Aflatoxin Absorbtion

J. TOXIC0L.
-TOXIN REVIEWS, 7 ( 2 ) , 95-120 (1988-89)
SKIN ABSORPTION AS A ROUTE OF EXPOSURE FOR AFLATOXIN

AND TRICHOTHECENES
B.W. Kemppainen*
Toxin Reviews Downloaded from informahealthcare.com by Kainan University on 04/03/15
Department of Pharmacal Sciences, School of Pharmacy

Auburn University, Auburn, Alabama 36849, U.S.A.
R.T. Riley
Richard B. Russell Agricultural Research Center
U.S. Department of Agriculture, Agricultural Research Service
Athens, Georgia 30613 U.S.A.
J.G. Pace
U.S. Army Medical Research Institute of Infectious Diseases
Department of Army
Fort Detrick, Frederick, Maryland 21701 U.S.A.
For personal use only.
ABSTRACT
Airborne occurrence of mycotoxins in agricultural and
residential environments has generated concern that these toxins
may be absorbed via the skin and respiratory tract. This paper
summarizes information available on the h viva and vitro
cutaneous permeability of several mycotoxins (aflatoxin, T-2 toxin
[T-21, diacetoxyscirpenol, and verrucarin A).
Published data is used to calculate the dose absorbed during
cutaneous exposure to fungal toxins in hypothetical agricultural
and research 1 aboratory environments. These estimated doses of
T-2 (0.004 and 0.041 pg/kg, respectively) were 25,000 and 2,439
times less than a reported oral dose (100 pg T-2/kg/day) which
caused immunosuppression in monkeys.
In general, exposure of human skin to aflatoxin and tricho-
thecenes results in slow absorption. The risk of systemic
toxicity resulting from dermal exposure increases in the presence
o f high toxin concentrations, occlusion, and vehicles which
enhance penetration.
95
Copyright 0 1989 by Marcel Dekker, Inc.

96 KEMPPAINEN, R I L E Y , AND PACE
TABLE OF CONTENTS
I. INTRODUCTION 96
11. METHODOLOGY USED TO STUDY PERCUTANEOUS ABSORPTION OF
FUNGAL TOXINS 98
A. In Vivo Studies 98
B. In Vitro Studies 98
I I I. RESULTS FROM STUDIES ON PERCUTANEOUS ABSORPTION OF FUNGAL
TOXINS 99
A. In Vivo Cutaneous Absorption 99
B. In Vitro Cutaneous Absorption 101
1. Relative Absorption of Mycotoxins Using Human Skin 101

2. Factors Which Affect Absorption of Mycotoxins 102
3. Distribution of T-2 in Skin Following Topical
Application 105
4. Metabolism of Mycotoxins Following Topical
Appl i cat i on 106
C. Comparison of In Vivo and In Vitro Cutaneous Absorption 109
D. Estimate of Absorbed Dose Under Hypothetical Conditions 111
IV. CONCLUSION 113
V. REFERENCES 114
I. INTRODUCTION
The presence of mycotoxins on grain dusts and other small
particulate materials indicates that people living or working in
agricultural environments are routinely exposed to mycotoxins via
the respiratory and dermal routes. Consumption of mycotoxin
contaminated foods and feeds has been a recognized problem for
many years. In 1960 the outbreak of Turkey "X" disease was found
to be due to ingestion o f peanuts contaminated with aflatoxins (1)
and perhaps other mycotoxins (2). Since that time a multitude of
symptoms have been observed in association with the consumption of
moldy feeds. Epidemiological studies in Africa, Asia and else-
where, have suggested a correlation between aflatoxin contamina-
tion of foods and liver cancer ( 3 , 4 ) . In the United States,
environmental monitoring during grain harvesting (5) and during
post-harvest grain management (6) has indicated that dust parti-
cles contain relatively high levels of aflatoxins. The respira-
tory route has received some attention as a possible route of ex-
posure to aflatoxins in industrial settings (7). The dermal route
has also been considered, but the results of the early studies in
rats were contradictory. Purchase and Steyn (8) reported that
SKIN ABSORPTION 97
internal accumulation of aflatoxin B1 (AFBI)~, applied between the

shoulder blades of rats, was primarily due to ingestion after
removal from the skin by the paws during grooming. This con-
clusion brought into serious doubt the earlier findings of Wei,
Liu & Lee (9) who reported significant uptake of [14C]AFB1 by rats
and the finding of Ungar and Joffe (10) who reported acute liver
lesions in rabbits attributed to percutaneous absorption of

aflatoxins. Joffe and Ungar (71) reported epidermal lesions in
rabbits following topical application of aflatoxins suggesting
that at the very least, aflatoxins have the potential to penetrate
the stratum corneum and elicit responses in the underlying viable
epidermis and dermis. It was the uncertainty of whether AFBl
could penetrate the skin and the knowledge of the presence of AFBI
on grain dusts which motivated us to determine whether or not AFBl
could penetrate human skin in vitro. At the same time that we

were commencing studies of the cutaneous penetration of AFB1, the
U.S. Army became interested in the potential for trichothecenes to
penetrate human skin. The U.S. Department of Agriculture’s
interest in trichothecenes stems from the fact that, like the
aflatoxins, trichothecenes are also common contaminants of cereal
grains (11). In addition to the U.S. Army’s concern about the
possible exposure of mi 1 i tary and ci vi 1 i an personnel to airborne
mycotoxins (12), trichothecenes have been isolated from fungal
contaminated duct work and insulation (13) and dermal exposure in
laboratory environments have been reported (14,15).
Research has been conducted in the United States, the United
Kingdom, Israel, Japan, and South Africa to determine the
potential of fungal toxins to penetrate skin. These studies have
involved skin absorption in animals and humans and have used h
-
vivo and in vitro techniques. The purpose of this paper is to
review the information available on skin absorption of fungal
toxins and use this information to estimate the potential for
percutaneous absorption o f mycotoxins under hypothetical
conditions. We recognize that the extrapolation of vitro
results to the in vivo situation is tenuous but we believe it i s a
98 KEMPPAINEN, RILEY, AND PACE
valid exercise to stimulate discussion and to serve as a point of

departure for future studies.
11. METHODOLOGY USED TO STUDY PERCUTANEOUS
ABSORPTION OF FUNGAL TOXINS
The following two sections are a brief summary of the methods
which have been used to estimate the cutaneous permeability of
fungal toxins. The readers interested in more detailed

methodological descriptions are recommended to consult the
appropriate references cited in the results section of this
review.
A. In Vivo Studies
-
In -vivo dermal toxicity studies are performed by applying
mycotoxins to the depilated dorsal skin surface of laboratory
animals and then measuring biologic responses (e.g., altered
weight gain, morphological changes in skin or other organs). In

percutaneous absorption studies the mycotoxins are applied to
depilated dorsal skin surface and the extent of skin absorption is
determined by measuring the quantity of fungal toxin in organs
(e.g., the disappearance of toxin from the skin) or biological
fluids. In both dermal toxicity and percutaneous absorption
studies it is important to prevent inadvertent ingestion of
mycotoxins which may occur during grooming (16).
B. In Vitro Studies
-
In -
vitro absorption has been assessed using both static and
flow through diffusion cells. In the static system, penetration
is measured by removing sequential samples from the stirred saline
solution (receptor fluid) which bathes the dermal surface of the
excised skin disc. In the flow-through system receptor fluid i s
continuously pumped through the chamber below the skin. The
vitro methods involve mounting discs of excised skin on diffusion
cells. Discs of full-thickness skin (epidermis and dermis) and
isolated epidermal sheets have been used. The isolated epidermal
sheets are prepared by immersing full-thickness skin in hot water
(6OOC) for one or two minutes, and then removing the epidermis
from the dermis. Epidermal surfaces can be nonoccluded (exposed
SKIN ABSORPTION 99
t o ambient temperature and humidity) o r occluded by p l a c i n g a

b a r r i e r t o water d i f f u s i o n above the epidermal surface. This
causes an increase i n t h e humidity above t h e s k i n and r e s u l t s i n
increased h y d r a t i o n o f the s k i n surface. Radiolabeled mycotoxins
are a p p l i e d t o t h e epidermal surface and t h e percent o f dose which
penetrates t h e s k i n can be e a s i l y c a l c u l a t e d . The percent dose
absorbed i s expressed as mycotoxin equivalents and o f t e n does n o t

take i n t o consideration t h a t some o f t h e r a d i o a c t i v i t y i n t h e
r e c e p t o r f l u i d s i s associated w i t h metabolites o f t h e mycotoxins.
The a s s o c i a t i o n o f the r a d i o a c t i v e l a b e l w i t h e i t h e r t h e parent
compound o r metabolites, i s p r e r e q u i s i t e t o equating r a d i o a c t i v i t y
t o mycotoxin equivalents, Cutaneous metabolism o f t h e mycotoxins
has been measured by i d e n t i f y i n g and q u a n t i f y i n g t h e r a d i o l a b e l e d
compounds i n t h e receptor f l u i d by t h i n l a y e r radiochromatography.
The advantages o f using b v i t r o techniques are: i)

p e n e t r a t i o n through human s k i n can be s t u d i e d w i t h o u t exposing
l i v i n g subjects and ii) inadvertent i n g e s t i o n o f t h e t e s t compound
i s eliminated. The disadvantages are: i)absence o f desquamation
v i t r o , ii) d i f f i c u l t f o r receptor f l u i d t o model t h e
p h y s i o l o g i c a l p e r f u s i o n o f dermis by blood, iii) decreasing
v i a b i l i t y o f excised tissue, and i v ) l a c k o f c e n t r a l c o n t r o l o f
p e r i p h e r a l t i s s u e (e.g. blood f l o w t o s k i n ) . The disadvantages o f
using i n v i t r o techniques can be minimized by comparing i n v i v o
and jn v i t r Q percutaneous penetration, and o p t i m i z i n g t h e i n v i t r o
methods i n order t o increase c o r r e l a t i o n between i n v i v o and i~
absorption.
I II. RESULTS FROM STUDIES ON PERCUTANEOUS
ABSORPTION OF FUNGAL TOXINS
A. I n Vivo Cutaneous AbsorDtion
The a b i l i t y o f trichothecene mycotoxins t o penetrate s k i n i s
demonstrated by t h e many b i o l o g i c a l e f f e c t s which occur f o l l o w i n g
dermal exposure t o these t o x i n s . Dermal exposure o f r a t s and
r a b b i t s t o small doses (10-120 ng) o f trichothecenes (T-2 t o x i n
[T-21, diacetoxyscirpenol [DAS]) consistently results i n a local -
non - spec if ic acute in f l a m a t o r y reaction characterized by
100 KEMPPAINEN , RILEY, AND PACE
hyperemia, edema and n e u t r o p h i l exudation, w i t h v a r i a b l e amounts

o f necrosis o f t h e epidermis ( 1 7 ) . T h i s inflammatory p r o p e r t y o f
trichothecenes l e d t o t h e development o f a s k i n t e s t (18) which
has been w i d e l y used t o screen l a r g e numbers o f samples o f g r a i n
f o r t h e presence o f these t o x i n s ( 1 9 ) . Dermal exposure o f mice
and r a b b i t s t o l a r g e doses o f T - 2 ( 5 t o 40 mg/kg) i n d i m e t h y l -
s u l f o x i d e (DMSO, a p e n e t r a t i o n enhancer) r e s u l t s i n systemic

e f f e c t s , i n c l u d i n g death (20,21).
Dermal exposure o f p i g s t o T-2
(15 mg/kg) i n DMSO r e s u l t e d i n changes i n body weight gain, r e c t a l
temperature, hematology, serum biochemistry and c e l l u l a r immune
response ( 2 2 ) . The systemic d i s t r i b u t i o n o f T-2 f o l l o w i n g dermal
exposure was confirmed by percutaneous absorption s t u d i e s i n
guinea p i g s i n which 82% o r 8% (DMSO o r methanol vehicle,
r e s p e c t i v e l y ) o f the dose was absorbed by day 28 ( 2 3 ) .
The a b i l i t y o f a f l a t o x i n s t o penetrate s k i n has been a

c o n t r o v e r s i a l subject because o f the i n c o n s i s t e n t observations
f o l l o w i n g dermal exposure t o a f l a t o x i n . Ungar and J o f f e (10)
reported severe epidermal necrosis and 1 i v e r l e s i o n s i n r a b b i t s
dermally dosed w i t h AFBl and a f l a t o x i n B2 (AFB2). Wei e t a l . (9)
r e p o r t e d uptake o f 80% o f a dermal dose o f [14C]AFB1 w i t h i n 24 h r
and a r e s u l t a n t decrease o f weight g a i n i n t h e t r e a t e d r a t s .
Purchase and Steyn (8) observed t h a t dermal exposure t o AFBl f o r
130 min r e s u l t e d i n percutaneous absorption o f AFBl i n unanes-
t h e t i z e d r a t s b u t n o t i n anesthetized r a t s . Purchase and Steyn
concluded t h a t t h e b i o l o g i c a l e f f e c t s observed i n animals dermally
dosed w i t h AFBl and AFB2 were a c t u a l l y due t o i n g e s t i o n o f t h e
t o x i n d u r i n g grooming. I n contrast, L l e w e l l y n (24) dermally dosed
g e r b i l s w i t h AFBl f o r 35 days and d i d n o t use any methods t o
prevent i n g e s t i o n . The l a c k o f l e s i o n s i n t h e g e r b i l organs and
absence of AFBl o r r e l a t e d metabolites i n f e c a l samples i n d i c a t e d
t h e r e was l i t t l e o r no AFBl absorbed o r consumed. The s h o r t
d u r a t i o n of dermal exposure i n Purchase and Steyn's study r e s u l t e d
i n inconclusive findings regarding t h e cutaneous p e r m e a b i l i t y o f
AFBI. The k i n e t i c s of s k i n p e n e t r a t i o n by chemical are charac-
t e r i z e d by an i n i t i a l l a g phase i n which t h e r a t e o f absorption i s
SKIN ABSORPTION 101
TABLE 1
-
In -
Vitro Absorption of Fungal Toxins in Human Skin
(Vehi c l e:Methanol ) .
Fungal Toxin Percent Dose Absorbed
T-2a 1.0
DASa 0.8
VCAa 0.2
AFB+ 0.05
aDiscs o f f u l l - t h i c k n e s s s k i n were exposed to 0.6 ug/cm2

trichothecene f o r 48 h r (27).
bDiscs o f i s o l a t e d epidermis were exposed t o 4.2 - 5.3
ug/cm2 AFBl f o r 46 h r (28).
slow (25). When d i s c s o f i s o l a t e d human epidermis were exposed t o

AFBl under nonoccluded c o n d i t i o n s t h e l a g t i m e was 18 h r (26). I n

order t o determine i f i n v i v o absorption o f AFBl occurs i t i s
necessary t o perform a percutaneous absorption study i n which t h e
p e r i o d o f dermal exposure t o A F B l i s longer (e.g., 24 t o 48 h r )
and appropriate methods are used t o prevent i n g e s t i o n o f t o x i n .
B. I n V i t r o Cutaneous AbsorDtion
1. R e l a t i v e absorotion o f various micotoxins u s i n q human
skin. AFBl penetrates i s o l a t e d s k i n very slowly r e l a t i v e t o o t h e r
t e s t e d mycotoxins and a l s o r e l a t i v e t o o t h e r organic compounds.
The r e l a t i v e p e n e t r a t i o n o f mycotoxins through excised human s k i n
was T-2 > DAS > Verrucarin A (VCA) > A F B l (Table 1). Comparisons
o f t h e r e s u l t s o f d i f f e r e n t i n v i t r o cutaneous absorption s t u d i e s
must be done c a u t i o u s l y because d i f f e r e n t methods are o f t e n used
i n d i f f e r e n t studies. The methods used t o prepare and s t o r e s k i n
d i s c s can e f f e c t the extent o f cutaneous p e n e t r a t i o n (29). For
instance, T-2 penetrates through i s o l a t e d epidermis f a s t e r than
through f u l l -thickness skin; and through once-frozen s k i n f a s t e r
than through s k i n which has never been frozen (Table 2). The
cutaneous p e n e t r a t i o n o f AFBl i s most l i k e l y an overestimate since
t h e s t u d i e s w i t h AFBl were conducted using i s o l a t e d epidermal
TABLE 2
Effect of Skin Preparation on T-2 Penetration Through Excised
Human Skin.
Absorption of T-2
Skin preparation Method of Storage (Percent of Dose)
Isolated epidermisa Frozen ( -20°C) 1.12
Full -thicknessa Frozen (-20°C) 0.33

Full -thicknessb Frozen (-60°C) 1.53
Full -thicknessb Refrigerated (4OC) 0.64
-
- -
aEoidermal surface was exDosed to TJH1T-2 adsorbed to corn dust
(p).
b[ HIT-2 dissolved in methanol was applied to the epidermal
surfaces (29).
discs from once-frozen skin whereas the cutaneous penetration of

T-2, DAS and VCA were determined using full-thickness skin which
had never been frozen. Franz (31) determined the j . ~vitro
penetration of 12 organic compounds using once-frozen full -
thickness human skin. The calculated total percent penetration of
AFB1 through isolated epidermal sheets in 48 hr was an order of
magnitude less than any compound tested by Franz (31).
f . There are
many factors which can affect the rate of penetration of
mycotoxins through skin. The effect of skin preparation and
storage has already been discussed. Other factors which can
effect the rate of penetration are: concentration, occlusion,
vehicle, species differences, and solubility. The amount of the
material which penetrates is affected by the concentration of the
dose applied. The effect of concentration of T-2 applied on
cutaneous absorption is shown in Table 3. As the dose increased
there was an increase in the percent dose absorbed up t o a dose of
5.8 ug/cm2, after which the percent penetrated decreased. More
importantly, as the dose increased there was an increase in the
amount (us) of T-2 absorbed.
SKIN ABSORPTION 103
TABLE 3
Effect of Applied Concentration of T-2 on It! Vitro Cutaneous
Absorption in Humans.a
Concentration Absorption of T-2°
(ug/cm2) Percent Amount (ug)

0.08 0.6 0.0005

0.58 1 .o 0.0058
5.8 1.2 0.0696
58.1 0.6 0.3486
581 .O 0.1 0.581
5,810.0 0.1 5.81
"From Kemppainen u.(32).
bDose applied in 6 - 68 ul methanol, absorption monitored for 48
hr.
Cutaneous penetration is often enhanced by occlusion (33).

Occlusion refers to covering the dosed skin site. For instance,
an agricultural or laboratory worker could begin working in a
contaminated environment and then decide to wear a shirt or gloves
to prevent further skin exposure. Occlusion increased the
cutaneous absorption of T-2 and AFBl by factors of 4.2 and 68,
respectively (Table 4). The cutaneous absorption of non-polar
compounds is less affected by hydration than polar compounds (34).
TABLE 4
Effect of Occlusion on In Vitro Absorption of Fungal Toxins.
Percent dose absorbed
Fungal toxin Nonoccl uded Occl udedC Factor increase
T-2d 0.37 1.55 4.2
AFB+ 0.05 3.41 68
"Discs of full-thickness human skin were exposed to 0.58 ug/cmL
T-2 for 48 hr (unpublished data).
bDiscs of isolated human epidermis were exposed to 4.2 - 5.3
ug/cm2 for 46 hr (28).
CThe epidermal surfaces were occluded by placing a glass coverslip
over the top of the donar chamber of the diffusion cell (28).
The vehicle which "carries" the fungal toxin to the skin

surface can affect the amount of toxin which will be absorbed. In
an agricultural setting it is likely that corn dust will be the
vehicle. Cutaneous exposure to corn dust containing 18 PPM T-2
resulted in absorption of 0.33% of the dose of T-2 (Table 5).
Laboratory workers frequently use sol vents to extract large
quantities of fungal toxins and then purify these toxins for use
in experiments. Cutaneous exposure to T-2, DAS, or VCA dissolved
in DMSO resulted in 29, 37, or 10% of the dose, respectively,
being absorbed after 48 hr. DMSO, which is frequently used in
laboratories because of its solvent properties, is a well known
enhancer of cutaneous penetration (35).
In order to relate in vivo studies with laboratory animals t o
-
in -vitro studies using human skin it is necessary t o compare &I
vStro absorption of mycotoxins using skin from laboratory animals

and humans. T-2 penetrates isolated monkey, guinea pig and rat
skin significantly faster than isolated human skin (Table 6).
Penetration through isolated animal skin provides estimates 2 t o
10 times greater than through isolated human skin. The relatively
slow penetration of T-2 through rabbit skin is surprising because,
in general, compounds are thought to penetrate through rabbit skin
more rapidly than through the skin of any other species studied
(36). The penetration o f T-2 and DAS through guinea pig skin was
2.8 and 6.8 times faster, respectively, than through human skin.
This demonstrates that the relative rate o f penetration o f
different compounds through human and animal skin varies,
depending on the compounds.
The j r ~vitro absorption of many lipophilic compounds can be
rate-limited due to low solubility in the receptor fluid which
bathes the dermal surface of the skin (37). The use of serum as a
receptor fluid does not increase the absorption of T-2 (38) or
A F B l (26) even though AFBl is 4 times more soluble in serum than
phosphate buffered saline.
SKIN ABSORPTION 105
TABLE 5
E f f e c t o f Vehicle on In Vitro Penetration o f Trichothecenes
Through Human Skin.
Percent dose absorbed
Trichothecene Corn dusta Methanol DMSOD
T-2 0.33 1.0 29

DAS ---c 0.8 37
VCA --- 0.2 10

aDiscs o f o ce-frozen, f u l l - t h i c k n e s s human s k i n were exposed t o 2
- 3 mg/cmF corn dust c o n t a i n i n g 18 PPM T-2 f o r 24 h r under
nonoccluded conditions, followed by 24 h r under occluded
c o n d i t i o n s (30).
bDiscs o f n ver-frozen, f u l l - t h i c k n e s s human s k i n were exposed t o
0.6 ug/cmf o f trichothecene f o r 48 h r under nonoccluded
c o n d i t i o n s (27).
CNot done.
TABLE 6
I n V i t r o Absorption o f Trichothecenes i n Human, Monkey, Rabbit,
Guinea Pig and Rat Skin.
Percent dose absorbedC
Trichothecene Human Monkey Rabbit Guinea Pig Rat
T-2d 1.0 2.5 3.0 2.0 9.7
DAS~ 0.8 ---d --- 6.8 ---
“From Kemppainen e t a l . (32).
bFrom Kemppainen u.
(27).
‘%iscs o f never-frozen, f u l l - t h i c k n e s s s k i n where exposed t o 0.6
ug/cm2 trichothecene i n methanol f o r 48 hr.
dNot done.
3. D i s t r i b u t i o n o f T-2 i n s k i n f o l l o w i n a t o o i c a l a w l i c a t i o n .
The d i s t r i b u t i o n o f T-2 i n t o once-frozen human s k i n and r e c e p t o r
f l u i d was studied by Maxwell e t a l . (38). The r e s u l t s are
summarized i n Table 7. I n Maxwell’s study (38) 7-10% o f a p p l i e d
r a d i o a c t i v i t y was recovered i n receptor f l u i d bathing t h e skin.
I n t h e s t u d i e s conducted i n our l a b o r a t o r y using never-frozen s k i n
d i s c s (27) t h e absorption o f T-2 was found t o be considerably l e s s
(approximately 1%o f applied dose penetrated i n 48 h r ) . It has
been shown (Table 2) that T-2 penetrates skin which has been
stored frozen faster than skin stored at 4OC (29). At the
termination of Maxwell et ale's T-2 penetration study, the skin
surface was blotted with filter paper to remove unadsorbed T-2.
The top 10 layers of stratum corneum were removed by stripping the
surface with tape and the layers analyzed for radioactivity to
determine amount of T-2 absorbed into outer layers of stratum

corneum. The largest amount o f T-2 was located in the 10 layers
of the stratum corneum and of the total dose applied, 44 to 73%
had either penetrated the skin or was in the process of pene-
trating the skin. This indicates that although less than 10% of
the dose was absorbed in 48 hr, a much larger percent of the dose
could be absorbed over a relatively long period o f time. The
stratum corneum can act as a reservoir, releasing material for
days and even weeks after the initial exposure period (39). A
cutaneous reservoir of T-2 could play an important role in chronic
toxicity following cutaneous exposure.
4. Metabolism of mvcotoxins followinq topical application.
Cutaneous metabolism is important in determining the toxicity of
surface-applied chemicals (40,41). It was therefore important to
evaluate the extent to which fungal toxins are metabolized during
TABLE 7
-
In -
Vitro Distribution of T-2 in Human Skin and Receptor Fluid
After 48 hr Penetration (Vehicle: Ethanol) .a
% Applied dose recovered in skin and receptor fluid
Dose Stratum Corneum Epidermis Dermis Receptor fluid TotalD

(uf/ (10 layers)
cm 1
1.0 15 16 9 9 44
1.8 30 3 5 10 48
2.6 55 3 8 7 73
"From Maxwell U.(38).
bTotal refers to total percent of dose absorbed into and through
skin discs.
SKIN ABSORPTION 107
cutaneous penetration. T-2 was e x t e n s i v e l y metabolized d u r i n g

p e n e t r a t i o n through human s k i n (42). When T-2 was a p p l i e d t o t h e
epidermal surface o f d i s c s o f excised human skin, more than 40% o f
t h e r a d i o a c t i v i t y i n t h e r e c e p t o r f l u i d was associated w i t h
metabolites o f T-2 (Table 8). This data i s c o n s i s t e n t w i t h t h e
hypothesis t h a t metabolism o f T-2 occurred d u r i n g p e n e t r a t i o n
through t h e s k i n discs, and d i d n o t occur i n the r e c e p t o r f l u i d

due t o enzymes leaching o u t o f the skin.
TABLE 8
Location o f S i t e o f T-2 Metabolism i n Cutaneous Penetration

System.
Metabol it e s d
Source o f metabol it e s T-2 t e t r a o l HT-2 T-2

P u r i t y o f [JH]T-2 preparation 0 1 96
H y d r o l y s i s by receptor f l u i d 0 5 88
H y d r o l y s i s by enzymes 0 2 92
leaching o u t o f s k i n i n t o
receptor f l u i d
Hydrolysis by enzymes 10 31 39
d u r i n g cutaneous p e n e t r a t i o n
“Expressed as percent o f r a d i o a c t i v i t y i n substance analyzed.
From Kemppainen e t a1 (42). .
TABLE 9
Comparison o f Mycotoxin Metabolism by Excised Human Skin (Vehicle:

Methanol ) .
Parent compoundC Metabol it e s C
T-2a 9 84
DASa 46 30
VCAa 77 21
AFB+ 83 --
s
aDisc o f never-frozen, f u l l - t h i c k n e s s s k i n were exposed t o 0.6
g/cm trichothecene f o r 48 h r (27,42).
!Discs o f once-frozen i s o l a t e d epidermis were exposed t o 4 - 5
ug/cm2 AFBl f o r 46 h r (28).
CExpressed as percent o f r a d i o a c t i v i t y i n r e c e p t o r f l u i d .
108 KEMPPAINEN, RILEY, AND PAGE
The T-2 metabolites identified in the receptor fluid (HT-2 toxin

and T-2 tetraol) are generally considered to be less toxic than
the parent compound (43,44) .
The in vitro cutaneous metabolism of several fungal toxins is
listed in Table 9. The extent to which these penetrants are
metabolized cannot be easily compared because different types of
skin preparations were used in the experiments. In the experiment

with [14C]AFB1, discs of isolated human epidermis were used. The
heat treatment which is used to separate the epidermis from the
dermis inactivates some of the cutaneous enzymes (45). This could
be the reason why most o f the radioactivity in the receptor fluid
was associated with the parent compound (AFB1). AFBl is meta-
bolized by the heme-protein cytochrome P-450 (46,47). P-450 has
been shown to be present in cutaneous tissue (48,49,50,51) and is
inducible by chemicals such as polycycl ic hydrocarbon carcinogens

and chlorinated hydrocarbons. Kao et al. (52) have shown that
induction of cutaneous enzymes can result in increased skin
penetration of xenobiotics. The extent of cutaneous metabolism of
trichothecenes increased from T-2>DAS>VCA. It is interesting to
note that the trichothecene which penetrates excised human skin
the fastest (T-2), is also metabolized the most extensively by the
skin.
TABLE 10
Comparison of Cutaneous Metabolism of 1 - 2 in Animals and Humans
(Vehicle: Methanol) .a
Metabolites in receDtor fluid (% radioactivity)
Species T-2 tetraol T-2 trio1 HT-2 T-2
Human 2 1 78 6
Rabbit 3 16 73 0
Monkey 0 0 10 85
Guinea pig 0 0 14 82
Rat 4 0 62 28
“Discs of never-frozen, full-thickness skin were exposed to 0.6
ug/cm2 T-2 for 48 hr (32).
SKIN ABSORPTION 109
TABLE 1 1
In Vitro and In Vivo Absorption of T-2 Through Guinea Pig Skin.a
Percent dose absorbedD
Static F1 ow-through
Vehicle -
In -
vivo --
in vitro --
in vitro
Methanol 22.5 3.9 14.6

DMSO 51.9 38.4 42.6
"From Kemppainen et a1 . (53).
bDuring 48 hr exposure at concentration of 1.2 ug/cm2.
In a previous section it was observed that monkeys, rabbits

and guinea pigs are the laboratory animals whose skin provides the
best model for in vitro penetration of T-2 in human skin. By
considering both cutaneous penetration and metabolism, a more
accurate conclusion can be drawn regarding which animal species is

the best model for evaluating the biological activity of surface-
applied chemicals in humans. In Table 10 the in vitro cutaneous
metabolism of T-2 in animals and humans is compared. Of the
monkey, rabbit and guinea pig, the rabbit is the only species in
which Cutaneous metabolism of T-2 is as extensive as in the human.
Excised rabbit skin consequently provides the best approximation
of human skin, both in terms o f penetration kinetics and metabolic
activity .
C. Comoarison of in vivo and in vitro cutaneous absorDtion.
An important question is whether or not vitro methods can
provide a reasonable approximation of in vivo absorption. The in
vitro and viva absorption of T-2 was compared in order to
assess the validity of using an vitro system to measure
cutaneous penetration of fungal toxins with excised skin (53). The
flow-through system provides a better approximation of in viva
absorption of T-2 than the static in vitro system. &
cutaneous absorption was 5.8 and 1.5 times greater than in vitro
absorption in the static and flow-through cells, respectively,
when methanol was the vehicle (Table 11). In vivo absorption was
1.4 and 1.2 times greater than in vitro absorption in the static
and flow-through cells, respectively, when DMSO was the vehicle.
In each group the u viva absorption was greater than in vitro
absorption, however the relative effect of vehicles on absorption
was similar in viva and for both of the in vitro systems. In
vitro techniques do not precisely predict in viva absorption
during exposure to large doses of T-2. In Table 3 the effect o f

applied concentration of T-2 on in vitro skin penetration is
shown. It is likely, however, that the amount of T-2 absorbed
during in vivo cutaneous exposure to large doses (>58.1 ug/cm2) of
T-2 would actually be much larger than the amounts shown in Table
3. When in vitro and in vivo cutaneous absorption of large doses
(2 - 9 mg/cm2) of T-2 were compared in guinea pigs, the in vivo
absorption was 100 to 167 times greater than the vitro
absorption (53). The applied dose in the large dose study was
more than 1000 times larger than the dose used in the low dose
study (1.2 ug/cm2, see Table 11). The large dose would further
increase the potential for problems with the limited solubility of
the penetrant in the aqueous receptor fluid, and could result in a
much smaller percent of the dose penetrating in vitro. The larger
discrepancy between in vivo and in vitro absorption in the large
dose study demonstrates that the size of the dose can be an
important factor when designing vitro cutaneous absorption
studies intended to correlate with in vivo absorption. Marzulli,
Brown & Maibach (54) reported that the degree of correlation
between in viva and in vitro cutaneous absorption in humans was
dose dependent.
The systemic toxicity which results from cutaneous absorption
of a compound depends on its rate o f absorption and the intrinsic
toxicity of the compound and its metabolites. In vivo studies
have indicated that the relative local and systemic toxicity of
trichothecenes (measured by skin irritation and lethality,
respectively, after dermal exposure) is T-2 > DAS ;=: VCA (55,56).
The results of the in vitro penetration studies (T-2 > DAS > VCA)
and in y&@ dermal toxicity studies are fairly consistent,
SKIN ABSORPTION 111
considering that the intrinsic toxicities (measured by lethality

after intravenous dose) of T-2 or VCA are greater than that of DAS
(57,58).
Skin application of extracts from a wide array of fungi
(Fusarium and other species) resulted in toxic effects in rats
(local necrosis, death) (59). However, other fungal extracts
caused severe toxicity when ingested by rats but not when
topically applied (59). These findings substantiate the

observation that dermal toxicity of fungal toxins i s dependant on
their intrinsic toxicity and their ability to penetrate skin.
D . Estimate of absorbed dose under hvoothetical conditions.
The available data has been used to calculate the dose absorbed
during cutaneous exposure to mycotoxins in hypothetical
agricultural or research laboratory environments (Table 1 2 ) . Two
methods were used to calculate the dose absorbed: i ) percent dose

absorbed in vitro adjusted for exposed body surface area and
duration of exposure, and i i ) h vitro rate of penetration
multiplied by surface area multiplied by exposure time. In each
case the body weight is assumed to be 70 kg. In the case of the
agricultural worker, the surface area of the head, thorax, and
arms (11,970 cm2) were assumed to be exposed to contaminated corn
dust during 8 hr of harvesting (60). In this hypothetical
situation, the exposed surface area was nonoccluded during the
first 4 hr, followed by 4 hr of occlusion (when the worker wore a
shirt). The concentration used for this calculation was 18 PPM of
T-2 in corn dust. By using the same concentration as was used in
the in vitro study (30), the percent dose absorbed in the in vitro
study can be used in these calculations. While 18 PPM is a high
concentration, T-2 concentrations in corn as high as 25 PPM have
been reported (61). The mycotoxin concentrations in airborne
grain dust particles have been reported to be equal to or many
times greater than the mycotoxin concentration in the whole grain
samples (5). Agricultural workers harvesting corn could be
dermally exposed to grain dust contaminated with mycotoxins in the
PPB or PPM range.
TABLE 12
Estimate of Absorbed Dose Under Hypothetical Conditions (Expressed
as Total ug Absorbed).
Dose Body Surface
Level Area xposed
Compound ( ug/cm2) 5
(cm 1 Case la Case 2b Case 3c
-
T-2 0.04 11,970 0.26'l --- ---

0.6 860 --- 2.9d,3.3e ---
DAS 0.6 860 --- 3.7d, 4.4e ---
VCA 0.6 860 --- 1 .od, 1. le ---
AFBl 4.8 860 --- --- 0.05d,
4.8 860 --- --- 0.10e
a70 k adult whose head, thorax and arms were exposed to 2 - 3
mg/cm?? corn dust containing 18 PPM T-2 for 4 hr under nonoccluded
conditions followed by 4 hr of exposure under occluded
conditions.
b70 kg adult whose hands were exposed to mycotoxin in DMSO for 1
hr under nonoccl uded conditions .

c70 kg adult whose hands were exposed to mycotoxin in methanol for
1 hr under nonoccluded conditions.
dDermal dose absorbed (ug) = Percent dose absorbed vitro
adjusted for exposed body surface area and duration of exposure.
eDermah dose absorbed (ug) = (In vitro rat8 of penetration,
ug/cm /hr) X (Exposed body surface area, cm ) X (Duration of
exposure, hr).
fNot enough information available for calculations.
In the case of the laboratory workers, the hypothetical situation

was that they spilled solvent containing mycotoxin on their hands.
The surface area (860 cm2) was exposed to the mycotoxin for one
hour under nonoccluded conditions (60). The doses used in these
calculations (ug/cm2) were the same as used in the in vitro
studies on the respective mycotoxins. The following additional
assumptions are made: i) presence of other chemicals in dose will
not affect penetration o f mycotoxin, i i ) extent of absorption in
48 hr will be proportional to absorption in shorter exposure
period, i i i ) elimination will not significantly effect sustained
dose, iv) extent of penetration in entire body will equal that o f
the abdomen, and v) temperature and hydration are similar to that
used in lab (62).
SKIN ABSORPTION 113
In Table 12, the largest amount of skin absorption (4.4 ug)

occurred when a laboratory worker's hands were exposed to DAS
dissolved in DMSO for 1 hr. This translates to 0.063 ug/kg. When
cancer patients were intravenously dosed with DAS (anguidine), 4.5
mg/m2/day (8.6 mg/70 kg/day), the patients experienced myelo-
suppression, manifested as leukopeni a and/or thrombocytopenia
(63). In some cases the myelosuppression was life threatening.
Cutaneous absorption of DAS in the hypothetical laboratory

situation was 136,500 times less than the dose administered to the
cancer patients. The minimum dose o f DAS or T-2 required to
produce immunosuppression or other toxic effects is not known. In
the agricultural environment the dose of T-2 absorbed was
calculated to be 0.26 ug (0.004 ug/kg). Monkeys dosed with T-2
(100 ug/kg/day, via gastric intubation) were immunologically
suppressed by the end of 4 wk of treatment (64). Cutaneous

absorption of T-2 in the hypothetical agricultural environment was
25,000 times less than the dose administered to the monkeys. The
estimated dose of T-2 absorbed under hypothetical laboratory and
agricultural conditions was calculated by using penetration
results published by Kemppainen et al. (27,30). The penetration
of T-2 through excised human skin when measured by Maxwell et a1 .
(38) was 7 to 10 times greater than the values published by
Kemppainen et al. (compare Tables 3 and 7).
IV. CONCLUSION
It is important to determine the amount of risk associated
with dermal exposure to fungal toxins because millions of people
all over the world are environmentally and/or occupationally
exposed to these toxins. Relative to other chemicals, mycotoxins
penetrate slowly through excised skin. However, this may not be
the case in vivo. The in vitro methods underestimate the in vivo
penetration, nonetheless the data that has been gathered provides
a point of departure for discussion and for designing future
studies. Considering the ability of mycotoxins to penetrate human
skin and their inherent toxicity and in some cases
c a r c i n o g e n i c i t y , i t would be wise t o l i m i t cutaneous exposure

whenever possible. This can be done by wearing p r o t e c t i v e
c l o t h i n g o r decontaminating surfaces exposed t o mycotoxins.
Solutions o f sodium h y p o c h l o r i t e have been used t o d e t o x i f y
l a b o r a t o r y equipment contaminated w i t h T-2 ( 6 5 ) . Solutions o f
sodium h y p o c h l o r i t e and acetone have been used t o d e t o x i f y
l a b o r a t o r y equipment contaminated w i t h AFBl ( 6 6 ) . Bunner ,
Wannemacher, Boobar e t a l . (67) have demonstrated t h a t

percutaneous absorption o f T-2 can be decreased by spraying and
scrubbing t h e contaminated s k i n surface w i t h a soap and water
s o l u t i o n . Vidal, Creach, Genton e t a l . (68) have shown t h a t t h e
cutaneous t o x i c i t y o f DAS can be decreased by decontaminating he
s k i n surface ( w i t h i n 15 min o f exposure) w i t h compresses soaked i n
a sodium h y p o c h l o r i t e s o l u t i o n containing 3% c h l o r i d e . Caut on
must be used when decontaminating s k i n surfaces because wash ng

s k i n surfaces can sometimes cause enhancement o f absorpt on
(69,70).
V. REFERENCES
1. Sargeant, K., Sheridan, A., O'Kelly, J. and Carnaghan,
R.B.A., T o x i c i t y Associated w i t h C e r t a i n Samples o f
Groundnuts, Nature, 192: 1096, 1961.
2. Cole, R.J., E t i l o l g y o f Turkey "X" Disease i n Retrospect: A
Case f o r t h e Involvement o f Cylcopiazonic Acid, Mycotoxin
Res., 2:3, 1986.
3. Keen, P. and Martin, P., I s A f l a t o x i n Carcinogenic i n Man?
The Evidence i n Swaziland, Trop. geogr. Med., 23:44, 1971.
4. Oettl'e, A.G., Cancer i n A f r i c a , E s p e c i a l l y i n Regions South
o f t h e Sahara, J. Natn. Cancer Inst., 33:383, 1964.
5. Burg, W.R., Shotwell, O.L. and Saltzman, B.E., Measurements
o f Airborne A f l a t o x i n s During t h e Handling o f 1979
Contaminated Corn, Am. Ind. Hyg. Assoc. J . , 43:580, 1982.
6. Zennie, T.M., Identification o f Aflatoxin B i n Grain
Elevator Dusts i n Central I l l i n o i s , J. Toxico\. Environ.
Health, 13:589, 1984.
SKIN ABSORPTION 115
7. Hayes, R.B., Van Nieuwenhuize, J.P., Raatgever, J.W. and ten

Kate, F.J.W., Aflatoxin Exposures in the Industrial Setting:
an Epidemiological Study of Mortality, Fd. Chem. Toxic.,
22:39, 1984.
8. Purchase, I.F.H. and Steyn, M., Absence of Percutaneous
Absorption of Aflatoxin, Toxicol. appl. Pharmac., 24:162,
1973.
9. Wei, R.-D., Liu, G.X. and Lee S.S., Uptake o f Aflatoxin B1 by

the Skin of Rats, Experientia, 26:82, 1970.
10. Ungar, J. & Joffe, A.Z., Acute Liver Lesions Resulting from
Percutaneous Absorption o f Aflatoxins, Path. Microbiol .,
33:65, 1969.
11. Joffe, A.Z., Fusarium Species: Their Biology and Toxicology,
John Wiley and Sons, New York, 1986.
12. Watson, S.A., Mirocha, C.J. and Hayes, A.W., Analysis for
Trichothecenes in Samples from Southeast Asia associated with
'Yellow Rain', Fund. appl. Toxic., 4:700, 1984.

13. Croft, W.A., Jarvis, B.B. and Yatawara, C.S., Airborne
Outbreak o f Trichothecene Toxicosis, Atmos. Environ., 20:549,
1986.
14. Brian, P.W. and McGowan, J.C., Biologically Active Metabolic
Products of the Mold Metarrhizium qlutinosum S. Pope, Nature,
Lond., 157:334, 1946.
15. Mortimer, P.H., Campbell, J., diMenna M.E. and White, E.P.,
Experimental Myrotheciotoxicosis and Poisoning in Ruminants
by Verrucarin A and Roridin A., Res. Vet. Sci., 12:508, 1971.
16. Wannemacher, R.W. Jr., Bunner, D.L., Pace, J.G. and
Dinterman, R.E., Efficacy of a Nonocclusive Barrier Model to
Study Toxin Pharmacokinetics after Dermal Exposure,
Toxi col ogi st, 6:245, 1986.
17. Hayes, M.A. and Schiefer, H.B., Quantitative and
Morphological Aspects o f Cutaneous Irritation by
Trichothecene Mycotoxins, Fd Cosmet. Toxicol., 17:611, 1979.
18. Wei, R.-D., Smalley, E.B. and Strong F.M., Improved Skin Test
for Detection of T-2 Toxin, Appl. Microbiol., 23:1029, 1972.
19. Chung, C.W., Trucksess, M.W., Giles, Jr., A.L. and Friedman,
L., Rabbit Skin Test for Estimation of T - 2 Toxin and Other
Skin-Irritating Toxins in Contaminated Corn, J.O.A.C., 57:
1121, 1974.
20. Schiefer, H.B. and Hancock D.S., Systemic Effects of Topical

Application of T-2 Toxin in Mice, Toxic. appl. Pharmac.,
76:464, 1984.
21. Wannemacher, R.W., Jr, Bunner, D.L. and Dinterman, R.E., The
Relationship between Concentration and Exposed Area on
Absorption and Excretion of T-2 Mycotoxin through Rabbit
Skin, Fed. Proceed., 45:575, 1986.
22. Pang, V.F., Felsburg, P.J., Beasley, V.R., Buck, W.B. and
Hascek, W.M., The Toxicity of T-2 Toxin in Swine Following
Topical Application, Fundam. appl. Toxic., 9:50, 1987.
23. Pace, J.G., Cant rbury, W.J. and Matson C., Fate and
Distribution of [fH]T-2 Toxin in Topically Exposed Guinea
Pigs, Toxicologist, 6:246, 1986.
24. Llewellyn, G.C., Toxic Responses in Gerbils Treated Topically
with Aflatoxin B and Dimethylformamide, Bull. Environm.
Contam. Toxicol, 14: 747, 1978.
25. Guy, R.H. and Hadgraft J., in Percutaneous AbsorDtion.

Mechanism-Methodolow-Druq Del iverv, edited by R.L. Bronaugh
and H.I. Maibach, p. 3, Marcel Dekker, New York, 1985.
26. Riley, R.T. and Kemppainen, B.W., in Percutaneous AbsorDtion:
Mechanisms-Methodoloqv-Druq Delivery, edited by R.L. Bronaugh
and H.I. Maibach, p. 387, Marcel Dekker, New York, 1985.
27. Kemppainen, B.W., Riley, R.T. and Biles-Thurlow S.,
C m p a r i s o n o f P e n t r a t i o n and M e t a o l i s m o f
f
[ H]Diacetoxyscripenol, ['jHI Verrucarin A and [$H]T-2 Toxin
in Skin, Fd. Chem. Tox., 25:379, 1987.
28. Riley, R.T., Kemppainen, B.W. and Norred, W.P., Penetration
of Aflatoxins Through Isolated Human Epidermis, J. Toxic.
Envir. Hlth., 15:769, 1985.
29. Kemppainen, B.W., Riley, R.T., Pace, J.G. and Hoerr, F.J.,
The Effects of Ski Storage Conditions and Concentration o f
9
Applied Dose on [ HIT-2 Toxin Penetration Through Excised
Human and Monkey Skin, Fd. Chem. Toxic., 24:221, 1986.
30. K mppainen, B.W., Riley, R.T. and Pace, J.G., Penetration of
LSHlT-2 Toxin Throu h Excised Human and Guinea Pig Skin
During Exposure to [IH]T-2 Toxin Adsorbed to Corn Dust, Fd.
Chem. Toxic., 22:893, 1984.
31. Franz, T.J., Percutaneous Absorption. On the Relevance of In
Vitro Data, J. invest. Derm, 64:190, 1975.
SKIN ABSORPTION 11 7
32. Kemppainen, B.W., Riley, R.T., Joyave, J.L. and Hoer F.J
-
In -
V i t r o Percutaneous Penetration and Metabolism o f f5H] T:i
Toxin: Comparison o f Human, Rabbit, Guinea P i g and Rat,
Toxicon, 25:185, 1987.
33. Scheuplein, R.J. and Ross, L.W., Mechanism o f Percutaneous
Absorption. V. Percutaneous Absorption o f Solvent Deposited
Solids, J. I n v e s t . Derm., 62:353, 1974.
34. Hawkins, G.S., Jr and Reifenrath, W.G., Development o f an In

V i t r o Model f o r Determining the Fate o f Chemicals Applied t o
Skin, Fundam. Appl. Toxicol., 4:S133, 1984.
35. Stoughton, R.B. and Fritsch, W., Influence of
Dimethylsulfoxide (DMSO), Arch. Derm., 90:512, 1964.
36. Grasso, P. and Lansdown, A.B.G., Methods o f Measuring, and
Factors A f f e c t i n g , Percutaneous Absorption, J. SOC. Cosmet.
Chem., 23:481, 1972.
37. Bronaugh, R.L. and Stewart, R.F., Methods f o r ,IJ Vitro

Percutaneous Absorption Studies: Hydrophobic Compounds, J.
Pharm. Sci., 73:1255, 1984.
38. Maxwell, S.A., Brown, R.F.R. and Upshall, D.P., The I n V i t r o

Penetration and D i s t r i b u t i o n o f T - 2 Toxin Through Human Skin,
Toxicology, 40:59, 1986.
39. Vickers, C.F.H., i n Percutaneous Absorption o f Steroids,

e d i t e d by P. Mauvais-Jarvis, C.F.H. Vickers and J. Wepierre,
p. 19, Academic, New York, 1980.
40. Holland, J.M., Kao, J.Y. and Whitaker, M.J., A Multisample

Apparatus f o r K i n e t i c Evaluation o f Skin P e n e t r a t i o n ,IJ
V i t r o : the I n f l u e n c e o f V i a b i l i t y and Metabolic Status o f the
Skin, Toxic. appl. Pharmac., 72: 272, 1984.
41. Kao, J., Patterson, F.K. and H a l l J., Skin Penetration and
Metabolism o f T o p i c a l l y Applied Chemicals i n S i x Mammalian
Species, I n c l u d i n g Man: an I n V i t r o Study w i t h Benzo(a)pyrene
and Testosterone, Toxic. appl. Pharmac., 81:502, 1985.
42. Kemppainen, B.W., Riley, R.T., Pace., J. G., Hoerr, F.J. and
L -
Joyave, J., Evaluation o f Monkey Skin as a Mo e l f o r I n V i t r o
Percutaneous Penetration and Metabolism o f [ HIT 2 Toxin i n
Human Skin, Fund. appl. Toxic., 7:367, 1986.
43. Bata, A., Vanyi, A and Sandor, G.S., Metabolization o f

Trichothecene Toxins ( T - 2 Toxin and Diacetoxysci rpenol) i n
Embryonated Hen’s Egg, Acta Vet. Hung., 31:189, 1983.
44. Cole, R. J., Dorner, J. W., Cox, R.H., Cunfer, B.M., Cut1 er,
H.G. and Stuart, B.P., The Isolation and Identification o f
Several Trichothecene Mycotoxins from Fusarium heterosporum,
J. Nat. Prod., 44:324, 1981.
45. Loden, M., The Vitro Hydrolysis of Diisopropyl
Fluorophosphate During Penetration Through Human Full -
Thickness Skin and Isolated Epidermis, J. Invest. Dermatol.,
85:335, 1985.
46. Decad, G.M., Hsieh, D.P.H. and Byard, J.L., Maintenance o f

Cytochrome P-450 and Metabolism of Aflatoxin B1 in Primary
Hepatocyte Cultures, Biochem. Biophys. Res. Commun., 78:279,
1977.
47. Neal, G.E. and Colley, P.J., Some High Performance Liquid-
Chromatographic Studies of the Metabolism of Aflatoxins by
Rat Liver Microsomal Preparations, Biochem. J., 174:839,
1978.
48. Akin, F.J. and Norred, W.P., Factors Affecting Measurement o f
Aryl Hydrocarbon Hydroxyl ase Activity in Mouse Skin, J .

Invest. Dermatol., 67:709, 1976.
49. Bickers, D.R., Kappas, A . and Alvares, A.P., Differences in
Inducibility of Cutaneous and Hepatic Drug Metabolizing
Enzymes and Cytochrome P-450 by Polychlorinated Biphenyls and
l,l,l-Trichloro-2,2-Bis (p-Chlorophenyl) ethane (DDT), J.
.
Pharmacol Exp. Ther., 188:300, 1974.
50. Bickers, D.R., Eiseman, J., Kappas, A. and Alvares, A.P.,
Microscope Immersion Oils: Effects of Skin Application on
Cutaneous and Hepatic Drug - Metabolizing Enzymes, Biochem.
Pharmacol., 24:779, 1975.
51. Pohl, R.J., Philpot, R.M. and Fouts, J.R., Cytochrome P-450
Content and Mixed Function Oxidase Activity in Microsomes
Isolated from Mouse Skin, Drug. Metab. Dist., 4:442, 1976.
52. Kao, J., Hall, J., Shugart, L.R. and Holland J.M., An J~J
Vi t r o Approach to Studying Cutaneous Metabolism and
Disposition of Topically Applied Xenobiotics, Toxic. appl .
Pharmac., 75:289, 1984.
53. Kemppainen, B.W., Pace, J.G. and Riley, R.T., Comparison o f
-
In -
Vivo and In Vitro Percutaneous Absorption of T-2 Toxin in
Guinea Pigs, Toxicon, 25:1153, 1987.
54. Marzulli, F.N., Brown, D.W.C. and Maibach, H.I., Techniques
for Studying Skin Penetration, Toxicol. appl. Pharmac.,
Suppl. 3:76, 1969.
SKIN ABSORPTION 119
55. Schiefer, H.B., Hancock, D.S. and B h a t t i , A.R., Systemic

Effects of T o p i c a l l y Applied Trichothecenes I. Comparative
Study o f Various Trichothecenes i n Mice, J. Vet. Med. A.,
33:373, 1986.
56. Ueno, Y., T o x i c o l o g i c a l Features o f T-2 Toxin and Related

Trichothecenes, Fund. appl. Toxic., 4:S124, 1984.
57. Bamburg, J.R., Marasas, W.F., Riggs, N.V., Smalley, E.B. and
Strong, F.M., Toxic Spiroepoxy Compounds from Fusaria and

.
Other Hyphomycetes, Biotechnol Bioengng., 10: 445, 1968.
58. Sato, N. and Ueno, Y., i n Mycotoxins i n Human and Animal
Health, e d i t e d by J.V. Rodricks, C.V. Hesseltine and M.A.
Mehlman, p. 295, Pathotox, Park Forest South, IL, 1977.
59. Abbas, H.K., Mirocha, C.J. and Shier, W.T., Mycotoxins
Produced from Fungi I s o l a t e d from Foodstuffs and S o i l :
Comparison o f T o x i c i t y i n F i b r o b l a s t s and Rat Feeding Tests,
Appl. Environ. Microbiol., 48:654, 1984.
60. Guy, R.H. and Maibach, H.I., Correction Factors f o r

D e t e r m i n i n g Body Exposure from Forearm Percutaneous
Absorption Data, J. Appl. Toxicol., 4:26, 1984.
61. Vesonder, R.F., i n Trichothecenes- Chemical, B i o l o g i c a l , and

T o x i c o l o g i c a l Aspects, e d i t e d by Y. Ueno, p. 210, E l s e v i e r ,
New York, 1983.
62. Brown, H.S., Bishop, D.R. and Rowan, C.A., The Role o f Skin
Absorption as a Route o f Exposure f o r V o l a t i l e Organic
Compounds (VOCs) i n D r i n k i n g Water, Am. J. P u b l i c Health,
74:479, 1984.
63. Thigpen, J.T., Vaughn, C. and Stuckey, W.J., Phase I 1 T r i a l
o f Anguidine i n P a t i e n t s w i t h Sarcomas Unresponsive t o P r i o r
Chemotherapy: A Southwest Oncology Group Study, Cancer
Treat. Rep., 65:881, 1981.
64. Jagadeesan, J., Rukmini, C., V i jayaraghavan, M. and Tulpule,

P.G., Immune Studies w i t h T-2 Toxin: E f f e c t o f Feeding and
Withdrawal i n Monkeys, Fd. Chem. Toxic., 20:83, 1982.
65. Thompson, W.L. and Wannemacher, R.W. Jr, D e t e c t i o n and

Q u a n t i t a t i o n o f T-2 Mycotoxin w i t h a S i m p l i f i e d P r o t e i n
Synthesis I n h i b i t i o n Assay, Appl. Env. M i c r o b i o l . , 48:1176,
1984.
66. Castegnaro, M., Friesen, M., Michelon, J. and Walker, E.A.,

Problems Related t o Use o f Sodium H y p o c h l o r i t e i n
D e t o x i f i c a t i o n o f A f l a t o x i n B1, Am. Ind. Hyg. Assoc. J.,
42:398, 1981.
120 KEMPPAINEN , RILEY, AND PACE
67. Bunner, B.L., Wannemacher, Jr., R.W., Boobar, L.R.,

Dinterman, R.E. and Broski, F.H., Chemical/Toxin Dermal
Decontamination Equipment using Soap and Water Spray,
Proceedings, 6 t h Medical Chemical Defence Bioscience Review,
Johns Hopkins U n i v e r s i t y Applied Physics Laboratory,
Columbia, MD, 1987.
68. Vidal, D., Creach, O., Genton, A., Beaudry, Y. and Fontages,
R., D e s t r u c t i o n and Cutaneous Decontamination o f
Diacetoxyscirpenol ( a Trichothecene Mycotoxin) by Sodium

Hypochlorite, C.R. Acad. Sc. Paris, 301:183, 1985.
69. Wester, R.C., Noonan, P.K. and Maibach, H.I., Frequency o f

A p p l i c a t i o n o f Percutaneous Absorption o f Hydrocortisone,
Arch. Dermatol., 113:620, 1977.
70. Wester, R.C. and Maibach, H . I . , i n Cutaneous T o x i c i t y , e d i t e d

by V. D r i l l and P. Lazar, p. 29, Raven, New York, 1984.
71. J o f f e , A.Z. and Ungar, M.D., Cutaneous Lesions Produced by

Topical A p p l i c a t i o n o f A f l a t o x i n t o Rabbit Skin, J. i n v e s t .
.
Dermatol , 52:504, 1969.
FOOTNOTES
A F B l = a f l a t o x i n B1
AFB2 = a f l a t o x i n B2
DAS = diacetoxyscirpenol
DMSO = d i m e t h y l s u l f o x i d e
T-2 = T-2 t o x i n
VCA = v e r r u c a r i n A

Aflatoxin Absorbtion

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Aflatoxin Absorbtion

Uploaded by

Copyright:

Available Formats

J. TOXIC0L.

-TOXIN REVIEWS, 7 ( 2 ) , 95-120 (1988-89)

SKIN ABSORPTION AS A ROUTE OF EXPOSURE FOR AFLATOXIN

Department of Pharmacal Sciences, School of Pharmacy

Copyright 0 1989 by Marcel Dekker, Inc.

1. Relative Absorption of Mycotoxins Using Human Skin 101

internal accumulation of aflatoxin B1 (AFBI)~, applied between the

lesions in rabbits attributed to percutaneous absorption of

could penetrate human skin in vitro. At the same time that we

valid exercise to stimulate discussion and to serve as a point of

fungal toxins. The readers interested in more detailed

weight gain, morphological changes in skin or other organs). In

t o ambient temperature and humidity) o r occluded by p l a c i n g a

absorbed i s expressed as mycotoxin equivalents and o f t e n does n o t

The advantages o f using b v i t r o techniques are: i)

hyperemia, edema and n e u t r o p h i l exudation, w i t h v a r i a b l e amounts

s u l f o x i d e (DMSO, a p e n e t r a t i o n enhancer) r e s u l t s i n systemic

The a b i l i t y o f a f l a t o x i n s t o penetrate s k i n has been a

aDiscs o f f u l l - t h i c k n e s s s k i n were exposed to 0.6 ug/cm2

slow (25). When d i s c s o f i s o l a t e d human epidermis were exposed t o

AFBl under nonoccluded c o n d i t i o n s t h e l a g t i m e was 18 h r (26). I n

Full -thicknessa Frozen (-20°C) 0.33

discs from once-frozen skin whereas the cutaneous penetration of

(ug/cm2) Percent Amount (ug)

0.08 0.6 0.0005

Cutaneous penetration is often enhanced by occlusion (33).

The vehicle which "carries" the fungal toxin to the skin

vStro absorption of mycotoxins using skin from laboratory animals

T-2 0.33 1.0 29

VCA --- 0.2 10

determine amount of T-2 absorbed into outer layers of stratum

Dose Stratum Corneum Epidermis Dermis Receptor fluid TotalD

cutaneous penetration. T-2 was e x t e n s i v e l y metabolized d u r i n g

through t h e s k i n discs, and d i d n o t occur i n the r e c e p t o r f l u i d

Location o f S i t e o f T-2 Metabolism i n Cutaneous Penetration

Source o f metabol it e s T-2 t e t r a o l HT-2 T-2

Comparison o f Mycotoxin Metabolism by Excised Human Skin (Vehicle:

The T-2 metabolites identified in the receptor fluid (HT-2 toxin

skin preparations were used in the experiments. In the experiment

inducible by chemicals such as polycycl ic hydrocarbon carcinogens

Methanol 22.5 3.9 14.6

In a previous section it was observed that monkeys, rabbits

accurate conclusion can be drawn regarding which animal species is

during exposure to large doses of T-2. In Table 3 the effect o f

considering that the intrinsic toxicities (measured by lethality

topically applied (59). These findings substantiate the

methods were used to calculate the dose absorbed: i ) percent dose

T-2 0.04 11,970 0.26'l --- ---

hr under nonoccl uded conditions .

In the case of the laboratory workers, the hypothetical situation

In Table 12, the largest amount of skin absorption (4.4 ug)

Cutaneous absorption of DAS in the hypothetical laboratory

suppressed by the end of 4 wk of treatment (64). Cutaneous

c a r c i n o g e n i c i t y , i t would be wise t o l i m i t cutaneous exposure

Wannemacher, Boobar e t a l . (67) have demonstrated t h a t

must be used when decontaminating s k i n surfaces because wash ng

7. Hayes, R.B., Van Nieuwenhuize, J.P., Raatgever, J.W. and ten

9. Wei, R.-D., Liu, G.X. and Lee S.S., Uptake o f Aflatoxin B1 by

'Yellow Rain', Fund. appl. Toxic., 4:700, 1984.

20. Schiefer, H.B. and Hancock D.S., Systemic Effects of Topical

25. Guy, R.H. and Hadgraft J., in Percutaneous AbsorDtion.

34. Hawkins, G.S., Jr and Reifenrath, W.G., Development o f an In

37. Bronaugh, R.L. and Stewart, R.F., Methods f o r ,IJ Vitro