DISTRIBUTION OF HEOD (DIELDRIN) IN MAMMALS:

I. P ~ L I M I N A R Y MODEL
F. T. LINDSTROM, J. W. GILLETT, and S. E. RODECAP
Environmental Health Sciences Center
Departments o f Agricultural Chemistry
and Mathematics
Oregon State University
Corvallis, Oregon 97331

A mathematical model for dieldrin (HEOD) distributions in mammalian tissues

is presented. The model is based upon lipid-phase transport of dieldrin, large tissue
region compartments and destruction of dieldrin via enzymes in the liver. Several
simple hypothetical cases involving a 300-gin mature male rat or a 68-kg mature
male human are given. Realistic estimates of compartment masses, blood flow, and
lipid content are provided, to illustrate the model's predictive potential. Discontinu-
ous dieldrin administration is considered to illustrate procedural effects upon both
blood and depot fat lipid-phase dieldrin concentrations.

Introduction
The literature on dieldrin (HEOD)I is very extensive and principally focused on toxi-
cology and metabolism. Of the several hundred references which the authors have in
hand, only seven deal with the pharmacokinetics of dieldrin. While considerable informa-
tion exists regarding the pharmacology and chemistry of dieldrin once it is in certain
mammalian tissues, the factors controlling the distribution of dieldrin have been investi-
gated only infrequently. On the basis of some generally accepted concepts and informa-
tion, we have developed a compartmental model to predict the concentrations of dieldrin
to be expected in various tissues of mammals under a variety of conditions.

Modeling in mammals. While mathematical models for large scale biological phe-
nomena have existed for several hundred years (see references in V. Volterra 1931, N.
Rashevsky 1938), significant advances in the use of models have been made only in the
past two decades. The reason for the slow buildup was the need for large-scale, high-speed
digital computers and numerical methods to simulate the very complicated difference
and differential equations which occur in models of most biological phenomena.

Mapleson (1963) successfully used an analog computer method to describe the uptake
and exchange of inert gases and other agents by mammalian tissues. Bellman et al. (1963)
have developed several computer simulators for blood flow and the distribution of drugs
injected intravenously. The simulators were based upon principles developed in the classic
Transport Phenomena by Bird et al. (1960).

Technical Paper No. 3423 of the Oregon Agricultural Experiment Station.

IDieldrin will be used to represent HEOD (1, 2, 3, 4, 10, 10-hexachloro-6, 7 -epoxy-l, 4, 4a, 5, 6,
7, 8, 8a-octahydro-1, 4:5, 8-endo, exo-dirnethanonaphthalene) which constitutes at least 85% of the
active ingredients in technical dieldrin.

Archives of Environmental Contamination

and Toxicology, Vot. 2, No. 1, 1974
© 1974 by Springer-Verlag New York Inc.
10 F . T . Lindstrom et al.

More recently analog and digital computer methods have been brought to the fore-
front and summarized by Bischoff and Brown (1966), who have reviewed the basic con-
cepts of compartmental models for mammalian systems from the engineer's point of
view and then investigated and described the pharmacokinetics of thiopental injected
intravenously into dogs (Bischoff and Dedrick 1968). Predictions from these mathemati-
cal models plotted on the same graphs as the experimental data for each major tissue
region show excellent agreement. Bischoff and Dedrick (1970) have summarized the two-
compartment model schemes for drug distribution and discussed how the method of
dosing or injection can affect subsequent distribution of the drug. The pharmacokinetics
of methotrexate, a cancer chemotherapeutic drug, have been extensively studied by
Bischoff et al. (1971) as a model of water-soluble drugs.
A recently published "state of the art" book on mathematical modeling via compart-
mental analysis (Jacquez 1972) provides a wealth of ideas on the modeling of numerous
drugs in mammalian systems together with the mathematics necessary to justify the com-
putational approach.
Dieldrin pharmaeokineties. Walker et al. (1969) and Robinson and Roberts (1969)
carried out intensive and thorough studies of the toxicology and pharmacology of
dieldrin in rats and dogs. These experiments covered the effects of dieldrin administra-
tion for two years or longer. At about the same time, Hunter and Robinson (1967) ana-
lyzed in detail the distribution of dieldrin in men of various ages and weights. Hunter
et al. (1969) followed up the intoxication segment of the experiments on man with an
eight-month post-exposure analysis. In all these programs efforts were made to interpret
the observed data by simple one- and two-compartment models or by empirical curve
fitting (not physical modeling from fundamental physical and chemical principles); and
various degrees of success were attained.

Finally, Baron and Walton (1971) conducted a very interesting and informative experi-
ment on the dynamics of dieldrin in adipose tissue of rats. Their data from experiments
with rats given radioactive dieldrin suggest a great dependence of the observed distribu-
tions on the method of administration, as well as on the time lapse between administra-
tion and analysis.

T h e lipid-phase m a m m a l i a n m o d e l
Dieldrin is highly lipid-soluble: the partitioning ratio is 300,000:1 between olive oil
and water (Wauchope 1972) and 34,000: I between hexane and water (Voerman 1969).
Most studies indicate that the dieldrin mass in a tissue region is roughly proportional to
its lipid content. The relationships of the compartment lipid fractions of an animal there-
by form the basis for a conceptual scheme of dieldrin distribution. To the extent that
other compounds have properties similar to those of dieldrin, such a concept should
serve to explain and predict their pharmacodynamics.
The model described herein allows prediction of lipid-phase concentrations of dieldrin
in major tissue regions of mammals. Bischoff and Brown (1966) formulated a conceptual
scheme with respect to a tissue region model (model e in Figure 4, p. 34) which is
adapted in Figure 1 to show the blood flows and large region compartments of the lipid-
phase mammalian model. The organ perfusion rates (liters of blood/hr) are translated
 Dieldrin Distribution Model 11

QB MB, U B (Heart + Lungs I ,.. Qg

+ Large Arteries and Veins) 1

QOT QM QF QBR QL QG! QK

~t r OF
1¢
MGI,UGt q Rex
MOT,
UOT
(other)
MM,
UM
(muscle)
MF,
UF
(fat)
MBR,
UBR
(brain)
l,,, !G,, 1 U~ } ~
• (kidr j
(Urinary
excretion)
!
QGI
~r

M c , UL
QF QBF "l (liver) ,,,,,,,P QK
QOT QM • (Metabolism)

--'IIQGI + QL
<

s ) I
I
(Lumen)
QBI
l
OF
(I-O)F
i ~ (Fecal
excretion)

Fig. 1. Diagram of blood lipid flows and tissue lipid compartments of the "mini-
mammal."
into equivalent blood lipid flows (Qx, kg of lipid/hr) for the tissue compartmental lipid
mass (Mx, kg lipid/tissue) containing the mass of dieldrin (Ux, g of dieldrin/tissue).
Regional blood lipid flows through and lipid masses of specific compartments are indi-
cated by subscripts: B (general blood pool, including the chambers of the heart, lungs,
arteries and veins), L (liver), K (kidneys), BR (brain), GI (gastrointestinal tract), F
(adipose tissue), M (muscle, including associated fat), and OT (other, including such as
skin, bone, other organs). The special terms, dieldrin source function ~_,and dieldrin de-
struction rate coefficient P will be elaborated later in the text.

The assumptions on which the model system is based are:

(I) Dieldrin is transported almost exclusively via lipids and interacts with other
molecules only weakly;
(2) Dieldrin transfer from blood to lipid-containing tissues is very rapid [A flow
limited model in the sense of Deddck and Bischoff (1968) is assumed] ;
(3) Metabolism of dieldrin is carried out exclusively in the liver at rates relatively
much slower than rates of transfer of dieldrin from one lipid compartment to
another;
12 F . T . Lindstrom et al.

(4) Dieldrin, being tied to lumen lipids, is almost totally absorbed from the intes-
tines via lipid absorption (Tidwell 1958) and not readily excreted in feces and
urine;
(5) Tissues can be assigned to large compartments of generally similar constitution;
(6) Each compartment behaves as a well-stirred chemical reactor;
(7) The complete open system is representable by a system of mass balance equa-
tions; and
(8) The blood lipid flow (Qx) and the compartment lipid mass (Mx) functions are
continuous functions of time (0 ~< t < ~o).

Note that these assumptions admit to the possibility of being relaxed, in certain instances,
without leading to disintegration of the model. Assumption (3) admits to the possibility
of liver enzyme induction or a time dependent destruction rate coefficient P. Assumption
(8) allows for time- or status-dependent changes in compartment sizes and flow rates
(e.g., a growing animal ingesting dieldrin). Additional assumptions will be noted in regard
to discussion of the source function ~ and destruction rate coefficient P.

Proceeding from these assumptions, the following system (1) of mass balance equa-
tions (Eq. la-i), derived in accordance with Bischoff and Brown (1966), provides the
basis for calculation of dieldrin tissue distributions (see Table I for compartment lipid
mass and lipid flow definitions).

F
dUG! = 0 ( ~ + ~ QBI UL UB - UGI
(a) dt ~ ) + QoI MB QcI MoI (G. I.)

(b) dUL UGI UB UL

dt = QGI --MGI + QL MB - (93 QBI + QGt + QL + IP) ~-~-
IVl L
(Liver)

dUK UB UK
(c) dt = QK ~ - (QK + Rex) G (Kidney)
(1)
dUBR UB UBR
(d) dt - QBR MBB- QBR MB
R- (Brain)

dUF UB UF (Depot fat)

(e) dt = QI+ ~-~B- QF

dU M UB UM (Muscle)
(f) dt = QM MBB- QM

dUoT UB UOT
(g) dt - QOT ~-7--- QOT (Other tissues)
lvlB MOT
 Dieldrin Distribution Model 13

dU B UL UK UBR UF
(h) dt (QG| + QL) MLL+ QK"~K + QBR MBR
-- + QF MF

UM UO----LT QB UB
+ QM MM + QOT Mo T ~ (Blood pool)

(i) QB = QG1 + QL + QK + QB R + QF + QM + QOT (Conservation of flow)

Looking briefly at Equation (a) in System (1), the reader will note that the instantaneous
time rate of change of dieldrin mass (gms dieldrin) in the gastrointestinal compartment
(dU G ffdt)is the sum of the new dieldrin mass in grams per unit time (OF) coming into ttle
system from the lumen and the mass flow of dieldrin QGffUB/M B being brought
back into the G. 1. tract from the general blood pool minus that mass of dieldrin

Table I. Physiologic and Anatomic Symbols Shown in the Flow Sketch

and Those Used in the Mass Balance Equations

Compartment Blood lipid flows Lipid mass

kg lipid/hr kg lipid
Blood chambers of heart, lungs, and
large veins and arteries (General
Blood pool) QB MB
Kidneys QK MK
G.I. QGI MGI
Liver QL ML
Brain QBR MBR
Fat (depot) QF MF
Muscle (includes marble fat) QM MM
Others (skin, bone marrow, etc.) Qo T Mo T

Special symbols:
QBI = Bile lipid mass flow (gms lipid/hour)
Rex = Renal dieldrin excretion coefficient (gins lipid/hour)
0 = Lumen-to-G. I. blood lipid dieldrin absorption coefficient
(dimensionless) 0 ~< 0 ~< 1
UL
F = 5 + R~QBI MLL = dieldrin mass transfer rate into and out of lumen (gms dieldrin/hr)

= dieldrin source function (defined in text)

,2t = effective dieldrin binding coefficient to bile matter (dimensionless when viewed
from lipid system)
14 F . T . Lindstrom et al.

(QGI-UGI/MGI) being lost to the liver via the portal vein. The other compartments fol-
low suit accordingly.

Compartment mass and blood flow estimation. In order to use the model system (1),
the values of all the blood lipid flows (Qx's) and the compartment lipid masses (Mx's)
must be known. Table II presents a summary of the various formulas used in estimating
the gross or total compartment mass (922x~S) and the whole blood flow rates (qx'S) through
the respective compartments. These estimates may be subject to considerable error due
to the scantiness of information in the literature on certain tissues; blood flow data
through adipose tissue are almost non-existent. Pending additional physiological findings
of the future, support for proceeding from current estimates is drawn from the statement
of Bischoff et al. (1971): "It was found in previous work that consistency of values for
the organ volumes [masses] is more important than absolute accuracy, which is the
reason for deriving the general [power taw] equations."

With estimates of compartment masses and blood flow rates it is now easy to calculate
the compartment lipid masses and the blood lipid flow rates. Define the respective com-
partment lipid masses as:

M x = fx • 2)~x

and the respective blood lipid flow rates as:

Qx = f13" qx

where fx is the lipid fraction (gms lipid/gm tissue) of that respective compartment or
tissue region. The subscript " x " assumes the same symbols as are shown in Table II.

For the remainder of the paper the words or phrases "coefficient(s)," "flow co-
efficient," and "blood lipid flow coefficient," will be used interchangeabty. The word
"coefficients" may at times be used to designate all (a set) of the compartment lipid and
blood lipid flow values.

Obviously these coefficients are highly dependent on species, age, sex, nutritional
status and other factors, but form a consistent system, capable of adaptation and limited
principally by the availability of data on the animal and of that status. For simplicity
these coefficients may be assumed to be constant (undergoing negligible change) in the
mature individual, such as an adult male rat or man. Alternatively, these coefficients in
any or all compartments may be allowed to vary continuously in a regular manner to
account for the physiologic changes relative to growth, diet, etc., and/or the toxicologic
effects of chronic exposure to an environmental agent. Furthermore, with additional
assumptions it would be possible to simulate the interaction of two (or more) agents
having properties consistent with this model through simultaneous, parallel representations.

It is also possible, by adding parallel compartments across the major blood lines, to
add important compartments such as the uterine/fetal system and the mammary glands
 Table II. Method of E~timating Compartment Masses and Blood Flows (B. W. = Body Weightin Kg).

Gross compartment mass Whole blood tissue flowa

Symbol (gms) (gms/hr)
Compartment (x) ~))~x qx

(Formula used) (Formula used)

Blood pool (blood B 77 (B. W.) o.99 , AA and HH 12,600 (B. W.) o,81 , BB
chambers of heart,
lungs, arteries,
and veins)
Kidney K 7.5 (B. W.) TM, AA and HH 2,038 (B. W.) o.89 BB
Gastrointestinal GI 49 (B. W.) TM, BB 2,312 (B. W.) o.87, BB
(small bowel)
Liver L 34 (B. W.) o.aT, AA and HH 2,820 (B. W.) o.aT, BB ©

Brain BR 8.0 (B. W.) 1,2, CC and DD 2,600 (B. W.) °.s2, CC and EE
©
Depot Fat F 180 (B. W.) 1.°, CC, DD, and II 124 (B. W.)O.84, CC
Muscle (includes M 400 (B. W.) 1.°, CC, FF, and II 1,484 (B. W.) o.Ss, BB
associated fat)
Other tissue OT 200 (B. W.) 1.o, CC and DD 371 (B. W.) T M , CC
(skin, etc.)

aAssumed blood density of 1.05 gm/cm 3 and an assumed hematocrit of 40% (Guyton 1964).
AA = Ad••ph(•949);BB=Bisch•ffetal.(•97•);CC=Auth•r`sestimati•n;DD=Bard(•96•);EE=A•t•nanandDittmer(•97•);
FF = Ashworth and Cowgill (1938); GG = Mapleson (1963);HH = Brody (1945); II = Pace and Rathbun (1946).
16 F . T . Lindstrom et al.

of female mammals. Because such additions require only estimation of lipid-phase

transfer to these compartments, they pose no special difficulty in the model. Each of
these particular compartments also constitutes a "sink" for dieldrin removal and becomes
increasingly significant in that regard as gestation proceeds to parturition and on to
lactation.
Dieldrin source function ~ and absorbtion coefficient O. Due to a current lack of
exact knowledge of the mechanism(s) of dieldrin movement from the lumen into the
internal environment, and consequently of the time-dependent, quantitative transfer
rates, it is necessary to assume a source function ~ (gg dieldrin/hr) to be the lumen input
of new dieldrin delivered from the stomach and a time-dependent lumen-G.I, tract
dieldrin absorption coefficient 0. For the remainder of this exposition 0 is assumed to be
synchronized with the feeding schedule of the concerned mammal. When lipids are pres-
ent in the lumen, it is assumed that 0 is -~ 1 and when lipids are absent 0 -~ O. This is
obviously a daily or periodic phenomenon going on regardless of dieldrin. 0 can assume
any real number value between zero and one. 0 then is seen to play a fundamental role in
the uptake and distribution of dieldrin in mammalian tissues.

As would be expected (Bischoff and Dedrick 1970) and as will be demonstrated again,
the character of ~ also has highly significant effects on the overall compartmental distri-
butions of dieldrin. For the purposes of this exposition ~ is assumed to be linear with
respect to dietary concentration and food intake rate.

One facet of dieldrin movement within the mammal is not treated in this model-the
salivary-gastric-jujenal secretion route (Cook 1970). By assuming almost complete ab-
sorption of dieldrin from the lumen (0 = 0.995, while lumen lipids are present, assump-
tion 5), the outputs through the bile and the other gastrointestinal secretions are assumed
to re-enter as a part of the GI source function OF. This cycle, which sends some dieldrin
back into the GI lumen to be added to the present dieldrin mass in the lumen, presents
many more questions relative to mechanisms of secretion and absorption than can be
answered on the basis of current knowledge or treated in this exposition. The assumption
that dieldrin absorption is almost complete is based upon knowledge (Heath and
Vandekar 1964) that a very small portion of any dose (less than a fraction of 1%) may be
carried out in the feces or urine by epithelial exfoliation or by entrapment or solution in
lipid or protein of bacteria.

Bile flow coefficient QBI and dieldrin-bile lipid binding 2. In most mammals the bile
fluid does not flow continuously, but rather flows intermittently throughout the day in
synchronization with the meals eaten and/or dietary input of food to the stomach. Differ-
ences in bile flow into the G.I. tract also would be seen between species without a gall
bladder (e.g., rat) and those with (e.g., man, pig). The bile lipid flow coefficient QBI
(gms bile lipid/hr) then has some definite fixed phase and amplitude relationship to food
intake times. The exact values of this phase and amplitude are obviously dependent on
species, age, sex, etc., and can at best be only approximated apriori.
The dieldrin-bile lipid binding coefficient ~3 is defined as an "effective" binding co-
efficient for dieldrin to the lipid (micellular?) portion of the bile fluid. Its value appears
to be in the range 50 ~< ~ ~< 200 for dieldrin and of the order of 100-1000 for endrin
 Dieldrin Distribution Model 17

(Cole et aI. 1970). That dieldrin and endrin differ in their ~ values is expected on the
basis of their fundamental chemical structure differences. ~ is therefore assumed to be
compound-specific.

Destruction rate coefficient P. The sole significant output of dieldrin considered by

this model is metabolic destruction [Eq. (1)b] at a rate determined by the destruction
rate coefficient F and the concentration of dieldrin in liver lipid (UL/ML). As for the
source function, the assumptions necessary to develop a basis for F must be derived from
fragmentary current knowledge and are critical in the resultant modeling of the com-
partmental distributions of dieldrin.

The metabolism of dieldrin is illustrated in Figure 2. There are quantitative differences

between mammalian species and distinct problems in interpreting the disposition of
metabolites and their conjugates by fecal and urinary elimination. Three basic routes of
breakdown are involved in the model: (1) microsomat, NADPH-dependent oxidation/

cm / ct \

\T / --</,o ':A2"o'°:'. o

III
uI
Diledrin
\ ~ (NI
I.A"
Ct "~
DPH,02)
"~icrosomes , t
(cytoplasm?) C l i o ~UDPGA
H20"~
CIt~Cl ~HO
£ Glucuronide
9-OH-Dieldrin conjugate
el OH el
Ct~--~-~ CI H C l ~ °
UDPGA~

Cl Glucuronide~ O Klein'smetabolite
Trans-6,7-dihydroxydihydroAldrin conjugate /

~ UDPGA

Glucuronide
conjugate
Fig. 2. Proposed major liver metabolic pathways for HEOD (chemical).
18 F . T . Lindstrom et al.

dehydrohalogenation/cyclization to form a hydroxy derivative which may be conjugated

to the glucuronide (Matthews and Matsumura 1969) or oxidized to the keto-compound
called Klein's metabolite (Klein et al. 1968, Daminco et al. 1968), the major urinary
product in rats and sheep; (2) hydroxylation by an NADPH-dependent microsomal
enzyme to form 9-hydroxy-dieldrin (Fell et al. 1970, Baldwin et al. 1970), which is the
major fecal metabolite in the rat (Richardson et al. 1968) and which may be conjugated
to the glucuronide (Matthews et at. 1971) or which might be oxidized to a 9-keto-dieldrin
(c.f. metabolite M-3 of Matthews and Matsumura 1969); and (3) hydrolysis of the
epoxide ring to form the optically-active aldrin-6, 7-trans-diol (Korte and Arent 1965),
which may be excreted in both the urine and the feces or may be conjugated as the
gtucuronide (Matthews and Matsumura 1969).

Any of a number of drugs, chlorinated hydrocarbon chemicals (including dieldrin)

and various natural products (including steroids) induce proliferation of the hepatic
endoplasmic reticulum (Conney 1967) and result in highly increased activity of diverse
microsomal mixed function oxidases. Many of these same inducers (DDT, phenobarbital,
3,4-benzpyrene) stimulate dieldrin metabolism and cause increased urinary excretion of
dieldrin metabolites (Street 1969). The question of the possibility (Durham 1969) of
chronic dieldrin exposure affecting the disposition of dieldrin residues therefore becomes
quite important in modeling considerations. Evidence (Eliason 1971) argues against auto-
induction of dieldrin metabolism, since placental transfer of dieldrin is decreased by
phenobarbital (a known inducer of dieldrin metabolism), but not by pre-treatment with
dieldrin itself. However, direct measurement of enzyme levels and activities has not been
reported for either the "normal" or inducer-treated animals.

On the basis of several studies with rats (Kinoshita et al. 1966, Gillett and Chan 1968,
Gillett 1968) and swine (Gillett et al. 1971), it has been concluded that the extent of
induction in short-term feeding trials is directly proportional to the dietary dosage of a
given chlorinated hydrocarbon insecticide. The slope of the dosage-response curve varies
between compounds (Gillett 1971) acting as inducers, for different enzyme activities,
and for different species. The problem is further compounded by variation in response
between different preparations of microsomes under the same regimen (c.f. Matthews
and Matsumura 1969).

From these facts it may be assumed that there are no concentration-dependent steps
intervening between movement of dieldrin into the rat from the diet and the dieldrin-
dependent rate of enzyme synthesis, presumably associated with genetic expression via
derepression in the nucleus. It is also apparent that, if dieldrin metabolism is not induced
directly, it will at least be affected by changes in microsomal structure associated with
induction of other microsomal activities by dieldrin. Holtzman and Gillette (1968)
established that phenobarbital inhibits phospholipid turnover in hepatic microsomes
without stimulating lipid synthesis, thus increasing the amount of lipid available. Protein
turnover on the other hand is unaffected, so that protein accumulates due to increased
synthesis. Davis et al. (1973) find a constant ratio of protein to lipid (0.55 gm lipid/gm
protein) in microsomal membranes regardless of the state of induction.
 Dieldrin Distribution Model 19

A number of studies have indicated the reversible nature of this xenobiotic-stimulated

induction. Cessation of exposure results in a decline of the induced activities back to
normal levels. Structural changes may persist beyond the point where activity is declining
(Ortega 1969, Gillett et al. 1971). However, Davis e t a l . (1973) note a maximum in mass
of microsomal protein after 20 to 30 days of exposure of rats to 150 ppm of DDT; the
protein level thereafter declines gradually to near normal levels. These same authors
suggest that inducers effect increased synthesis of individual proteins or groups of pro-
teins at different rates, enriching the membranes in a time-dependent manner.

A schematic model of dieldrin metabolism (shown in Figure 3) involves several assump-

tions: (1) the 3-hydroxylation/cyclization (El) is an inducible microsomal enzyme,
(2) 9-hydroxylation (E2) is an NADPH-dependent, microsomal enzyme but not inducible
(see Matthews and Matsumura 1969 and Matthews et al. t971 regarding the peculiar
properties of this reaction), (3) cyclodiene epoxide hydratase (E3) is cytoplasmic and
non-inducible, (4) the dieldrin metabolites are neither inducers nor inhibitors of other
reactions affecting dieldrin metabolism, (5) dieldrin does not induce any totally new
activities nor does it (allosterically) alter the rate constants of any reaction involved in
dieldrin metabolism, (6) adequate amounts of amino acids and lipid are available for
synthesis of new membranes, (7) membranes are formed with a constant phospholipid/
protein ratio (Davis el al. 1973), and (8) dieldrin does not affect catabolic rates of
proteins in the membranes.

Liver Microsomes
kc! I I t ~ , 1 V Breakdown
QL L

QGI 12 Die,dr,n
I koI / metabolite
QL QG! Lv UL !
4 ! ! !
l
m
9 H-Died . . . .

- ' , \ Fir ! 0 '

I •
I i"
.... N, ..........
1
-' ÷

Fig. 3. Schematic diagram of assumed liver dieldrin movement and metabolism.

....... movement of dieldrin, - - - - induction control, and . . . . metabolite exit
from model.
20 F . T . Lindstrom et al.

In view o f the preceding assumptions on liver dieldrin metabolism, let the time rate of
change o f the ith enzyme be defined as

dPi _ UL(t)
dt kcpiPi + kcpiPi(O) + ~TmP ML(t ) (2)

where it is assumed that Pi(0) =/= 0, i = 1, 2, 3 . . . N (see Table 1II). The solution to equa-
tion (2), for each i, in convolution form is

Pi(t) = Pi(O) + ~ o m p do t M ~ ~ exp(- k~ p i (t- r))dz (3)

Table III. Destruction Rate Coefficient Nomenclature

Liver Cell Protein Masses; Pi , i = 1, 2, 3, . . . , N* (gms)

P1 inducible**, active***, microsomal enzyme; P2 non-inducible, active, microsomal
enzyme; P3 non-inducible, active, cytoplasmic enzyme; P4 inducible, non-active, micro-
somal protein; Ps non-inducible, non-active, microsomal protein (remaining microsomal
protein)

Liver Cell Lipid Masses (gms)

MMs (microsomal); MCy (cytoplasmic); MN (nuclear); MMc (mitochondriat)

Compound Specific Induction Coefficients; (gm protein)

gm dieldrin
hr
gm lipid
•comp
Pi ,i = 1, 2, 3 , . . N; also **** below.

Liver Cell Protein Catabolism Coefficients: (1/hr)

kcpi, i = 1, 2, 3 . . . N; also **** below.
1
Compound Specific Proportionality Coefficients; ( )
gm protein
• hr
gm lipid
kDi , i = 1, 2, 3 . . . N; also **** below.

*N = 5 for dieldrin. N may be larger or smaller for other compounds.

**Inducible is defined here as a direct increase in the protein synthesis rate.
***Active means dieldrin-metabolizing ability.
****Note that in view of the available literature on dieldrin metabolism in mammalian
tissues (liver mainly), only/3p I and ~P4 =~:0. Likewise, only kDi, i = 1, 2, 3 4: 0.
 Dieldrin Distribution Model 21

The total microsomal protein is simply

PMs (t) = Z Pi(t) (4)

(summing microsomal
proteins only)

As noted by Davis et al. (1973), the microsomal lipid mass MMs(t) is in fixed proportion
to the microsomal protein PMs(t) so that

PMs(t)
MMs(t) a, a constant (5)

The concentrations of the various microsomal enzymes Ei, (i ranging over microsomal
proteins) is simply

Pi(t)
Ei(t) - M--~-s(t) (6)

and the cytoplasmic enzyme concentration Ej (j ranging over cytoplasmic proteins) is

Ei(t) _ Pj(_~t) (7)

mC y

The total liver lipid mass is defined as the sum

ML (t) = MMs (t) + Mcy + MN + MMc (8)

Postulating the microsomal destruction rate to be proportional to the active enzyme

masses present yields

I?Ms (t) = Y]. kDiMMs (t)Ei(t) (9)

O)
(active microsomat)
or

FMs(t) = Z kDiPi(t) (t0)

0)
(active microsomal)

The form that PMs(t)assumes then is simply

rMs(t) = (11)
0)
(active microsomal)
22 F . T . Lindstrom et al.

Since the cytoplasmic destruction rate coefficient Pcy is assumed to be proportional to

the total active cytoplasmic enzyme mass, the cytoplasmic destruction rate coefficient is
represented by

rcy = ~] kDiPi(t ) (12)

(i)
(active cytoplasmic)

The total liver dieldrin destruction rate coefficient F(t) is

P(t) = l'cy (t) + FMs (t) (13a)

or

N N

r(t)= kDiPi(0) + Z kDiPPi

ocompfo t{UL(~')}exp(_kcpi(t_7))d
~M 7
i= 1 i= 1
(active protein) (13b)
Upon substitution of P(t) into equation (lb) (the liver compartment mass balance
equation) the model system becomes a coupled system of non-linear integro-differential
equations which, because of the complicated non-linear nature of the system, appears
unsolvable in closed form (i.e., a sum of known exponential functions). We now turn our
attention to a numerical simulation method approximating system (1) subject to
equation (13b).

Simulation of the model system. Because equation system (1)subject to equation (13b)
is actually a non-linear integro-differentiat system, the classical solution methods, such
as linear superposition of eight exponential functions together with an additive constant
term (Bischoff and Brown 1966, Jacquez 1972) or Laplace transforms, are no longer
valid. The methods of perturbation theory, while valid for small deviations from the
linear case, are not exploited here, since we have no a priori knowledge of the magnitude
of induction coefficients in the definition of P for all combinations of lipid-soluble drugs
and mammalian systems. To be sure though, if there are no induction effects (i.e., all/3's
are zero), then the system reduces to the linear model, which, if the mass and flow co-
efficients are well-behaved functions of time, is in principle solvable exactly by classical
methods.

However, let us approximate System (1) by a finite difference simulator. The mass
balance equations are then approximated (in matrix equation form) by the equation

AU
- AU + S(t) (14)
At

where
 Dieldrin Distribution Model 23

f "h

0 0 0 0 0 0
o o o o

0 0 0 0 C~ g~ 0

0 0 0 0
c'l= 0 o O'l;~
i

0 0 o ~ 0 0 o ~

0 0 0 0 0 0

4--
,._]
0 0 0 0
o +o,
O'
+~

~2 0 Q 0 0 0 0

J
II
<
24 F . T . Lindstrom et al.

U = (UGI , UL, UK, UBR , U F , UM, UOT , UB)T

and

~ ( t ) = (0~(t), 0, 0, 0, 0, 0, 0, 0) T

w4th T denoting transpose. The numerical simulation is now accomplished by using a

backward difference in time on AU]At. This is one of the classical Pade approxima-
tions, which for F > 0 yields an unconditionally stable implicit difference scheme (Varga
1962, p. 265). It is also of the order of At2 accurate locally [O(At:)] and O(At) globally
(Varga 1962, p. 263).

U(tn) = U(nAt) = U n, n = 0, 1, 2 . . .

provides

U n + 1- U n
- - - A(n)Un+l 4~n+1 (16)
At

which upon solving for Un+l yields

Un+: = (I-A(n)At)-I{Un +~n+l At} (17)

Note that the superscript n on the A and U matrices means that the explicit time depen-
dent elements of A are evaluated at the point tn+ 1 = (n+l)At, and the implicit elements
are evaluated at t n = n a t in time. Equation (17) is then the actual simulator, in matrix
form, of the dieldrin distributions in the respective compartments. In addition to
equation (17) the difference analog of equation (2) together with equations (4), (5), (8),
and (13a) must be used to characterize the changing destruction rate coefficient F(t). Any
growth changes, either in compartment lipid mass Mx or blood lipid flow Qx, must also
be accounted for by evaluating new mass and flow parameters at each iteration step.
This latter type simulator is readily programmable for any high speed digital computor of
reasonable storage capacity and access time.

The explicit listing of the difference analogs mentioned above for dieldrin are:

comp U ~
pn + kcpi Atpo + flPi At
pn+ 1 = , i = 1, 2 . . . . ,5 (18)
1 + kep i At

5
pn+l
Ms
= E p--in + l _ p~+l (19)
i=t
 Dieldrin Distribution Model 25

Mn+l 1 p~+l (20)

MS =~-

M~+ 1 -raMs
_ un+ 1 +M N +Mcy+MMc (el)
(assumed constant)

3
pn+l = tMsrm+l+ tCyr~n+l= E k D i Pn+l (22)
i=l

Equations (17) to (22), together with the initial condition Uo = U(t = 0) known,
constitute the mammalian lipid-phase dieldrin distribution model.

Discussion
The purposes of developing the mammalian lipid-phase model of dieldrin pharmaco-
kinetics are directed toward interpretation of available data and application of that data
for subsequent experimental design, testing of hypotheses, and projection of potential
hazard to human populations. The suitability of the model for these purposes therefore
is derived from the validity of the assumptions and a priori knowledge of the relationships
implicit in the structure and function of the model.

The lipid-phase mammalian model of highly lipid soluble compounds can now be
exploited for a myriad of purposes relating known body physiology, dieldrin bio-
chemistry, and the dieldrin administrative method to body dieldrin distribution and
toxicological status. The extent to which it provides a reasonable approximation of ob-
served experimental dieldrin distributions in mammals will be considered in subsequent
papers. Additionally, the model may be used to explore the effects of (1) continuously
changing compartment sizes, e.g., growth, disease or nutritional status; (2) alterations in
the schedule and/or route of administration, e.g., dietary dieldrin as opposed to a single
gelatin capsule of dieldrin; (3) co-exposure to an agent affecting dieldrin metabolism or
the metabolism of which might be affected by dieldrin, e.g., dietary dieldrin plus pheno-
barbital; and (4) contributions by the several mechanisms of intestinal uptake of dietary
dosages and their relationships to internal cycles, e.g., enterohepatic and salivary-gastric-
jejunal secretory.

Application of the model to several of the simplest systems will serve to illustrate some
of the types of information that can be derived, thereby demonstrating its general
applicability. Simple a priori physiological considerations and data allow calculation of
the compartment and flow rate parameters for a given case.

Case I. Mature male rat, no auto-induction. A typical 300-gm mature male rat, pre-
viously brought up on a dieldrin-free diet, is the first example. Assume all Q's and M's
are fixed in time and that the dieldrin destruction rate coefficient (P) is constant (all
/Ys = 0). Suppose that this hypothetical rat is eating about 20 gms food daily (Baron
26 F . T . Lindstrom et al.

1972) over a 12 hour period and that the dietary dieldrin concentration is 25 ~g
dieldrin/gm food, giving® = 41.67 #g/hr over the 12-hr feeding period. During the re-
maining 12 hours the rat essentially eats nothing so<~ = 0 for this portion of the day.
This scheme is assumed to be repeated day after day as long as the administrator desires
(periodic function of time). The tissue lipid fractions (fx) are derived from Long (1961),
Williams and Landsford (1967), and Spector (1956). The calculation of compartmental
lipid masses (~-~)~x " fx = Mx) is shown in Table IV along with the flow parameters
of the tissue (Qx)-

After about 4 weeks on this particular dieldrin administration scheme the adipose
tissue residues have built up in a dynamic oscillatory fashion to almost the "steady state"
value. Figure 4 shows example curves of this oscillation about a steady mean value for
both the blood lipid dieldrin and depot fat lipid dieldrin concentrations. In order to be
able to generate the curves shown in Figure 4 an estimate of P (constant in this example)
must be obtained, as explained below.

At "steady state" conditions, following feeding of rats for several weeks, Walker e t al.
(1969) and Baron and Walton (1971) found depot fat dieldrin concentrations of about
50 t~g dieldrin/gin adipose tissue at a 25 ppm dietary dieldrin concentration. Assuming a
lipid fraction fF of 0.637 and knowing that the depot fat lipid phase dieldrin concentra-
tion C~, (/ag dieldrin/gm lipid) is related to the depot fat tissue dieldrin concentration C v
(gg dieldrin/gin adipose tissue) via the simple formula

CF
C~ fv

a value of 78.5 #g/gm lipid is calculated for the depot fat lipid phase dieldrin concentra-
tion C{. It is assumed that C!~ -- 78.5 /Jg/gm lipid is the mean value about which the
depot lipid dieldrin concentration oscillates. Also, it is assumed here that the liver lipid
mean dieldrin concentration is about the same value. Since this value is then constant,
the liver must be destroying the daily 500 ttg intake (neglecting the small amount not
absorbed, but passed out of the lumen via the feces). Hence, we estimate F from the
destruction formula

UL
#g dieldrin destroyed/day = F - -
ML

UL
with ~ = 78.5 (by assumption) and 500 ~g being the daily amount destroyed. This
simple ~omputation gives a r value of 6.37 gms lipid/day or 17= 0.27 gms lipid/hr.

The appropriate elements of the A matrix are calculated from this value of P and the
other parameters (Table IV), using a At of 0. I hr. The inverse iteration matrix is calculated
and substituted into equation (17), forming the simulator. Assuming no dieldrin present
initially (UO = 0, zero column matrix), the iteration scheme generates the typical blood
lipid and adipose lipid phase dieldrin concentration curves (Figure 4) over the first 40
 Table IV. Compartment Mass, Blood Flow, and Other Important Parameters
For Hypothetical 300-gin Mature Male Rat
Lipid fraction Lipid mass Blood flow rate Blood lipid
Mass fx Mx qx flow rate Qx
Symbol ~,}~x (gins lipid~ (gms b lood~ (gins blood lipid.)
Compartment x (gins) .g~ue~ (gins lipid) hr hr

Blood pool (blood chambers B 23.4 0.004 0.09 4,751.6 18.21

of heart, lungs,
arteries, and veins)
Kidney K 2.7 0.02 0.05 698.0 2.79
Gastrointestinal (small bowel) G.I. 15.8 0.07 1.11 811.1 3.24
Liver L 11.9 0.06 0.72 989.3 3.96 ~.
o-~
Brain BR 1.9 0.09 0.17 680. t 2.72 =
e-b

Depot fat F 54.0 0.64 34.56 710.1 2.84 c

Muscle (includes M 120.0 0.07 8.40 533.3 2.13 o
associated fat)
Other tissue (skin, etc.) OT 60.0 0.11 6.60 133.3 0.53

Rex = 0.0 (Heath and Vandakar 1964)

~ = 100.0 [estimated from data of Cole et al. (1970)]
0 = 0.995, 0 ~< t < 12 h°urS}'rep eatablet daily (periodic function)
0 =0.050, 1 2 ~ < t < 2 4 h o u r s ~
QBI = 0.0032 gm bile lipid/hr, 0 ~< t < 12 hours}
[ periodic function (24-hour period)
QBI 0.00032 gm bile lipid/hr, 12 ~ t < 24 hours~
NJ
--.1
 b3
oo

I00 Blood

oo?
-E
-S="--~ 10

13
'F,

; 1; 1; 2; 2; 3~ ~ 2o 4; 5'o & 60
T i m e (days)

Fig. 4. Lipid phase dieldrin concentration in adipose tissue and blood buildup and decay curves for a 300-gm male rat fed 25 #g
dieldrin/gin food in the diet and eating 20 gms food daily.
 Dieldrin Distribution Model 29

days while dieldrin is being fed. On Figure 4 the blood lipid phase dieldrin concentration
is seen to rise very rapidly in the first several days to give a maximum value of about
33 og/gm lipid at day 5 with a corresponding minimum value of about 25/~g/gm lipid,
then averaging about 30 /~g/gm lipid. By day 40, it appears that residues have about
reached a "steady state." The blood lipid dieldrin concentration has a maximum of about
80 og/gm lipid, a minimum of about 65 #g/gm lipid, and an average of about 72 #g/gm.

On the other hand, the adipose tissue lipid phase dieldrin concentration at day 5 has a
maximum of about 32 ~g/gm, a minimum of about 26/~g/gm, and an average value of
about 28/1g/gin. The "swing" on the adipose tissue lipid phase dieldrin concentration is
not as pronounced as in the blood lipid, understandably due to the large "inertia, poor
perfusion" effect that the adipose compartment presents to the system as a whole. On
day 40 the lipid phase dieldrin concentration in the adipose tissue has a maximum of
about 75 og/gm lipid, a minimum of about 72/~g/gm lipid, and an average value of about
73/ag/gm lipid. From the viewpoint of the adipose tissue we have approximately reached
the "steady state" residue level. It can be shown mathematically that under the present
dieldrin administrative scheme (12 hours on - 12 hours off) the average adipose residue
level of 78.5 #g/gm lipid (Walker et al. 1969, Baron and Walton 1971) will never be
reached. It would be possible to reach this value if,~, 0 and QBI were all fixed constants
for all time.

Turning our attention to the decay portion of the curves in Figure 4 we obtain the
following information. At day 50 (10 days after cessation of dietary dieldrin) the blood
lipid phase lipid phase dieldrin concentration has dropped to the value 19/Jg/gm lipid,
while the adipose tissue lipid phase dieldrin concentration has dropped to 21 /~g/gm
lipid. This gives an adipose tissue (and hence body burden) half life of about six days for
dieldrin in our hypothetical rat. Equivalently, a value of F = 0.27 gms lipid/hr gives a
destruction rate of about 1 l%/day of the remaining dieldrin. Though we have not shown
them, the other six compartments follow the blood decay curve very closely.

If in the generation of the curves in Figure 4 we had fed the rats a diet containing a
different dietary dieldrin concentration (e.g., 12.5 /~g dieldrin/gm food) instead of the
25 ppm HEOD diet shown in Figure 4, then th~ "steady-state" (average value of daily
"swing") would be altered accordingly. However, since the ~'s (induction coefficients)
are all assumed equal to zero in this case and the compartment sizes and blood flow rates
are supposed dieldrin independent, it is easy to show that the model system is linear and
hence any multiplication or division of the dietary dieldrin concentration directly
multiplies or divides the ordinant values of the residue curves shown in Figure 4.

A more realistic example to consider would be that of auto-induction, i.e., not all
¢l's = O. In that case the model would be non-linear. In particular it can be shown with a
one-tank simulator (Bird et al. 1960) that, depending upon the magnitude of ¢~(only a
single, non-zero induction coefficient needed), the long time ("steady-state") concentra-
tion in the tissue is approximately proportional to the dietary feeding rate raised to some
power whose limits are between 0.5 and 1.0. That is, if after several weeks of dieldrin
administration Ceq is the tissue lipid phase dieldrin concentration (/lg HEOD/gm tissue
30 F . T . Lindstrom et al.

fipid) and® is the dieldrin intake rate (~g HEOD/hr) then Ceq = o~Sn(~), (or is a constant
and n(/~) is the E-dependent power). In the linear case under consideration ~ = 0 with
n(0) = 1. If ~ -+ large, then n(~) -+ 1/2. This approximation is obtained directly from the
model with auto-induction effects alone. It is also in good agreement with the experi-
mental evidence of Walker et al. (1969).

For the reader's convenience now and for comparison purposes later on, Figure 5 is
given to demonstrate the effect of different dietary dieldrin concentrations on the
adipose lipid phase dieldrin residues. Only average (daily oscillation) values are plotted.
The magnitude of the "swing" is linearly multiplied or divided accordingly.

Case II. Tissue residues in a mature male human fed dieldrin. Hunter and Robinson
(1967) fed several male human subjects daily doses of 211 /lg of dieldrin in gelatin
capsules for several hundred days. Although those authors supplied some data (Table V)
about these subjects, the parameters necessary for the model are not sufficiently well-
known for any individual to permit explicit modeling of the resultant residues. However,

lO01
0 ® 0 Q
®

.o
.o A A Z~
c o &

o
c
8 o
.c_ ~
®
A
X O
Z~
a='~- 10
"o~_ ~ x
'~_ "o X o 25 ppm dieldrin in diet
=~ 12.5 ppm dieldrin in diet
& ®
x
x 6 . 7 5 ppm dieldrin in diet
&X

0
Q. X A
~5

1 -- 5' - 1 'o '

15 2'o . . . . . . . .30
25 . . . . . . . . .35
. 40 4'5 5'0 5 60'

T i m e (days)

Fig. 5. Lipid phase dieldrin concentration buildup and decay curves for the 300-gm male
rat, with a dieldrin destruction rate constant F = 0.27 gm lipid/hr and the rat eating
20 gms food/day, as a function of the dietary dieldrin concentration. (Same dieldrin
administration scheme as is shown in Figure 4.)
 Dieldrin Distribution Model 31

information does exist on average values, so that a representation of expected residues in

blood and depot fat may be made for a hypothetical mate human, 30-39 years old,
weighing 68 kg and having a cardiac output of 5.4 liters/min (Bard 1961). Estimates of
initial dieldrin mass data for the hypothetical 68 kg mature male human are given in
Table VI, based upon the depot tissue averages in Table V. The compartmental masses
and blood flows, estimated by formula (Table II) and the compartment lipid fractions
(fx) are given in Table VII. The numerical simulation of the model from these values
yields the tissue residue curves (drawn from average values of daily oscillations) shown in
Figure 6, comparable to the data points of Hunter and Robinson (1967). It is important
to note that relationships of the residues in depot fat or blood are not directly correlated
in the individuals with respect to a single paraaneter (age, body fat, etc.). Rather, the
inter-relationships of many parameters come into play, so that the model representation

Table V. Pre-exposure Data of Hunter and Robinson (1967}

HEOD
Body Body cone. Whole Oral
Subject Age Height weight fat (ppm) blood dose
number (yrs.) (cm) (kg) (kg) (depot fat) ppm ~g/day

6 43 181 78.8 11.6 0.145 .00094 211

9 38 181 61.8 4.3 0.199 .00139 211

11 52 180 80.5 13.7 0.270 .00155 211

Table VI. Initial Dieldrin Mass Data For

the Hypothetical 68 kg Mature Male
Human Based Upon Depot Tissue
Averages in Table V

Compartment U ° (~g dieldrin)

G.I. 23.2
Liver 74.3
Kidney 5.2
Brain 63.3
Depot fat 2406.4
Muscle 1101.4
Others 762.3
Blood pool 6.5
 t~
tO

Table VII. Compartment Mass, Blood Flow, and Other Important Parameters
For a Hypothetical 68 kg Mature Male Human

Lipid fraction Lipid mass Blood flow rate Blood lipid

Mass fx Mx qx flow rate Qx
Symbol ~ x (gms lipid, Ggms blood~ gms blood lipid)
Compartment x (gms) "gm tissue ) (gms lipid) ~ hr " hr

Blood pool (btood chambers B 5,020 0.004 20 384,329 1,532

of heart, lungs, arteries,
and veins)
Kidney K 271 0.053 14 87,124 348
Gastrointestinal (small bowel) G.I. 2,587 0.065 168 90,839 363
Liver L 1,336 0.06 80 110,798 443
Brain BR 1,265 0.104 132 23,328 93
Depot Fat F 12,240 0.637 7,797 4,293 17
Muscle (includes M 27,200 0.110 2,992 53,588 214
associated fat)
Other tissue (skin, etc.) OT 13,600 0.150 2,040 13,397 54

Rex = 0.0 [Assumed from data given by Hunter and Robinson (1967)]
~3 = 100.0 (Assumed same as rat, Table IV)
0 =0.995, 0 ~ < t ' < 1 2 h o u r s ~
£ repeatable daily
0 0.050, 12 ~< t <~ 24 hoursy
QBI = 0.085 gm lipid/hr, 0 ~< t < 12 h°urS~p e r i o d i c , z with 24 hour period
QBI 0.0085 gm lipid/hr, 12 <~ t < 24 hours J
 Dieldrin Distribution Model 33

provides an excellent guide to anticipated residues, with predictions certainly within the
limits required by analytical methods. The value of 1~ = 3.5 gms lipid/hr was used in the
computation, equivalent to a half life of 125 days in the 68 kg man.

Case III. Effects of experimental dosage methodology on residues. The exact manner
in which an experiment is carried out, often for reasons of financial exigency beyond the
control of the experimentor or analyst, can present difficulties in the ultimate inter-
pretation of the results. For instance, Keane and Zavon (1969) administered daily doses
of dieldrin in a corn oil solution in gelatin capsules to dogs. The dosage for the first five
days was five times that used subsequently for the remaining 55 days, but dieldrin was
administered only Monday through Friday. Blood samples were taken on Mondays and
Thursdays prior to dosing, and fat samples were taken monthly. They observed that
(a) total body burden of dieldrin tended to decrease between the first and second
months' adipose tests in all but the most obese animals, and (b) the concentrations of
dieldrin in adipose and whole blood tended to be inversely correlated with obesity. They
emphasized (and rightly so) the lack of a simple relationship between body burden, blood
and adipose residue concentrations, obesity, and level of dosage.

An examination of the effect of this administration regimen in the hypothetical

300-gm mature male rat is instructive. Figure 7a illustrates the effect of this discontinuous
administration schedule on blood and adipose dieldrin concentrations over a 35-day ex-
perimental period. The source function ~ (one gelatin capsule of dieldrin in oil/day) is
characterized as follows:

10 ] I ] I r-

Depot fat dieldrin concentration

O
ppm Symbol Hunter & Robinson Subject
J
O o 6
0.1-- zx 9
[] 11

Blood dieldrin concentration zx O __

0.01 -
LX

D
B

o.ooto--t- I t t I I I 1 1 _1_ I I 1 t I _ _ 1 1 1 _ I I I J l L ]
o 50 100 150 200 250
Time (days)
Fig. 6. Typical computed model curves for depot fat and arterial blood dieldrin levels in
man for feeding rates, flows, and compartment sizes for 68-kg man. Data points plotted
are from Hunter and Robinson (1967). lP = 3.5 gm lipid/hr, a constant.
34 F.T. Lindstrom et al.

100"0t 1
t A A Blood pool lipid phase dieldrin concentration /
] A A [t [~--~ 9ePOt fat lipid phase dieldrin concentration [

o 10.0
E
c
O
o

1 .O0 5 10 15 20 25 30
Days
100.01

Blood pool lipid phase dieldrin concentration

.~" Depot fat lipid phase dieldrin concentration

g
"~ 10.0
io

O
O

1.0~ 5 10 15 20 25 30
Days
100.0

Blood pool lipid phase dieldrin concentration

E

g
.'~ 10.0

c)

1"C0 5 10 15 20 25 30
Days

Fig. 7. Blood and d e p o t lipid-phase dieldrin c o n c e n t r a t i o n s in h y p o t h e t i c a l 300-gm

m a t u r e male rat according to simulation of the dose s c h e m e o f Keane and Zavon (1969).
1~ = 0.27 gm lipid/hr.
 Dieldrin Distribution Model 35

= 750/~g dieldrin/hr, 0 ~< t < 1 hour

= 0, 1 -G<t < 24 hours.

Assuming the animal is freely eating a standard grade of lab chow (one containing a few
percent lipids, enough to supply his daily needs), we characterize the absorption co-
efficient 0 and the bile lipid flow QBI the same as is shown in Table IV. These characteri-
zations are assumed repeatable each day, i.e., ~ and 0 are periodic functions of time.
After the first week on dieldrin (5 doses, one per day Monday-Friday, no dieldrin
Saturday and Sunday), the dosage level is cut to 150/~g dieldrin in oil in each gelatin
capsule. This reduces the magnitude of Usaccordingly, while leaving 0 unchanged.

As a consequence of administering dieldrin for five days at one level and then at 20%
of that level for the remaining days, the "preloading" would tend to overshoot the mean
concentration expected at equilibrium. Furthermore, the administration method would
be expected to introduce relatively marked swings in blood dieldrin concentration,
particularly during the "preloading" phase of exposure and over weekends. The amplitude
of these perturbations are related to compartment flows, lipid compositions, and hepatic
activities of individual animals, thus complicating the calculation of averages among
animals at any given sampling point.

Use of the model (Figure 7b) would predict that "preloading" with a daily dose only
twice that given after the initial five day period of Figure 7a would bring the animal
somewhat more quickly to approximately the same state of "equilibrium" as an
administration (Figure 7c) not using any "preloading." But this "equilibrium" terminal
residue would be below that expected for the same 150/ag/day dose absorbed con-
tinuously ( 5 = 6.25/~g/hr), since the relatively rapid loss would not be occurring over the
weekend.

Case IV. Parallel models of two similar compounds. Aldrin, heptachlor, and isodrin are
readily oxidized to the respective epoxides dieldrin, heptachlor epoxide, and endrin
by a variety of plant and animal species, and particularly by the hepatic microsomes
in mammals. Assuming that (1) the physical properties of each pair of cyclodiene
insecticides (e.g., aldrin-dieldrin) are not markedly dissimilar (both highly lipid-soluble),
(2) the loss of the parent cyclodiene is equal to the rate of formation of the epoxide, and
(3) neither compound affects the metabolism of the other, then two parallel sets of mass
balance equations (one for aldrin and one for dieldrin) can be set up analogous to
equation system (1). The output (UL/ML) for aldrin becomes the source for the dieldrin
model by adding this term to the dieldrin liver compartment mass balance equation and
setting the dieldrin source function'~ to zero. The destruction rate coefficients PA and
FD for aldrin and dieldrin, respectively, can be assumed to be independent for short time
intervals and are known to differ by about two orders of magnitude.

The blood lipid-phase aldrin and dieldrin concentrations resulting from a single intra-
venous injection of one mg of aldrin into a hypothetical 68-kg mature male human
(c.f. Case II) are shown as functions of time in Figure 8. Aldrin epoxidation (PA) is
 36 F . T . Lindstrom et al.

assumed to proceed at 100 times the rate of dieldrin degradation (I'D). Curves of similar
shape (Figure 9)have been obtained for heptachlor/heptachlor epoxide in blood following
single injections into the vena cava of 60-80 kg young swine (Gillett et al. 1971). Taking
into account the greater circulation rate and possible interspecific difference in rates of
degradation, and noting that heptachlor is epoxidized at a slightly lower rate than aldrin,
whereas heptachlor epoxide is metabolized at a greater rate than dieldrin, the results in

100.0~

Blood dieldrin (lipid phase)

(K = 3.517 gm/hr)

10.0

1.0
Blood aldrin (lipid phase)
(K = 351.7 gm/hr)
g
g
C3

0.1"

I I I I I I ! ! I I a I I I
0.01
0 20 40 60 80 1oo 120 140
Time (min)

Fig. 8. Aldrin and dieldrin blood (lipid-phase) residue curves for a single intravenous
injection of I000 #g aldrin at time t = 0 into venous blood of a hypothetical 68-kg
mature male human; FD = 3.5 gm lipid/hr.
 Dieldrin Distribution Model 37

swine are quite consistent with the model for man. Thus, although the lipid-phase
mammalian model is constructed, through the implicit assumptions, to represent the
chronic, long-term pharmacokinetics o f compounds such as dieldrin, it has reasonable
validity in application to shorter time spans and other compounds. To the extent that
representative simulators can be developed for any set o f interacting chemicals for which
the assumptions o f this model are valid, multiple parallel analyses can be accomplished.

Theoretical m a x i m u m
initial concentration

50.0

200.0

150.0

_o

m
100.0 -
Ol Heptachlor O
O
0
] HeptachlorEpoxide
= \ O
e.
r~

Q.
so o 10.0 (3.

×
O
c
8 \
t-
z \ o
,a

T
7- \
\

/ `4

100

/
/

t/
5.0 I i I I 1.0
o 5 lO 20 40
Time (min)

Fig. 9. Whole blood heptachlor and heptachlor epoxide concentrations plotted as log
concentration against time for two 68-80 kg pigs, each one receiving a 1000/~g/kg body
wt injection of heptachlor at time t = 0. Solid ( ) and dashed ( - - - - ) lines indicate
the values for each individual. (from Gillett e t al. 1971)
38 F . T . Lindstrom et al.

Conclusions
This model illustrates well how the pharmacodynamics of dieldrin and similar com-
pounds may be better understood simply by using existing knowledge of the physiology
and chemistry of the mammalian body. That knowledge plus the simplifying assumptions
and postulates constitute the model, which is demonstrably applicable to analysis of
data, testing of hypotheses, and examination of experimental design. No claim can be
made regarding the broader applicability of this mammalian lipid-phase model to com-
pounds which (1) have relatively low partition coefficients (less than 1000:1, oil vs.
water), (2) are membrane-transport, rather than flow-limited, and (3) have complex
metabolic steps producing metabolites interacting with the rate(s) of metabolism of the
chemical to be modeled. Nevertheless, a priori knowledge of such characteristics may be
employed in a model for such compounds, yielding a corresponding set of differential
equations simulating metabolism and distribution to represent the pharmacodynamics
within those constraints. Exposure of special tissue compartments (e.g., spleen, bone
marrow) not necessarily represented in this exposition can be predicted by knowledge of
the lipid mass and blood perfusion rates and by information from whole body exposures,
blood analyses, or fat biopsies, rather than elaborate and costly experimental procedures.

Application of the model is especially instructive as to the limitations on investigators

and interpretations of results. The demonstration cases point out potential pitfalls in
experimental design and in analysis of both the compound's distribution and the test
subject's physiology. Those examples also indicate certain deficiencies in the current
model: why, for instance, do the three physiologically diverse human subjects in Case II
have residue values so similar to that of the hypothetical man? In part the deficiencies
stem from inadequate knowledge of two especially critical areas requiring further work
for a more explicit and fundamental representation of dieldrin behavior in man and
other mammals.

The mechanism(s) of absorption of dieldrin and similar highly lipid-soluble chemicals

and drugs are poorly known, particularly in relation to normal and pathological alterations
in physiological status of the intestinal epithelium, liver, and serum as affected by diet,
diurnal cycles, and growth. Such knowledge is crucial in analysis of acute or short-term
effects of exposure as well as in application of prophylactic or chemotherapeutic pro-
cedures relative to contaminated individuals. As a corollary, information must also be
obtained on the salivary/gastric/jejunal secretory processes recirculating dieldrin through
the gastrointestinal tract regardless of initial mode of exposure.

Equally important to explicit knowledge of the source terms are the mechanism(s) of
dieldrin metabolism and possible interactions, either by dieldrin or other agents, leading
to possible induction and/or inhibition of that metabolism. The ~'s in Equation 11 are
compound/species/sex/organ/enzyme - specific and probably are regulated hormonally
or physiologically, so that age, diet, and diurnal changes may be reflected to greater or
lesser degrees. Although the task of developing such information for even one compound
and one species would appear overwhelming, the model fortunately provides for inclusion
and consideration of that knowledge as it develops. Moreover, the simplifying assump-
 Dieldrin Distribution Model 39

tions may be relaxed partially without loss of overall integrity of the model, thus adjusting
the application of the model to the needs of the investigator.

Acknowledgments

The authors gratefully acknowledge the advice and encouragement of their colleagues,
especially V. H. Freed, Director of the Oregon State University Environmental Health
Center and I. J. Tinsley, Professor of Chemistry, in preparation of this paper.This re-
search was supported in part by U.S. Public Health Service grant ES-00040 from the
National Institute of Environmental Health Sciences.

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Manuscript received July 9, 1973; accepted for publication September 15, 1973