when used as a mothproofing compound in wool fabricat a deposition of U.5 percent (w/w), Since the weight of wool fabricto be used has not been specified it is impossible to estimate the degreeof safety.