AD-780 010
THE T0XIC0L06Y OF CYCLOTRIMETHYLENETRINITRAMINE (RDX)
AND CYCLOTETRAMETHYLENETETRANITRAMINE (HMX)
SOLUTIONS IN DIMETHYLSULFOXIDE (DMSO), CYCLOHEXANONE,
AND ACETONE
Edgewood Arsenal
April 1974
DISTRIBUTED BY:
National Technical Information Senrin
U. S. DEPARTMENT OF COMMERCE
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UNCLASSIFIED
SECURITY CLASSIFICATION OF this PACE (WJwn Dmim Enertd)
REPORT DOCUMENTATION PAGE
READ INSTRUCTIONS
BEFORE COMPLETING FORM
V REPORT NUMBER 2. GOVT ACCESSION NO.
EB-TR-73040
I. RECIPIENT'S CATALOG NUMBER
A3- 7!?A A/O
« title r-d jhe TOXICOLOGY OF CYCLOTRI-
METHYLhNETRINITRAMINE (RDX) AND CYCLOTE-
TRAMETHYLENETETRANITRAMINE (HMX) SOLUTIONS
IN DIMETHYLSULFOXIDE (DMSO), CYCLOHEXANONE,
AND ACETONE
5 TYPE Of REPORT a PERlOO COVERED
Technicr1 Report
Mar-Sept 1970
6 PERFORMING ORG. REPORT NUMBER
7. authors Bcrnard p McNamara, Ph. D., Harold P. Avenll,
Ph. D, Edmund J. Owens, John F. Callahan, David C. Fairchild,
MAJ, VC, Henry P. Ciuchta, Ph. D., Roy H. Rengatorff, LTC
ami Donald K. Biskup
0 CONTRACT OR GRANT NUMBERfaJ
9. PERFORMING organization name ano aooress
Commander, Edgewood Arsenal
Attn: SAREA-BL-T
Aberdeen Proving Ground, Maryland 21010
10. PROGRAM ELEMENT. PROJECT, TASK
AREA 6 WORK UN"? NUMBERS
Picatmny Arsenal Customer
Order No. RA02850202GGF4
¦ 1 CONTROLLING OFFICE NAME ANO AOORESS
Commander. Edgewood Arsenal
Attn: SAREA-TS-R
Aberdeen Proving Ground, Maryland 21010
12. REPORT OATE
April 1974
NUMBER OF PAOES
112
M MONITORING AGENCY NAMe 6 AOORESSC" dllltrmnl from Cantra/l/n* Olllcm)
IS. SECURITY CLASS, fot th!» fpoti)
UNCLASSIFIED
IS«, DECL ASSIFI CATION/DOWNGRADING
SCHEDULE
NA
16. DISTRIBUTION STATEMENT (o 1 1/1/• Report)
Approved for public release, distnbution unlimited.
17 DISTRIBUTION STATEMENT (of fdo <6«(>*cl orland In Black 20. If rflHoronl from Roport)
19 KCV WOROS (Contlnv* on rivwi* «id* it nmcmtmmry mtd identify by 6JocA nuoi6«r;
KDX DM SO Intravenous administration
Cyclotrimethylenetnnitramine Dimethylsulfoxide Toxicology
HMX Cyclohexanone LD50
Cyclotetramethylenetranitramine Acetone Skin
20 ABSTRACT (Confirm* on r*v«r<« i(d« If ntetHiff and Identity by block number)
A study of the toxicology of the explosives cyclotrimethvlenetrinitramine (RDX) and
cyclotetramethylenetetranitramine (HMX) in acetone, cyclohexanone, and pure and technical grade
dimethylsulfoxide (DMSO) was initiated to establish whether there is any danger to plant personnel
that handle such mixtures. This report contains a review of the existing literature on each explosive
and on each solvent. It also describes tests that were conducted to establish the intravenous toxicity
of the explosives in DMSO, the skin effects, the pharmacological effects, the sensitization potential,
(Continued on reverse side)
DD , jSJTti 1473 EOITION OF I NOV 68 IS OBSOLETE j UNCLASSIFIED
SECURITY CLASSIFICATION 0? THIS PAGE Ott* Ent»r»d)
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UNCLASSIFIED
SICUBITV CL AS8IFICATIOM OF TmH PAOEPnin Dim Bnlmnd)
19. KEYWORDS (Contd)
bxplosives Dermatitis Ocular administration
Topical application Intradermal application Skin sensitization
Cataracts
20: ABSTRACT (Contd)
and the ocular effects of the explosives in each solvent. All of these tests were conducted on
animals. In mice, the intravenous LD50 for RDX in DMSO is 18.7 mg/kg and for HMX in DMSO in
28.9 mg/kg. In guinea pigs, the intravenous LDSO Mr RDX in DMSO is 25.1 mg/kg and for HMX in
DMSO is 28.2 mg/kg. The LDSO's of RDX and HMX in other solvents were not established. RDX
and HMX in the three solvents did not penetrate the skin, as evidenced by the lack of physiological
responses in dogs and unchanged blood component values in rabbits. From the intravenous studios
in dogs, it was shown that acetone and cyclohexanone alone exert a depressant effect on the
cardiovascular system. Cyclohexanone also causes changes in the electroencephalogram pattern and
produces a semicomatose to comatose condition. DMSO had relatively little effect. Therefore, the
majority of these studies were done with the explosives in DMSO. It was found that the immediate
effects of RDX and HMX differ, RDX affecting the CNS immediately and HMX producing a
circulatory collapse initially, with delayed CNS disturbances. Topically and intradermally applied
RDX and HMX in the three solvents did not produce usually any greater skin damage than the
solvents alone, but there were several exceptions. Repeated,topical applications caused dermatitis
without fissures, eschars or scabs, but intradermal injection caused severe skin damage. Topical or
intradermal application of the solvents or of KDX and HMX in the solvents 3 days j week lor 3
weeks, followed in 2 weeks by topical or intradermal challenge, gave no evidence of sensitization.
Ocular administration showed that RDX and HMX are no more damaging than the solvents alone,
but the solvents themselves cause cataracts in guinea pigs. From these studies, it is evident that strict
precautions should be taken to avoid skin and ocular contact with HMX and RDX in the solvents
studied.
, UNCLASSIFIED
SECURITY CLASSIFICATION or This PAQl(Tfi*n Om« Enimnd)
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SUMMARY
A study of the toxicology of the explosives cyciotrimethylenetrinitramine (RDX) and
cyclotctramcthylcnctetranitraminc (HMX) in acetone. cyclohexanone, and pure and technical grade
dimethylsulfoxide (DMSO) was initiated to establish whether there is any danger to plant personnel
that handle such mixtures.
This report contains a review of the existing literature on each explosive and on each
solvent. It also describes tests that were conducted to establish the intravenous toxicity of the
explosives in DMSO. the skin effects, the pharmacological effects, the sensitization potential, and
the ocular effects of the explosives in each solvent. All of these tests were conducted on animals.
In mice, the intravenous LDSO for RDX in DMSO is 18.7 mg {tg and for HMX in
DMSO is 28.9 mg/kg. In guinea pigs, the intravenous LDSO for RDX in DMSO is 2S.I mg/kg and for
HMX in DMSO is 28.2 mg/kg. The LD50's of RDX and HMX in other solvents were not established.
RDX and HMX in the three solvents did not penetrate the skin, as evidenced by the
lack of physiological responses in dogs and unchanged blood component values in rabbits.
From the intravenous studies in dogs, it was shown that acetone and cyclohexanone
alone exert a depressant effect on the cardiovascular system. Cyclohexanone also causes changes in
the electroencephalogram pattern and produce:* a semicomatose to comatose condition. DMSO had
relatively little effect. Therefore, the minority of these studies were done with the explosives in
DMSO. It was found iliat the immediate effects ot RDX and HMX differ. RDX .ifleetinj; the CNS
immediately and HMX producing a circulatory collapse initially, with delayed CNS disturbances.
Topically jnd intradermully applied RDX and HMX in the three solvents did not
produce usually any greater skin damage than the solvent alone but there were several exceptions.
The animals treated with single or multiple I .0-ml doses of RDX in DMSO conMStently had
dermatitis while those receiving the same doses of DMSO alone did not. Some of the rabbits that
received single or multiple doses of RDX in acetone or cyclohexanone had dermatitis and the
solvent controls did not. HMX in acetone and in cyclohexanone, applied repeatedly, caused
dermatitis but the solvents alone did not. Repeated topical applications of the mixtures caused
dermatitis without fissures, eschars, or scabs, but intradermal injection caused severe skin damage.
Topical or intradermal application of the solvents or of RDX and HMX in the solvents
3 days a week for 3 weeks, followed in 2 weeks by topical or intradermal challenge, gave no
evidence of sensitization.
Ocular administration showed that RDX and HMX are no more damaging than the
solvents alone, but the solvents themselves cause cataracts in guinea pigs.
From these studies, it is evident that strict precautions should be taken to avoid skin
and ocular contact with HMX and RDX in the solvent studies.
3
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PREFACE
The work described in this report was requested by Picatinny Arsenal and authorized
by Customer Order Number RA02850202CCF4. The work , was started in March 1970 and
completed in September 1970. Experimental data arc contained in notebooks MN 2242, MN 2130,
and MN 2377.
In coi.iucting the research described in this report, the investigators adhered to the
"Guide for Laboratory Animal Facilities and Care" as promulgated by the Committee on the Guide
for Laboratory Animal Resources, National Academy of Sciences - National Research Council.
Reproduction of this document in whole or in part is prohibited except with
permission of the Commander. Edgewood Arsenal, Attn: SAREA-TS-R, Aberdeen Proving Ground,
Maryland 21010; however, DDC and the National Technical Information Service are authorized to
reproduce the document for US Government purposes.
The information in this document has been cleared for release to the general public.
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CONTENTS
Page
I. INTRODUCTION 9
II. REVIEW OF LITERATURE 9
III. EXPERIMENTATION. STUDIES OF THE TOXICITY OF RDX AND HMX
IN DMSO. CYCLOIIEXANONE, \ND \CETONE CONDUCTED AT
EDGEWOOI) ARSENAL 24
A. Intravenous Toxicity 24
1. Mice 24
j. Procedures 24
b. Results ... 15
c. Discussion 25
2. Guinea Pigs ->5
a. Procedures 25
b. , Results 26
II. Cutaneous l-'llciis 2(>
1. Rabbits 26
a. Procedures 26
h Results 29
c. Discussion 31
2. Guinea Pigs 31
C. Pharmacology of RDX and HMX in Unanesthetized Dogs . . 32
I. Experimental Procedures 32
a. Suigical Preparations 3j
b. Restraint 33
c. Monitoring of Physiologic Parameters 33
d. Tests Employed 33
2 Exposures .... 34
3. Discussion ^
4. Conclusions
5
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CONTENTS (Contd)
Paye
D. Sensitization Potential ol KDX and HMX 41
1. Procedures 41
2. Results 42
3. Discussion 49
E. Cataracts Found in Guinea Pigs Following Cutaneous and Intradermal
Applications of Solvents und Solutions of RDX and HMX .... 52
1. Procedures 52
2. Results 52
3. Discussion 54
IV. RESUME 56
A. Intravenous Effects of RDX, HMX and Three Solvents 56
B. Local Effects of Topical Applications 56
C. Systemic Effects Following Topical Skin Application 57
D. Sensitization • 58
V. CONCLUSIONS 58
LITERATURE CITED 60
APPENDIXES 65
DISTRIBUTION LIST 10P,
LIST OF TABLES
Table
I Single-Dose Toxicity of DMSO 12
II Toxicity of DMSO in Mice and Rats 13
III Highest Doses of DMJO Not Producing Deaths When Administered 3-7 Days
Per Week for 2-26 Weeks 14
6
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LIST OK TABLES (Contd)
Table Page
IV Summary ot Results of Inhal.iiion ol Acetone in Some Animals 20
V The Intravenous Toxicity ol RDX in DMSO to Mice 25
VI Intravenous Toxicity ol' Solution* ol' RDX and HMX in DMSO in G;unea Pigs 26
VII Gradation of Skin Effects 27
VIII Cumulative 1.0-MI Doses Received by KaDbits in Repeated Topical Application
Studies 28
IX Dermatitis Produced in Rabbits Ircatcd With Single and Repeated Topical
Doses ol RDX and HMX in DMSO. Acetone, and Cyclohexanone ... 30
X Eflectsol RDX and HMX Solutions in DMSO Applied to Backs of Guinea Pigs . 31
XI Exposure of Unanesthetized Dogs to RDX. HMX and Solvents 3(,
XII Residual Skin Effects Caused by tlic Repealed Topical Application of 0.5 Ml
RDX and/or Several Solvents on the Ba^ks ji Clipped Guinea Pigs (Sensiti-
/.dtion Penod) .... .... -43
XIII Residual Skin Effects Cjuscd by the Topical Application of 0.5 Ml HMX and/or
Several Solvents on the Bjcks 01" Clipped Guinea Pigs (Sensitization Period) . . 44
XIV The Skin hi loots of 0.0f Ml ol TI Saline Mixtures of Acetone, Cyclohexanone.
and Pure and Technical Grade DMSO. With and Without RDX and HMX.
Applied Intradcrmally to the Clipped Dorsul Thorax of Guinea Pigs . . 45
XV The Determination of a Suberythenial Dose of Intradermal^ Administered
RDX 111 Acetone. Cyclohexanone and Technical Grade DMSO in Clipped
Guinea Pigs 4*"
XVI The Determination of a Suberyilieinai Dose of RDX in Acetone', Cyclohexanone.
Pure and Technical Grade DMSO When Administered Topically 111 PEG 200 111
Clipped Guinea Pigs .47
XVII The Determination of a Suberytiiemal Dose of Iniradermally Administered HMX
in Acetone. Cyclohexanone. and Pure and Technical Grade DMSO in Clipped
Guinea Pigs 48
7
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LIST OF TABLES (Contd)
Tabic Page
XVIII The Determination of a Suberythemal Dose of HMX in Acetone, Cyclo-
hexanone, and Pure and Technical Grade DMSO When Administered
Topically in Polyethylene Glycol 20° in Clipped Guinea Pigs 50
XIV Cataracts Found in Guinea Pigs 53
LIST OF FIGURES
Figure
1 Slit lump Photograph of Guinea Pig Eye 40 Days After Receiving Intradermal
Application of 0.05 Ml Pure DMSO in Saline Three Times a Week for Three
Weeks 54
2 Photomicrograph of the Crystalline Lens shown in Figure 1 55
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TUB TOXICOLOGY OF CYCLOTRIMETHYLENETRINITRAMINE (RDX) AND
CYCLOTETRAMETHYLENETETRANITR AMINE (HMX) SOLUTIONS IN
DIML I HYLSULFOXIDi: (DMSO), CYCLOHEXANONE. AND ACETONE
I. INTRODUCTION.
The object of this report is to present the results of a toxicologic^! study performed at
Edgewood Arsenal of the explosives cyclotrimethylenetrinitramine (RDX) and cyclotetramethyl-
enetclranitraininc (HMX) dissolved in dimethylsulfoxide (DMS), cyclohexanone, and acetone.
In September 1969 the Toxicology Division was asked by Picatinny Arsenal to perform
the study because cf concern for the safety of plant personnel who handle RDX and HMX in
various solvents. A proposal to conduct single and repeated dose tests of rabbit skin, sensitization
tests in guinea pig*, and mechanism of action studies in dogs was approved. In addition to this work,
the intravenous toxicity of the compounds was tested in mice and guinea pigs; eye effects in guinea
pigs were evaluated, and an extensive review of the literature was prepared.
II. REVIEW OF LITFRATURE.
A. RDX.
1. Chemical and Physical Characteristics.1,2,3
The formula for RDX is-
H,
r
c
/ \
0-,N-N N-NO-
\
H:c
CH'
V
i
N
0-
RDX is a solid having a melting point of 200-203°C. It is insoluble in water, ether, and carbon
tetrachloride, but soluble in some organic solvents such as dimethylsulfoxide (DMSO). acetone,
cyclohexanone, hot aniline, phenol, and nitrobenzene.
2. Biological Effects.
The most prominent effect of RDX appears to be central nervous system excitation.
Thus, RDX is pharmacologically different from nitrites and nitrates, which act on the cardiovascular
system, causing hypotension.
9
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Sundcrman. ci at* showed that the acute oral LDSO for nonfasting rats was about 200
nig/kg and for fasting animals was SO-100 mg/kg. When they gave pentobarbital to rats (he
mortality from a given dose of RDX decreased. RDX given in intravenous doses of 18 mg/kg and
intraperitoneal doses of 10 mg/kg killed rats.
The oral LDSO for mice was reported by Slanskaya and Pozharsky5 to be aoout S00
mg/kg. They found a dose of 100 mg/kg caused no signs in mice hut was lethal to cats.
Patty1 showed that daily oral doses of IS, SO, and 100 mg/kg for 10 weeks killed I,
17, and IS rats, respectively, in groups of 35 animals. The animals became irritable and vicious, and
convulsed frequently. The lungs and gastrointestinal tracts of the dead animals were congested.
There were no pathological changes in the survivors.
In the experiments of von Oettingen et al.3 one of seven dogs that were fed 50
mg/kg/day of RDX. 6 days/week for 6 weeks, died. The animals became excited and irritable within
a few hours after the first dose. Within 1 week, reflexes were hyperactive, salivation was evident,
and convulsions and collapse occurred. After the first week, the animals lost weight even though
their appetites were good. There were no blood changes, no methemoglobinemia, and no
microscopic pathological changes. After acute exposure of animals, Slanskaya and Pozharsky5
noted changes in the fibrous material of the walls of the blood vessels in the central nervous system
and degeneration of the nerve cells: after chronic exposure, tne liver, lungs, and heart were also
affected.
Woikers in RDX plants in Italy developed epileptic-like seizures: followed by amnesia,
weakness, fatigue, and malaise.6 These effects occurred most frequently in men performing tasks
where inhalation of the dust was possible. Recovery was complete when the individuals were
removed from contact with the compound.
In other plants where the RDX was handled in a moist state, systemic toxicity was not
seen. Primary irritant dermatitis and skin sensitization did occur during the nitration process. This
was possibly caused by unidentified component of the fumes emanating from the reaction
mixture.7,8 The final purified material did not produce dermatitis.7 Patch tests with moistened
RDX did not cause irritation.3
li. HMX.
1. Chemical and Physical Characteristics.9
The formula for HMX is:
NO->
I ~
N-CH?
/ \"
ch2 n-no2
I I
CbN-N CH2
\ /
ch2-n
no2
10
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HMX is j solid having a melting point of 279°C. It is soluble in some organic solvents such as
DMSO, acetone 9 and cyclohe iione.
2. Biological I fleets.
No information w. a available from the published literature on the biological actions of
HMX.
C. DMSO.
1. Chemical and Physical Characteristics.
DMSO is a colorless liquid with the following formula:
O
II
CH-p S — C'H3
DMSO has a molecular weight of 78.13, a boiling point of I 89°C, a freezing point of 18.55°C, and a
density of l.0958,gm/inl at 25°C. It is miscible with water and many organic solvents. It has been
widely used for its solvent properties.10,11
2. Biological Effects.
DMSO has been studied extensively. It has been used as a skin penetrant and vehicle to
carry drugs through the skin 12 It protects a variety of cells from the damaging effects of freezing
and thawing. When present before and during exposure, DMSO protects against radiation damage to
cells and whole mammals.13 It is anti-inflammatory, analgesic, bacteriostatic, diuretic, sedative, and
fibrinolytic. DMSO has been reported to be helpful in rheumatoid arthritis, inflammatory
conditions, neuroskeletal injuries, and scleroderma. It is beneficial in dermatological, urological, and
ophthalmological disorders as well as in diseases of the ear, nose, and throat.12
A number of undesirable toxicological effects have also been reported.11 12 DMSO is
metabolized to form dimethyl sulfide, which gives the breath a garlic-like odor. Occasional allergic
reactions have been noted. Topical application has caused local and general dermatitis. Prolonged
administration of large oral or dermal doses produces lens opacities in swine, dogs, and
rabbits.'41 8 Tcmtogcnic effects have also been reported.19 DMSO docs not sensitize the skin of
guinea pigs,20 and does not seem to be carcinogenic.21,22
3. Toxicology of DMSO.
a. Single Doses.
There is a close similarity of the signs produced by DMSO in various animal spe..es and
by various routes of administration. For example, the LD50's for single, intravenous injections in
II
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mice, rats, cats, and dogs ranged Irom 2.5 to 8.9 gm/kg.1 8 The signs observed at lethal or near lethal
doses were similar in all species (deceased activity, tremors, weakness, dyspnea, stupor,
convulsions, hypothermia, and hematuria).18 However, Fcinman et al. 23 infused 4.0 gm/kg
intravenously into two monkeys without causing death or signs.
Single oral doses also produced similar signs in various species. The signs were like
those seen in the intravenous studies except that emesis was also noted 1 °
Subcutaneous and intramuscular injections caused local areas of inflammation, edema,
hemorrhage, and necrosis.1 K
DMSO applied to the skin produced transient erythema. When the bodies of mice and
rat* were immersed in DMSO. most animals died within 24 hours.18
Ocubr jJminislrjiion produced lacrimation and edema and erythema of the orbital
tissues.1 8
LDSO values and toxicity of DMSO by various routes of administration in several
;itiimal species ate shown in table I as reported by Smith ft a/.18 and in table II as reported by
Caujolle ci at.19
Table I. Singli-Dose Toxicity of DMSO
LDSO
Specm
Intravenous
Oral
Subcutaneous
intraperitoneal
Dermal
Mouse
J .8 - 8.9
(18,24,25,27.281*
16.5 - 24.6
(18,25.28)
gm/kg
13.9 - 20.5
(18.24)
14.7 - 17.7
(18.27)
<50(31)
| Kai
5.2 - H.l
(18.24.25)
17.4 - 28.3
(18.25)
12.0- 20.5
(18,24)
13.0(18)
<40 (18)
Ciuincj Ply
>11 (29)
-
>55 (29)
-
Ciiickcn
-
-14 (29)
-
-
-
Cat
<1
-
-
-
-
l
IA>|1
-•2.5 (24)
>10
-
-
>11 (18)
Monkey
>4(23)
>4(23)
-
-
>11 (18) j
* Ilk- numbers in pjren theses jfe reference*.
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Tabic II. Toxicity of DMSO in Mice and Rati
l-.tteel (24 hr)
Dose
1 ill ravenous
'Intraperitoneal
Subcutaneous
Oral
gm/kg
Mue
MDNI- JJ
5 0
14.0
12.2
>14.0
LD5U
11.0
20.1
16.0
N.E. W
MDAF H
14.8
>22.1
22.5
14.0
Kjlb
MDNK
> () 0
11 1
10.0
>15 0
LD50
N.h.
13.7
j 3.7
N E.
MDAF
> 6.0
15.0
15.0
>15.0
¦j] MUNI" - mjMinum dnv m-vor Ijljl
MOAI - imniniuni Ui"*- jlw i> - IjIjI
I. I - not cvjlujti'd
b Repeated Doses.
( I) Intravenous
l'lie intravenous LD50 DMSO for dogs is about 2.5 gm/kg.24 Rosenkrantz i'l al24 gave
two dogs steadily increased intravenous doses of DMSO daily. They survived 1.86 gm/kg but died
when dose was increased to 2 95 gm/kg.
Intravenous doses of 0.3 to 2.4 gm/kg of DMSO (both undiluted and diluted) were
given to dogs by Willson i'l ul25 6 days per week for 4 weeks. Anemia, hemoglobinuria,
bilirubintiru. increased serum glutamic o\alacctic transaminase activity, iron-positive pigment in the
liver, spleen, and kidney, as well as cloudy swelling in the liver resulted. The four dogs that received
the 2.4 gnvkj: dose died.25
(2) Oral.
Oral doses of 2.5 gm/kg/day given by Caujolle and coworkers (cited by Smith et ai1 8 )
for 6 weeks to mice caused degeneration of the liver and indications of nephritis. When Caujolle
gave oral doses of 1. 2 and 5 gm/kg/day for 6 weeks to rats, these doses also caused changes in the
liver and kidneys. Smith18 (unpublished data) found reduction in body weight, and some organ
13
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weigh Is. bill no gioss adverse- eflccts when I. 3. and 10 gin/kg were administered orally to rats lor
59 consecutive days. Rats survived oral doses of 0.4 and 7.0 gm/kg lor 13 wee'ks in the experiments
of Sommcr ami Tauberger (cited by Smith rt al 18). The lowc; doses caused some atrophy of the
spleen. Doses of 14.1 gni/kg caused sedation and death. Hematological findings and urinalysis were
normal. Necropsy revealed generalized hyperemia, gastrointestinal hemorrhage, and splenie ehanges.
Smith cr al 1 8 reported that oral doses between 2.5 and 10 gm/kg given for 14 days
caused death in one ol llircc dogs. Continued dosing of the other two animals caused sedation,
ulceration of the oral mucosa, injected sclera, muscle tremors, elevated hemoglobin and hematocrit,
increase iii transaminase activity, fatty degeneration of the liver, and hemorrhages in the
^astro-intestinal tract. These doses also caused some changes in the ocular lens after 48 days.
Monkeys tolerated live daily oral doses of 4.0 gm/kg without adverse effects.23
Daily oral administration ol I or 3 ml/kg, or dermal administration of 1. 3, or 9 ml/kg
ol DMSO to rhesus monkeys lor IK months produced no toxicological effects in body weifjit.
blood pressure, heart rate, icspiratory rate, body temperature, water consumption,.neurological
reflexes, electiocardiogiams. hematology, and urinary constituents."6 There were no pathological
iiiangcs or lenticular cllects attributable to DMSO. Some animals receiving oral doses of 9 ml/kg
uicd between IS and 53 days of study. Atelectasis and emphysema were the only pathological
Jiangcs.
Repeated doses of DMSO that did not produce deaths, as reported by Smith ct al.1 8
..re slmwn in table III.
Table III. Highest Doses of DMSO Not Producing Deaths When
Administered 3-7 Days Per Week for 2-26 Weeks
Dose
^ Species
Intravenous
Oral
Subcutaneous
Intraperitoneal
Dermal
gm/kg
! ' Kat
2.5(24)*
1 1.0(29)
10.0(24)
8.2(29)
10(18)
Dog
48(18)
10.0
-
-
11
Monkey
4.0(23)
4.0(23)
-
-
11.0(18)
* The numbers in parentheses ure references.
c. I'l'iccts on Skin.
(I) Animals.
Brown el al.29 painted neat DMSO on hairless mice twice a week for 30 weeks with no
noticeable effects. They also found no gross or microscopic signs of damage when the liquid was
applied to clipped backs of guinea pigs daily for 28 days.
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Undiluted DMSO and ftO',< solutions were applied to the shaved skin of dogs and
monkeys hv Smith ct at 1,1 in doses of 3.3 to 33.0 gm/kg/week for ft months.18 Application
produced transient warmth and reddening of the skin. Purfuraceous jnd membranous desquamation
of the epidermis started within 3 weeks jnd persisted throughout the experiment but microscopic
examination revealed no involvement ol the deeper layers. Cutaneous application to rats produced
hyperkeratosis, parakeratosis, and Ibcal ulceration.111
When applied topically to anesthetized dogs, increased temperature of the skin,
subcutaneous tissues, and underlying muscles wa, noted '.>y Bradham and Sample.30 DMSO did not
produce dermal sensitization in guinea pigs tested by (JolJman el a 12,1
(2) Man
Kilgnian31 applied 9 ml of 90% DMSO to the torsos of 20 people for 2ft weeks. He
noted a transient erythema, mild scaling, and d:Tuse erythematous dermatitis Skin biopsies of
other subjects who received twice as much DMSO showed j mild perivascular lymphocytic
infiltration, acanthosis, absence of a granular layer, and parakeratosis
Various 'nvcstigators have described dermal effects of DMSO Erythema3* heat,20-33
local irritation.2" -'4 35 burning and tingling36,37 are often noted immediately or soon after
application. l'he burning sensation lasts 10 to 30 minutes.36 and the erythema disappears within 1
hour.32 These efleets are less intense with continued use.
With repeated use and occasionally alter a single application, a variety of skin effects are
noted32 These include redness, lashes.'6 3 "•^peeling.3''scjluig.3 2-3h local dermatitis3,J 1 general
dermatitis.3 2 and vesicul.ition 32, "* I'lie site ol application sometimes becomes sensitive to sunlight4()
Sulzberger4 2 reported on wheals and Hares following application 10 scratches or
intracutaneous injection DMSO may cause liberation ol histamine 3h
Urticaria.2"'35 angioneurotic edema, and swelling ol the tot ue have been mentioned
as consequences of using DMSO 35 Bad breath is a frequently mentioned effect noted after dermal
application.3 2-34 •3h'37 -il>
-------
When Klebergcr'6 administered 9 ml/kg/day (50% DMSO) to oeagle dogs by stomach
tube, it caused vomiting. Marked refraction changes toward myopia were noted in I week. A slight
opalescence of the lens developed alter 2 months.
Lenticular changes and myopia are not readily seen upon ophthalmoscopic gross, or
microscopic examination ot' the lens. lixamination by rctinoscope or slit-lamp is required. Smith ct
al 17 found lenticular changes and myopia in dogs alter oral administration of 5 or 10 gm,'kg/day of
DMSO. Without slit-lamp examination, no abnormalities were noted in dogs following intravenous
doses of 1.6 gm/kg/day for 62 days, or in one dog that was given 2.4 gm/kg/day for 33 days,
followed by 4.8 gm/kg/day for 28 days. Dermal doses of 6.6 gm/kg given 5 days/week for 6 months
did not reveal any abnormalities
Burnett and Noll44 found changes in the refractive index of the crystalline lens of
seven out of eight rhesus monkeys receiving 9 ml/kg/day orally and suspected such changes in six of
eight animals dosed with 3 ml/kg/day. An increased brightness first appeared at the central or
nuclear zone of the lens which later became clearly demarcated from the peripheral zone. However,
these changes were not seen in monkeys receiving intravenous infusions of 4 gm/kg/day 5 days j week
lor 09 days."J oral doses ol 5 gm/kg/day lor 100days,4S topical application up ;o 33 gm/kg/,veek for
6 months,' "or following daily doses tor 18 months of I or 3 ml/kg orally or 1,3, or 9 ml/kg dermally.26
Gordon46 reported that a 16-year old boy with severe bilateral uveitis exhibited a
small subcapsular lens opacity for approximately 5 months of treatment with eye drops of DMSO.
The opacity was not increased with continued administration of DMSO and was believed to be a
consequence of the uveitis and not the therapy. Other patients received the eye drops for 1 to 15
months.46
Numerous studies in man have not revealed any actions on the lens.32,3s ,37 •39-40'4 ' -46
e. Teratogenic Effects.
In 1965, Caujolle ct al.41 found that low doses of DMSO in different species produced
some abortions but no malformations. In 1966, these investigators" gave oral or intraperitoneal
doses of 5 to 12 gm/kg to mice during the 6th to 12th day of gestation. Doses of S to 10 gm/kg
were given to rats by the same route and for the same time course. The animals were sacrificed 1 to
3 days before parturition. DMSO produced malformations in the mice afterthe intraperitoneal doses,
but not after oral administration. Both orjl and intraperitoneal doses caused abortion and
malformations in the rats.
Kabbits received 5 gm/kg orally or 4 gm/kg subcutaneously from the 6th to the 14th
day of gestation.' 9 The animals were sacrificed on the 20th to 24th day of gestation. Only one
malformation was found among 83 fetuses.
Malformations were produced in the chick embryo when doses approaching the LDS0
(10.3 mg/embryo at 72 hours and 12.2 mg/embryo at 96 hours) were employed.19
Fcrm48 found DMSO to be teratog' ic when administered in high doses to hamsters.
A single oral dose (10 ml/kg) of DMSO given during the period of organogenesis (7 th
day of gestation) was teratogenic to hamsters.49
16
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I. ruiiiurit'tnu.- Actions.
Research hi Ilk area indicates thai DMSO is not tumorigenic. The influence of the
daily ingestion of 50 ppin of DMSO on the production of breast tumors was studied in
Sprague-D.iwley rain The DMSO was started 3 days before or alter gavage with
7,1 2:dimetliylhcnx|(/1 anthracene. Die gavage alone produced tumors in almost all animals in 4 to 8
weeks. Atter 18 months DMSO had no beneficial or deleterious effect on the num!>er of rats that
developed tumors, or on mortality Hals that received DMSO had fewer tumors than control
animals.21
DMSO was more cytocidal to leukemic lymphocytes than to normal lymphocytes.
Cells from patients with acute granulocytic leukemia, cells of spontaneous lymphomas ol AKR
mice, and cells of transplantable rat lymphoma also were relatively sensitive to DMSO
DMSO wjs louud to be ol vjlue in research on cancer of the cervix V'lien Avre and
LeGuerrier5" applied the compound to prvc.ulcerous lesions or to iii .situ carcinomas, no structural
changes were seen When dccardron. an adrenal steroid, was applied in DMSO to such tissue,
regressive cylologic.il changes occurred in 2 to 3 weeks Barium chloride changed normal epithelial
cells of the iervix to bizarre multinucleated cells characteristic of premahgnant dysplasia. DMSO
potentiated this <:ttect ot turium chloride
D. Acetone.
I Clieni'i ai and Physical Characteristics.
Acetone < 2-|impaiN;ne. ili iu-ihyl ketone) is j colorless liuuid with the lollowing
fornutlj
O
C»3 CH3
it has a iiiolvt'il.ir weight of S8.08, a boiling point of 56.1°C, a freezing point ol -95 fi°C and a
density of 0 791 i gm/ml at 20°C. It is miscible with water and other organic solvents.51 5 *
it is widely used as a solvent lor resins, fats. oils, collodion cotton, celluloid, cellulose
acetate, etc li ilk. artificial leather, and lubricating oils.5 3
Acetone is prepared commercially by the destructive distillation of wood, by
distillation of. calcium acetate, by fermentation of corn products using selected bacteria, and by
catalytic oxidation of isopropyl alcohol, cumene. or natural gas.5 *
Because of its low flash point, the lire and explosive hazard of acetone is a major factor
in its handling.5 *
17
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2. Hazard jnd Physiological Considerations.
Acetone has a more rapid narcotic action than methyl alcohol when received
intravenously, orally, or by inhalation, but is less toxic. Its effect is similar to but greater than that
of ethyl alcohol. Due to its volatility the most likely route of administration is by inhalation. When
inhaled, acetone is readily absorbed in the blood stream because of its solubility in water and thus
transported rapidly throughout the body.53 liyc and skin contact may occur, but ingestion is not
likely because of its sharp and bitter taste 52 When absorbed through the skin, acetone penetrates
more slowly than ethyl ether or chloroform because its high solubility in water results in slow
penetration through the epidermal cells.54 The danger of skin absorption is small and unlikely to
occur in normal industrial operations.51 The principal hazards to health arc directly related to the
inhalation of vapors at very high concentrations and to repeated and prolonged extensive skin
contacts because of the potent solvent effects of the compound.52 Acetone also h-s an irritating
cflcct on mucous membranes.1*3
,.j Some early investigators considered acetone to have some toxic action on the kidneys
when inhaled as well as when ingested. Liter experiments have shown the toxicity to be much lower
(almost identical to the toxicity of ethyl alcohol! than (hey reported. However, kidney damage has
been demonstrated when acetone is taken orally.5 3
Acetone is excreted rapidly, mainly by the lungs. In excessive exposures, some is also
excreted through the skin and in the urine 52
Data on metabolism of jeetone suggest that much of it is split to formate and acetate,
to acetoacctate, and to the J-carbon intermediates of glycolysis.5 2
3. Toxicity in Animals.
a. Single Oral Dose.
The toxicity of acetone in animals by this route is low. The lethal and narcotic doses in
rabbits are reported to he 10 ml/kg and 7 ml/kg, respectively. In dogs, the lethal and narcotic doses
were determined to be 8 ml/kgand4 ml/kg, respectively.5 2
b. Single Intravenous Doses.
Lethal doses for rabbits and rats are 4 ml/kg and 6 to 8 ml/kg respectively. The
narcotic dose was determined to be 2 ml/kg for both species.5 2
c. Single Intramuscular Doses.
KabOits were depressed but not made unconcious when given 5 ml/kg
intramuscularly.5 2
d. Skin Irritation.
Acetone may produce IocjI dermatitis due to its defatting action on the skin if
repeated prolonged contact occurs. An occasional short exposure should not cause skin irritation.5 •
18
-------
e Skin Mwirplioii
l.j/j icw (7 i//54 teportcd that I lie amount ui acetone absorbed through the skin was
slight when lliey measured the amount ..xlialcd and the amount in llu blood following immersion of
animal's lout.
I. Inhalation
I he inhalation elleci.s ol .ic clone are shown in table IV 5 ' As with (he other routes ol
administration. the inhalation toxicity ol acetone hi .mini .In is low
g. Lye Irritation
C'aipcitlcr and Smyth5 s reported th.it sin.ill doses ol .icetone caused- moderate
uritation to the eyes ol labbits. L.nson and co-wiiikets''*' deni.msiraied mild edema. (lomer57
su^ested that iLnulr.ition ol the scleia liy Ihe juIimk icsulUJ m gelatinous llocculation and
opacity ol Hie sJeia. Iniuiies so incuired would he expected lo resolve completely
4 I.IIc'cIs hi Mj.i.
a. Repeated Oral Duscs
Albc-none* * iC|>orted that .icetoiic taken orally in doses ol i«> 20 gin daily for
seveial days produced no ill«. Ikcts other ih.in slight drowsiness.
b. l ye I Ii>-c is
Nelson .inJ eo-wwik,.is5v n-poricd thai |>crsons nol ,iuuslome«l t»» atinosplierc's
containing acetone vapor experienced eye n.is.il. .,nd thio.it irritation • lien exposed to
concentrations ol 50u ppm. However, v>jilest>y •'.//<•> cited in I'atiy5 - I Inund ili.il accluua: d
peisons .Muld toierate as nuiel) as 250l) ppin Willi niii> minor irrita.ion ol the nose and throat, that
2(Ju lo 4l)l) ppin was detectable only up*mi immediate contact. jnd that altei j short time. 700 ppm
was undetectable
Inhalation.
l';.t(y5- repotted that Kagjn. ih experiments v.i. hinr.. .. lomul lh.il it was impossible
lo mil.ile acetone concentrations ol 22 nij»/ inei I''.<00 ppm i lor iiiueir ilian 5 minutes because ol'
acute irritation ol Hie tliroal. In jddiiion to determining the mtoieraiile concentration. kjgan also
determined Ins absorption of mli.ileil vapor lo be 71',. lor uie '"-uiimiie e\posme Two other men
exposed by Kuwait to I i nig, I l-k>50 ppm) lor I 5 minutes absorbed i(< and 77'./ iespcct:uly.
tfnggs and Scilallef, as cited 0y I'.itt^3". ic^itcd th.it ihe cOetfieient ol distribution
ol ucc'ionc' between alveolar air and hiood or water was I $3*. expiessed i.i mg/liter Thus, a
workman breathing 1000 ppm t 2 3 uig.'liici I ol acetone ui an would icacli ei|iiilihiiuin when he had
attained a blood concentration of 0.77 gm/nicr Under ilicse exposuic conditions, a man of average
weight would accumulate' 40 gm ol' acvlonc thioughoui the bod> Once ihis level wjs attained, the
only acetone absorbed would We that textured u> lepi.ne mc .imount incuboli/.ed or excreted and
sufficient to equilibrate water consumed lla^gaid. cf al also cited m Patty5 * demonsti yted that
this equilibrium is never actually reached even alter several day^ ol continuous exposure.
-------
Table IV. Summary of Results of Inhalation of Acetone in Some'Animals
Animal Species
Concentration
Duration of Exposure
Effects
mg/liter
ppm
hr
Mice
20
48
8.300
20,000
7-3/4
1-1/2
Side position after 4 to 7-3/4 hr: deep narcosis
in only a few animals after 7-3/4 hr.
Side position after 60-70 min; deep narcosis
after 1-1/2 hr.
110
46.000
1
Side position after 20-30 niin;deep narcosis after
40-60 min: death a few minutes after end of
experiment.
Cats
8-10
3,370-
4,220
5
Initial salivation, irritation of nose and eyes: slight
stupor and drowsiness after 5 hr.
20-50
8,440-
21,100
3-4
Usually drowsy in first 1/2 hr; later sleepy, increased
sensitivity to pain.
80-ICO
33,700-
42.200
4
No drowsiness, marked irritation of central nervous
systen.; giddiness. ataxia, narcosis, twitching and
convulsions during narcosis.
125
52,750
1-1/3
As above.
40
17.000
4-1/2
Side position after 3-3/4 to 4 hr; recovery.
114
48,000
3
Side position after 1-1/2 hr. no deep narcosis,
recovery.
178
75.000
1-1/2
Side position after 1/2-lhr.deep narcosis after 1 to
1-1/4 hr with preliminary convulsions.
Guinea pigs
50
21.000
25 min
Lacrimation only.
50
21.000
4 to 8-:/2
9 to 23-1/2
Loss of auditory reflex; side position.
Coma, death.
-------
I'jit) cited I'armcggiam and Sjssi5* who showed that excretion ofacetone in liumuns
alter a single oral dose is rapid lor 8 hours, but is not complete in 24 hours. They reported that
under conditions ol' light worl' and normal urination, the proportion of acetone excreted was
approximately 40 to 70% in the breath. 15 to 30% in the urine, and 10% through the skin
tl. Skin Hlects
Acetone is only slightly irritating to the skin even after rather severe exposure. It may
be slowly absorbed through the skin but this appears to be of little practical significance. The
principal lia/ard to the skin involves the strong solvent action of acetone on skin lipids and other
skin constituents Damage of this kind only occurs with repeated, prolonged, and extensive skin
contact.5 2
5. .Summary.
I he incidence ol acute jcetone poisoning is very low when the popularity of this
solvent .ind the huge world-wide yearly consumption are considered. Although nonfatal industrial
poisonings have been reported, they have resulted from-the inhalation of high concentrations of
acetone.5 3
Reports of po. onings due to repeated exposures have usually involved ate tone as the
solvent in combination with other materials. It is doubtful that the toxic symptons reported by
workers who were affected could be attributed to acetone.5 2
l'jtiy ;i- cites the extensive studies conducted by Oglesby and co-workers over j
15-year penod that icpresent 21 million man-hours ol experience (average exposure concentrations
up to 2000 ppm). and the 10-year studies pertorined by l-'assett. Both groups have demonstrated
that no individual was injured by chronic acetone exposures. It is obvious from these data that the
toxicity ot acetone is low. the lethal dose tor humans cannot be estimated.52
0 Hygienic Standards ot IVrmissible Exposure.
The American Conlerence of Governmental Industrial Hygienists60 has recommended
a threshold limit of 1000 ppm (2400 mg/cu m) for acetone. This level is in good agreement with the
results reported by Oglesby.5 2
t. Cycloliexanone.
1 Chemical arid Physical (.'luraUcnsiics.
(. yclohexanone is a colorless, neutral liquid ot peculiar ketone-likc odor, with the
following formula
0
II
Hyf^YH2
H/'s>s^H2
1 *¦
h2
21
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The molecular formula is CgHjQO and the molecular weight is 98.14. The boiling point is 155.6°C,
the melting point is -45°C, the specific gravity 0.9478 20/4°C, and the vapor pressure 4.5 mm Hg at
25°C. Cyclohexanone is only slightly soluble in water but miscible with organic solvents. It is
manufactured by catalytic oxidation of cyclohexanol and distillation of pimelic acid salts.3 2 ,5 3
Like many of the ketones, cyclohexanone is used both as a solvent and as a chemical
intermediate. It has many applications in the lacquer, paint, and pi>ting-ink trades, its excellent
solvent properties have made it desirable as a spot remover in the dry cleaning and textile industries.
It is used in relustering and spray-painting fabrics. It is used as a degreaser, expecially in removing
grease from nickel sheets. It is used in the leather industry as a thinner for fast-coating finishes on
light and fancy leathers and for wet and dry degreasing in this trade, it improves the adhesion of
varnishes, expecially on greasy leathers.5 2 ,s 3
It is stable and should not present a problem of flammability unless handled at elevated
temperatures.5 2
2. Hazards of Physiological Action.
In the handling of cyclohexanone in industrial applications, skin and eye contact and
inhalation of the vapors are most likely to occur. Ingestion or absorption of dangerous quantities
through the skin are unlikely unless excessive exposures are encountered.5 2
Cyclohexanone is both an irritant and a narcotic agent. Death is thought to be due to
respiratory failure. At high dosage levels, Treon el a/.61 found the organic sulfate and glucuronic
acid output in the urine of rabbits increased.
No specific lesions were found in rabbits following exposure to lethal doses. Toxic
effects were general vascular injury and inflammation. In repeated exposures of rabbits to smaller
concentrations (190 ppm), liver or kidney changes were barely demonstrable and no significant
blood changes were observed.61
Due to its low volatility, the possibility of obtaining hazardous levels during industrial
handling is slight unless the process is conducted at elevated temperatures. The compound has
strong warning properties (eye, nose, and throat irritation) at low concentrations; thus,
overexposure to concentrations that may cause systemic injury are not likely to be tolerated
voluntarily by most humans. Cyclohexanone has a low acute oral toxicity; occasional skin contact
with the liquid should not be irritating but prolonged or frequent skin contact will cause defatting,
irritation, or dermatitis. Eye contact may result in marked irritation and some transient corneal
injury.5 2
3. Toxicity in Animals.
a. Single Oral Doses.
Jacobi, Hayashi, and Szubinski62 reported the minimum lethal dose of cyclohexanone
for mice to be 1.3 to 1.5 g/kg. Treated mice were seen to develop paresis of the hind quarters,
narcosis, and deep, slow respiration before death. Treon, Crutchfield, and Kitzmiller63 reported the
LDIOO value for rabbits to be 1.6 to 1.9 gm/kg and observed an increased excretion of organic
sulfates and glucuronic acids in the urine. Some lung damage was seen at high dosages.
22
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b. Single Intravenous Dose
Pally5 2 cited Caujolle and coworkers who reported that 630 mg/kg of cyclohexanone
administered intravenously to anesthetized dogs caused death in 60 minutes. Accelerated
respiration, vasodepression, and hypotension were noted.52
c. Single Intraperitoneal Injection.
Intraperitoneal injections of 0.5 ml/mouse were reported, by Fillipe, as cited in
Patty.52 to cause excitation, paresis of hind quarters, marked hypothermia, and convulsions
followed by death. One of the metabolic products found was adipic acid, presumably due to the
oxidjtion of cyclohexanone.
d Skin Irritation.
No published data are available on the effects of cyclohexanone on animal skin.
e Skin Absorption.
Treon ct a/.63 determined the LDI00 by skin absorption in the rabbit to be 10.2 to
23.0 gm/kg. Tremors, narcosis, and hypothermia were reported prior to death. These effects are the
same as those reported for other routes of administration but the dose required is larger.52
f. Eye Irritation.
Cyclohexanone applied to rabbit eyes caused marked imtation and some corneal
damage. Therefore, liquid cyclohexanone may be expected to cause marked irritation and possibly
some transient corneal injury when in contact with the human eye.55
g. Inhalation - Acute Exposures.
A 6-hour exposure of guinea pigs to 4000 ppm of cyclohexanone. as reported by
Speclit and coworkers,64 caused typical narcotic symptoms: laenmation, salivation, depression of
body temperature and respiratory heart rates, and opacity of the cornea. Recovery from the
narcosis was slow. Patty52 cited Smyth, who found that a 4-hour exposure o! rats to 8000 ppm
resulted ic anesthetic death but that a 4-hour exposure to 4000 ppm caused no deaths. When Gross
(cited by Patty52 I exposed mice, guinea pigs, and cats to 3800 ppm of cyclohexanone. the signs
seen in the guinea pig by Specht*4 were noted. No abnormalities were found in the urine
h. Inhalation - Repeated Exposures.
Patty5 2 reports that monkeys and rabbits were exposed for fifty 6-hour penods to 190
ppm with no detectable effects other than very slight kidney and liver injury. At 309 ppm, slight
eye irritation was seen; at 773 ppm, salivation and eye irritation were noted; and at 3082 ppm. the
highest level used, light narcosis, labored breathing, incoordination, and a slightly increased
mortality were seen. As in the single-dose oral studies, increased amounts of organic sulfate and
glucuronic acid were found in the urine of the rabbits.5 2
23
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4.
fclTccts in Man.
a. Skin Effects.
Although no published data are available on the effects of cyclohexanonc on human
skin, it is reasonable to assume that frequent, repeated, or prolonged contact may possibly cause
some irritation or dermatitis. This assumption is based on the fact that cyclohexanonc is an
excellent fat solvent and could dissolve the skin lipids and other constituents of the dermis. This
hazard is considered to be of a low degree except for unusual situations where proper safety
practices are not followed.5 2
b. Inhalation and Eye Effects.
Nelson anu coworkers5' exposed men to concentrations of 25, 50, and 75 ppm to
determine the tolerable level for prolonged exposures. They reported 50 ppm was definitely
objectionable, and that 75 ppm caused pronounced eye, throat, and nose irritation. A level of 25
ppm was thought by most volunteers to be the highest tolerable concentration for an 8-hour
exposure.
5. Summary.
The principal hazard to health in handling cyclohexanone is inhalation of the vapors.
However, because it is capable of defatting the in, prolonged or frequently repeated skin contact
may logically be expected to result in irritation or dermatitis.
6. Hygienic Standard of Permissible Exposure.
A threshold limit value of 50 ppm has been recommended by the American Conference
of Governmental Hygienists.60 This level should prevent narcosis but may be somewhat high, based
on the work of Nelson et a!,59 If comfort is to be attained, the concentration of cyclohexanonc in
the air may have to be maintained below 50 ppm.60
111. EXPERIMENTATION. STUDIES OF THE TOXICITY OF RDX AND HMX IN DMSO.
CYCLOHEXANONE. AND ACETONE CONDUCTED AT EDGRWOOn ARSFNAI
A. Intravenous Toxicity.
I. Mice.*
a. Procedures.
Ten percent (wt/vol) solutions of RDX in DMSO and HMX in DMSO were prepared at
room temperature and injected into the-caudal vein of20'gm mice.The mice, in groups of six,
received 5, 10. 15, 20, 25, or 30 mg/kg of RDX in DMSO, or 15,25, 30, 35, or 50 mg/kg of HMX
in DMSO. All mice were observed for 4 hr after injection and daily for 30 days.
* This part of the investigation was conducted by Edmund J. Owens, Toxicology Division and MAJ David G.
Fuirchild, VC, Veterinary Medicine Division.
24
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b kl'sulls
(1) RDX in DMSO.
Hie 1.1)50 of RDX in DMSO administered intravenously to m .c was calculated to be
18.71 (15.(>o-22.24) mg/kg Irom the data in Table V. Deaths occurred within 5 to 10 minutes and
were preceded by mild convulsions and labored breathing. The survivors displayed lethargy which
pcisisted for up lo 2 hr; all appeared normal in 24 hr.
Ijhlc V 1 lie Inlijvonom Toxicity ol KDX in DMSO to Mice
bxpcriinenijl
Bliss Analysis
Dose
Moilalily
Morialiiy
Dose
ing/kg
/O
„ mg/kB
5
0/1
i
11.0(5.9-20.4)
10
U/b
16
14.9(10 7-20.8)
15
1/0
30
16.6 (13 0-21.2)
:o
4/6
50
18.7(15.7-22.3)
:s
i
5 lb
84
23.5 (18 4-30.1)
i
i v
i
b/b
31 9(18.9-53.8)
Slope 10.0
e. Discussion.
Solutions of DMSO containing eilhc>* 109'< HMX or RDX arc of the same order of
toxifiy when administered intravenously to the mouse. It is estimated that between 10 and 20 ml
of either solution would be required to present a lethal hazard to man if accidental injection
occured.
2. Guinea I'ius *
a. Procedures
Solutions of RDX and HMX (each 33% wt/vol) in DMSO were prepared and given
intravenously to guinea pigs, two animals per dose. Controls, two per dose, received 0.20, 0.23, and
0.25 ml of DMSO alone.
t-.Jmund J. Owens Jnd DjviJ (J l-jn\hild conducted this investigation.
25
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b. Results.
Death occunred within 5 min after injection of either RDX or HMX solution. No signs
were noteJ /» the guinea pigs that received DMSO alone. The LD50, based on the data in table VI is
25.1 mg/kg of RDX and 28.2 mg/kg ol HMX.
Table VI. Intravenous Toxicity of Solutions of RDX and HMX in DMSO in Guinea Pigs
Experimental
LD50 by Bliss analysis
Compound
Dose
Mortality fraction
(95% C.L.)
mg/kg
mg/kg
RDX
15.8
0/2
25.1 (20.0- 31.6)
20.0
0/2
25 1
"1/2
31.6
2/2
HMX
20.0
0/2
28.2(20.0-39.8)
25.1
1/2
31.6
1/2
39.8
2/2
B. Cutaneous Effects.
I. Rabbits.*
a. Procedures.
(I) Single Dose.
The backs of rabbits were clipped free of hair: those animals with observable skin
abnormalities (abrasions, scratches, etc.) were not used. One milliliter of the following mixtures
(wt/vol) were applied to six rabbits per mixture: 33% RDX or HMX in DMSO; 75% in
cyclohexanone. and 5.4% in acetone. The HMX formed a suspension in cyclohexanone and acetone.
Control animals, in groups of three, received I-ml applications of the solvents alone. After all
applications a polyethylene sleeve was taped to the back of each rabbit and left in place for 24 hr.
When the sleeve was removed, each rabbit's back was examined for skin irritation and any irritation
was graded according to FDA standards (table VII). The rabbits were observed daily for 30 days for
evidence of skin irritation and systemic toxicity.
Blood samples were drawn trom each rabbit and the following parameters were
analyzed: red blood cell count, white blood cell count, hematocrit, hemoglobin, alkaline
phosphatase, serum glutamic oxalacetic transaminase, blood urea nitrogen, creatinine, sodium,
potassium, chloride, and carbon dioxide.
* This part of the study was performed by Ronald K. Biskup and Hubert L. Snodgrass, Toxicology Division
-------
Table VII. Gradations of Skin Effects (Draize Test)
l.rythcma
No erythema
Mild eryihenia
Moderate erythema
Severe eiytliemj
I-rytlierna with edema
Necrosis
No necrotic tissue
Leys thjn 50% necrotic tissue
50% to 100% ncciotic tissue
I GOV; necrotic tissue wtli well-defined eschar formation
L)ehy Jrjlion ^nd Desquamation
No dehydration or desquamation
Mild dehydration or desquamation
Moderate dehydration or desquamation
Severe dehydration or desquamation
I'wo rabbits from each dose group and one contro! rabbit from each solvent group
were sacrificed for pathological examination at I hr, 3 days, and 30 days.
(2) Repeated Doses.
file (allowing solutions were prepared for repeated topical application to the clipped
backs of rabbits: KDX -.33% in DMSO; 7.5% in cyclohcxanone; and 5.4% in acetone;HMX - 33%
in DMSO: 2.5'/ m cyclohcxanone. and 2.0% in acetone. The compounds were applied in doses of
1.0 or 0.1 ml to m\ rabbits per mixture and volume; 5 days/week for 4 weeks. Control animals in
groups of three icceived applications ol 1.0 or O'l ml of the solvents alone. All rabbits were
observed daily during the study for eut.meous or systemic effects. Skin irritation was graded as
shown in table VIi. The cumulative doses jre listed in table VIII.
Ui«.ih1 samples were drawn trom each rabbit and the same blood constituents analyzed
as cited in the single dose stu-ly.
I'wo ubbits from each dose group and one control rabbit from each solvent group
were sacrificed for pathological examination at 7 days (2 days after the fifth dose). 14 days (2 days
alter the 10th dose), and 28 days (2 days alter the 20th dose).
(3) Supplemental tests With UMX OY/> in DMSO).
One-milliliter doses of HMX (33¦/<> in DMSO) were applied topically to the clipped
backs of five sets of rabbits (two animals per set) daily for 1, 2, 3, 4. or 5 days. They were observed
daily lor 30 days for cutaneous and systemic effects.
27
-------
Table VIII. Cumulative I.O-MI Doses Received by Rabbits in Repeated Topical Application Studies
Cumulative dose
No. of
doses
33% RDX or
HMX/DMSO
7.5 RDX/Cyclohexanone
5.4% RDX/Acetone
2.5% HMX/Cyclohexanone
2.0% HMX/Acetone
1
2
3
4
5
10
20
165
330
495
660
825
1650
3300
37.5
75.0
112.5
150.0
187.5
375.0
750.0
mg/kg
27
54
81
108
135
270
540
12.5
25.0
37.5
50.0
62.5
125.0
250.0
10.0
20.0
30.0
40.0
50.0
100.0
200.0
-------
h. Results.
(1) Single Dose.
Single doses ol I 0 ml RDX and HMX and the different solvents and (he solvents jlone
produced no iiross evidence ol cutaneous irritation or systemic el I vets throughout the 30-day
observation period. No changes in blood constituents thai could bv attributed to either the solvents
or the dissolved explosives were noted. However, microscopic examination of the dosed areas
showed iliat KL)X in all solvents caused dermatitis that persisted for as long us 30 days. The solvent
system causing (he most pronounced effect was 330' KDX in DMSO. Dermatitis was not seen
microscopically in any of the animals receiving doses of HMX solutions.
(2) Repeated Doses.
The icpcalcd doses (daily 5 days/week lor 4 weeks) ol RDX in DMSO produced .10
^rovN evidence ol cutaneous irritation throughout the 30-day observation period. Although no gross
eflcds could Ik seen, a death occurred alter the eighth application of the 1.0 m! dose ol RDX in
cyclohexanone (1 Otli day of test). one death after the tilth application of the 0 I ml dose of RDX
in .icctonc (7th day), and another death after the 10th application of 1.0 ml dose of RDX in
acetone (13III day).
Repeated doses (daily 5 days/week lor 4 weeks) of 0.1 ml HMX in DMSO, 0.1 and 1.0
ml HMX in cyclohcxunone and acetone produced no gross evidence of cutaneous initation or
svsteniK toxicif. thioughoul the 30-day observation period Of the solvents, only the I ,U-inl dose of
ci.ide DMSO had .m> <:lfeci. and this was a slight desquamation of the skin in the 2d week of
application.
Repealed doses of i.O ml HMX in DMSO produced a mild desquamation of skin at 7
days. Three deaths occurred, one alter the second application (2 days), one after the sixth
application : applications appeared weak and dehydrated. Two
deaths occurred alter two applicahoits (2d and 4th days), and one death after the lllth application
i8tli uay i. Alt survivors appeared normal upon gross examination 30 days alter dosing.
(4) I'ail'olorficai Findings.
Lesions which could be attributed to the compounds tested were confined to the -ate
of application. When skin was alfooted, it was olten reddened or thickened and there was
microscopic evidence of inflammation. The incidence of dermatitis, as noted by the pathologists on
necropsied animals, is shown in table IX. When minimal dermatitis occurred in animals that received
the mixtures, there was dermatitis of a similar degree in the corresponding solvent control animal,
wilh these exceptions. The animals treated with either 1. 10. or 20 1-ml doses of RDX in
DMSO consistently had dermatitis at the time ol neciopsy while those receiving the same doses of
DMSO alone did not. Two rabbits that leceived one 1.0-ml dose ol RDX in acetone and two that
received 20 0.1-mMoses of RDX in eyclohexanone had dermatitis and the solvent controls did not
29
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Table IX. Dermatitis* Produced in Rabbits Treated with Single and Repeated
Topical Doses of RDX and HMX in DMSO, Acetone, and Cyclohexanone
o
Number of rabbits showing dermatitis
Compound
Dose
Number of
Time of
DMSO
Acetone
Cyclohexanone
Cbntrol
applications
observation
Experimental
Experimental
Control
Experimental
Control
ml
33%
5.4%
7.5%
RDX
1.0
1
1 hour
—
2/2
1/1
0/2
0/1
3 days
2/2
1/1
2/2
0/2
2/2
1/1
30 days
2/2
0/1
0/2
0/1
0/2
0/1
0.1
5
7 days
0/2
0/1
0/2
0/1
0/2
0/1
10
14 days
0/2
0/1
0/2
0/1
0/2
0/1
20
30 days
0/2
0/1
0/1
0/1
2/2
0/1
1.0
5
7 days
0/2
0/1
0/2
0/1
0/2
0/1
10
14 days
2/2
o/l
0/2
0/1
0/2
0/1
20
30 days
2/2
0/1
0/1
0/1
0/1
0/1
339?
5
4%
7.
5%
HMX
1.0
1
1 hour
0/2
0/2
0/2
0/1
0/2
0/1
3 days
0/2
0/1
0/2
0/1
0/2
0/1
30 days
0/2
0/1
0/2
0/1
0/2
0/1
2.0%
2.5%
0.1
5
7 days
0/2
0/1
2/2
0/1
2/2
0/1
10
14 days
0/2
0/1
0/2
0/1
0/2
0/1
20
30 days
0/2
0/1
0/2
0/1
0/2
0/1
1.0
5
7 days
2/2
1/1
2/2
1/1
2/2
1/1
10
14 days
0/2
0/1
1/1
1/1
2/2
1/1
20
30 days
0/2
0/1
-
1/1
2/2
1/1
* Noted by pathologists on necropsicd animals.
-------
There was no dilicicncc in test and control animals receiving HMX in DMSO Two rabbits Hut
received live applications ol 0.1 ml I1MX in acetone and two that received live applications ol 0.1
ml HMX in cyclohexanone hud dermatitis while the controls were normal.
No lesions were found in the livers, kidneys, spleens, lungs, tracheas. hearts, intestines,
bladders, muscles, bones, or bone marrows of the rabbits which died or were sacrificed following
repeated topic.il applications of RDX or HMX in the three solvents, or the solvents alone. Gross
examination of eyes revealed no cataracts. (Eye effects are discussed further in paragraph VI of this
report )
When presented for sacrifice and necropsy, three animals had signs of posterior leg
weakness or posterior let; paralysis (possibly attributed to broken backs). They had been treated
with ten 1.0-ml doses of HMX (2.5% in cyclohexanone), ten 1.0-ml doses of HMX (33% in DMSO»,
jnd five 1.0-ml doses of RDX (33''. in DMSO), respectively.
c. Discussion.
The most serious hazard incident to handling the test solutions appears to be that of
repeated skin conljct with 33% HMX in DMSO <3 deaths), 5 4% RDX in acetone (2 deaths), and
7.5% in cyclohexanone (I death). It is recommended, however, that workers avoid skin contact
with any of the solvents to avoid damage to the human skin that might not be readily predicted
from the response in rabbits.
2. (iuinea Pigs.*
Solutions of RDX or HMX (33% wt/vol) were prepared and 316 to 1,000 mg/kg were
applied to the clipped backs of guinea pigs in groups of four animals per dose Observations are
given in table X.
lablc X kttccis ol RDX and HMX Solutions in UMSO Applied to Backs of Guinea Pigs
!
Dose oi
explosive i
mg/kg
.Uo
510
l,000
2,000
1,000
Number of
applications
Observations (4 animals)
KDX
HMX
'¦r
No eiieet
No effect
Slight erythema
Slight erythema
Slight eryihema after
iu>t application, later
application showed no
further erythema
i !
Applications of 2 ml DMSO alone produced no effects.
No eftect
No effeel
Slight erythema
Slight erythema
Slight erythema after first
application, later applications
showed no further erythema.
Skin spongy and absorbent after
each application Alter 3rd dose,
apprehension, loss ol weight,
and loss of normal skin color
were observed
* Ronald K. Biskup and Hubert L. Snodgrass, Toxicology Division, conducted this investigation.
31
-------
1'hurriKicnloj^y til RDX anil IIMX in Unaiicsihcli/.id l)up.*
I. r.xperiiiicntal I'rocedtncs
I orty-live healthy beagle dogs. averaging I 1.4 kg. were used in ;i study lo delinc the
eiiects ol KDX and IIMX in various vehicles u|h>ii physiologic paramcteis. especially the central
iieivous sysloiii. ol iinancsihcti/.ed Jog-, siller .kuIc jnd chronic percutaneous application. In
.iJuilioii. j iiiiulci1 si mi y ol' the toxic signs and the mechanism ol action ol these explosives alter
>.iii avenous administration was conducted.
a Surgica' 1'rep.n.ilion
About 2 weeks belore iestin.\ electrodes were implanted under pentobarbital
..i.^llicsia into the skull ol each dog above I lie cortical area for recording electroencephalograms
ii r.tii. A longitudinal incision jbout 2-1/2 in. long was made across the scalp exposing the muscles,
v.iikIi were 'enacted nvci jii area ul the skull approximately 2 in. square. A limited .iniounl of
camei i/.atr.m was used io redact the muscle and control bleeding. The exposed area of the skull was
then St..i|icd clean.
I wo types oi electrodes were used during the test series. The lirsi consisted of a
.Wi-iniii-loiig nylon bolt wuh an 18-gage stainless steel core. The second was a round nylon plug
< 1/2 null diameteri through which four silver-platinum wires were passed and extended underneath
u>i altotu 1/4 to 1/2 inch.
lo implant the lirst type of electrode, four holes were drilled through the skull to the
ditia. I'hc holes weie lalcral to the midline, two being posterior to the external Irontalcrest and two
in the central p.metal area (figure A-1 in the appendix). The holes were then threaded and the
electrodes were screwed in (two to three turns) and cemented in place with cpoxy or acrylic. Alter
tlie cement hardened, the skin and subcutaneous tissue were sutured back together and holes were
.ui n.iv/ ilu skin so thai the electrodes could be passed through.
I'o implant the second type ol elc-ciiod.1. lorn small holes (0.04 m.). slightly larger than
ilu diameter of the wne. were drilled into me skull in the same arc:> .is those for the lirst type of
niuioiic. I lie wires and nylon plug were cemented ui place and allowed to di>. The skin and
sunuiianeous tissue were pulled hack llrinly around the plug and sutured: tuc skin that bulged up
.¦loiind tile electrode was excised.
I lie second type of electrode was found to be more Jumble lor long term studies
i.iv.in>e me first type wjs more liable to be loosened when the Uoy-. were hi the holding cages. Both
t, pes iccorded equally well.
because of the length of the study (approximately 6 months), precautions had to be
t.ikcn u> prevent sickness or death from ini'ection. l-lach animal was maintained on .mtibiotic
literacy (Uuraullin or Bicillin) for about 10 days alter surgery and put back on therapy if any
•ni'cciioii occurred during the holding time. The implant area was cleansed daily for 2 weeks with
hydrogen peroxide, and any necrotic tissue was sw jbbed away. In addition, some dogs had to be
maintained on Hydrocortisone injections for several days alter the operative piocedure to control
niilamalioii and swelling.
" 'I'hi-. investigation wjs conducted by Henry P Ciuchlj, l'h. D.. jnd SP4 J. Denny, Toxicology Division.
32
-------
11'*-" administration ol j)v. niiilurtjilji (Nembutal), 30 ing/kgr iv, lo duns lor surgical
implantation ur I l.li electrodes pioduced typical changes in the wave pattern. Iigure A-l shows a
.cpicscntalive iiacing Iron) .1 Joy under 1 lit.- inlJuencc ol the barbiturate compared to a tracing from
tlu' alert or ui,..iK\llieli/ed animal (typical ol ail dogs prior to exposure).
b Kesti.iint
l>og> u.sed lor the subacute percutaneous study were restrained in a stall-type holder
consisting ol two movable sheets ol I '4-m. pegbo..ui (I 2 X 3(> in.). I:ach animal was placed between
two sheets ami ihe pegboard was adjusted 10 lit snugly against the dog's Hanks. Rods were then
insetted i:i.ou;:Ii ilie holes (<> keep die dog in any desired position. Most of the dogs were relaxed
and not bothered by this type ol icstraint
c. Moiuioiiii;; oi Physiologic Parameters.
Mood pressure was measured by means ol a catheter inserted into the femoral artery
•iiid attached to .1 Matliam piessine li..usdticer and earner preainphlier. hlectrocardiograin il-.KC.)
.11 in ileal 1 and uspiralory rates hk icc-onlcd lioin needle electrodes placed 111 either side of the
ciiesi wall, taped, and attached lo an impedance pneumograph pieaiiipliticr and then to a cardiac
I'Kainphliei I In I.I.Ci was momiorcd oy the implanted electrodes winch were connected to a
swio.li box and lueii to high gain pre.n.ipniiers. All the physiologic preamplifiers were then coupled
to a six-channel I-AM Physiograph Kecoidcr 01. m some instances, to a Sanborn 350 system. As
depicted in ligure A-l. a total ol six l-.r.li leads could be monitored, two at u time, by adjustment of
inc switch box. Leads I and II weie used i>ninarily tor prolonged recording while Leads III through
Vl iiioinioicd oliier areas to determine whether similar activity was being shown at those points,
1 c. 11 cpilcpiiloim discharges occunmg hi the tionial-parietal area were also occurring in the trontal
01 occipital aic.is. ,tc.
d. Tests l.mptoyed
Dou-. w.111 implanted clcctiodes weie given an uitrasiiort-acting barbiturate (Surital)on
tlie Hioiiiiiig ol me experimeiii to peuuii placement 01 the catlieter in the femoral artery. The
.niiiiiais rciovvied within 3u to o(> minutes .nid ^*e re then placed in the resitauiers. Physiologic
luiaiiieteis tl-l.(j, I KG. blood pressiue. lespnalion, and hc-rt rate) were monitored in order to
csixiiiisu contiol values Tncse same paiaiuctcis were then assessed after a batUry of 10 tests were
pi,, settled (t) eucli jiiiitial to jvsure initiiei mat the animal's physiologic responses were normal prior
to ...iy exposure I licsc tests were as tollow*.
(1) Atulitoi^ I lie normal responses ot the animal to noise were
..VM.SV.J T lie • iin.uliis was geneiaicd by a bone .lud.oplione (Sonolone Audiometer; placed near the
eji 01 (lie dog wan a i'.ieney selling ol _'.S0 eps lollowed by 7Sl) c;>s.
(2) Visual. A beam of light was passed in a vertical and then a
hoi 1..0111.1I line across the dog's field ol vision.
(3) Paul. A pair ol electrodes was placed against the inner aspect of a
moistened cai and a Grass stimulator was set 10 discharge 25-voit shocks lasting 7 5 msec with a
-'-msec interval between shocks The stimuli were repeated until the animal's head jerked or the ear
iwiteiicd.
33
-------
(4) l-'yolid. I he eyelid was touched Willi a modified Von I'rey hair
< vCimiIcvs steel 27-gygc needle cleaner wall u blunted end) until a blink occuncd.
(5) Corneal. The cenicr 01 the cornea was touched with the same wire
uniit blinking occtiricd.
(6) Nasal. An ammonium hydroxide-impregnated cotton swab was
l<.i.Yv.'d undei the nose lor I to 2 seconds.
(7) Pupillary i. oi neck, :iiul neural response was noted.
(V) Light. A o-v, (>-amp bulb was ilaslud lor I second in Iront ol the
; uoj' >. e>es and t'NS iespouse was noted
(IUI Vibiatory (Kap>. I he rest, jiner holding llie dog was rapped or hit
mkii a metut object, and neural response was noted.
The responses to all these stimuli weie recordeJ on the physiograph and arc illustrated
hi ngures A-2 to A-G in Appendix A.
2. Exposures.
KDX and MMX were prepared hi DMSO, acetone. jiiJ c^ciohexanone but the
concentrations were not all the same because ol dillcrvnccs in solubility. Ihe dogs were exposed
.ulier hy topical application or intravenous injection (tabic XI)."
The vehicles alone were adnimistered topically in volume* ui I nil and injected
intravenously (0.125 ml/kg) into several dogs to assess activity
The topical exposures were made by dropping I ml ot KDX and MMX solutions onto
On clipped backs ot dogs. The animals were clipped weekly during the su.d)
Intravenous injections were given to two ui-js through an indwelling caliictci in (lie
icmoral vein. All other injections were made direuiy into the cephalic vein. Table XI show* how
many dogs were exposed to each compound and to each vehicle alone.
The acute studies consisted of applying the test compounds iu the dorsal area only
uncc and recording blood pressure, respiratory and heart rates, EKG, and LLC at exposure time,
llie same parameters, except for blood pressure, were monitored weekly i. r the next 4 weeks,
iiiood pressure was recorded during the fourth weex. The subacute studies wue similar to the acute
with the exception that the test compounds weie applied daily 5 days per week lor 4 weeks
Massive chronic exposures were an afterthought and entailed the percutaneous application of RDX
.nid HMX in DMSO to two dogs each. The animals were exposed to 480 mg/kg daily for 3
consecutive days. The animals' EEC's, respiratory and heart rates, and EivG's were recorded at the
time of each administration and at I week. The dogs were aiso obs«.i\ed grossly tor any
hyperrcflexiu. The intravenous studies were conducted pnnurily to determine potency and type
34
-------
Table XI I \poiuic of UiiancMl en/ed Dogs to RDX. HMX ani! Solvent
Ago in
Peicuuneous
IniMveneows
33',? RDX
in DM SO
%Aci RDX
in acetone
7.5% RDX in
cyclohevanone
AiUle
Dose No of dogs i
mg/kg |
289.0 2
47 3 2
65 7 2
Cliionk"2/
~r
Dose | No ol" dogs
nig/kg |
289.0
7 3
65 7
— .. Masme chronu.^
Dose No of dogs
mg/ky
480 0
Do
No of dogs
mg/kg
400
20 0
6 75
3 37
9 40
4 70
I
i
,
3 37
1
! 33% HMX
289 0 1
289.0
* 480 0
2
40 0
6
{ in DMSO
J
20 0
2% HMX
17.5 I 1
17.5
i
i
2.5
3
in acetone
»
!
•
i 5.4% HMX^/
>
! 6.75
2
in acetone
I
2.5% HMX in
21.9 1
21.9
1
*>
3 10
.
cyclohexanone
i
1 55
1
f
Solvents alone
ml ;
ml
i
\
i
i
! ml
| (total dose)
i
DMSO
1.0 ; 1
1 0
i
¦>
! io
4
Acetone
1 0 | 1
i
1.0
¦> i
i
1.0
4
Cyclohcxjnone
'
1.0 j 1
1
1.0
i
i
¦> ,
I
! i-o
3
j I
Applied djil> 5 djys per ucck lor 4 »cck\
k. i
J Applied dailt for 3 tonsciutivt Jj>s •
^ Not a ilcar solution
-------
pharmacologic activity. 'I'll.- dogs thai were exposed by the iv route were observed subjectively lor
signs and symptoms such as twitching, convulsions, labored respiration, heart rate, salivation,
lacrimution, cyanosis, prostr.uion. and dejth.
II jny of tile animals in any phase exhibited subconvulsivc jerking, twitching, etc.. they
wcie observed in (he holding area alter expcnnicntation and were monitored periodically lor
hypcrrellcxia.
j. Topical Application.
(i ( Acute.
t able B-l in appendix B shows the values obtained for blood pressure and heart and
inspiratory rates, before and alter acute exposure, over the 4-wcek period. No consistent increase or
uev.ie.isc in any of the physiologic parameters was noted, and the high values for respiratory rate
observed in some dogs reflected panting due m changes in the ambient temperature. Tables b-l 1 and
U-lll depict irends in responses to test stimuli belore and jitcr acute exposure. The predominant
responses were variations in respiratory and heart rates. No consistent enhancement or blockade of
uiiy ui the responses inomtoicd was noted. It also should be mentioned that all animals in this phase
ol the study showed a normal response to the lid. corneal, and visual tests before and after
exposure. Responses to these tests were not tabulated.
(2) Subacute.
Table B-IV shows the values for blood pressure, hearl rate, respiratory rate during the
control-recording and the four subsequent weeks. No appreciable or consistent changes were
observed at any time. Tables B-V and B-VI depict the trends in responses to the test stimuli before
anu after exposure. All annuals in this phase also gave a normal response to the lid, corneal, anJ
visual tests at all times. No blockade or enhancement of any physiologic response to a test stimulus
was noted duriag the period. During the second or third week of application of DMSO alone or 111
combination with RDX and MMX some dogs exhibited slight erythema and desquamation of the
back.
(3) Massive Chronic.
The administration of 480 nig/kg of RDX and 11MX in DMSO to four dogs (two with
cuch test compound) for 3 consecutive days produced no consistent tiross noticeable changes in the
ainmals. Although one of the beagles receiving RDX-DMSO appeared to be sligluly more irritable
and hyperactive for 20 10 30 minutes after the application of the first and second doses, no
disturbances were noted in the litG. Animals were held for 2 weeks after exposure jnil appeared
no: ilia!.
b. Intravenous.
To assess the type of activity that RDX, HMX, and the vehicles possessed once the
compounds entered the circulation, a number of dogs were given the test compounds intravenously.
36
-------
(I) Vehicle Controls.
Four experiments were conducted with I-ml injections of DMSO. Two dogs showed
no apparent changes in uny parameters while two dogs demonstrated appreciable decreases in blood
pressure (25 to 35 mm Hg), but only for 5 to 10 sec. Compensatory tachycardia occurred with the
hypotension. Recovery was prompt (figure A-7). Four experiments conducted with 1-ml injections
of acetone demonstrated decreases in blood pressure ranging from 10 to 50 mm Hg and being 5 to
60 sec in duration. Bradycardia, followed by tachycardia, was observed in 3/4 animals. A typical
response to acetone is shown in figure A-8. The administration of 1 ml of cyclohexanone to three
dogs produced a marked and immediate cardiovascular collapse with cardiac arrest lasting
approximately 10 sec. Respiration, was inhibited and pulse pressure became narrow during this
phase. Figures A-9 to A-l 1 demonstrate these effects. Note the high-voltage, low-frequency activity
of the EEC both at the 5-sec and 5-min intervals post injection. Animals during this phase were
comatose or semicomatose and had a dulled pain response. Recovery occurred 20 to 120 min after
injection.
(2) RDX-DMSO (40 mg/kg).
Two dogs given this dose expired within 45 min and 90 min. Figure A-l2 shows a
typical response to RDX-DMSO at this dose. Within 15 to 30 sec animals demonstrated
subconvulsive jerking, twitching, and convulsions. The seizures were cyclic and the latter phase of
the experiment was marked by inadequate respiratory movements, decreased blood pressure, and a
flat line on the EEC.
(3) RDX-DMSO (20 mg/kg).
Two dogs were given this dose. One animal demonstrated CNS hyperactivity within 15
sec after injection and exhibited hyperreflcxia lor at least 1 hr. Other parameters were not affected
appreciably. The other dog convulsed within 90 sec after the injection and did not recover until 16
hr later (figure A-l 3).
(4) RDX-Acetone (3.37 and 6.78 mg/kg).
One dog was given the smaller dose and three the larger. The parameters monitored
were generally unaffected except for decreased blood pressure and erratic EEC disturbances. All
animals appeared normal after removal from the restrainer.
(5) RDX-Cyclohcxanone (4.7 and 9 4 ing/kg).
One animal was tested with each dose and in both cases the effects were similar:
marked decrease in blood pressure, cardiac arrest, and respiratory inhibition occurring in the
presence of high-voltage, low-frequency EEC discharges. Animals were semicomatose to comatose,
eyes were dilated and the pain threshold was elevated.
(6) HMX-DMSO (40 mg/kg).
Six dogs were given HMX-DMSO. Four were given a single dose of 40 mg/kg while two
others received the same amount in two separate and equal doses. The single dose produced a severe
cardiovascular collapse in all four animals. This occurred concurrently with a narrow pulse pressure.
37
-------
bradycardia, and respiratory alterations (figure A-14). The EEC was characterized hy high-voltage,
low-frequency discharges. Two of the four animals died at I and 3 min. while two others survived
the circulatory collapse. Vomiting was observed prior to their removal from the restrainer. Both
expired in approximately I4i 2 hr.
Two animals receiving two separate doses of 20 mg/kg each exhibited a severe
cardiovascular depression and one animal died in I min. The other dog recovered from the
cardiovascular embarrassment but demonstrated EEC hyperactivity, vomiting, and extreme
sensitivity to light and stroke stimuli at 1 to 2 hr alter injection. The animal A'""* ¦" yA+ 3 hr (figures
A-15 to A-19).
(7) HMX-DMSO (20 mg/kg).
Two dogs were given a 20-mg/kg dose. In one animal (figures A-20 to A-22) there were
minimal changes, although at 15 min the animal vomited. The dog then stabilized for 2 hr. at which
time hypcrrcflcxia to vibratory and light stimuli occurred. Visual perception and lid and corneal
reflexes were normal. At 5-hr postinjection the dog became-extremely hyperactive and had
convulsive seizures. Recovery did not occur until 5 days after exposure.
In order to observe the toxic signs of HMX-DMSO without the influence of the restrainer,
a 20 mg/kg dose of the compound was given to a second dog which was unrestrained and unoperated
upon. The HMX-DMSO was injected in a volume of 0.6 nil into the cephalic vein of a 10-kg dog at
10:30 am. At 10:32 the animal became hyperpneic and retched. Defecation and salivation were
observed at 10:37 am. At 10:47 am the dog became cyanotic: heart rate was 60 and respiratory rate
was 120. At this point the animal was still standing although its pulse was becoming faint. At 10.53 am
the dog was prostrate but capable of getting up. At 11 • 14 am the heart rate was 72 and respiratory rate
was 120. For a short period of time the eyes reflected increased circulation and the pulse was getting
stronger. At I 1:20 am the dog was cyanotic again. Retching occurred at 11:56 am and again at
12:22 pm. At 1:40 pm breathing became labored. At 1:4! pm there was an onset of subconvulsive
jerking, especially in response to the auditory stimulus. Clonic-tonic convulsions occurred at 1:47
pm and again at 2:01 pm. Opisthotonus was noted at 2:02 pm. At 3.00 pm the animal was
convulsing. The dog sat up at 3:20 pm but hyperreflexia was pronounced. Convulsions occurred
again at 4:07 pm. The animal was prostrate at 4:45 pm and died between 10:00 pm and 3:00 am.
(8) HMX-Acetone (2.5 mg/kg of 2% and 6.75 mg/kg of 5.4%).
The administration of the lesser dose to a dog produced transient hypotension. The
EEG demonstrated a sleep-like wave pattern during the 12- to 40-min postinjection period, but the
animal could be aroused easily. No irregularities were noted the following day. The larger dose
produced a blood pressure decrease along with high-voltage, low-frequency EEG discharges. Upon
removal from the restrainer the animals were lethargic, but all appeared normal the next day.
One dog was given 6.75 mg/kg (5.4%) HMX-acetone after receiving a prior injection of
2.5 mg/kg of the 2"A solution. The second dose produced a drop in blood pressure, cardiac arrest,
and a depressed respiratory rate. The EEG was not adversely affected. At various intervals the dog
vomited and periodically fell asleep but could be aroused (figures A-23 to A-27). The animal
appeared normal the next day.
38
-------
() I IMX-CycloliexaiioiK- (1.55 and 3.1 ing/kg).
Oik* dog w;is used lor each d«se lioth demonstrated cardiovascular collapse, onset ol ;i
co ina I ike stale, and elevation ol' the pain threshold. The animal receiving the larger dose retched,
had tremors, and vonntei!. No convulsive seizures weie noted. Both dogs recovered completely in 2
to 3 days.
3. Discussion.
The absence of consistent changes in blood pressure, heart rate, respiratory rate. I:KC».
and lillG over a 4-week period after topical application of KDX and HMX in the vehicle indicates
that the mixtures do not penetrate the skin at the doses and concentrations tested in the acute and
subacute experiments. This is supported by the fact th t intravenous administration of these
materials causes changes in the physiologic parameters
One of the two dogs receiving the massive dose of RDX-DMSO appeared to
demonstrate slight hyperexcitabihty approximately 15 to 30 inin alter the second and third
application of the compound. Sine-.' these particular animals were held for so long, they even
appeared anxious in the control phase and a more involved study'would have to be conducted for
substantial evaluation.
The majority of the intravenous studies were conducted with RDX and HMX in
DMSO. Although iv injections of the .agents in acetone and cyclohexanone were also evaluated, it
was found that thesv* vehicles themselves (figures A-8 to A-l 1) exerted a depressant action on the
cardiovascular system, and cyclohexanone induced changes in tlu- l:l:G pattern. Animals exposed to
cyclohexanone also demonstrated a semicomatose to comatose condition in some instances.
RDX-acetone did cause some liEG disturbances, but these were not well defined. Further difficulty
was encountered because the concentrations of the active agents in these vehicles was considerably
less in f)MSO. DMSO itself was shown (figure A-7) to produce relatively little effect when
administered intravenously, and no gross complications were observed during a holding period ol 2
weeks.
It is evident from these studies that the immediate effects of RDX and HMX differ.
RDX affecting the CNS immediately after injection and HMX producing a circulatory collapse
initially, with delayed CNS disturbances.
Our RDX studies seem to bear out Sunderman's work66 which suggested that RDX
itself and not a breakdown product was responsible for the CNS effects. He demonstrated a rapid
onset of symptoms after intraperitoneal and iv injections, and we experienced similar results with iv
administration. Von Oettmgen^ had previously considered that an amine may be degraded and cause
CNS disturbances because it is known that certain aliphatic amines may cause a toxicologic response
similar to those tha: he observed with continued RDX feedings.
Von Oettingen's studies !iave also indicated that oral feedings of RDX to dogs (I Of/
acacia mixture) produced questionable, or no physiological effects when the dose ingested was 5 to
15 mg. However, those dogs were ^retreated with ainytal and undoubtedly the barbiturate
influences the response to any compound that may have a CNS effect. In addition, the dose buildup
in the stomach was not comparable to our iv dose. Sunderman66 has shown that CNS disturbances
39
-------
can lie abolished by dcccicbrjtion or administration of Nembutal. thereby indicating a CNS
involvement .it a higher love!.
RI)X according to Von Oct lumen3 and Sunderman,66 produced no
pathologic changes r the brain, although nonspecific lesions were observed in the renal tubules, the
liver, and the hcait muscle. Slanskaya and Po/harsky,5 however, observed pathologic changes in
various organs as well as the CNS alter acute and chronic feedings. In acute cases the CNS vascular
supply appeared to be impaired because of changes in the vessel walls. Nerve cell degeneration was
also observed. The areas most affected in descending order were: spinal cord, brain stem, and
coitex.
From the pharmacologic activity of HMX in these limited iv studies, one could assunn*
that this agent is more of a "nitritelike" compound, unlike RDX. Nitrites, however, arc known to
produce methemoglobinemia, which may induce respiratory embarrassment and. in conjunction
wall a collapsed circulation, may bring about convulsive seizures. However, we did not observe the
lypical chocolatc-coloied blood indicative ot metheinoglobm in a small sampling of dogs.
At this point several possibilities might explain HMX activity
1. HMX produces a nitritelike effect on the cardiovascular system: if
respiration and circulation are severly impaired and methenioglobin is formed, CNS disturbances
result:
2. Because of some inherent chemical property, it takes longer for HMX to
exert CNS effects than it docs for RDX.
J. HMX produces a circulatory effect initially, followed by recovery and then
CNS effects because of some specific action of the agent which may be dependent upon some
tune-consuming metabolic pathway.
One has to bear in mind that CNS disturbances were still apparent in some of our
HMX-poisoncd dogs even alter their blood pressure and respiration recovered to what one would
lunik were adequate levels.
In order to be more definitive in the evaluation of HMX, pharmacologic activity studies
snould be conducted in enough animals to assess the cardiovascular, respiratory, and hematologic
pictures during various stages of response
4. Conclusions
There was no evidence that either explosive agent (RDX or HMX) had any physiologic
cflcct upon the dog when acute and subacute exposures were made topically. The intravenous
administration of KDX. depending 011 the dose administered, produced CNS hyperactivity, tremors,
convulsions, and death. HMX, on the other hand, initially produced cardiovascular collapse, which
111 some cases led to immediate death or. depending on the dose, to recovery and eventually to CNS
disturbances.
40
-------
D. Sensitization Potential of RDX and UMX.*
I. Procedures.
Sensitization studies of RDX and HMX in acetone, cyclohexanone, and DM SO were
conducted using the experimental methods of Landstciner67"71 and others who followed his classic-
procedures. Sensitization effects were evaluated using the Draize test as recommended by the US
Department of Agriculture. Food and Drug Administration (table VII, page 27 ). Young adult guinea
PUj ,'f both sexes were exposed to the test compounds by topical application to the skin and by
intradermal injection.
For each topical application, O.S ml of test compound was applied. Acetone and
cyclohexanone solutions were applied as free-falling drops dispensed (through a 20-gage needle)
from a 2.5-ml hypodermic syringe. DMSO, pure and technical grade, was dispensed in the same way
but it was necessary to spread it out with a sterile, cotton-tipped applicator because (he material
was quite viscous and tended to bead on the skin surface. All animals were clipped "-everal hours
before exposure to encourage maximal skin penetration. The skin was not covered alter exposure .11
order to simulate the conditions of a "spill exposure."
For intradermal injection, 0.05-ml volumes of test solution were given with a 0.5-ml
hypodermic syringe attached to a 26-gage needle, hach exposure site was dipped several hours
before administration and sterilized with ethyl alcohol immediately before injection.
These sensitization studies were divided into three areas, the senrilizing phase, the rest
phase; jnd the challenge phase
During the sensitizing phase the guinea pigs were exposed to the test compounds 3
days per week for 3 weeks. Either topical or intradermal doses were applied to the clipped dorsal
thorax and observations for skin effects at the site of the exposure as well as for systemic toxicity
were made daily.
During the rest phase (2 weeks) no compounds were administered and the animals were
observed daily. Concurrent with this phase, experiments were conducted to determine the
maximum suberythemjl dose to use during the challenge phase. Serial dilutions of the sensitizing
stock solution were made in saline or polyethylene glycol (PEG 200) and injected into a group of
naive guinea pigs until the dose was found that caused no skin effects. In this study of RDX and
HMX, considerable time and ettorl were devoted to obtaining this subetfective dose because of the
activity of the solvent used.
In the challenge phase, the clipped thighs of the animals were exposed intradermally
and topically to single doses of test material at the predetermined maximum subeffective level.
Although one route per animal was used in the sensitization phase, either intradermal or topical,
both routes were used to challenge, one on each thigh at different times. The animals were observed
daily for 3 days after dosing for signs of skin effects at the application site. The challenge doses of
RDX and HMX were prepared in I I (v/v) solvent-saline mixtures (intradermal) and PEG (topical)
Using the nonirntating PEG eliminated the need for excessive dilution allowing substantial amounts
* John P CaUahin. Toxicology Division, performed the »rnsiti/Jtion investigation
41
-------
uf the compounds to he ;ipplicd. The erythematous effects of undiluted cyclohexanone and DMSO
were also eliminated ;ind the contact persistency of the challenge materia' was increased. Some
animals wcti' also exposed to diluted (intradermal) and undiluted (percutaneous) doses of the three
solvents alone and then challenged.
2 Results.
Initial experiments lor sensitization with topieal HMX in TMSO were conducted using
0.5 nil of 33% HMX/DMSO solutions. Deaths occurred after the first and second sensitization
exposure (5/12 animals). Two animals showed signs of hyperirritability and intermittent
convulsions within 24 to 48 hours after one or two exposures. One of these animals died within 24
iw 48 hours after the first treatment: the other survived and received all of the scheduled exposures.
I lie other deaths occurred overnight and toxic signs were not observed. These deaths were
apparently due to HMX poisoning since other animals receiving DivlSO alone showed no toxic signs
jiid Old not t ie.
When die topical sensitizing doses were reduced to the following, none of the animals
died during the sensiti/ing period:
2% HMX in acetone
2.5% HMX in cyclohexanone
3.3% HMX in pure and technical grade DMSO
100'/) acetone, cyclohexanone, and DMSO
5.4% RDX in acetone
7.5% RDX in cyclohexanone
33% RDX in pure and technical grade DMSO
Because the solvents used are known skin irritants, the animals were carefully observed
throughout the sensitization period for skin effects. Th<* differences in skin effects between RDX-
aiul HMX-solvent mixtures and the effects from the solvents alone were evaluated. The results are
shown in table XII and XIII.
It was readily seen that acetone, with or without RDX or HMX. cause no skin effects. This
(notably is related to us high volatility. Significant skin elI'ects occurred in animals receiving repeated
topical exposure to cyclohexanone or DMSO, with and without RDX or HMX. Most animals had
Mgmfieant skin effects trom DMSO by the end of the first week. Similar effects were not seen in most
animals until the end of the second week of application of cyclohexanone. The intensity of effects of
both solvents were generally increased by the end of the third week. Although not shown in the tables,
these effects, with or without RDX and HMX. diminish when treatment is slopped. Visual inspection
of the exposure sites showed absence of effects 18 to 24 days after the final treatment.
No significant differences between the skin effects caused by the two solvents alone
and those caused by RDX- and HMX-mixtures were evident. It is apparent, therefore, that DMSO
42
-------
Table XII. Residual Skin Lffects Caused by the Repealed Topical Application
ol 0 5 ml KDX and/or Several Solvents on ihc Backs of Clipped
Guinea Pigs (Sensitization Period)
Krjction ol group showing residujl topical skin effects"
Treatment
Cumulative number of treatments^
1
2
3
4
5
6
7
8
9
27 Mg of KDX in
acetone
(S 4% RDX)
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
37.5 Mg of RDX in
cyclohexanone
(7 5% RDX)
0/6
0/6
0/6
1/6 s
1/6 s
1/6 S
1/6 se
2/t, S
1/6 Sc
2/6 s
2/6 s
4/6 Sc
1/6 s
4/6 S
1/6 Sc
5/6 S
1/6 Sc
165 Mgof RDX in
pure DMSO
(33.0% RDX)
0/6
0/6
1/6 s
4/6 S
3/6 S
2/6 S
2/6 S
4/6 S
1/6 s
4/6 S
1/6 5c
4/6 S
1/6 Se
1/6 Sc
1/6 s
3/6 S
1/6 S
1/6 Se
1/6 i
2/6 S
3/6 S
165 Mgof RDX in
tech grade DMSO
(33 0% RDX)
3/6 S
3/6 S
2/6 S
3/6 S
3/6 S
1/6 Se
2/6 Sc
1/6 S
1/6 Se
1/6 S
2/6 Sc
1/6 Se
1/6 S
4/6 Sc
1/6 Se
1/6 S
4/6 Sc
2/6 S
4/6 Sc
2/6 S
4/6 S
Acetone
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
Cyclohexanone
0/6
0/6
0/6
1/6 e
1/6 e
4/6 s
1/6 se
3/6 S
1/6 Se
1/6 Sc
4/6 S
1/6 S
2/6 S
4/6 Sc
Pure grade
DMSO
3/6 S
6/6 S
1 2/6 S
3/6 S
2/6 S
2/6 Se
1/6 St
3/6 S
2/6 Se
1/6 s
2/6 S
2/6 S*
1/6 Sc
4/6 S
1/6 Se
1/6 Sc
1/6 s
1/6 Se
1/6 Sc
1/6 s
4/6 S
1/6 S
Tech grade
DMSO
-
1/6 S
4/6 S
6/6 S
3/6 S
3/6 Se
3/6 S
2/6 Se
1/6 S
1/6 S
2/6 Sc
1/6 Se
1/6 Sc
1/6 S
2/6 S
1/6 Se
1/6 Se
1/6 Sc
1/6 S
4/6 S
1/6 Sc
2/6 s
4/6 S
U Based on observations made 48 to 72 hi alter each treatment
^ IVrcutaneous treatments were given 3 timet per week over a 19-djy period.
Cyclohexanone and both types of DMSO. with jnd without KDX. had immediate ettects after a single applicaUon
Mild erythema wjt observed in all animals exposed to cyclohexanone and lasted from I lo 6 hrs Both typei of DMSO
induced a moderate erythema which subsided to a mild erythema by 6 hr and was usually absent at 24 hr.
LJXiTNJD s: Slight scaling ol the skin
S. Moderate scaling ol the skin.
S Heavy scaling of skin.
Se Moderate scaling ot skin v.niti mild ciythema.
SE Moderate scaling ot skin with modruie erythema.
Sc Moderate scaling ot skin with ouckinf.
Sc Heavy scaling ol skin with cracking
43
-------
Table XIII. Residual Skin Effects Caused by the Topical Application
of 0.5 mJ HMX and/or Several Solvents on the Backs of Clipped
Guinea Pigs (Sensitization Period)
Fraction of group showing residual topical skin effects^
Treatment
Cumulative number of treatments^
1
2
3
4
5
6
7
8
9
10 Mg IIMX in
acetone
(2% HMX)
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
12.5 Mg IIMX in
cyclohexanone
(2.5% HMX)
0/6
0/6
0/6
0/6
0/6
4/6 S
6/6 S
5/6 S
1/6 Sc
2/6 S
4/6 Sc
10.5 My HMX in
pure grade DMSO
(3.3% HMX)
0/6
3/6 S
2/6 S
5/6 S
1/6 Se
6/6 S
5/6 S
3/6 S
2/6 Sc
2/6 S
2/6 SC
1/6 Sc
2/6 s
2/6 S
165 Mg HMX in
pure grade DMSO
(33% HMX)
0I5CJ
0/4
0/4
1/4 S
3/4 S
4/4 S
4/4 S
1/4 S
3/4 Sc
3/4 S
16.5 Mg HMX in
tech grade DMSO
(3.3% HMX)
0/6
2/6 S
1/6 S
6/6 S
6/6 S
6/6 S
5/6 S
1/6 Sc
5/6 S
1/6 Sc
3/6 s
105 Mg HMX in
tech grade DMSO
(33% HMX)
0/3^
0/3
0/3
2/3 S
2/3 S
2/3 S
3/3 S
3/3 S
3/3 s
Acetone
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
0/6
Cyclol.. tanone
0/6
0/6
0/6
C/6
0/6
5/6 S
6/6 S
6/6 S
2/6 S
4/6 Sc
Pure grade
DMSO
0/6
0/6
1/6 S
2/6 S
3/6 S
6/6 S
5/6 S
5/6 S
1/6 Sc
2/6 s
3/6 S
Tech grade
DMSO
0/6
0/6
1/6 S
3/6 S
5/6 S
5/6 S
6/6 S
5/6 S
1/6 Sc
3/6 s
2/6 S
iJ Bused on observations mjdc 48 lo 72 hi after the second treatment. Cyclohexanone and both types of DMSO. with and
without IIMX, had immediate effects after a single appkcation. Mild erythema was observed in all animali exposed to
cyclohexanone und lasted from I to 6 hr. Both types of DMSO induced a moderate erythema which subsided to a mild
erythema by 6 hr and was usually absent at 24 hi.
^ Percutaneous treatments were given 3 times per week over a 19-day period.
^ One jmmul died within 24 lo 48 hr after the first treatment and a second animal died within 24 to 48 hr after the
second treatment.
^ Three aninuls died within 24 to 48 hr after the fust treatment.
J,KC£NI): s: Slight scaling of the ilun.
S. Moderate scaling of the skin.
S: Heavy scaling of the skin.
Sc. Moderate waling of the sktn.with mil] erythema.
SR: M-vlerate scaling of the skin with m< derate erythema.
Sc: Moderate scaling of the sktn with cracking.
Sc . Heavy scaling of the skin with cracking.
44
-------
and cyclohcxanone produce .skin irritation when administered topically and thai R1)X and HMX
nuke little or no contribution.
Similar evaluations were made of the effects caused by RDX- and HMX-solvent
nuxtuics and the solvents alone when the*: materials were administered intradermal^ (table XIV).
As would be expected, the severity ol°skin effects was much greater alter intradermal injection than
after topical application. All solvents caused moderate to severe erythema, edema, and necrosis
during the course of repeated intradermal exposures. Just as in the topical exposures, RDX and
HMX apparently contributed little to these effects. The results shown in tabic XIV indicate that
cyclohcxanonc caused the most severe effects, acetone the intermediate, and DMSO the least severe
eltects. All three solvents are considered unsafe for intradermal injection at the concentrations used
in this study.
Tables XV and XVI show the work done to determine suberythcmal doses for topical
and intradermal challenges with RDX-solvent mixtures. Tables XVII and XVIII show similar
determinations for HMX-solvent mixtures by both routes.
The suberythcmal doses lor RDX in acetone an<1 in DMSO for intradermal challenge
were I 32 jnd 1:16, respectively, saline dilution-, of the stock solution used in ihe sensitization
pnase. For the RDX-cyclohexanone mixture, it wjs j 1:64 dilution. The intradermal suberythcmal
dose of HMX in acetone and DMSO was 116 and 1'32. respectively, saline dilution and for
-------
I..lilt \JV 2 IK ^klll I lUi i i . wojtlliwi i i ^ WtNlUK* »il Av««»l.lC.
i ) i Iimiv %jii»»cu\ jiiJ l'ui« 4nii i wviiiuvj» ly.»uO, Wi»i» j«J Wiilioni
KUA «iij IIMXi u J U»isil lhur«ix
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rii'jiiiiciM
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14
10
k9>H fl))i til i z
I uu
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U.I 25 Kir HOX in
.Kt'lOllw*
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4i t 2> mj lito* n» (
4.cionc
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23 4 ni|t '>i
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lycluhcxjnwtic
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'i 125 Mi, HMA nt
1 OtluhCAuilOllC
(U.25XIIM\«
2V ink* <>i
, |M»K UM.|J
0 125 inn KMX :n
pure UMSO
<0.25'4> IIMXl
27 4
uch (UJiic
UMSO
0.125 mg KUX «.«
lech ^rudc UMSu
(0.25', RDX,
0 I 25 mp iivix id
u\h judv UMSu
10 25- HMa)
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l-«S
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1-eS
l-od
5-cS
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2 -..S
I- S
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1 cS
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b Sc^r 'i^jc Uiiinjnuii
4u
-------
I'jble XV I'lic Determination ol j Sulxrythciual Dom: of Intradermully Administered
KOX in A ('yiloliexanune, jnd 'lecbniMl Crude DMSO
in Clipped Guinea Pigs
Skin etfectsa
Dilution
RDX Acetone-'
RDX.Cyclohcxanone^
RDX.DMSO (Tech Grade)JV
of
No of
Observation
line
No. of
Observation time
No. of
Observation time
stock
Animals
24 hr
48 In
72 hr
animals
24 hr
48 hr
72 hr
animals
24 hr
48 hr
72 hr
Stock
2
2E4
2-k4
J-L4
'2
2-E4 .
2-E4
2-E4
4
4-E4
4-E4
4-E4
I-N2
I-N2
I-N2
2-N2
2-N2
2-N2
1-N2
I-N2
1:2
4
4-E4
4-E4
4-t4
2
2-E4
2-N2
2-E4
2-N2
2-E4
2-N2
4
2-E4
l-EI
2-EI
0
1.4
4
4-E1
4-El
l-EI
2
2-E4
I-N2
2-E4
I-N2
2-E4
2-N2
4
2-EI
2-EI
1-E4
l.X
4
l-EI
0
l-EI
2
2-E4
2-E4
2-E4
4
2-El
2-EI
i r.i
116
4
2-EI
0
0
4
4-E4
2-E4
l-EI
2-E4
2
0
0
0
I.J2
4
0
0
U
4
4-EI
2-EI
2-EI
2
0
0
0
1:64
4
0
0
0
2
0
0
0
2
0
0
0
•^"lhe Draue Test was used to evaluate skin ettects (table VII, p. 27 ).
-^'n 05 nl of 0.25% RDX in 1:1 solvent, sjline solution.
Table XVI. The Determination of a Suberythcmal Dose ol RDX in Acetone. Cyclohexanone.
Pure and Technical Grade DMSO When Administered Topically in PEG 200 in Clipped Guincj Pigs
Compound
Solvent
Dilution in
No. of animals
Skin effect ^
PEG 20U
exposed'1'
4-5 hr
24 hr
' 48 hr
72 hr
'PtG 200(100%)
-
-
16
0
0
0
0
1
RDX (5.4%)
Acetone
1.10
3-
0
0
0
0
RDX (5.4%))
Acetone
1:100
3
0
0
0
0
RDX (7.5%)
Cyclohexjnone
1 10
3
0
0
0
0
RDX (7.5%)
Cyclohexanonc
1 100
3
0
0
0
0
RDX (33 0%)
Pure DMSO
1 10
3
0
0
0
0
RDX (33.0%)
Pure DMSO
1.100
3
0
0
0
0
RDX (33.0%)
Tech grade DMSO
1 10
3
0
«
0
0
RDX (33.0%)
Tech grade DMSO
rioo
3
0
0
0
0
¦if 0.5 ml
^ The Draize TEst was used to evaluate skin effects (table VII, p. 27).
47
-------
Tibic XVIi Tl* U it'iniiuliua ot j ^ubcit ilKmjl Pu« uf IniudiinuU) Adminnirrrd liMX id Acttonc.
<~)tluhr\4nnnL' and Pliic and 1*.* hr.n DMSO in Clipped (•uinr* Pn*»
"
Skid f((ii u"
Dilution
0 25-; HMX
accionc^
0 23'.' HMX <.)<-lohcwnunA'
0 25.
HMX oun' DMSOW
0 25? HMX UMSOttCfch padc) —
of
Stock
No of
Otnrc
»4tion Um
c
No ot
Otxi\4iton ttmi
No uf
Obwiwttun time
No ol
Oforrwtion timr
ammjK
2- 3 hi
24 hi
48 hi
72 hi
cvahijie skin cffcui ((able VU.p27)
y 0 05 ml
-------
Table XVIII.Tin- Determination o! a Suhcrytltcmal Dose of HMX in Acetone, Cyclohexanone, and
Pure jnd Technical Crude DMSO When Administered Topically in Polyethelcne Glycol 200
in Clipped Guinea Pigs.
1
Compound
Solvent
Dilution
in
IM G 200
No. ul
animals
exposed &
Skin effects^
3 hr
24 hr
48 hr
72 hr
No. animals/elTccts
l'l.C. 200 (100%)
-
-
16
0
0
0
0
HMX (2.0%)
Acetone
1 10
3
0
0
0
0
HMX (2.0%)
Acetone
1 100
3
0
0
0
0
HMX (2.5%)
Cyclohcxanonc
1 10
3
0
0
0
0
HMX (2.5%)
Cyclohcxanonc
1 100
3
0
0
0
0
| hmx o.y/t >
Pure DMSO
1 10
3
0
0
0
0
HMX (3.3%)
i
Pure DMSO
1 100
3
u
0
0
0
HMX <3.?.3',0
Pure DMSO
1 1.0
3
0
0
0
0
HMX (33.3%)
Pure DMSO
I 100
3
u
0
0
0
i
HMX (3.3%)
Tech grade DMSO
1 10
-t
J
0
o
0
0
HMX (3.3'/1
Tech grade DMSO
1 100
3
0
0
0
0
HMX (33.3'?)
Tech grade DMSO
1:10
3
0
0
0
0
HMX (33.3%)
Tech grade DMSO
1 100
3
u
0
0
0.5 ml
The Drill/c Teat wj* used to evulujlc bkin 1»«_•».i.-> i ublc VII. |> 27 )
-------
i Discussion.
Sensitization usually involves I he entire skin surtaic. It m:iy occur .inywliere 011 the
Uitiy luii is must e.i ily induced in an urea that l\u.> been inflanr.-d because of primary irrituiion.
.uleclion. hums, and oilier causes. ll has been postulaicd ilui (lie conccntiauon ol lymphocytes hi
s>l^ li an area greatly aids or is even necessary loi (lie pioductiou of sciisitizalKm. It is not the simple
dicniical haptcnc ilsell that act:, as the antigen, hut its combination with the proteins ol° the skin
ioiiii the liapiene-protein antigen conjugate. I Ins conjugate then acts as tile stimulus that produces
.li. immune 1 espouse in the host. I lie antibodies lormed are not only directed against tl'.c haptcnc
ol the conjugate luit also against the protein carrier. Wln.11 a sensitized animal is challenged
1.. die haptcnc ilsell leitlier by topical application or intiadei..ial mjeciion) an inflammatory
espouse is manifested within 24 to 48 houiv
On this basis, it is reasonable lo assume that a simple chemical which is a potential
..neigen will induce sensitization mote icadily it a is a primary irritant. I here are. however, cases
t.iuie nonirriiating substances have produced sensitization, anil vice versa, where highly potent
,miliary iriilauts have tailed lo pioduce sensitization Ko'.libci.:'2 showed that primary nritants
.a.1 e sensitization in his studies with guinea pi;'s which were designed lo determine the sensitization
i'.-Kii(iat dI a-broiiio-u-tolunitrilc i( A). I0-ciiloio-5.)0-dihydrophcii.nsa/iiic 1 DM),a-chloro-aceto-
.iiieuoiie tC'Nl and u-chloiobcnzylidniciualoiioiiilrile (CSi. In these studied Doth CN and CS were
.Miiiij lo be printaiy irritjiits. causing extensive erythema, edema, neciosis. and sometimes eschar
urination wli-.il administered mtradermally or percutaneously during the sensitization period. They
tvnc also louiid 10 be sensitizers.
In our studies, both cydolic-xaiionc jnd DM SO. with and without KDX and MMX.
vwie I on i)0 to be irritants when applied topically, 'topically applied DM SO was also observed to
i.iiise a "smoking" action, an cflccl obviously due to a chemical reaction with the skin When
.iuiiiinotcicd intraderuully. with or without the explosive components, cyclohexanonc. DMSO. and
.icetone caused skin damage at the concentrations used. Yel none of these solvents or solvent
UDVIIMX combinations demonstrated a sciiMiizanon potential when the animals were later
challenged by either route. These findings confirm that a compound can be a skin irritant without
living .1 sensitizer.
Since all solvents invlovcd in these studies except topically administered acetone
...1. *.d skin 11 illation j 1 die concentration* used, it is dillicult to determine the contribution of
i\ij\ .ind ItMX to iiritaucy. By the intradermal route any contribution uy KDX or HMX was
.1...emeu by (hesolvent eiiects. The order ot solvent iirituiicy uy tins route is. (I) cyclohexanonc,
t , ..-ctoue. and (J) DMSO. This order does not appear to be related to weight of solvent
.¦a iiMeied per injection oecause the least cttectivc, DMSO. was administered in the greatest
i.i..iiiuy
The contribution made to skm irritancy by topically administered KDX and HMX in
c.icn solvent is also difficult to determine because of tnc initial erythematous and residual
desquamation effects of cyclohexanonc and DNiSO. Since acviouo was not irritating and the
¦ csidual amounts of kDX and HMX lett on the skin alter each treatment caused no topical
iiiiiaiion. it would appear that RDX and HMX themselves do not cause irritation. riiisciiccl could
be due to poor skin absorption of the neat compounds. Patty1 i.,'orts thai mere is no evidence of
skm absoipuon of KDX. Intormation on the skin absorption of HMX is not available.
50
-------
In the topical sensitization exposures reported here, RDX and HMX were used with the
solvents at the highest concentrations obtainable. RDX and HMX were also tested by intradermal
injection which is even more direct and reliable than topical testing.
Our data show that neither RDX nor HMX cause sensitization by either exposure route
when tested by the method of Landsteiner and Chase.69,70,71 Also, none of the solvents used in
these studies produce sensitization.
In a report of primary irritancy and sensitization dermatitis, particularly of the face
and eyelids during the nitration process of RDX. it was shown that an unidentified component in
the fumes from the reaction mixture was responsible.7 McConnell8 attributed some dermatitis to
the manufacture of RDX, but this was probably related to intermediates because significant
dermatitis was not observed in individuals handling the purified material. This observation was
corroborated by Von Oettingen's3 findings that patch testing with the moistened solid did not
produce irritation.
Sunderman66 showed that powders of both RDX and HMX caused primary irritancy
in humans when patch tested as described by Swartz and Tulipans.73 With this method he also
demonstrated a negative human sensitization potential for RDX but a positive sensitization
potential for HMX. RDX is manufactured by the nitration of hexamethylcnetctramine which is
obtained by the reaction of formaldehyde and ammonia.74 Formaldehyde is liberated from this
reaction and is oxidized by the nitric acid if the mixture is allowed to stand. If the formaldehyde
remains in the spent acid after drowning, there is difficulty in recovering the nitric acid from the
spent solution. As a result of the rupture and degradation of the hexamethylenetetramine molecule,
numerous aliphatic and cyclic nitro compounds are present in crude RDX. The most important is
cyclotetramethylenetetranitramine, or HMX. Since HMX is a byproduct of RDX and has the same
basic molecule and group types, it is difficult to understand why RDX would not, and HMX would,
cause sensitization. As a general rule, if a parent compound is a true sensitizing agent, most of the
intermediates involved in its synthesis are also sensitizers. A good example of this is the
intermediates involved in dye production.7 5
It was not within the scope of this study to determine the intradermal or cutaneous
toxicity of RDX or HMX in guinea pigs. However, since several animals (5/12) died from the topical
application of one or two 0.5-ml applications of 33% HMX in pure or technical grade DMSO, a
comment is in order Even though five animals died from HMX-DMSO exposures, seven others were
able to withstand, without symptoms, nine such treatments administered at 48 to 72-hr intervals
over 19 days. Also, in the studies shown in table XM1 a single dose of 2 gm/kg and three 1 gm/kg
doses did not kill any guinea pigs.
The lowest single dose to cause death was 0.465 gm/kg. If the human were comparable
to the guinea pig in toxicologic response to HMX-DMSO administered topically, the lethal dose
would be approximately 33 grams HMX (70 kg man) in 100 ml of DMSO. This dose is based on the
assumption that HMX-DMSO was uniformly spread over a large surface area of skin and that no
effort to remove it was made. This is a considerable amount of HMX and when compared to other
percutaneously potent compounds, it must be considered relatively nontoxic.
Thirty-three percent RDX-DMSO showed no toxic effects when applied in the same
amounts and over the same period of time.
51
-------
4. Summary of Topical. Intradermal and Sensitization Studies of
Solutions of RDX and IIMX. and ol Solvents.
Under the experimental conditions Ascribed in this study, the following can be said.
1. No deaths occured in guinea pigs given repeated topical doses of 5.4%
KDX-acelone. 7.5% KDX-cycloliexanone, or 33% RDX-DMSO.
2. No Jcaths occurred in guinea pigs given repeated topical doses of 2.0%
IIMX-awctoiie. 2.5% HMX-cydohcxanone, or 3.3% HMX-DMSO. Several deaths occurred after one
cr two exposures to 33% HMX in DMSO. The deaths were attributed to HMX because no deaths
woe pioduccd by neat DMSO.
3. Topically applied DMSO caused greater skin damage than cyclohexanone.
Solutions of KDX and HMX in these solvents did not produce noticeably greater skin damage than
Uk-solvents themselves. No visible damage was caused by topical applications of solutions of RDX
jiui HMX in acetone.
4. C'yelohi-Nanonc. acetone, ami DMSO; administered mtradcrmally, with or
without KDX and HMX, caused severe skin damage. This prevented a reliable assessment of the
ctieL'ts of KDX and HMX by this route.
5. Acetone, cyclohexanone, and DMSO did not produce skin sensitization in guinea
pigs when administered intradermal^ or topically.
6. Solutions of RDX or HMX did not produce skin sensitization in guinea pigs
when administered intradermally or topically.
£. Cataracts Found in Guinea Pigs Following Cutaneous and Intradermal Applications
of Solvents and Solutions of KDX and HMX.*
1. Procedures.
The eyes of 210 guincj pigs were examined after they had received cutaneous or
intradermal applications of DMSO (pure and technical grade), acetone, or cyclohexanone In sortie
of the applications, the solvents contained KDX or HMX. F&r intradermal injections, uie solvent or
a >uluiion of explosive in solvent was mixed 1:1 (v/v) with saline (table XIX).
The materials were administered three times a week for 3 weeks to 116 male and 94
lemale 9- to 18-week old animals. Examinations were made 26 to 110 days after application.
2. Results.
Microscopic examination, using an ophthalmoscope, revealed cataracts in 20% of the
.uiimals. The appearance of one typical cataract as revealed by a slit-lamp photograph and a
photomicrograph is shown in figures I and 2.
• This investigation wai conducted by MAJ Roy H. Rcngstortf, Medical Rejeurch Division, John K Caliuhan,
I'uMcology Division, and PFC William Webb, Medical Research Division
52
-------
Table XIX. Cataracts Found in Guinea Pigs
Route
Cutaneous
Solutions used
0 5 ml solvent mixed with
HMX(
-------
i
Figure I. Slitlamp Photograph of Guinea Pig Eye 40 Days after Receiving Intradermal
Application of 0.05 Ml Pure DMSO in Saline Three Times a Week for Three Weeks.
Vacuoles (V) can be seen in the crystalline lens periphery, partly covered
by the iris (I). Magnified 12X.
Of the 98 animals that received HMX and RDX in solvents, 16% developed cataracts.
The remaining 112 animals received only solvent, and 23% had cataracts. The common factor for
both groups was the solvents, and the explosives did not appear to increase the probability of
cataracts.
3. Discussion.
A variety of substances, fed or injected, can cause cataracts in experimental animals.
These include naphthalene, iodoacetate, Myleran (1-4-butanediol dimethanesulfonate), Mimosine
(3-hydroxy-4-oxo-l(4H)-pyridinealanine), dinitrophenol, certain quinoid substances, ergot,
thallium, adrenaline, morphine-like drugs, and DMSO. In a cursory search of the literature, no
reports of cataracts caused by acetone and cyclohexanone have been found.
In a separate study in this laboratory, small multiple doses of acetone, cyclohexanone,
and dimethyl sulfoxide administered either topically or subcutaneously on the backs of guinea pigs
over a period of 3 to 8 weeks caused cataracts in 29 out of 120 animals. Lens changes began as early
as 8 weeks and as late as 6 months. They consisted of subcapsular focal or extensive vacuolated
areas extending from the periphery towards the center of the lens. The histological appearance of
the lenses was similar to that of senile cataracts and some forms of diabetic cataracts.65
54
-------
ANTERIOR SURFACE
CA
./ \
\ /
V
POSTERIOR SURFACE
Figure 2. Photomicrograph of the Crystalline Lens Shown in figure 1.
The abnormality appears anteriorly and posteriorly as exaggerated tissue vacuoles (V)
involving the entire subcapsular region. CA - capsule. Magnified 80X.
55
-------
I\ . Kl MJ.Vll-
liHiawnotis I'lleets ol KDX. IIMX. .md I liree Solvent'.
I. Intiavenous Toxicity ol KDX .nul UNIX in Muc jiuI (»uinca hgs
I lie intravenous Ll)50 lot mice ol KDX in DMSO in 18.7 ( 15.7-22 3) my/ky and of
11<.l.\ hi DMSO is 2«.1^ (25.1-33.3) ing/kg. Death* occurred in 5 to 10 niinuics and were preceded
i loiiswl-mni* .nut l.ibnred breathing. I lie survivors were lethargic lor several hours thereafter.
in.i iwu1 m.iinal within 24 liouis.
Ilu intravenous LDSO's m guinea pigs !oi KDX and HMA in DMSO .ire 2S.I
• i, ti ¦ i i>i n>w-iiei|(;u)i.y patterns in Hie IT 'i Tins is
. i'a;i: ol ventral nervou* uepiession. oi ol sle».p During tins piuse. tlie amni.ils were seinieomatose
|/.nii i espouses weie dcpicssed I hese w»Uent ittccts obscured the action;, ol dissolved KDX or
ii.ti.s I lie wuly elleet ol (I I ml/kg DMSO was an appreciable but transient decrease m blond
•Ji .Still..
4.u :nii adunnibii jiiou ol 4uing/kgoi KDX iii DM SO was tollowed in less i!ian
I.) sekoikis by an epileptic-type, spike-and-dome ITAI pattern and concomitant convulsive
ii.iino. iliaiigei in blood pressure and he.ui rate were negligible jt this time. Two dogs died,
.lently in cardiovascular collapse, at 45 and 90 minutes alter injection ol the matciial I wenty
u; ot lui not death « I ne convulsive-type I I (i pattern
.••iu ..m>ile activity could be ptccipitated by pnolk or tactile stimuli.)
IIic intravenous aUininisiralion ol 40 nig/kg ot ilM.\ in DMSO caused deatli in
w..iiiikiv.iseiilui° collapse hi I .ind 3 minutes in two Jo.:-. isvo othcis died in about 14 lir. When two
Jo mg/kg uoscs UvK' given >0 nnnutcs .ip.nt to tu.i ....Jilional dogs, there wjs jn iniinediate lall in
i><.i.m [Hosing .nui one dog died in I uini. L^ikpiic-i/.K' lT;(i and muscle movements developed in
uog alter seveial hours. It died 14 hums .titer an apparent recovery lioin (lie central and
>. .ilOIOka.Mtil.il elicits
. prominent diiieronce between KDX and II.ViX m the dog was the time oi appearance
ious system ellecls. I he epilepuioiin aclivit) occuned alinoit immediately with
KDX nut Jiei a delay with KMX.
li I ocal I Meets ol I opical Applications
I he three solvents and solutions ol Kl)\ and HMX m me solvents ov..asionally cause
eiyihema and signs ol' imtaiion or pain when applied 10 the skin ol' iahbiis. guinea pigs, or dogs.
56
-------
DMSO in the least volatile .1...: |>ossil>iy the iiirsl jitivcon tlu-skin Acetone wiiiili is
the most volatile was the least jwtsvc. DVtM) sometimes caused Ink1 wisps ol smoke to rise Ironi the
skin surface. Also, the animals vocah/.cu. squirm, d. licked the site ol application, and exhibited
reactions indicative of pain. I liese skin re.u turns .ne 111 accord with those previously described lor
DMSO in animals and man
Hollowing repeated application. DMSO jnd cydohexjnouc caused llie skin of rabbits,
dogs. and guinea pigs to heroine diy and turd A sligh. Daking or desquamation occurred in the
outei layers. There wore no lissures, eschars, or scab formations.
C. Systemic Hffecls l-ollowing I 0pK.1l Skm Application.
1. Blood Values 111 Kahbiis
Single or repeated topic.il skm .tppiicjiioii to rjbl.it-, 01 I 0-n>l doses ol DMSO.
cyclohcxanonc. or acetone, or solutions ui !11.111 Iheie were no .ihnonnal gioss or M:(>
response., to visual, auditory, tactile, milaiii. or panitul stnnuli. Doses ui KDX and IIMX were as
great as 480 mg/kg repeated on .t consecutive days
Since intravenous doses ol 20 10 4U mg/kg 01 KDX 01 IIMX producejl marked
loxicological responses in dogs, it is indicated that the ^.innie skm i\ |>oorl\ penetrated by these
explosives.
4. Lenticular Opacities.
As mentioned in our review ->! the lueratuie. lenticul.u opacities have been produced
by DMSO administered by various mines of adiuiuisti.iiioii. including percutaneous: 111 various
annual specie-..114 ¦' *7 but not in mai».J 5 ¦J,< 4" 4 '"4* I his oc.iut ellccl was noted lollowmg
topical and nuradcimal application ol O1Y1SO. .icvioiie and cycloticxanoue 10 guinea pigs in llie
present study. To the knowledge ol the present investigators, lenticular elle.ts have not been
attributed previously to these solvents llie nuiuDcr ol .mimals wu-> loo small to rank the solvents
lor tins cflect. however, it was indicated 1I1.1I DMSO was uie miosi uam.iiMiig and cyclohexauone was
the least injurious. Also, it wj> iihIk iKd thai the presence ol KDX or IIMX was 110 more damaging
than the solvents alone. Subsequent studies m these laboratories 10 muiice lenticular opacities in
rabbits with acetone have not been sticicssiui. *
* Rengstorff, R. H., Pel rail, J. P , jnd Sim, V M. Unpublished Dau
57
-------
Although lenticular actions due to these solvents have not been seen in m;m. it would
he injudicious to mimmi/.e human exposure.
5. Toxic Si^ns and Death hi Kjbhils. (iuinea I'igs. and Dogs.
No toxic signs or deaths resulted 1/1 rabbits, guincj pigs, or dogs Iroin the single or
ie|>eated topical skin application of
DMSO. acetone, or cyclohexanoiK'
KDX in l)MS« >
IIMX in acetone or cycloliexanone
I'aralysis in rabbits, convulsions in guinea pigs, and some deaths in both species
iKiiiiivil following lopKul .ipplication. One rabbi' died alter the fifth 0-1 -ml dose and one al ter the
tenth _|>|iiicjiion ol die I .(J-uil dose ot 5.4'/, KI)X in acetone. Another rabbit died alter the eighth
I 0-nil application ol 7.5% KDX in cycloliexanone Six rabbits died after repeated 1.0-ml doses of
.13.; IIMX in DMSO: three alter the >ccond dose, one after the filth dose, one alter the sixth dose,
ami one alter the twentieth dose. I;ivc ol 12 guinea pigs died 24 to 48 hr alter topical exposure to
33% HMX hi DMSO. lour alter single doses ol 4(>5. 477. 507, jnd 546 mg/kg. and oAe alter two
vioses ol I I l(t mg/kg.
In another abineviated test to find the percutaneous LD50. no deaths in guinea pigs
occurred wuh KDX oi IIMX in DMSO at doses iroin 3W> to 2.000 mg/kg. Thus, although some
ilea Ills occurred i" »'imei pigs at single doses of 40 5 to 546 mg/kg. the I D50 may be al>ove 2000
mg/kg. Following three lu()0 mg/kg doses of 33'/ IIMX in DMSO, guinea pigs became apprehensive
and lost weight, their skin uecame spongy and absorbent alter each application and lost its normal
color.
Convulsions preceded death in guinea pigs. Lethality could possibly be attributed to
c.xcilaiion of the cciiual nervous system. It is not known whether or not deaths in rabbits were
preceded by signs of excitation ol the central nervous system. I'aralysis. attributed to broken backs,
preceded death in rabbits It is possible iliat the backs were broken during convulsive episodes
which occuivd during the night or during other periods when the animals were not being observed.
It is questionable that the ucaths were attributable to the agents
Ce.itral excitation has been noted in the electroencephalographs pattern in dogs. This
information indicates that IIMX in DMSO can penetrate the skin ot some mammals in sufficient
dosage to produce to ic signs and death. Very high doses are needed
D. Sen-.ili/a lion
Skin application of the three solvents or of KDX or HMX in the solvents, followed by
topical skin or intradermal challenge with the sjme material, yielded no evidence of sensitization.
V. CONCLUSIONS.
I. Acetone and cycloliexanone in sufficient dosages caused some drying and
Hardening of the skin (locally), and central and cardiovascular depression systemically DMSO
58
-------
caused tlie above mentioned cutaneous eneus All three solvents produced sonic lcnliciil.il cli.i:it£cs.
Nunc of tlu* solvents influenced blood values or histological appearance of varum* tissi.ev None of
the solvents caused sensitization.
2. DMSO was llic strongest and acetone the weakest in producing local skin cllccts
when applied topically.
3. DMSO was the strongest and cyclolicxauone the weakest in causing lenticular
opacities.
4. MMX in DMSO pcnctiated the skin ot rabbits in sufficient (|uaiilities to produce
convulsions, paralyses, and death only when applied in very large doses.
59
-------
LITERATURE CITED
1. Patty, F. A., Editor. Industrial Hygiene and Toxicology, Second Revised
Edition. Vol 11. Toxicology, pp 2097-2099. Intersciencc Publishers, (a division of John Wiley &
Sons), New York, New York (19633).
2. Kaplan, A. S., Berghout, C. F., and Peczenik, A. Human Intoxication from RDX.
Arch. Environ. Health 10, 877-883 (1965).
3. Von Oettingen, W. F., Donahue, D. D., Yagoda, H., Monaco, A. R., and
Harris, M. R. Toxicity and Potential Dangers of Cyclotrimethylenenitramine (RDX). J. Ind. Hyg.
Toxicol. 31, 21-31 (1949).
4. Sunderman, F. W., Clark, J. K., and Bills, E. S. Compilation of Informal Monthly
Reports on Hazards to Health of Individuals Working with RDX, May 1943-June 1944. National
Defense Research Committee of the Office of Scientific Research and Development (NDRC).
Contract No. OEM sr-962, 1944. UNCLASSIFIED document.
5. Slanskaya, R. M., and Pozharsky, F. I. Toxicity of Hexogen, Farmakol i
Toksikol, 7,43-47 (1944) (Russian) C. A. 39:3073, (1945)
6. Barsotti, M., and Crotti, G. Epileptic Attacks as Manifestations of Industrial
Intoxication Caused by Trimethylenetrinitroamine (T4). Med. lavoro, 40,107-112 (1949).
7. Sterner, J. H. Eastman Kodak Co. Personnel Communication, 1961 (Cited by
Patty).
8. McConneil, W. J., Flinn, R. H„ and Brandt, A. D. Occupational Medicine J, 551
(1946).
9. McCrone, W. C. Cyclotetrair.ethylene tetranitramine. Anal. Chem. 22,
1225-1226 (1950).
10. MacGregor, W. S. The Chemical and Physical Properties of DMSO. Ann. N. Y.
Acad. Sci. 141, 3-12 (1967).
11. Leake, C. D. Introductory Remarks - N. Y. Acad. Sci. Symposium on the
Biological Actions of Dimethyl Sulfoxide. Ibid. 1-2.
12. Leake, C. D., Rosenbaum, E. E., and Jacob, S. W. Summary of the N. Y. Acad,
of Sci. Symposium on the Biological Actions of Dimethyl Sulfoxide. Ibid. 670-671.
13. Ashwood-Smith, M. J. Radioprotective and Cryoprotective Properties of
Dimethyl Sulfoxide in Cellular Systems. Ibid. 4S-62.
14. Rubin, L. F., and Barnett, K. C. Ocular Effects of Oral and Dermal Application
of Dimethyl Sulfoxide in Animals. Ibid. 333-345.
15. Wood, D. C., Sweet, D., Van Dolah, J., Smith H, J. C., and Contaxis, 1. A Study
of DMSO and Steroids in Rabbit Eyes. Ibid. 346-380.
60
-------
16. Klcbcrgcr, Kurt-I.hcrhard. An Ophthalmological I-valuation of DMSO. lb til
381-385.
17. Smith. H. R., Mason. M. M.. and hpstcin, K. The Influence ot Dimethyl Sulfoxide
on the Dog with Linphasison the Ophthalmologic hxainination. Ibid. 386-391.
18. Smith, H. R., Madidian, Z., and Mason, M. M. The Single .ind Repeated Dose
Toxicity ol Dimethyl Sulfoxide. Ibid 96-109.
19. Caujolle, F. M. L., Caujollc, D. H.. Cros. S. B., and Calvct, M.-M.-J. Limits of
Toxic and Teratogenic Tolerance of Dimethyl Sulfoxide. Ibid 110-125.
20. Goldman, L., Igclmaii. J. M., and Kitzmillcr, K. Investigative Studies with DMSO
in Dermatology. Ibid. 428-43
-------
31. Kligman. A. M Topical Pharmacology and toxicology ol Dimethyl Sulfoxide. J.
Am. Med. Assoc. IW, 796-804. 923-928 (IW»5).
32. Steinberg. A. The Employment ol Dimethyl Sulfoxide as jn Anti-inflammatory
At'vni and Steroid-Transporter in Diversified Clinical Diseases. Ann. N. Y. Acad. Sci. 141. S32-SSO
(I9(>7).
33. Brcchnc V. L., Cohen, D. I)., and Pretsky, I. Dermal Anesthesia by the Topical
Application of Tetracaine Base Dissolved in Dimethyl Sulfoxide. Ibid. 524-531.
34. Lockie. L. M., and Norcross. B. M. A Clinical Study on the Effects of Dimethyl
Suboxide in 103 Patients with Acute and Chronic Musculoskeletal Injuries and Inflammations. Ibid
599-602.
35. Demos, C. II.. UecUolT. G. L.. Donm. M. N., and Oliver, P. M. Dimethyl
Sulloxide m Musculoskeletal Disorders. Ibid. 517-523.
36. Zuckner. J.. Uddin, J. and Gaudier. G. L. Local Application of Dimethyl
Sulfoxide and DMSO Combined with Triamcinolone Acetonide in Rheumatoid Arthritis. Ibid
>.*>5059.
37. Scherbcl, A. L„ McCormack. L. J., and Layle, J. K. Further Observations on the
Lnect ot Dimethyl Sulfoxide in Patients with Generalized Scleroderma. (Progressive Systemic
Scleiosis) Ibid. 613-629.
38. Persky, L.. and Stewart. B. H. The Use of Dimethyl Sulfoxide in the Treatment
ol Genitourinary Disorders. Ibid. 551-554.
39. Blumenthal, L. S . and Fuchs. M. The Clinical Use of Dimethyl Sulfoxide on
Various Headaches, Musculoskeletal and Other General Medical Disorders. Ibid. 572-585.
40. Paul, M. M. Interval Therapy with Dimethyl Sulfoxide. Ibid. 586-598.
41. John, H. and Laudahn. G. Clinical Experience* with the Topical Application of
DMSO in Orthopedic Diseases: Evaluation of 4180 Cav;s. Ibid 506-516.
42. Sulzberger, M. B., Cortese, T. A. Jr., Fishman, L.. Wiley, H. S.. and Peyakovich,
I'. S. Some Effects of DMSO on Human Skin hi Vivo. Ibid 437-450.
43. Weston, J. K. The Development of Drugs and the Responsibilities Involved. Ibid
J4-34.
44. Barnett, K. C., and Noll, P. R. B. Dimethyl Sulfoxide and Lens Changes in
i'limatcs. Nature-7/-#. M 15-1116(1967).
45. Rubin, L. F., and Mattis. P. A. Dimethyl Suboxide Lens Change in Dog* During
Oral Administration. Science 153, 83-84 (1966).
62
-------
46. Gordon, D. M Dimethyl Sulfoxide in Ophthalmology with Especial Reference
to Possible Toxic Effects. Ann. N. Y. Acad. Sci. 141, 392-402 (i967).
47. Caujollc. F. M. E., Caujollc, D. H., and Cros, S. B.,Culvet, M.-M. J., and Tollun.
Y. Pouvoir Teratogene du Dimethyl Sulfoxide et du Dicthylsulloxide. C. R. Acad. Sci. (Paris) 2h0,
327-330( I9(i5).
4S. Ferm, V. H. Teratogenic Effects of DMSO, Lancet 7430, 208 (1966).
49. Kobens, J. F. Teratologic Studies of Carbaryl, Diazinon, Norea, Disulfiran and
Thinim in Small Laboratory Animals. Toxicol. Appl. Pharmacol. 15 152-163 (1969).
50. Ayre, J. E., and LeGuerrier, L. Some (Regressive) Effects of DMSO
Dcxamethasone (Jpon Cervical Cells in Cervical Dysplasia and Carcinoma in Situ Ann. N Y. Acad.
Sci.74/, 414-422 (1967).
51. Handbook of Chemistry and Physics. 44th Edition. The Chemical Rubber
Publishing Co., Cleveland, Ohio.
52. Patty, F. A.Jbid. p. 1719-1770.
53. Browning, E Toxicity of Industrial Organic Solvents. Chemical Publishing Co.,
Inc., New York. pp. 320-334 (1953).
54 Lazarew. N. W., Brussilowskaja, A. J. and Lawrow, J. N. Quantative
Untersuchungen uber die Resoiption cinigcr organischer Gifte durch die Haut ins Blut. Arch.
Gewerbcpathol. Gewerbehyg., 2, 641 I) 931).
55. Carpenter, C. P., and Smyth, H. F. Jr., Amer. J. Opthalmol. 29, 1363 < 1946).
56. Larson, P. S., Finnegan, J. K., and Haag. H. B. Observations on the Effect of
Chemical Configuration on the Edema-Producing Potency of Acids, Aldehydes, Ketones, and
Alcohols. J. Pluirmacoi. Exp. Therap. 11ft. 1 19-122 (1956)..
57. Comer, J. J. Z. Hyg. Infektionskrankh, 130, 680 (1960). Ind. Hyg. Digest 15.
Abstract No. 168(1951).
58. Albertoni, P. Die Wirkung und die Verwandlungen einiger Stoffe im Organismus
in Beziehung zur Pathogenese der Acetonamie und des diabetes. Arch. Exp. Path. Pharmakol. 18,
219(1884).
59. Nelson.K. W., Ege.J. F., Ross, M., Woodman, L. E. and Silverman, L. Sensory
Response to Certain Industrial Solvent Vapors. J. Ind. Hyg. 25, 282 (1943).
60. American Conference of Governmental Industrial Hygienists. Amer Ind. Hyg.
Assoc. J. 22, 325-328 (1961). Threshold Limit Values for 1961.
63
-------
61. Treon. J. F.. Crutchfield. W. I:. Jr.. and Kitzmillcr, K. V. The Physiological
Response ol° Animals to Cyclohexane, Mcthylcyclohcxane. and Certain Derivatives of 'llmse
Compounds. II. Inhalation. J. Ind. Hyg. Toxicol. 25, 323-347 (1943).
62. Jacobi. Huyashi. and S/.ubinski. F. Arch. Exp. Pathol. Pharmakol. 50,
IW (1903).
63. Treon, J. F., Cmtchfield, W. I-. Jr.. and Kitzmiller, K. V. The Physiological
Response of Rabbits to Cyclohexanc, Methylcyclohexane. and Certain Derivatives of These
Compounds. I. Oral and Cutacnous Application. J. Ind. Hyg. Toxicol. 25,199-214 (1943).
64. Spccht, H., Miller, J. W., Valaer. P. J., and Sayers, R. R. Acute Response of
Guinea Pigs to the Inhalation of Ketone Vapon>. National Inst. Health Bull. No. 176, U. S. Public
Health Service, 1940.
65. Kcngstorff, R. H., Petrali, J. P., and Sim. V. M. Cataracts Induced in Guinea Pigs
b> Acetone, Cyclohexanone. and Dimethyl Sulfoxide. Amer. J. Optom. 49, 308-319 (1972).
66. Sunderman, F. W. OSRD Report No. 4174. Hazards to the Health of Individuals
Working with RDX. September 1944. UNCLASSIFIED Report.
67. Landsteiner, K., and Chase, M. W. Studies on the Sensitization of Animals With
Simple Chemical Compound*. J. Exp. Med. 61, 643 (1935).
68. Landsteiner, K. and Jacobs, J. Studies on the Sensitization of Animals With
Simple Chemical Compounds. J. Exp. Med.64,625 (1936).
69. Landsteiner, K„ and Chase, M. W Studies on the Sensitization of Animals With
Simple Chemical Compounds. J. Exp. Med. 66, 337 (1937).
70. Landsteiner, K. and Chase, M. W. Studies on the Sensitization of Animals With
Simple Chemical Compounds. J. Exp. Med. 71 237 (1940).
71. Landsteiner, K. and Chase, M. W. Studies on the Sensitization of Animals with
Simple Chemical Compounds. J. Exp. Med. 73,431 (1941).
72. Rothberg, S. EATR 4219. Skin Sensitization Potential of the Riot Cont'd
Agents, CA. DM, CN, and CS in Guinea Pigs. March 1969. UNCLASSIFIED Report.
73. Swartz, L., and Tulipan, L. Occupational Diseases of the Skin. First Edition. Lea
Febiger. Philadelphia, Pennsylvania. 1939.
74. Department of the Army Technical Manual 9-1919, Department of the Air Force
Technical Order 11 A-1 -34, Military Explosives, pp 177-182. April 1955.
75. Swartz, L., Tulipan, L., and Birmingham, D. J. Occupational Diseases of the
Skin. Third Edition. Lea Febiger, Philadelphia, Pennsylvania. 1957.
64
-------
APPr.NDIX A
IIG URLS
AAV\f/\|Vv^-VvvnVAr\JV^^
» ANESTHETIZED
unanesthetized
LEFT (LEAD
5 SECONDS-
LEADS electrodes
anesthetized
RIGHT (LEAD III
UNANESTHETI7E0
Figure A-l. Diagrammatic Sketch ot Brain Showing Approximate Placement ot Electrodes.
Sample Tracings ot EEG in Anesthetized and Unanesthetized Dog.
65
-------
bb a
4
EEC
Ji < I I I II ¦' I 1111(1111'
EKG
250 cpa
J
Auditory Response
Figure A-2. Typical Responses Obtained from Various Phynotogic
Panmeten of Dog after Noise (2S0 cps)
66
-------
BEG
||~ ¦ ^ ^ " -I r i " ** III « - * . ¦ ttn I » I II11 I * , >1 M I >1 irm m '< fr. , ,, _ n.i*-_
BEG
LUJ.J I | | I I | 1 I 1 I
EKG
RESP
750 cps
r
Auditory Response
fs/
CT5
ft-
cn
Wk.'
cv
Figure A-3. Typical Responset Obtained from Various Physiologic
Parameters of Dog after Noise (750 cps)
67
-------
EEC
.1.1 I. I. .1 ¦) .1 .1.1 ,1 I.I. I IJ
Nasal Stimulation !
(
i
Figure A-4. Typical Responses Obtained from Various Physologic
Paramel.rs of Dog after Nasal StunuUtion
68
-------
_1
Figure A-S. Typical Responses Obtained from VariouJ Phyuologic
Parameters of Dog after Stroking
69
-------
Ugtit Response Vibratory Response
Figure A-6. Typical Responses Obtained from Various Physiologic
Parameters of Dog after Exposures to Light and Rapping
70
-------
VA/'vwww
RESP
1 ml DfcBO Oners venous)
25 mm/sec
bMiW^WWl/WW\/Wvw\AA/W
1 ml DKCO (Intravenous)
1—i i t—i—r
2. S mm/sec
Figure A-7. EfTecli of DMSO Upon Physiologic Panmetrn of Unannihetized Oogi
71
-------
-------
SBC
Ml I i
EEC
IIUIIHI111 ll/lk
BKC
,/. .;¦¦•¦. 1
Ikv.VKJ.
rrrrf / /; /—r
KVKTvjV-
-v-—
1 ml Cyclohexanonc (Intravenous)
1 mln post Injection
Figure A-9 kllecti of Cycloho*anomr Upon Phvuo logic Parameter*
ol jn UiijiU'^ilu'iiud Dogf I mini
73
-------
EKU
iixii I
VP
—i 1—111111111111 m 11111 1111111
2 mln post injection 30 minute poet injection 60 mln post
1 nii'Cyclonexanone (intravoious)
Hisuk A-10 Effects of Cycluhexjnonc Uiw» I'tiyiiologic Parunii-ii-rs of jh
Unjnesthelized Dog 12. JO. jnd 60 mm)
74
-------
ELG
' wE1r~A/'~Vyv v vVv-l'V
ik
nirt^rJhnrii
j -144 - ^r-I~r\\A^c\A\\A\~riJA'^r'r(j: ^ y\
*• \ J-„v)\j\Jv^\JVNJvJVJMMM^NJVXJ^MVjv.^
Br r
U'
/ { 1 I I 1 I
I m) (."ycloliLv^norw (ini, ivtrituM
I n In post injcs rion 5 mtn post Lnjcvtlon
I ir«iu A II I fK¦ i< <>i < > > nt'i «jii' 1. I in•<) l*|i\ Pjijfiirit r\ ol
intWuMhi il »»»~ * nun and?hf)
2 hr poit ln)ecaoo
-------
Upon Physiologic Parameter* of
-------
BEG
VI
RESP
nWWU^MJWIMVjVJ^^
J x I l —L
EEC
Before Injection
20mgAg RDX-DMSO (33%)
TP
IS sec post
Vibratory Response
5 mtn post Injectton
jr
Fifure A-13 EfTecti of Intravenous Administration of 20 mg/kg RDX-DMSO
Upon Physiologic Puameten of an Unaneithetued Dog
77
-------
EEC
-4^1-4-4-^4—^— ~\lj— f|^> ration at 40 os|/k| HMX-OMSO
Upon Phyoolopc Pmmctrn of an Uoinolbtfiud Dog
-------
BBO
EKG
20 mg/kg HMX-DMSO <33%)
after reflushlng catheter
j-n.
Figure A-l 5 Effects of Intravenous Adnunislrjlion of ?0 mg/kg HMX-UMSO
Upon PhyootogK- Parameters of an Unanoslhcti/fd Dog (after rellushing catheter*
79
-------
EEC
EKG
20 m#/kg HMX-DMSO (33%) 30n)ln after first injection
Figure A-16 Hff.-cli ol' IntrjM'iiom AJmnmirjt on oi ;0 mg/kg HMX-DMSO Upon
Phyuologic Pjranu'tcrb of .111 (¦njiuMhi'ii/ai Dog (30 mini
80
-------
^KMWU\>
10 in la post Injection
1 hr post
1 hr post injection
J I I I I I I I ) lU
R 'I
Figure A-17. Effects of Intra vtnouj Adrumstration of 20 mg/kg HMX-OMSO Upon
Physiologic Parameters of an Unanesthetized Dog (10 nun and t hr)
81
-------
EEC i I
—*iv—^—*j a—*^iv^—
EKG
Stroking Response
BP
F esponse to Light
1 hr post Injection
Spontaneous Discharge
-n.
Figure A-18. Effect! of Intravenous Administration of 20 mg/kg HMX-DMSO Upon
Phi siolo&ic Parameter* of in Unaneithttiied Dog (stroking and light at I hi)
82
-------
Arvvv*v"*vvVVw>riV
^^
*1*. 1 !¦>:..
BEG
EEC
EKG
ArlVVwV
A/VIAaAA
r
^aaAAAaAaa^^^
RE3P
1-1/2 hr post Injection
2 hr pcjt injection
Figure A-19 Effects of Intnvtnout Administration of 20 mg/kf HMX-OMSO Upon
Physiologic Pinmetrn of in (JruncDhclucd 0o|< 1-1/2 and ? hr)
83
-------
PhyuoiogK Pjnmcicn ol Jn Uninmhrti/cd Dog I JO xc)
84
-------
Figure AO I ttkct> ol Inirjv .nous AdminiMtjiion ol 20 mg/kp MMX-OMSO Upon
Phyuologk Parameter* of an Unum-klhiMizrd Do# (I nun. S hrl
-------
I
I
I
rx
Figure A-2? Effect! of Intravenous Adnmiitiration ot TO mg/kg HMX DMSO Lpon
Physiologic Parameter* of on Unanmhctucd Dog 11 and 4 days)
86
-------
-¦n fv rOn'Aki' j,/
physfog
RESP
Control 6. 75 mg/kg HMX-Acettne (5. 4%) 1 mln post Injection
Fil>urt' A-23 fcftc.isor Iqirjvtnoua Adminisirjiion ol 6 75 nig/kg HMX-Aceionr Upon
Physiologic Puimetert of jn Unjnetthelizcd Dog (I mini
87
-------
Kigure A-.4. Iiu-cii of lmra»enom AJinin^irjiion of ft 7S nig/kg HMX-Acvtonv Upon
Physiologic Parameters ol jn liiunmhriizcd Dog i2 inmi
88
-------
2-1/2 min post Injection
4 mln post injection
l-igurc A-25 LftcU» of Intf-vamns Administration of 6 7S mg/l>g H\1X-A«lonc Upon
Physiologic Pjrameter» ol jii Unanttthrtizrd Dog (2-112 and 4 mint
89
-------
Kffi<
K
; i!11;
11
KKC
hM
'i'!:i;!!!;l!lll!
Hill
|f
;l!: 1
A
J
»ii
: i
H-
RESP
7 nun pose Injection
NiMIMiiMjjM
45 min post injecrton
i i ¦ ¦ ¦
Fi|iurc -\-^f> I ifrtfH ol lnlrjw"0 Aiininii^irjlioii hi <¦ 7< HMX-Ah'Mii. Lp.'ii
I'iuuologK PjrjiiK'U'r* 01 .hi LiijikNllu*ti/it! Dou* < ~ 111J 4S nun)
90
-------
EEQ
i iltbin
il
f
III
II
I
2 hr post injection
EEC
! I
\fA a^A-JVjfA
EKQ
RESP
BP
1 day post Injection
¦n
rifufc \-Zl bltcusui Inlrjvtfuoiis AJtniiiisirjinjii ol
-------
AI'I'KNDIX B
TABLHS
Table U-l. Values lor blood Procure. Heari Rjte, and Respiration During 4-Week Observations
in L>t«g> Alter Acute Percutaneous Applicatiun (I ml)ul KUX. HMX.and Solvents
Treatment
Cuutiol
1st Week
2d Week
3d Week
4th Week
HPJ
IIRb
Resp
l(R
Resp
MR
, Reip
HR
Resp
BP
HR
Resp
DMSO
I
180
9 5
M2
16
96
lb
104
24
96
30
187
100
114
24
| Acetone
275
100
102
12
100
38
108
30
72
50
180
100
120
24
| Acetone
IVO
110
108
12
-
-
120
48
120
-
-
-
-
1
Cyclo
200
125
15(>
18
-
-
182
36
120
36
225
100
180
36
RDX-DMSO
187
140
14
136
14
96
36
96
36
190
100
114
24
RDX-DMSO
184
120
165
27
104
30
96
24
120
24
180
100
144
26
RDX-Acetone
IK7
100
144
13
102
24
120
28
84
36
190
100
96
30
KDX-Aceione
iyo
90
78
30
78
30
72
38
72
48
180
100
84
50
RDX-Cyclo
170
45
144
24
108
24
84
18
108
30
180
100
108
24
RDX-Cyclo
:is
125
176
28
110
36
96
36
96
36
165
75
130
30
IIMX-DMSO
2U0
100
156
36
108
48
120
48
168
60
175
75
140
52
HMX-Acetone
180
60
144
24
98
48
108
30
108
36
121
75
84
24
IIMX-Cyclo
ISO
90
102
24
«0
54
90
30
84
42
I7S
100
108
30
JUP = I.'moral arterial blood pressure (mm Hg)
bHR ° heart rate (beats per minute)
¦"Kesp = icspiratory rate (breaths per minute).
92
-------
Tjblc B-ll Pjiii Threshold and Pup'l Kc->pun->c id L(^ii Before and Ai'ier Acuie
Fciiuijncuub Applicjtiun (I ml) ol RDX. HMX. and Solvents
(. oilllol
1 si Week
2d Week
3d Week
4tl> Week
Treatment
I
Pjmij
Plipil
rebpuit*c
j Pjiii
Pupil
res|x>nse
Pain
Pupil
response
Pain
Pupil
response
Pain
Pupil
response
i
v
nim
V
mm
V
mm
V
mm
V
mm
DM.SO
I
10
-
-
-
10
10 0-1 5
15
100-40
10
12.5-3 5
Acetone
11 5-3.5
37
11 0-3.0
34
11 0-3.0
24
11.0-3.0
30
11 5-3.5
Aw lime
:s
12 0-3 5
50
110-2 5
31
11 0-3 0
-
-
_
j
Cyclo
4»
9 0- * 0
"
38
10 0-3 5
44
10.5-3.5
2K
11.0-2 5 i
'
KDX-DMSO
3K
10 0-3.5
.
9 5-3 5
40
100-25
-
-
*
loa-i.o i
KDX-DMSO
32
I00-*0
40
10 0-2 0
4b
9 5-2.0
40
10.0-2 5
26
I
10.0-2 0
KDX-Au'i.ine
5«
1 1 0-3 5
45
1 1 0-3 0
30
1 2 0-3.5
47
11 5-2 5
12.0-3 0 !
KDX-Ateioiie
.>(»
36
11 5-:5
37
12 0-2 5
23
9 5-2 0
-7
12 0-2.5
RDX-Cyclo
£
') 0-3 0
46
10.0-2 5
34
90-2 0
2')
11.0-2.0
40
10 0-2.5 i
KI)X-C'v>.lo
<>0
1 1 0-» 0
50
10 0-10
60
110-4 0
60
10.5-2 5
54
s>5-2 0
HMX-DMSO
60
i o o-: u
40
10 0-2.5
28
110-2 5
26
11 0-2.0
20
0 0-2 0
HMX-Al'CIKIIC
(.4
100-2 5
33
10 0-^ 0
38
KJ0-2 5
2K
110-3 0
20
ro.o-2.0
HMX-Cvel..
'
40
9 0-2 o
31
10.0-2 0
23
9 0-2 0
34
10 0-2.0
22
9 0-2 5
J Pjiii (v) = the jinounl of vultjye needed to eliwii u positive response.
^l*upil iespouse = the rclj'ivc m/c • >! the pupil helorc Jiid jfler li^iii stimulus.
Appendix B
93
-------
Tjblc B-lll. l-.liects of Certain Test Stimuli Upon Physiologic Parameters After Acute
Percutaneous Application (I ml)ol KIJX. HMX, and Solvents
N.isjI
Vibratory
Stroking
Li glit
Auditory
Tieat litem
stiiniil.iiion
stimulation
response
Hash
response
111'
UK
K.'sp
UP
1 IK
Resp
UP
lll<
Resp
UP
I1K
Rcsp
BP
HR
Resp
DM SO
0
0
I
u
0
-
-
-
-
0
0
0
-
-
-
Acetone
i
0
I
i
1
i
0
0
0
0
0
0
0
0
0
Acetone
0
1
1
0
0
0
0
0
0
0
0
0
0
0
0
I'yclo*
0
0
0
0
0
0
0
0
a
0
0
0
0
0
KDX-DMSO
u
u
I
-
-
-
0
0
0
-
-
0
0
0 I
KDX-UMSO
i
u
1
i
-
-
-
0
0
0
0
0
(J
-
0
1
i
1
1
KDX-Acetoue
i
1
1
0
1
0
0
0
-
-
-
0
0
0 j
KDX-Aeetono
-
-
0
u
I
0
0
i 1
1
RDX-Cyclo
1
1
I
i
I
i
t
0
0
0
0
0
0
0
1 !
RDX-Cyclo
-
-
J
I
I
I
0
0
0
0
V
0
0
0
0 1
' MMX-DMSO
t)
1
I
0
I
t
0
1
I
0
0
0
1
I
i
1 HMX-Accione
l
0
0
1
0
0
I
i
1
0
-
-
-
i
I
;
IIMX-Cyclo
0
i
f
0
0
1
0
I
I
0
0
0
NOIL: 0 = no change
- = not monitored
t = increase
1 = decrease
•Cyelo = cyclolicxaiione.
Appendix B
94
-------
Ijhk' B-IV Vjlue> lui Bluod Piessuie IIljiI Rjte jnd Re\pirjtiun Dunnj; --Week Obseiv.n miii
in Dogs Alter Stibjuile Peitiu mcuus Application (I ml/d.iyInl KDX. IIMX. jnd Solvem-.
(out rul
l-l Week
2d Week
3d Week
4th Week
liutiueiii
Bl,J
lllS
8U
KDX-IJMSU
|9Q
100
1 1 ^
,0
*0
108
24
108
24
:oo
"ioo
108
15
RDX-DMSO
_
100
25
100
25
100
25
%
20
-
104
25
RDX-DMSO
-
104
50
120
25
100
24
120
25
"
1 16
25
KDX-Accioih?
I"0
100
90
84
90
66
108
*6
84
78
180
90
X4
40
RDX-Aci'ioni'
I-'S
100
1 .lis
I2t.
50
IOa
4X
1 14
M)
; /ii
"s
1 2o
42
KDX-( vi lo
1X0
(>0
9(>
:o
XO
2J
7-'
88
40
190
100
84
RDX-Cy.lo
l'>5
100
1 -iS
30
126
50
108
48
1 14
30
191)
75
120
HMX-DMSO
1x0
Xj
1 38
33
96
C>0
120
48
144
75
1 9(J
1 20
102
50
HMX-DMSO
IVO
11:
130
36
84
24
80
36
84
3S
I 65
100
7-
36
HMX-DMSO
130
40
72
25
9»>
25
:s
"
72
25
HMX-DMSO
-
9(>
(>0
100
90
25
9„
25
-
90
25
HMX-Aicumc
175
90
ioo
28
90
90
30
*>
36
1 9()
100
9<)
3o
HMX-Acetoue
iss
100
144
24
108
60
120
38
i:o
38
185
T3o
84
36
HMX-Cydu
1X0
40
1.32
48
120
24
144
3(i
1 5o
1 80
100
1 50
!
UMX-Cyclo
|6<)
"
I0K
28
IOO
i
2(>
120
24
120
3o
l_80
105
1 SO
aBP = fcmuidl arterial blood pressure (mm Hg)
^HR = liejrl rale (beats per minute)
cResp = respiratory rate (breaths per minute)
Appendix B
95
-------
Table B-V. Pain Threshold and Pupil Response to Light Before and After Subacute
Percutaneous Application (I inl/day)uf RDX. HMX,and Solvents
Treat men!
Control
1st Week
2d Week
3d Week
4th Week
Paul*1
Pupil
response
Pain
Pupil
response
Pain
Pupil
response
Pain
Pupil
response
Pjin
Pupil.
response
v
mm
v
inm
V
mm
V
mm
V
mm
DM SO
48
9.0-1.5
50
9.5-2.0
32
9.0-2.5
41
11.0-2.5
-
-
DM SO
57
11.0-4.0
46
11.0-2.0
38
110-3 0
40
10.0-2.5
42
11.0-3.0
Acetone
28
10.0-3.0
18
9.0-3.0
24
11.0-3 0
-
10.0-2.5
-
10.0-2.5
Acetone
(>2
10.0-3.0
38
10.0-3.0
47
11.0-3 0
42
110-3 0
38
10.0-3.0
Cyelo
4V
11.0-3.0
-
10.0-2.5
30
11.0-2 5
24
10.5-3.0
24
10.5-3 0
Cyclo
34
11.0-3.5
61
11.0-3.0
43
10.0-2.5
34
10.0-2.0
30
9.0-2.0
RDX-DMSO
47
12.0-3.0
52
11.5-3.5
28
12.0-3.5
26
12.0-3.5
24
11 0-3.5
KDX-DMSO
40
10.0-3.0
55
11.0-3.0
38
11.5-3.5
41
11.0-3.5
20
10.0-3.5
KDX-Acetone
43
11.0-3.0
20
12.0-3.5
-
10.0-2.5
38
11.0-2.5
20
12.0-3.0
RDX-Acetone
37
11.0-3 0
42
10 5-2 5
45
9 5-2 0
44
10.0-2.5
40
10.5-2.5
RDX-Cyclo
34
11.0-4.0
35
9.0-2.5
41
9.0-3.0
44
9.5-3.0
-
-
RDX-Cyclo
5:
10.0-2.0
32
9.0-2.0
34
10.0-2.5
24
9.5-2.5
48
10.0-3.0
HMX-DMSO
28
10.0 \0
15
9.0-2.0
26
9.0-2.0
44
10.0-3.0
35
10.0-3.0
HMX-DMSO
29
10.5-3.5
24
10.0-2.0
40
10.0-2.0
27
9.0-2.0
24
9 0-2.0
HMX-Acctone
40
10.0-3.0
54
10.0-3.0
55
11.0-3.5
49
11.0-3.5
36
11.0-3.0
HMX-Acctone
42
11.0-3.0
20
9.0-2.0
32
10.0-2.5
3S
10.0-3.0
38
11.0-3.0
HMX-Cyclo
IS
11 0-2.0
-
10.0-2.0
49
9.0-2.0
25
10.0-3.0
31
10.0-3.0
HMX-Cyclu
43
11.0-3.0
50
9.0-2.0
40
10.0-2.0
25
11.0-2.5
33
9.5-3.5
aPain (v) = the amount of voltage needed to elicit a positive response.
''Pupil response 3 the relative size (in mm) of the pupil before and after light stimulus.
Appendix B
9(6
-------
Ijblc U-VI. l:llcilsol Ccitjin Test Stimuli Upon Physiologic Paranuclei* Alter Subacute
I'cicutaiicuu* Application (I ml/day I ol KDX, HMX. and Solvent*
Treatment
NumI
-.lunulatioii
Vibralo.y
stimulation
Stroking
response
Light
llubll
Audiloiy
rcbponsc
BP
MR
Rc*p
BP
HR
Rtip
BP
HR
Rcbp
BP
HR
Rcsp
BP
HR
Rcsp
DM SO
0
1
I
0
4
4
0
u
0
0
4
0
0
4
DM SI)
J
J
4
4
4
4
-
-
-
4
4
4
0
i
0
Acetone
0
4
4
0
0
0
-
-
-
0
0
It
0
0
41
Acetone
0
0
it
U
0
41
-
-
0
4
4
4
4
4
Cyclo *
0
0
4
0
J
i
0
0
0
0
0
it
0
4
0
Cyclo
0
0
4
0
«
I
-
-
-
4
4
4
i
i
4
RDX-DMSO
0
0
4
0
'
0
0
0
0
0
0
0
0
0
0
RDX-DMSO
0
4
0
u
;
0
u
0
0
4
i
-
RDX-DMSO
-
RDX-DMSO
-
RDX-Acetonc
1
4
4
0
4
0
0
4
4
0
4
4
0
0
i
RDX-Acetone
0
0
4
0
4
0
I
*
4
0
4
0
0
I
4
RDX-Cydo
0
0
4
0
0
It
-
0
0
i
0
o
0
RDX-Cyclo
0
4
4
0
0
0
4 t
0
0
4
0
'
i
HMX-DMSO
0
1
i
0
0
*
0
0
o
0
4
it
0
1
4
IIMX-DMSO
u
4
4
0
0
0
0
0
u
4
0
0
i
4
HMX-DMSO
HMX-DMSO
HMX-Aecione
0
4
4
0
1
4
0
0
41
0
4
it
0
0
41
HMX-Acetone
0
4
4
0
4
4
0
0
4 t
0
0
41
0
i
4
HMX-Cyt-lo
0
0
i
0
0
41
0
0
4 t
0
0
41
0
4
0
HMX-Cyclo
0
4
4
0
4
4
0
0
0
0
4
4
0
i
4
NOTK 0 = ho change
— = not monitored
t = mcrcubt;
I = decrease
•Cyclo = cyclolicxanone.
Appendix B
97
-------
-o Table B-VII The Sensitization Potential in Clipped Guinea Pigs of Intiadermally Administered RDX in Acetone.
1| Cyclohexanone, and Pure and Technical Ciade DM SO When Challenged by the Percutaneous Route
o.
Sensitization exposure3
Challenge exposure'*
Skin effects'
Compound
Strength of
solution
Solvent
period
Compound
Solvent and
dilution
4-5
hr
24
hr
48
hr
78
hr
%
days
RDX
0.25
1:1 Acetone -saline
80
RDX-S A% acetone
1.10 PEG
0
0
0
0
RDX
0.25
1:1 Cyclohcxanone-saline
80
RDX-7.5% cyclohexanone
1:10 PEG
0
0
0
0
RDX
0.25
1 1 Tech grade DVUJ-saline
80
RDX-339J tech grade
DMSO
1:10 PEG
0
0
0
0
Acetone
0.50
S?!:.ie
80
RDX-5.4% acetone
1.10 PEG
0
0
0
0
Cyclohexanone
O.SO
Saline
80
RDX-7.5% cyclohexanone
1.10 PEG
0
0
0
0
Pure DMSO
0.50
Saline
80
RDX-339J pure DMSO
1 10 PEG
0
0
0
0
Tech grade DMSO
0.50
Saline
80
RDX-33% tech grade
DMSO
1.10 PEG
0
0
0
0
a0.05 Ml given intradermal^ (dorsal thorax) 3 limes per week for 3 weeks to six animals.
^0.5 Ml given topically (thigh).
cThe Drji/e test was used to evaluate skin effects (table VII, p. 27).
-------
Table B-VIII The Sensitization r'otcnlial in Clipped Guinea Pigs ol" Intradermal!} Admjnistere. tone-saline
1 64 Saline
0
0
0
1
0
CyclohexanoiK
05
Saline
IS
RDX-0 25',< 1 1 c\clohe\anonc-
saluie
1 64 Saline
1-tl
l-fcl
0
• ;
Pure DMSO
05
Saline
IS
RDX-0.25?? 1 1 lech grade
DMSO-soline
1 64 Saline
0
0
0
0
Tech grade DMSO
05
Saline
18
I
RDX-0 25™ I.I techgiade
DMSO-sjIme
1 64 Saline
l-EI
l-EI
0
0
1
JU05 Ml given intradermally (dorsal lliorax) 3 limes pci week for 3 weeks id m\ animals
**0.05 Ml givm iniradermally (thigh).
cTiie Dtji/c icsi was used to evaluate skin effects (table VII, p. 27 )
-------
Tabic B-IX Tlic Sensitization Potential in Clipped Gumca Pigs of PctculaneousK Applied RDX in Acetone.
Cyclohexjiiunc. and Pure and Technical Grade DMSOas Determined b\ Topicjl Challenge
Using Polyethylene Glycol 200
Sensitization exposure3
Rest
period
Challenge exposure'1
Skin eflectsc
Compound
Strength of
solution
Solvent
Compound
Solvent and
dilution
5
hi
24
hi
48
hr
72
hi
%
days
RDX
5.4
Acetone
25
RDX-5.4^ aceiuuc
1:10 PEG
0
0
0
0
RDX
7.5
Cyclohexanone
25
RDX-7.5^ cyclohexanone
1:10 PEG
0
0
0
0
RDX
33.0
Pure DMSO
25
RDX-333 pure DMSO
1:10 PEG
0
0
0
0 i
i
RDX
33.0
Tech grade DMSO
25
RDX-33% lech grade DMSO
1:10 PEG
0
0
0
0
Acetone
100.0
-
25
RDX-5.4% acetone
1:10 PEG
0
0
0
0
Cyclohexanone
100.0
-
25
RDX-7.5% cyclohexanone
1 10 PEG
0
0
0
0
Pure DMSO
100.0
-
25
RDX-33% pure DMSO
I .JO PEG
0
0
0
0
Tech graJe
DMSO
100.0
—
25
RDX-33% tech grade DMSO
1.10 PEG
0
0
0
0
a0.S Ml given topically (dorsal thorax) 3 limes per week for 3 weeks to six animals.
**0.5 Ml given topically (thigh).
cThe Draize test was used to evaluate skin effects (table VII, p 27)
-------
Table B-X. The Sensitization Potential in Clipped Guinea Pigs of Percuianeously Applied RDX in Acetone,
Cyclohexanone, ind Pure and Technical Giadc DMSO as Determined by Intradermal Challenge
Sensitization exposuies3
Ri'ii
period
Challenge exposure''
Skin effects' I
Compound
Strength of
solution
Solvent
Compound
Solvent and
dilution
4
hr
24
hr
48
hr
72
hr
%
days
RDX
S.4
Acetone
18
RDX-0 2S9S 1 ) acetone and
saline
1:32
0
0
0
0
i
RDX
7.4
Cyclohexanone
18
RDX-0.259£ 1 1 cyclohexanone
and saline
1.64
0
0
0
0
1
RDX
33.0
Pure DMSO
18
RDX-0.25"* 1 1 pure DMSO-
saline
1.32
0
0
0
i
0
RDX
33.0
Tech grade DMSO
18
RDX-0.25% I.I tech grade
DMSO saline
1:32
0
0
0
0
Acetone
100.0
—
18
RDX-0 25% 1.1 acetone and
saline
1:32
0
1 -El
0
0
Cyclohexanone
100.0
—
18
RDX-0.25% 1.1 cyclohexanone
and saline
1:64
0
0
0
0
Pure DM SO
100.0
—
18
RDX-O.25% 1.1 pureDMSO-
saline
1.32
0
0
0
0
Tech grade DMSO
100.0
—
18
RDX-0 25% 1:1 tech grade
DMSO-saline
1:32
0
0
0
0
a0.5 Ml given topically (dorsal thorax) 3 times per week for 3 weeks lo six animals.
^0.05 Ml saline given inlradermally (thigh).
cThe Oraize test was used lo evaluate skin effects (table VII, p. 27)
-------
Table B-XI The Senhiii/jimn Potenti.il in Clipped Guinea Pigs of liiKadctmally Administered UNIX in Acetone.
Cyrlohexanuiic. and Pure and Technical Grade OMSO Wlien Chjllenjvd hj lIk- Perc> taneous Rome
Sensitization exposuies3
Rest
period
Chjllcniv cxposuieb
Skin effects0
Compound
Strength of
solution
Solvent
Compound
Solvent and
dilution
5-6
hr
24
hr
48
hr
72
hr
t
djys
HMX
0.25
1 1 Acetone-saline
25
HMX-2/? acetone
1:10 PEG
0
0
0
o
HMX
0.25
1:1 Cyclohexanone-saline
25
HMX-2.5% cyclohexanone
1:10 PEG
0
0
0
0
KMX
0.25
1:1 Pure DMSO-salmc
25
HMX-3 3% pine DMSO
1:10 PEG
0
0
0
0
HMX
0.25
1:1 Tech grade DMSO-saline
25
HMX-3.3% tech grade DMSO
1:10 PEG
0
0
0
0
Acetone
0.50
Saline
25
HMX-2^ acetone
1:10 PEG
0
0
0
0
Cyclohexanune
0.50
Saline
25
HMX-2.5% cyclohexanone
1.10 PEG
0
0
0
0
Pure DM SO
0 50
Saline
25
HMX-3.395 pure DMSO
I.I0PFG
0
0
0
0
Tech grade
DM SO
0.50
Saline
25
HMX 3 3% tech grade DMSO
1 10 PEG
0
0
0
0
a0.05 Ml given intradermally (dorsal thorax) 3 times per week for 3 weeks to six animals.
^0.5 Ml PfcG given topically (thigh).
cThe Draiic test was used to evaluate skin effects (tabic VII. p. 27).
-------
Compound
Table B-XII. The Sensitization Potential in Clipped Guinea Pigs of Intudermjlly Administered HMX in Acetone.
Cyclohcxanonc. and Pure and Technical Crjde DM SO as Determined by intradermal Cha! enge
Scnsili/alion expo>uresa
HMX
HMX
HMX
HMX
Acetone
Cyclohexanonc
Pure DM SO
Tech grade DMSO
Strength of
solution
0.25
0.25
0 25
0 25
0 50
0.50
0.50
0.50
Solvent
I I Acetone-saline
I I Cyclohexanone-sahne
I I Pure DMSO valine
I I Tech DMSO-salinc
Saline
Saline
Saline
Saline
Ron i
period
days
20
20
20
20
20
20
20
20
Challenge exposure**
Compound
Solvent and
dilution
HMX-0 259? I I acetone-saline
HMX-0 259? I I cyclohe>tanone-
saline
HMX-0 25% I I pme DMSO-
saltne
HMX-0 25% I I tech DMSO-
saline
HMX-0 25% 1.1 acetone-saline
HMX-0 25% 1.1 cyclohexanone-
saline
HMX-0 25% I I pure DMSO-
saliuc
HMX 0.25% 1 1 lech DMSO-
saline
I 32
I 64
1 32
I 32
1:32
1.64
1.32
1.32
Skin effects1"
5-6
hr
0
0
0
24
hr
0 j 0
0 I 0
0
a0 05 Ml given iniradcrmally (dorsal thorax) 3 times per week for 3 weeks to six animals
^0 05 Ml saline given iniradcrmally (thigh)
cThe Draize test was used to evaluate skin effects (table VII, p. 27)
-------
Table ll-XIII The SciiMiizaticn Potential in Clipped Guinea Pijs of PcicuuncoiiNh Applied HMX in Acetone.
Cyclohcxuimne. and Pure and Technical Gude DM SO as Determined b\ Percuuneous
Challenge Using Polyethylene Glycol 200
Sensitization e\posuie>J
Rest
period
Challenge exposure''
Skin effects'"
Compound
Strength of
solution
Solvent
Compound
Solvent and
dilution
6
hr
24
hr
48
hi
72
hr
/*
days
HMX
2.0
Acetone
25
HMX-2 09! acetone
1.10 PEG
0
0
°
0
HMX
2.5
Cyclohcxanone
25
HMX-2 57t cyilohexanone
1:10 PIG
0
0
0
0
HMX
3.3
Pure DMSO
25
HMX-3 37' pure DMSO
1 10 PEG
0
0
0
0
HMX
33.0
Pure DMSO
25
HMX-33 07. pure DMSO
1 10 PEG
od
od
0d
0d
HMX
3.3
Tech grade DMSO
25
HMX-3.3S U\h gwde DMSO
1 10 PEG
0
0
0
0 ;
HMX
33.0
Tech grade DMSO
25
HMX-33.C/S tech grade DMSO
1.10 PEG
od
od
0d
od
Acetone
100.0
-
25
HMX-2.0S?"acetone
1:10 PEG
0
0
0
0
Cyclohcxanone
100.0
-
25
HMX-2.5% cyclohcxanone
1 10 PEC
0
0
0
0
Pure DMSO
100.0
-
25
HMX-3.391 pure DMSO
1:10 PEG
0
0
0
0
Tech grade OMSO
1000
-
25
HMX-3 3% te It grade DMSO
1:10 PEG
0
0
0
0
a0.5 Ml given topically (dorsal thorax) 3 times per week for 3 weeks to six animals.
^O.S Ml PEG given topically (tlugh)
cThe Drai/c test was used to evaluate skin effects (table VII. p 27)
dTwo animals from the 33% HMX/pure OMSO group and three animals from the 33% HMX/tech grade DMSO group died during sensitizing prixess
-------
TaHe B-XIV. Tin. Siiiiin/jiion K.uuujl in (lippni Cuiikj Pig* of PcicuIjikoiisIn Applied UNIX 111 Acetone
C>cl()hc\jnunc, and Pure and Ti.Jinu.al Grade DMSOas l)cteinii;ied b> Intuderinal Clijllenge
Sensitization e\po->uie\J
R(.-t
Challenge exposure^
Skin elicits
<
Compound
Strength of
solution
[ Solvent
pt nod
Compound
Dilution
(i
hi
24
hi
4 K
hr
72 i
hr !
C-
iiuS >
I
HMX
2.0
Acetone
18
HMX-0 257-' 1 1 ace tune-saline
1 32
°
0
0
°
HMX
2 5
f)dohc\„i:onc
1 H
HMX-0 25'- 1 1 cyclolie\anone-
salinc
1 64
0
0
0
1
1
HMX
3.3
Pine DMSO
IS
HMX-0 2<', 11 puio DMSO-
salim
1 32
0
0
0
0
HMX
33 3
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IX
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1 .¦>"!
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od
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HMX
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1 32
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1 .b
-------
Tabic B-XV. The Sensitization Potential in Clipped Guinea Pigs of IniiaderinalK Administered Acetone.
Cyclohexanone. and Pure and Technical Crude DMSO When Challenged bj the Peicutaneous Route
Sensiii/uiion cxposuies"
Rest
penod
Challenge exposure'1
Skin effects'"
Compound
Strength of
solution
Solvent
Compound
Solvent and
dilution
4
hr
24
hi
48
hr
1- ¦
1-
hr
9f
days
Acetone
SO
1:1 Acetone-saline
24
Acetone
1.10 PEG
0
0
0
0
Cyclohexanonc
SO
1 1 Cy Johexaiioitc-salinc
24
Cyclohexanonc
1.10 PEG
0
0
u
0
Pure DMSO
50
1 1 Pine DMSO-saltne
24
Pure DMSO
1:10 PEG
0
0
0
0
Tech grade DMSO
50
1 1 Tech grade
DMSO-saline
24
Tccli giade DMSO
1 10 PEG
0
0
0
0
a0.05 Ml given iniradermally (dorsal lliorav) 3 dines per week for 3 weeks to four an'mals
^0.5 Ml PEG given lopieally (thigh)
cThe Drane test was used to evaluate skin effects (table VII. p. 27).
-------
Table B-XVI The Schmii'jiioii Pukuiial in Clipped Guinea Pigs ol Intr. dennall) Administered Acetone.
Cyclohexanone, and I'uie and Icchniial Grade DMSO as Deteimined b\ lulradeimal Challenge
Sensili/.alion exposures3
Rest
period
Challenge exposure'5
Skin e
fleets
C
Compound
Strength of
solution
Solvent
Compound
Solvent and
dilution
6
hr
24
hr
48
hr
72
hr
rA
da> s
I
Acetone
SO
I.I Acetone-saline
17
Acetone 50/r 1 1 saline
1 64 saline
0
0
0
0
Cyclohexanonc
5C
I.I fyclohcxanone-salnic
17
C>ilohe\anone 50'^ 1 1 saline
1 64 saline
0
0
0
0
Pure DMSO
50
1 1 Pure DMSO-saline
17
Pure DMSO 509c' 1 1 saline
1 64 saline
0
o
1
0
0
Tech grade DMSO
50
1 1 Tcili grade
DMSO-salnu
P
Tech grade DMSO 50'V
1 1 saline
1 64 saline'
0
1
0
0
0
a0.05 Ml given •ntradermally (dorsal thorax) 3 lime* per week foi 3 weeks to tour animals
b0 05 Ml given intradermally (thigh)
^The Drai/e test wjsused to evaluate skin effects (table VII, p 27)
-------
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